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Efficient plant regeneration via organogenesis in winter squash (Cucurbita maxima Duch.)
Abstract
Using cotyledonary explants excised from seedlings germinated in vitro, efficient plant regeneration via organogenesis was established for two winter squash (Cucurbita maxima Duch.) cultivars. To establish optimal conditions for adventitious shoot induction, a variety of explants were prepared from seedlings of different ages and these were cultured using media containing different concentrations of 6-benzylaminopurine (BA). For both cultivars, plant regeneration was optimal when the proximal parts of cotyledons from 4-day-old seedlings were cultured on induction medium composed of Murashige and Skoog (MS) medium with 1 mg/l BA. After 3 weeks of culture in induction medium, 82 and 92% of explants from the two cultivars regenerated shoots. Adventitious shoots were subcultured on elongation medium composed of MS medium with 0.1 mg/l BA and the elongated shoots were successfully rooted in MS medium without growth regulators for 2 weeks. Flow cytometric analysis revealed that most of the regenerated plants were diploid.
... Previous studies have demonstrated that organogenesis in the genus Cucurbita is influenced by explant type, genotype, induction media, and other factors [22][23][24][25]. Lee et al. reported the cultivation of cotyledon explants from two winter squash varieties, both of which successfully produced complete plants, and investigated several factors affecting organogenesis, such as the size of the explants and the type of culture media [26]. The critical factor in inducing plant organogenesis is the use of hormones on explants with regenerative potential at a certain growth stage [27]. ...
... We achieved a regeneration rate of 52.62% for the 5 mm long hypocotyl. However, Lee et al. reported that shoots cannot be regenerated from winter squash hypocotyls [26]. We speculate that differences in the specific location of hypocotyl sampling and genotype may account for this discrepancy in results. ...
... However, we can still draw some insights from previous research. Lee et al. induced shoot formation from two genotypes of winter squash on 6-BA media supplemented with four different concentrations of salt [26]. The results showed that the different genotypes had distinct regeneration rates on the same medium and that the same genotype had varying regeneration rates on different media, indicating an interaction between these two factors. ...
Improving plant regeneration ability and shortening regeneration time can promote the development of genetic transformation breeding technology for horticultural crops. We optimized several culture conditions, including explant type, mother plant genotype, and medium, to improve shoot formation in winter squash (Cucurbita maxima Duch.). Histological analysis of the occurrence of shoots was also carried out. The results indicate that cotyledon was the most suitable explant for inducing the shoot regeneration of winter squash. We found that ‘Jin-li’ had a shorter shoot induction time and a higher average number of shoots. The highest induction rate of 95.23% among the five lines. The average shoot induction rate of five lines was the highest (84.85%) on Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 6-benzylaminopurine (6-BA) and 0.2 mg/L indole-3-acetic acid (IAA). We also found that there was an interaction between genotypes and induction media, and their interaction had a greater impact on the shoot induction rate than individual effects. Histological observation revealed that the induced shoots of winter squash cotyledons originated from subepidermal cells. We also found that the optimal medium for de novo root regeneration was 1/2 MS. We acclimatized and cultivated regenerated plants and harvested their fruits, which maintained the characteristics of mother plants. These findings lay an important foundation for further research on direct shoot regeneration and accelerate its application in winter squash genetic transformation.
... Moreover, different explant types expressed differently in both embryogenesis and organogenesis system of squash tissue culture (Obembe et al., 2017;Dursun et al., 2019). Some other factors like explant age also influenced in plantlets regeneration potentiality of squash (Lee et al., 2003;Stipp et al., 2012;Nanasato et al., 2013). However, silver nitrate (AgNO 3 ) has evidenced to be a very effective inhibitor of ethylene action and is widely used in plant tissue culture of C. sativus (Venkatachalam et al., 2018) but limitedly reported for C. pepo. ...
... In this research several factors of somatic embryogenesis of hybrid C. pepo genotypes were studied. Different genotypes of C. pepo have been regenerated via somatic embryogenesis (Chee, 1992;Gonsalves et al., 1995;Urbanek et al., 2004) and organogenesis (Lee et al., 2003;Kiss-Baba et al., 2006;Stipp et al., 2012;Mookkan, 2015;Obembe et al., 2017;Dursun et al., 2019) to date. A wide range of genetic variation within C. pepo species could contribute different regeneration potentiality for different genotypes (Gonsalves et al., 1995;Kathiravan et al., 2006;Stipp et al., 2012). ...
... This result can be compliant with the results of Gonsalves et al. (1995) stated that, cotyledons are the most efficient and dependable source of somatic embryogenesis. Additionally, most of the previous organogen-esis protocols of squash (Lee et al., 2003;Kiss-Bába et al., 2010;Stipp et al., 2012;Obembe et al., 2017;Dursun et al., 2019) have been developed using cotyledon explant. However, it was found that the height 66.7% shoot regeneration frequency obtained from 6 days cotyledon explants. ...
Squash (Cucurbita pepo L.) is highly polymorphic in nature which could contribute its different regeneration potentiality for different genotypes. The present investigation was carried to know the effects of plant growth regulators, explant types, explants age and AgNO3 on in vitro regeneration potentiality of hybrid squash genotype and to optimize regeneration protocol. In this protocol, cotyledon and hypocotyl segments were grown on MS medium supplemented with 1.5 mg/l 6-Benzylaminopurine (BA) and 0.2 mg/l α-Naphthalene acetic acid (NAA), successfully induced callus and subsequently shoots for C. pepo L. cv. First Runner. The height percentage of shoot production frequency was 66.7% from 6 days old cotyledon explants in MS + 1.5 mg/l BA + 0.2 mg/l NAA combination. The shoot regeneration frequency was increased up to 86.67% when 2.0 mg/l AgNO3 was added with MS + 1.5 mg/l BA + 0.2 mg/l NAA media combination by using 5 days old cotyledon explants. Shoot regeneration potentiality of four other C. pepo genotypes were also investigated with this system and showed a substantial amount of regeneration potentiality for different genotypes. Regenerated shoots induced the height 93.33% rooting frequency which promoted long, thick roots on 10 to 12 days in MS medium supplemented with 0.1 mg/l NAA. The regenerated plantlets with long and thick roots were acclimatized in pot soil and eventually grown in natural environment with culture facility. The present study describes a simple and efficient protocol for in vitro plant regeneration of hybrid C. pepo genotypes and may be utilized for further transgenic development.
... Valková, V. 1,2 , Ďúranová, H. 1 , Ivanišová, E. 3 , Tóthová, Z. 1 , Gabríny, L. 1 1 Výskumné centrum AgroBioTech, Slovenská poľnohospodárska univerzita v Nitre 2 Ústav záhradníctva, Slovenská poľnohospodárska univerzita v Nitre 3 Ústav potravinárstva, Slovenská poľnohospodárska univerzita v Nitre veronika.valkova@uniag.sk ...
... Valková, V. 1,2 , Ďúranová, H. 1 , Ivanišová, E. 3 , Tóthová, Z. 1 , Gabríny, L. 1 1 Výskumné centrum AgroBioTech, Slovenská poľnohospodárska univerzita v Nitre 2 Ústav záhradníctva, Slovenská poľnohospodárska univerzita v Nitre 3 Ústav potravinárstva, Slovenská poľnohospodárska univerzita v Nitre veronika.valkova@uniag.sk ...
... moschata) a tekvica obyčajná (C. pepo), ktoré sú variabilné tvarom, veľkosťou a farbou [1]. Nakoľko čerstvé plody tekvice po nakrájaní veľmi rýchlo podliehajú mikrobiálnemu znehodnoteniu (3-5 dní), a to dokonca i v chladných podmienkach, je vhodné ich spracovať do formy zabezpečujúcej predĺženie ich trvanlivosti [2]. ...
Tekvica je druh zeleniny pochádzajúci z rodu Cuburbita (C.; čeľaď: Cucurbitaceae). Vo
všeobecnosti patria medzi celosvetovo najviac konzumované tri druhy tejto plodiny, a to
tekvica veľkoplodá (C. maxima), tekvica muškátová (C. moschata) a tekvica obyčajná (C.
pepo), ktoré sú variabilné tvarom, veľkosťou a farbou [1]. Nakoľko čerstvé plody tekvice po
nakrájaní veľmi rýchlo podliehajú mikrobiálnemu znehodnoteniu (3-5 dní), a to dokonca
i v chladných podmienkach, je vhodné ich spracovať do formy zabezpečujúcej predĺženie ich trvanlivosti [2]. Jedným z inovatívnych spôsobov riešenia tejto problematiky je výroba
tekvicovej múky, ktorá disponuje trvanlivosti cca. 1 rok [3]. Použitie tejto netradičnej múky je veľmi žiadané z hľadiska jej príjemnej chuti a oranžovo-červenej farby [4]. Navyše, takto
pripravená múka je cenným zdrojom mnohých biologicky aktívnych látok vrátane karotenoidov a kyseliny askorbovej. Taktiež disponuje vysokým obsahom sacharidov, škrobov, vlákniny, bielkovín a popolovín, a naopak je charakteristická pre svoj nízky obsah lipidov a hrubej vlákniny [5]. Vzhľadom na vyššie uvedené benefity bola pozornosť vedeckej komunity, v mnohých výskumných prácach, zameraná na výrobu zmesných tekvicových múk, ďalej aplikovateľných do pekárenských a cukrovinárskych výrobkov [6,7].
... However, decreasing in number and height of new shoot were found on the treatment of BA at lower and higher than 1 and 0.5 mg L -1 , respectively (Table 1; Figure 2A). These results were in accordance with previous researches that percent shoot induction (Lee et al., 2003;Rai et al., 2012;Bekele et al., 2013), number of new shoots explant -1 (Compton and Gray, 1993;Ahmad and Anis, 2005;Ntui et al., 2009), new shoot length (Haque et al., 2008;Sangeetha and Venkatachalam, 2014;Mali and Chavan, 2016), and average growth rate of new shoot (Abdul-Awal et al., 2005) of many cucurbitaceous plants decreased when BA was used at improper level. Furthermore, a high concentration of cytokinins can reduce, delay, and inhibit shoot regeneration (George et al., 2008). ...
... This BA concentration produced the highest new shoot numbers without breaking height of new shoot off (Table 1; Figure 2B). One mg L -1 BA was also used as the optimum concentration for in vitro propagation of various cucurbitaceous plants (Ananthakrishnan et al., 2003;Lee et al., 2003;Sangeetha and Venkatachalam, 2014) and also some herbaceous plants (Chuengpanya et al., 2014). ...
... Usage of PGR-free MS agar medium for reducing cytokinins effects is one of the effective in vitro rooting methods (George et al., 2008). This medium could stimulate root formation in some herbaceous plants (Chuengpanya et al., 2014) as well as some cucurbitaceous plants (Ananthakrishnan et al., 2003;Lee et al., 2003;Ntui et al., 2009). After transferring explants onto BA-free MS agar medium for 4 weeks, roots were significantly increased when compared to the 4 th week of culture (Table 1). ...
The genus Zehneria belongs to the Cucurbitaceae family and includes plant species that contain glycosides and sphingolipids in roots and leaves. Some species of Zehneria are used in local folk medicine for curing skin diseases. It also shows antimicrobial and antitumour activities. In Thailand, Zehneria platysperma is classified
as an endemic plant which is found only in northern and south-western regions. This plant may risk extinction due to the deforestation problem. Therefore, this study established an in vitro propagation protocol for conservation of Z. platysperma. Detached seeds were surface sterilized using sodium hypochlorite and inoculated onto Murashige and Skoog (MS) agar medium for initiating the in vitro culture. Sixteenweek-old seedlings were selected for use as plant materials. The first and second nodes from base of seedling were excised and used as explants. Nodes were cultured on MS agar medium supplemented with different concentrations of N6-benzyladenine (BA; from 0 to 4 mg/L) before transferring onto BA-free MS agar medium. After 8 weeks, the highest shoot numbers (9.30 shoots/explant) with an average shoot height at 9.45 cm was obtained from Z. platysperma that had been cultured on MS agar medium supplemented with 1 mg/L BA. This medium also provided an average root number/explant at 12.40 with the longest root length (20.67 cm). Increasing of BA concentration in culture medium reduced shoot and root regeneration. Moreover, hyperhydric symptom was found from explants that were cultured on 2 and 4 mg/L of BA. In addition, flowering and fruiting were found during in vitro condition. Vigorous plantlets with well-developed roots survived in natural condition after acclimatization process. This protocol can be a useful tool for rapid proliferation and conservation of Z. platysperma which could be used for future phytochemical or medical studies.
... There are only two in vitro regeneration protocols have been reported for C. maxima) (LEE et al [2]) and C. moschata (ZHANG et al [3]). Otherwise, efficient plant regeneration protocols have been developed and generated in Cucumis sativus (VASEDUVAN et al [4]; GROZEVA and VELKOV [5]), Citrullus lanatus (WANG et al [6]; ZHANG et al [7]), Cucumis melo (CURUK et al [8]; RHIMI et al [9]; TEKDAL and CETINER [10]) and Cucurbita pepo (LEE et al [2]; ANANTHAKRISHNAN et al [11]; STIPP et al [12]; MOOKHAN [13]). ...
... There are only two in vitro regeneration protocols have been reported for C. maxima) (LEE et al [2]) and C. moschata (ZHANG et al [3]). Otherwise, efficient plant regeneration protocols have been developed and generated in Cucumis sativus (VASEDUVAN et al [4]; GROZEVA and VELKOV [5]), Citrullus lanatus (WANG et al [6]; ZHANG et al [7]), Cucumis melo (CURUK et al [8]; RHIMI et al [9]; TEKDAL and CETINER [10]) and Cucurbita pepo (LEE et al [2]; ANANTHAKRISHNAN et al [11]; STIPP et al [12]; MOOKHAN [13]). ...
... Furthermore, the physiological conditions and hormonal contents of explants are crucial points for regeneration potential (ZHANG et al [3]; MENDI et al [14]; PETROVA et al [15]). Proximal cotyledon part with hypocotyl segment collected from in vitro germinated seedling was found as the best explant type for the regeneration in winter squash (LEE et al [2]) and pumpkin (ZHANG et al [3]). Although clonal plants were successfully propagated in aforementioned research, only 2 winter squashes and 1 pumpkin genotype were used for in vitro regeneration. ...
A productive direct regeneration protocol for a wide range of winter squash (Cucurbita maxima Duch.) and pumpkin (Cucurbita moschata Duch.) lines Abstract The success of Cucurbit transgenic technology depends on regeneration protocol and recovery of transformed plants. Therefore, an efficient regeneration protocol was established for in vitro shoot regeneration from cotyledon explants to optimize the regeneration of multiple adventitious shoots formation in winter squash and pumpkin lines. Proximal parts of cotyledon explants were excised from in vitro seedlings and were cultured on solid MS medium supplemented with various concentrations and combinations of BAP and TDZ. The number of shoot per regenerated explants was ranged from 2.1 (in line 14BO01) to 7.8 (in line 57Sİ21) in MS medium by addition of 0.5 mg/l BAP and 1 mg/l BAP + 1 mg/l TDZ, sequentially. The cultures were incubated at 25 ± 1°C and 16-h photoperiod for 4 to 5 weeks. Donors were effective on shoot formation, and 57Sİ21 in winter squashes and G9 in pumpkins were superior lines. Regenerated shoots were rooted and elongated on MS medium combined with 0.01 mg/l IAA. Elongated and rooted plants were acclimatized with 84% survival rate. Overall, 122 plants (91 of winter squash and 31 of pumpkin) were obtained during the regeneration process. The ploidy analyses revealed that 118 plants were diploid and 4 plants were tetraploid. Keywords Regeneration, cotyledon explants, winter squash, pumpkin. To cite this article: KURTAR ES, BALKAYA A, KANDEMIR D. A productive direct regeneration protocol for a wide range of winter squash (Cucurbita maxima Duch.) and pumpkin (Cucurbita moschata Duch.) lines. A productive direct regeneration protocol for a wide range of winter squash and pumpkin lines 1349
... There are only two in vitro regeneration protocols have been reported for C. maxima) (LEE et al [2]) and C. moschata (ZHANG et al [3]). Otherwise, efficient plant regeneration protocols have been developed and generated in Cucumis sativus (VASEDUVAN et al [4]; GROZEVA and VELKOV [5]), Citrullus lanatus (WANG et al [6]; ZHANG et al [7]), Cucumis melo (CURUK et al [8]; RHIMI et al [9]; TEKDAL and CETINER [10]) and Cucurbita pepo (LEE et al [2]; ANANTHAKRISHNAN et al [11]; STIPP et al [12]; MOOKHAN [13]). ...
... There are only two in vitro regeneration protocols have been reported for C. maxima) (LEE et al [2]) and C. moschata (ZHANG et al [3]). Otherwise, efficient plant regeneration protocols have been developed and generated in Cucumis sativus (VASEDUVAN et al [4]; GROZEVA and VELKOV [5]), Citrullus lanatus (WANG et al [6]; ZHANG et al [7]), Cucumis melo (CURUK et al [8]; RHIMI et al [9]; TEKDAL and CETINER [10]) and Cucurbita pepo (LEE et al [2]; ANANTHAKRISHNAN et al [11]; STIPP et al [12]; MOOKHAN [13]). ...
... Furthermore, the physiological conditions and hormonal contents of explants are crucial points for regeneration potential (ZHANG et al [3]; MENDI et al [14]; PETROVA et al [15]). Proximal cotyledon part with hypocotyl segment collected from in vitro germinated seedling was found as the best explant type for the regeneration in winter squash (LEE et al [2]) and pumpkin (ZHANG et al [3]). Although clonal plants were successfully propagated in aforementioned research, only 2 winter squashes and 1 pumpkin genotype were used for in vitro regeneration. ...
... Somatic embryogenesis is a significant method for mass propagation, plant regeneration for successful transformation and to develop genetically uniform plants (Etienne et al., 2002;Thomas and Sreejesh, 2004;Thakare et al., 2008;Gatica-Arias et al., 2008;Lee et al., 2003;Martin, 2004). It is also used when evaluating growth, differentiation, de-differentiation and development of plant cells (Ikeda-Iwai et al., 2003;Martin, 2004). ...
... Several protocols have been established for plant regeneration of cucurbits via somatic embryogenesis from different explant type (Chee, 1990;Choi et al., 1994;Nakagawa et al., 2001;Chaturvedi and Bhatnagar, 2001;Lee et al., 2003;Thomas and Sreejesh, 2004;Park et al., 2005;Pal et al., 2007;Selvaraj et al., 2007;Cho et al., 2008;Shah et al., 2008) nevertheless, there are no reports on somatic embryogenesis of Cucumis melo var. ...
... Based on the colour of the seeds, the origin of pumpkin has been attributed to Guetmala, Central Mexico or Columbia. The name pumpkin originated from a Greek word Pepon which means large melon [25].Among the ingredients in pumpkin are Telfairia occidentalis, Cucurbita moschata, Cucurbita pepo, Cucurbita maxima, Cucrbitamixta, and Cucurbita facifola [26].The three most widely cultivated pumpkin species worldwide are Cucurbita pepo, Cucurbita maxima, and Cucurbita moschata [27]. It is grown all over the world, and depending on the region and traditions, it is used in various ways. ...
... Pumpkins are also highly perishable or can be stored for months without much change in quality [28].Worldwide production of pumpkin, squashes, and gourds is 24.62 million metric tones from an area of 5,10,0000 ha and in India, the total production is 49,00,000 tones from an area of 45,000 ha. The weight of the fruit varies from 8 to 10 kg on average, and or sometimes even up to 20 kg [29].Three varieties of pumpkin are widely grown all over the world Curcurbita pepo, Curcurbita maxima, and C. moschata [27]. In India, pumpkin is also commonly referred to as "Sitaphal," "Kashiphal," or "kaddu." ...
This research work was carried out to explore the possibility of utilization of underutilized but highly nutrient rich pearl millet and pumpkin pulp in cookies. Cookies are generally made from Maida flour that provides high energy but low in fibre. Bakery products are often high in energy and fat and the most consumed bakery products is cookies. This research work proposed a healthy composition of cookies by incorporating pumpkin pulp while replacing usual principal ingredients like Maida flour with Pearl Millet (Bajra) and refined sugar with honey. The quality cookies were prepared from 50% pearl millet flour and 50% pumpkin pulp,60% pearl millet flour and 40% pumpkin pulp, 70% pearl millet flour and 30% pumpkin pulp and 80% pearl millet flour and 20%pumpkin pulp. There was decrease in carbohydrates and fat and increase in moisture, vitamins, ash and dietary fibre .Good quality cookies can be prepared by substituting pumpkin pulp(40%) with pearl millet flour (60%).This study evaluated the health benefits of millet pumpkin cookies based on their nutritional value and sensory analysis. pearl Millets are rich in all nutritious aspects like vitamins, fats, protein and also high in micronutrients such as folic acid, niacin and Vitamin B6, B complex vitamins that help in providing various medical and health benefits, millets into diet will reduce the risk of cardiovascular problems helps in treating constipation, diabetes ,obesity, hyperlipidemia. Pumpkin flesh is rich in fibre, vitamin C, vitamin E, Mg, K and a variety of carotenoids being the important sources of these amazing phytonutrients and its prevent various diseases, such asantidiabetic, antioxidant, anticarcinogenic, and anti-inflammatory.
... An efficient plant regeneration system is therefore necessary for genetic transformation and propagation of gherkin. During the past years, considerable efforts have been made for in vitro plant regeneration through indirect organogenesis of Cucumis sativus using hypocotyl (Selvaraj et al. 2006) and cotyledon derived calli (Kim et al. 1988;Selvaraj et al. 2007), direct regeneration from cotyledon and hypocotyl explants of Cucurbita maxima (Lee et al. 2003), C. melo and C. sativus (Curuk et al. 2003). In spite of the availability in various reports on cucumber organogenesis, several problems are still encountered to regenerate cucumber species in tissue culture. ...
... The age of seedling is reported to be an important factor for obtaining high frequency adventitious shoot (Compton et al. 2000;Lee et al. 2003;Krug et al. 2005;Stipp et al. 2012). For melon organogenesis, explants of 4-5-day-old seedlings are active and respond efficiently to exogenous hormones (Fang and Grumet 1990;Curuk et al. 2003). ...
An efficient protocol for in vitro organogenesis was achieved from callus-derived immature and mature petiole explants of West Indian gherkin (Cucumis anguria L.). Calluses were induced from immature petiole explants excised on 7-day-old in vitro seedlings and mature petiole explants of 40-day-old in vivo plants. The maximum frequency of immature petiole explants (98.0 %) and mature petiole (91.5 %) produced green, compact organogenic callus in Murashige and Skoog (MS) medium with Gamborg (B5) vitamins containing 30 g l −1 sucrose, 8.0 g l −1 agar and 4.0 μM naphthalene acetic acid (NAA) with 2.0 μM benzyl amino purine (BAP) after two successive subculture at 11 days interval. Adventitious shoots were produced from the organogenic callus when it was transferred to MSB 5 medium supplemented with 3.0 μM TDZ, 1.0 μM NAA and 0.05 mM L-glutamine with shoot induction frequency of immature petiole 45 shoots and mature petiole 40 shoots per explant. The shoots were excised from callus and elongated in MSB 5 medium fortified with 3.0 μM gibberellic acid (GA 3). Then elongated shoots were rooted in half strength MSB 5 medium supplemented with 3.0 μM indole 3-butyric acid (IBA). Histological analyses of the regeneration process confirmed the indirect organogenesis pattern. Plantlets with well-developed shoot and root systems were successfully acclimatized (95 %) in winter season and exhibited normal morphology and growth characteristics. The survival percentage differed with seasonal variations.
... An efficient plant regeneration system is therefore necessary for genetic transformation and propagation of gherkin. During the past years, considerable efforts have been made for in vitro plant regeneration through indirect organogenesis of Cucumis sativus using hypocotyl (Selvaraj et al. 2006) and cotyledon derived calli (Kim et al. 1988;Selvaraj et al. 2007), direct regeneration from cotyledon and hypocotyl explants of Cucurbita maxima (Lee et al. 2003), C. melo and C. sativus (Curuk et al. 2003). In spite of the availability in various reports on cucumber organogenesis, several problems are still encountered to regenerate cucumber species in tissue culture. ...
... The age of seedling is reported to be an important factor for obtaining high frequency adventitious shoot (Compton et al. 2000;Lee et al. 2003;Krug et al. 2005;Stipp et al. 2012). For melon organogenesis, explants of 4-5-day-old seedlings are active and respond efficiently to exogenous hormones (Fang and Grumet 1990;Curuk et al. 2003). ...
An efficient protocol for in vitro organogenesis was achieved from callus-derived immature and mature petiole explants of West Indian gherkin (Cucumis anguria L.). Calluses were induced from immature petiole explants excised on 7-day-old in vitro seedlings and mature petiole explants of 40-day-old in vivo plants. The maximum frequency of immature petiole explants (98.0 %) and mature petiole (91.5 %) produced green, compact organogenic callus in Murashige and Skoog (MS) medium with Gamborg (B5) vitamins containing 30 g l −1 sucrose, 8.0 g l −1 agar and 4.0 μM naphthalene acetic acid (NAA) with 2.0 μM benzyl amino purine (BAP) after two successive subculture at 11 days interval. Adventitious shoots were produced from the organogenic callus when it was transferred to MSB 5 medium supplemented with 3.0 μM TDZ, 1.0 μM NAA and 0.05 mM L-glutamine with shoot induction frequency of immature petiole 45 shoots and mature petiole 40 shoots per explant. The shoots were excised from callus and elongated in MSB 5 medium fortified with 3.0 μM gibberellic acid (GA 3). Then elongated shoots were rooted in half strength MSB 5 medium supplemented with 3.0 μM indole 3-butyric acid (IBA). Histological analyses of the regeneration process confirmed the indirect organogenesis pattern. Plantlets with well-developed shoot and root systems were successfully acclimatized (95 %) in winter season and exhibited normal morphology and growth characteristics. The survival percentage differed with seasonal variations.
... The genus Cucurbita, indigenous to the western hemisphere, is comprised of five domesticated species viz., Cucurbita moschata, C. maxima, C. pepo, C. ficifolia, and C. turbaniformis. Among these, Cucurbita moschata Duchesne ex Poiret, C. maxima Duchesne ('winter squash') and C. pepo L. (squash) are the worldwide commonly grown species of pumpkin (Lee et al., 2003) [21] . C. moschata and C. maxima are the most frequently consumed species of pumpkin in countries like India, Pakistan, Bangladesh etc., whereas, C. pepo is consumed up to lesser content in comparision to the other two species in these countries. ...
... The genus Cucurbita, indigenous to the western hemisphere, is comprised of five domesticated species viz., Cucurbita moschata, C. maxima, C. pepo, C. ficifolia, and C. turbaniformis. Among these, Cucurbita moschata Duchesne ex Poiret, C. maxima Duchesne ('winter squash') and C. pepo L. (squash) are the worldwide commonly grown species of pumpkin (Lee et al., 2003) [21] . C. moschata and C. maxima are the most frequently consumed species of pumpkin in countries like India, Pakistan, Bangladesh etc., whereas, C. pepo is consumed up to lesser content in comparision to the other two species in these countries. ...
Present study was conducted to evaluate the various physico-chemical attributes of different portion of ripe pumpkin. Ripe pumpkin comes under the category of underutilized fruits and is cultivated all over the World. Ripe pumpkin also called as lal kaddu in India is considered as good source of phyto-chemicals such as carotenoids, vitamins and minerals. For analysis of quality attributes of different portions (flesh, brains/fluffy portion and peel) of ripe fruits, two species of pumpkin viz. C. moschata Duchesne ex Poiret and C. maxima Duchesne were selected. Among different species of pumpkin, highest ascorbic acid and titratable acidity were found in flesh portion of C. maxima and minimum titratable acidity, ascorbic acid and beta carotene were obtained in peel portion of C. moschata. Brains/fluffy portion of C. maxima species was found with highest amount of beta carotene and total fiber among all portions of both the species. However, peel portion of C. maxima showed highest values for total soluble solids, total phenols and fiber among all portions of both species of ripe pumpkin. Thus, ripe pumpkin of C. maxima species was found with better quality attributes in comparison of C. moschata.
... It is extensively cultivated across India and other warm regions in world, with its center of origin traced to Central Mexico. The genus Cucurbita includes about 27 species, both wild and cultivated, with Cucurbita moschata, Cucurbita pepo, and Cucurbita maxima being the most commonly grown species worldwide (Lee et al., 2003). For example, the 'Arka Chandan' variety of C. moschata is known for its medium-sized fruit, bright orange flesh, and superior nutritional qualities compared to local strains (Gupta & Srinivas, 1979). ...
Pumpkin (Cucurbita moschata L.) is an economically important crop cultivated for its nutritional value and diverse uses. The present study focuses on understanding the relationships among yield and yield attributing traits in identifying key contributors to yield improvement. Eighteen diverse genotypes of pumpkin were examined in Randomized Complete Block Design with three replication for correlation and path coefficient analysis. Correlation analysis showed that genotypic correlation coefficients were generally higher than phenotypic ones, highlighting a strong inherent relationship between the traits. Fruit yield had a significant positive correlation with fruit polar diameter, followed by pericarp thickness and fruit weight, at both genotypic and phenotypic levels. Whereas path coefficient analysis elucidated that the fruit polar diameter had the highest positive direct effect on fruit yield (0.771), followed by peripheral thickness (0.375), male/female bud ratio (0.234). Fruit yield showed a positive and significant correlation with polar diameter and fruit weight, which also exhibited a positive direct effect on yield. These traits are, therefore, key targets for improvement to enhance overall yield potential.
... Dayanıklı olarak belirlenen kestane kabağı hatlarının gerek çeşit ıslahında ve gerekse anaç ıslahında değerlendirilmesi büyük önem taşımaktadır. Literatürde bazı çalışmalarda su kabakları ve kestane kabaklarının, türler arası melez ve bal kabaklarından daha az dayanıklı olduğu bildirilmiştir(Lee 2003; Yücel ve ark. 2022). ...
Hıyar yetiştiriciliğinde Fusarium oxysporum f. sp. cucumerinum J.H. Owen (FOC) ürün kaybına neden olan toprak kökenli önemli bir fungal bitki patojenidir. Günümüzde kabakgillerde özellikle karpuz, kavun ve hıyarda aşılı fide kullanımı Fusarium solgunluğu mücadelesinde etkili ve yaygın kullanılan bir yöntemdir. Aşılı hıyar fidesi için anaç olarak, en çok türler arası kestane kabağı (Cucurbita maxima) × bal kabağı (Cucurbita moschata) melezleri ile tür içi bal kabağı melezleri kullanılmaktadır. Bu çalışmada, önce aşılı hıyar üretiminde anaç ıslah programı için 49 kestane kabağı genotipinden oluşan bir gen havuzu oluşturulmuş, daha sonra kestane kabağı hatlarının F. oxysporum f. sp. cucumerinum’a karşı dayanıklılık durumlarının belirlenmesi ve FOC ile muamele edilmiş kestane kabağı hatları ile kontrol bitkilerinin vejetatif büyüme özellikleri yönünden incelenmesi amaçlanmıştır. Pozitif kontrol olarak Nun 9075 ticari kabak anacı ve negatif kontrol olarak Çengelköy hıyar çeşidi kullanılmıştır. Araştırma sonucunda, 20 genotipin yüksek düzeyde dayanıklı, 23 genotipin ise orta düzeyde dayanıklı ve 5 genotipin düşük düzeyde dayanıklı olduğu belirlenmiştir. Patojenle enfekteli olan ve kontrol uygulamasında incelenen bitki boyu, gövde çapı, yaprak sayısı, kuru kök, gövde ve yaprak ağırlığı özellikleri yönünden istatistiksel olarak çok önemli düzeyde farklılıklar olduğu saptanmıştır. Araştırma sonucunda; hem hastalık dayanımı hem de vejetatif büyüme parametreleri yönünden öne çıkan hatlar yeni hibrit kabak anaçlarının geliştirilmesine yönelik ıslah programında ümitvar kestane kabağı ebeveynleri olarak seçilmiştir.
... The genus Cucurbita comprises five domesticated species: Cucurbita moschata, Cucurbita maxima, Cucurbita pepo, Cucurbita turbaniformis and Cucurbita ficifolia. Among these, Cucurbita species namely C. moschata, C. maxima and C. pepo, are universally grown species [2]. C. moschata, originating in Central America, is now widely scattered over the globe. ...
Summary
Worldwide, vegetables play a very crucial role in human nutrition but their non-traditional edible parts like peels, seeds, leaves, stems are discarded as waste. Despite being enriched with healthful nutrients, vegetable by-products are not utilized efficiently. In the present scenario, pumpkin has gained extensive attention because of its rich nutritional composition as well as the health-defensive properties of its by-products (peel and seeds) that comprise 18-21% of pumpkin. Pumpkin seeds, which are often discarded, are loaded with protein, fat, fiber, and minerals that are vital for good health. Pumpkin seeds are also a significant source of unsaturated fatty acids and phytochemicals that exhibit a nutraceutical effect on human health. On the other side, peel of the pumpkin is a significant source of minerals, dietary fiber, tocopherol and carotenoids as well as an economic source of pectin. The nutritional and phytochemical compositions of pumpkin by-products along with their uses in functional food developments as a nutraceutical are highlighted.
... Adventitious shoots were sub-cultured on elongation medium composed of MS medium with 0.1 mg/l BA and the elongated shoots were successfully rooted in MS medium without growth regulators for 2 weeks. Flow cytometric analysis revealed that most of the regenerated plants were diploid (Young et al., 2003). In order to study the in vitro callus induction and plant regeneration in the asparagus plant and indirect organogenesis in explants, it was found that the presence of two plant growth regulators NAA And BAP are necessary for seedlings regeneration and callus induction in asparagus. ...
Growing regulators have enabled humans to track plant growth and have proven to be the most powerful tool in agriculturist hands for increasing yield and improving crop quality. As Indian agriculture becomes more mechanized and science Plant
growth regulator technologies that have an immediate impact on crop enhancement projects and are less time intensive would result in quantifiable gains for the customer. Plant growth regulators in vegetables equips clinicians and researchers with the
knowledge and expertise needed to effectively use these adaptable techniques to increase vegetable production extends the possibilities for using inputs to increase productivity and food safety, the role of plant growth regulators becomes more important. The majority of physiological processes and development in plants are regulated by the activity and association of hormones and some naturally occurring inhibitors. Plant growth regulators help to get better yield.
... of Nasarudin and Mansor (2021) who reported that BAP was essential for the growth of shoot or shoot proliferation in Momordica charantia. Also, Lee et al. (2003) reported that adventitious shoot bud regeneration was effectively achieved on MS medium supplemented with BAP alone for Cucurbita maxima. ...
... 19 Because of the abundance of health-promoting compounds and phytochemicals, it contains, pumpkin is used to treat a wide range of metabolic disorders. 20,21 Pumpkins are members of the Cucurbitaceae family, which has a wide variety of edible plants and is largely distributed across several countries. 22,23 Pumpkin seeds are healthy because they contain proteins, triterpenes, lignans, phytosterols, polyunsaturated fatty acids, and antioxidative phenolic components. ...
Background of the Study
The increase in the therapeutic use of tramadol in the management of moderate to severe pains in some disease conditions and its unregulated access has led to its associated toxicity and there is little or no information on the protection against its associated toxicity.
Aim of the Study
Considering the medicinal value of pumpkin seed oil, its availability, and neglected use, it becomes necessary to evaluate the possible potential of the seed oil in tramadol-induced oxidative stress in Wister Albino rats.
Methods of the Study
This study used fifty-six (56) albino rats to determine the impact of Cucurbita pepo seed oil (CPSO) on tramadol-induced oxidative stress. The rats were grouped into 7. After a week of acclimatization, rats in group 1 (normal control) had access to water and food, while rats in group 2 received 5 mL/Kg (b.w) of normal saline. 100 mg/kg of tramadol (TM) was delivered to groups 3–6 to induce toxicity. The third group (TM control) received no treatment, whilst the other 3 groups (TM-CPSO treatment groups) received 5, 2.5, and 1.5 mL/Kg of CPSO, respectively. Group 7 received only 5 mL/kg CPSO (CPSO group). Similarly, groups 2 through 7 had unrestricted access to food and water for 42 days and received treatments via oral intubation once per day. Indicators of oxidative stress were discovered in the brain homogenate.
Results
TM toxicity was demonstrated by a considerable increase (P < .05) in the brain MDA level and a significant drop (P < .05) in the brain GSH level, as well as a significant reduction (P < .05) in GPx, catalase, SOD, GST, and quinone reductase activities.
Conclusion
The dose-dependent delivery of CPSO was able to restore not only the activity but also the concentrations of the altered markers.
... Phytoestrogen supplementation with pumpkin seeds extract has been reported to increase uterine weight, mammary gland, bone density, and prevent hyperlipidemia, the indication of estrogen-like activities in ovariectomized female Sprague dawley rats (Gossell- Williams, et al., 2008). Pumpkin seeds oil contains rich vitamin E such as a-tocopherol and gtocopherol that exhibited positive health effects (Rabrenovic, et al., 2014) 3 . The researchers have so far focused particularly on the three major components of fatty acids, phytoestrogens, and tocopherol in pumpkin seeds oil because they gained attention due to the several health benefits such as antioxidant, anti-inflammation, antidiabetic, anticancer, anti-cardiovascular, antihyperlipidemia, and estrogenic-like effect. ...
The pumpkin belongs to the family of Cucurbitaceae, is a well-known edible plant that has been frequently used as functional food or herbal medicine. Pumpkins contain rich unsaturated fatty acids, phytoestrogens and vitamins E in their seeds that have potential pharmaceutical, nutraceutical, and cosmeceutical properties. Information regarding their nutritional components and therapeutic properties of pumpkin seeds has expanded dynamically in the recent years and this review focus on the three main components of pumpkin seeds that described before. Several types of unsaturated fatty acids are the dominant component in pumpkin seeds which can play a role in the disease prevention and promote health. Pumpkin seeds also contain the important phytoestrogen compounds, i.e., secoisolariciresinol and lariciresinol that have estrogenic-like effect such as preventing hyperlipidemia and osteoporosis for menopausal women. Phytoestrogens in pumpkin seeds also could be related to a reduced hormone-dependent tumor. Pumpkin seeds are rich in vitamin E contents as an emerging free radical scavenger, anti-aging and antioxidant such as a-tocopherol and g-tocopherol. Findings of these studies prove that patents field for the innovation product of pumpkin seeds holds promise for the future along with their immense nutraceutical properties.
... Pumpkin belongs to the genus Cucurbita, which is in the cucubitaceae family. Cucurbita pepo, Cucurbita mixta, Cucurbita maxima, and Cucurbita moschata are a few varieties of pumpkin plant species that have flourished as a vegetable in different regions across the globe [5]. Seeds and nuts are receiving more attention in recent years due to the significant medicinal and nutraceutical benefits of their bioactive compounds [6]. ...
Pumpkin, belongs to the family Cucurbitaceae and genus Cucurbita, is gaining appeal across the world for a number of reasons. Pumpkin seeds have always been discarded as waste even with containing essential pharmaceutical micro and macro constituents such as proteins, antioxidative phenolic compounds, tocopherols, triterpenes, saponins, phytosterols, lignans, and carotenoids as well as these compounds D-chiro-inositol, trigonelline, and nicotinic acid. Pumpkin seeds are also rich in fibre, polyunsaturated fatty acids, vitamins, and minerals including zinc, iron, magnesium, calcium, manganese, and copper that may be used in the food industry. Pumpkin seeds are now generally used in traditional medicine in treatment of many diseases, including hypertension, rheumatoid arthritis, hyperglycemia, inflammation, dyslipidemia, bacteria, fungal infections, and tumours. As a result, pumpkin seeds are now often utilised as a herbal treatment or health-improving agent for both people and animals, and food scientists are providing the pumpkin-infused products to the food and health industries. Hence, food manufacturers have worked to broaden the range of appetiser, baking, and snack uses for pumpkin seeds. This review article provides insights into the pharmacological activities of pumpkin seeds and the possible processes which, might reduce the chance of a wide range of problems.
... Pumpkin belongs to the genus Cucurbita and family Cucurbitaceae, which includes cucumber and squash, grown in tropical and subtropical countries. In the world there are three most common varieties of pumpkin named as Cucurbita maxima, Cucurbita moschata and Cucurbita pepo (Lee et al., 2003). The family of pumpkin is well known for its chemical composition including proteins, fibers, polysaccharides, fibers and minerals (Abuelgassim and Al-Showayman, 2012). ...
Present study was planned and conducted at Institute of Food Science and Nutrition, University of Sargodha and Food Technology Section, AARI Faisalabad during 2020, to develop and analyze pumpkin flesh powder (PFP) and to prepare, standardize and evaluate biscuits with 0, 5, 10 and 15% replacement levels of PFP. Fully ripe pumpkins were sliced, dried and grinded to obtain PFP. Straight grade flour (SGF) and PFP were analyzed for physico-chemical properties. PFP exhibited greater (6.15 ± 0.08) a* (redness) and (36.94 ± 0.19) b* (yellowness) values, significantly higher contents of moisture (14.29 ± 0.05%), ash (1.36 ± 0.03%), fat (1.24 ± 0.03%) and fiber (1.82 ± 0.01%), as compared to SGF. Upon analysis of physical characteristics of developed biscuits, as replacement level of PFP was increased to 15%, with SGF to develop biscuits, a* and b* values of biscuits were significantly increased to 5.06 ± 0.079 and 28.01 ± 0.14 respectively, thickness was also significantly increased to 48.50 ± 0.75 mm, while width was significantly decreased. Chemical analysis of biscuits revealed a significant increase in ash, fat fiber, Na, K and Fe contents of biscuits with increasing level of replacement of PFP. Whereas, non-significant results for moisture and protein contents were found. The contents of Ca, Mg and P were significantly decreased in added PFP biscuits. Sensory evaluation of biscuits resulted that 5 to 10% replacement level of PFP got good scores close to the control so up to 10% replacement level of PFP with SGF was acceptable to develop good quality, nutritional biscuits. By keeping in view, the nutritional profile of pumpkin an effort was made to highlight the nutritional significance of pumpkin, pumpkin powder and biscuits developed with addition of pumpkin powder.
... In the world there are three most common varieties of pumpkin named as Cucurbita maxima, Cucurbita moschata and Page 4 of 14 Hussain et al. Food Production, Processing and Nutrition (2023) 5:22 Cucurbita pepo (Lee et al. 2003). Pumpkin is available in many shapes, sizes and colors depending on the variety and climate. ...
Progression of today’s world has been given setback due to the adversity of a novel, viral, deadly outbreak COVID 19, which raised the concerns of the scientists, researchers and health related officials about the inherent and adaptive immune system of the living body and its relation with healthy diet balanced with pharma foods. Now world is coming out of the destructive pandemic era, the choice of right food can help to build and boost adaptive immunity and pumpkin due to excellent profile of functional and nutraceutical constituents could be the part of both infected and non-infected person’s daily diet. Vitamins like A, C and E, minerals like zinc, iron and selenium, essential oils, peptides, carotenoids and polysaccharides present in pumpkin could accommodate the prevailing deficiencies in the body to fought against the viral pathogens. In current post COVID 19 scenario adequate supply of healthy diet, balanced with pharma foods could play a basic role in boosting immune system of the populations. This review covers the pharmacological activities of pumpkin functional constituents in relation with COVID 19 pandemic. Pumpkins are well equipped with nutraceuticals and functional bioactives like tocopherols, polyphenols, terpenoids and lutein therefore, consumption and processing of this remarkable vegetable could be encouraged as pharma food due to its antihyperlipidemic, antiviral, anti-inflammatory, antihyperglycemic, immunomodulatory, antihypertensive, antimicrobial and antioxidant potential. Need of healthy eating in current post COVID 19 period is very crucial for healthy population, and medicinal foods like pumpkin could play a vital role in developing a healthy community around the globe.
Graphical Abstract
... Pumpkin belongs to the Cucurbitaceae family, and Curcurbita pepo L., Curcurbita maxima Duchesne and Cucurbita moschata Duchesne ex Poir are the three most common species available worldwide [1]. These species are the most important in terms of quantity and spread in the world [2]. ...
In this study color, texture, starch–pectin, total antioxidant capacity, microbial count, and microstructure of HPP-treated Violina pumpkin cubes were evaluated. Samples were treated at six different pressures (100 to 600 MPa–HPP100 to HPP600) for 3 min. Moisture, total soluble solids, and pH showed no significant differences between untreated (UNTR) and treated samples. Pumpkin tissue showed great structural modifications as changes in cell size and shape, cell wall damage, increased cell wall thickness, cell detachment and dehydration, and calcium ions deposition mainly from HPP300 to 600. UNTR samples showed the highest value of maximum and minimum cell elongation, perimeter segment, and a more regular cell wall thickness whereas HPP600 showed the lowest values for all these parameters. A noticeable difference was observed in HPP600 samples, with a difference in terms of color (ΔE 11.3 ± 1.9) and hardness (87.4 ± 27.8 N) compared to the UNTR ones (194.9 ± 37.9 N) whereas treatments at other pressures changed less markedly the color and texture. HPP200 could ensure a higher amount of starch and pectin availability while HPP200 and HPP400 showed the highest total antioxidants capacity. High-pressure treatment from HPP400 to 600 gave the highest destruction of microorganisms but negatively influenced the structural quality as well as texture and microstructure.
... The pumpkin belongs to the wide family of cucurbits (Cucurbitaceae), and the three most common varieties of pumpkin are called Cucurbita maxima, Cucurbita moschata, and Cucurbita pepo [1,2]. Europe is the second-largest producer with approximately 4.9 million tons of pumpkin, which is about 17.5% of world production [3]. ...
Innovations in food drying processes are usually aimed at reducing drying time and improving the overall properties of dried products. These are important issues from an economic and environmental point of view and can contribute to the sustainability of the whole process. In this study, the effects of ultrasonic treatment on the drying kinetics of pumpkin pulp are investigated, and mathematical models to predict the drying kinetics are analyzed and optimized. The results show that ultrasonic pretreatment significantly reduces drying time from 451 to 268 min, with optimal processing parameters at 90% of the maximum ultrasonic power and a processing time of 45 min. The total color change of the samples was the lowest at the obtained optimal processing parameters. Based on the values (RMSE and R²) of the investigated mathematical drying models, it was found that the Weibull model is the best fit for the experimental data and is considered suitable for the drying kinetics of ultrasonically pretreated pumpkin samples. In this study, an artificial neural network with 15 neurons in hidden layers was also used to model the drying process in combination with ultrasound pretreatment. The network had a performance of 0.999987 and the mean square error was 8.03 × 10⁻⁵, showing how artificial neural networks can successfully predict the effects of all tested process variables on the drying time/moisture ratio.
... It includes squash and cucumbers which are grown throughout the tropical and subtropical countries. There are three common types of pumpkin worldwide, namely curcurbita pepo, curcurbita maxima and C. moschata (Lee et al., 2003). Pumpkin can be found in many shapes, sizes and colors. ...
... Plant growth regulators are used in horticulture to improve crop growth by increasing fruit number, fruit set and size. Improved vegetative growth and production traits can increase crop productivity [11][12][13]. The productivity in the horticultural system often depends on the manipulation of the physiological activities of the crops by chemical means [14]. ...
The present experiment was carried out during Zaid 2021. In order to study the “Effect of Naphthalene Acetic Acid and Indole Acetic Acid on Growth, Yield and Quality of Muskmelon (Cucumis melo. L). The trail was conducted at Biotechnology polyhouse, Department of Horticulture, Naini Agriculture Institute and Prayagraj. The foliar spray two growth regulators like Naphthalene Acetic Acid and Indole Acetic Acid were spray separately at different concentration (NAA @10ppm, 20ppm, 30 ppm, 40 ppm, 50 ppm, 60 ppm, 70 ppm, 80 ppm. IAA @ 100 ppm, 200 ppm, 300 ppm, 400 ppm, 500 ppm, 600 ppm, 700 ppm, 800 ppm. The result revealed among all the treatments foliar application of NAA (80 ppm) is significantly gave positive impact on growth, yield and quality parameters. i.e. Vine length (153.96 cm), number of days for 1st male flower (33.6 Days), No. of days for 1st female flower (38.83 Days), No. of Node at which 1st Male flower appears ( 4 . 1 ), No. of Node at which 1st Female flower appears ( 5 . 83) No. of Male flowers (207.33), No. of Female flowers(5.3) fruits per plant (3), diameter of the fruit (22.53 cm), length of the fruit(13.4cm), fruit yield per plant (1.4 kg), fruit yield per hectare (12.46 t), total soluble solids (11.30 Brix°), gross returns (3,73,800 Rs/ha), net returns (2,39,844 Rs/ha) and Benefit: cost ratio (2.79) were obtained among all other treatments with foliar application of NAA @80 ppm compared to other treatments.
... Pumpkins (Cucurbita) belong to the Cucurbitaceae family, which is comprised of many species consumed as food and is mostly found in China, Pakistan, India, Yugoslavia, Argentina, the Mexican regions, America, and Brazil (Jia et al., 2003;Andrade-Cetto and Heinrich, 2005). Three main types are cultivated: Cucurbita pepo, Cucurbita maxima, and Cucurbita moschata (Lee et al., 2003). The health benefits of pumpkin seeds are attributed to their macro-and micronutrient composition; proteins, triterpenes, lignans, phytosterols, polyunsaturated fatty acids, antioxidative phenolic compounds, carotenoids, tocopherol, and minerals (Kim et al., 2012). ...
This study evaluates the potential effects of pumpkin seeds protein on blood pressure (BP), plasma adiponectin, leptin levels, and oxidative stress in rats with fructose-induced metabolic syndrome. Twenty four male Wistar albino rats were divided into four groups and fed a 20% casein diet, 20% casein diet supplemented with pumpkin protein, 20% casein diet with 64% D-fructose, or 20% casein diet with pumpkin protein and 64% D-fructose for 8 weeks. Contin-uous fructose feeding induced an increase in plasma insulin/glucose ratio, BP, insulin and glucose, aspartate aminotrans-ferase, alanine aminotransferase (ALT), alkaline phosphatase (ALP), creatinine, urea, and uric acid levels, and a decrease in the liver and muscle glycogen stores. In addition, elevated levels of total cholesterol (TC), triglycerides (TG), and leptin and lowered adiponectin levels were observed in rats fed a fructose-enriched diet. These groups also exhibited lower plasma levels of ascorbic acid and glutathione, higher thiobarbituric acid-reactive substances, hydroperoxide, carbonyl, and nitric oxide in both the liver and kidneys than rats fed the control diet. Interestingly, pumpkin seed protein treatment significantly counteracted alterations induced by fructose improving glucose, insulin, BP, TG, TC, ALT, and ALP levels, increasing liver and muscle glycogen stores, adiponectin level, and adiponectin/leptin ratio, and reducing plasma leptin lev-els. In addition, rats fed pumpkin protein with a high-fructose diet improved oxidative stress in the liver and kidneys. In conclusion, proteins from Cucurbita pepo L. seeds effectively improve metabolic parameters and protect against oxidative stress induced by a high-fructose diet.
... Agarwal and Kamal obtained shoot induction on MS medium fortified with BAP alone in Momordica charantia [12] . Lee et al., affirmed that adventitious shoot bud regeneration was effectively achieved on MS medium augmented with BAP alone for Cucurbita maxima [13] . ...
Tissue culture technique was developed for mass production of multiple shoots from valuable medicinal plant of Ammannia baccifera Linn. High frequency of direct shoot proliferation was induced in shoot tip explants cultured on Murashige and Skoog's medium supplemented with 6-benzylaminopurine (BAP). Among the various cytokinins tested (BAP, kinetin and Zeatin), BAP proved to be the most effective. The most suitable medium for shoot regeneration was MS medium fortified with 2.0 mg/l 6-benzylaminopurine (BAP), on which highest percentage of shoot response (80±1.90), the maximum number (7.8±1.40) of shoots per explant, having a shoot length of (2.8±1.01cm) were produced. The highest number of shoots multiplication response (85±1.68), the maximum number shoot per explants (28.6±1.63) and shoot length (3.0±1.12 cm) was documented on MS medium supplemented with 2.0 mg/l BAP and 0.5 mg/l NAA. Further shoot elongation was achieved on MS medium containing 0.5 mg/l GA3. The elongated micro shoots were rooted on MS medium fortified with 2.0 mg/l IAA and 0.8 mg/l IBA on highest percentage of rooting response (80±2.16), the maximum number of root per explants (18.4±1.50) and root length (4.2±1.34 cm) was recorded. The well developed shoot and rooted plantlets were successfully transferred into paper cups containing vermiculite, sand and soil in the ratio of 1:2:1 and subsequently they were established in the greenhouse. The results of this study provide the first successful report on in vitro direct plant regeneration from shoot tip explants of A. baccifera L.
... An efficient plant regeneration system is therefore necessary for genetic transformation and propagation of gherkin. During the past years, considerable efforts have been made for in vitro plant regeneration through indirect organogenesis of C. sativus using hypocotyl and cotyledon derived calli (Kim et al., 1988;Selvaraj et al., 2007), direct regeneration from cotyledon and hypocotyl explants of Cucurbita maxima (Lee et al., 2003), C. melo and C. sativus (Curuk et al., 2003). In spite of the availability in various reports on cucumber organogenesis, several problems are still encountered to regenerate cucumber species in tissue culture. ...
Gherkin (Cucumis anguria L.) is an important vegetable that belongs to the family of cucurbitaceae. The fruit of the vegetable contain very high amounts of protein, calcium, phosphorous, iron and vitamin C. The gherkin is also used for traditional medicinal purpose such as stomach ache, jaundice, hemorrhoids and preventing stone formation in kidney. It was previously reported that there are many useful phytochemical constituents from gherkin, for example cucurbitacin B, cucurbitacin D and cucurbitacin G. Cucurbitacin B have potential to be used as a favorable phytochemical for cancer prevention. The pickled gherkins ensure worldwide demand and many food companies have started to develop opportunities for large scale production of gherkin. We have established the in vitro culture and Agrobacterium mediated genetic transformation of C. anguria. This study was involved in the optimization of conditions for in vitro multiplication of important vegetable and medicinal Cucumis anguria. Specifically micropropagation, organogenesis and somatic embryogenesis (using different explant types) were employed to achieve large-scale multiplication from elite cultivars. We used shoot tip and nodal explants for micropropagation. Direct organogenesis from leaf and petiole explants of C. anguria. We established the somatic embryogenesis using suspension culture of geherkin. Additionally, the standardized protocols were also tailored for genetic transformation using disarmed Agrobacterium tumefaciens for transformants. Finally, the transgenic plants were confirmed by PCR and southern blotting. We have published many international papers for the evidence of my Ph.D work.
... Pumpkin (Cucurbita) seeds are traditionally known around the world for their remedial effects on urinary tract complications, such as nocturia, urinary frequency, and stress urinary incontinence [7]. Pumpkins belong to the Cucurbitaceae family which includes various species such as Cucurbita pepo, Cucurbita moschata, and Cucurbita maxima [8]. Cucurbita pepo seed oil consists of high amounts of free fatty acids serving as a natural source of vitamins, proteins, trace elements, and polyunsaturated fatty acids, such as omega 3, 6, and 9 [9]. ...
Background
Benign prostatic hyperplasia (BPH) is very common in aging men. We aimed to compare the effects of tamsulosin and pumpkin (Cucurbita pepo) seed oil on BPH symptoms.
Methods
This single-blind randomized clinical trial included patients with BPH aged ≥ 50 years referred to the Urology Clinic of Shahid Beheshti Hospital, Hamadan, Iran, from August 23, 2019 to February 19, 2020. Patients were randomized into two groups. One group received 0.4 mg tamsulosin every night at bedtime and the other received 360 mg pumpkin seed oil twice a day. Patients’ age, weight, height, and body mass index (BMI) were recorded. The International Prostate Symptom Score (IPSS) was filled out by the patients at baseline and then 1 month and 3 months after the initiation of treatment. The BPH-associated quality of life (QoL), serum prostate-specific antigen, prostate and postvoid residual volume, and maximum urine flow were also assessed at baseline and 3 months later. Drug side effects were also noted.
Results
Of the 73 patients included in this study with a mean age of 63.59 ± 7.04 years, 34 were in the tamsulosin group and 39 in the pupkin seed oil group. Patients were comparable with respect to age, weight, height, BMI, and baseline principal variables in both groups. Also, there was no significant difference between groups in terms of principal variables at any time point. However, there was a significant decrease in IPSS and a significant improvement in QoL in both groups. Although the decrease in IPSS from baseline to 1 month and 3 months was significantly higher in the tamsulosin group compared to the pumpkin group (P = 0.048 and P = 0.020, respectively), the decrease in IPSS from 1 to 3 months was similar (P = 0.728). None of the patients in the pumpkin group experienced drug side effects, while dizziness (5.9%), headache (2.9%), retrograde ejaculation (2.9%), and erythema with pruritus occurred in the tamsulosin group.
Conclusions
Pumpkin (Cucurbita pepo) seed oil relieved BPH symptoms with no side effects, but was not as effective as tamsulosin. Further studies are required to confirm the role of pumpkin seed oil as an option for the treatment of BPH symptoms.
Trial registration Iranian Registry of Clinical Trials, IRCT20120215009014N340. Registered 19.02.2020. Retrospectively registered, https://en.irct.ir/trial/45335.
... In the current work, although the Fe-EDDHA had a more benefical effect on meristem regeneration than AgNO 3 and H 3 BO 3 (Fig. 4D), none of the medium combinations containing Fe-EDDHA, AgNO 3 or H 3 BO 3 was the optimum medium composition. These results almost resembled previous works on in vitro regeneration of Lagenaria siceraria (Saha et al. 2007), Cucurbita maxima (Lee et al. 2003), T. vulgaris (Ozudogru et al. 2011). ...
Thymus cilicicus is an endemic Eastern Mediterranean element that has aromatic-medicinal properties. Its natural population spreads across gravelly ground and open rocky areas of South and Southwest Anatolia. The current study on in vitro propagation of T. cilicicus focused deeply on environmental applications such as the development of an optimum medium composition for efficient and genetically stable micropropagation and improved preservation procedures for long-time conservation of elite germplasms for further studies. For this purpose, MS and OM media were used individually and in combination with cytokinins, charcoal, AgNO3, Fe-EDDHA, and H3BO3. The raw data were statistically analyzed via the graph kernel method to optimize the nonlinear relationship between all parameters. The optimal proliferation medium for T. cilicicus was OM supplemented with a combination of 10 g L-1 charcoal and 1 mg L-1 KIN and the calculated averages of the best regeneration rate, the best shoot number and the best shoot length were 96.89%, 3 and 1.24 respectively on this medium. The determination of genetic stability of in vitro grown plants on the optimum medium compositions obtained by the graph kernel method was carried out with the use of the ISSR-PCR technique. All the ISSR primers produced a total of 192 reproductive band profiles, none of which were polymorphic. Furthermore, the micropropagated plants were successfully rooted and acclimatized to greenhouse conditions. In this study, we present a graph kernel multiple propagation index which considers all the possible parameters needing to be analyzed. Such an index is used for the first time for the determination of the optimum proliferation medium.
... Adventitious shoots were sub-cultured on elongation medium composed of MS medium with 0.1 mg/L BA and the elongated shoots were successfully rooted in MS medium without growth regulators for 2 weeks. Flow cytometric analysis revealed that most of the regenerated plants were diploid [112] . The presence, NAA and BAP is necessary for in vitro seedlings regeneration and callus induction in asparagus and indirect organogenesis in explants. ...
Cucurbits are group of cross pollinated, warm season fruit vegetable
which are vine in nature and holds a very important place with respect to
nutrition and economy. As climbing in nature these plants requires support to
grow vertically for maximum vegetative growth, fruit yield and quality.
Research findings suggested that trailing of vines on trellis structures
improved better vegetative growth, higher fruit yield and superior quality
over the ground trailing. Again regular pruning of the excessive leaves and
non-productive vines has shown great improvement with respect to
increasing the plant productivity, optimal use of resources and reducing the
disease and pest pressure. Traditionally cucurbit farmers follow the
indigenous training methods which may increase the fruit yield over the
ground trailing, but the productivity can be enhanced further by adopting
newly developed, cost effective trellis methods. This work covers the various
importance of training and pruning practices in cucurbits and the different
scientific research findings in recent years on different trellis structure and
pruning methods in different cucurbitaceous vegetables. Dissemination of
these training practices on cucurbits will surely benefit a large number of
growers. Moreover a small architectural modification can bring more yield,
improved fruit quality, better profitability and also adds aesthetic beauty in
production system.
... Agarwal and Kamal obtained shoot induction on MS medium fortified with BAP alone in Momordica charantia [12] . Lee et al., affirmed that adventitious shoot bud regeneration was effectively achieved on MS medium augmented with BAP alone for Cucurbita maxima [13] . ...
Tissue culture technique was developed for mass production of multiple shoots from valuable medicinal plant of Ammannia baccifera Linn. High frequency of direct shoot proliferation was induced in shoot tip explants cultured on Murashige and Skoog's medium supplemented with 6-benzylaminopurine (BAP). Among the various cytokinins tested (BAP, kinetin and Zeatin), BAP proved to be the most effective. The most suitable medium for shoot regeneration was MS medium fortified with 2.0 mg/l 6-benzylaminopurine (BAP), on which highest percentage of shoot response (80±1.90), the maximum number (7.8±1.40) of shoots per explant, having a shoot length of (2.8±1.01cm) were produced. The highest number of shoots multiplication response (85±1.68), the maximum number shoot per explants (28.6±1.63) and shoot length (3.0±1.12 cm) was documented on MS medium supplemented with 2.0 mg/l BAP and 0.5 mg/l NAA. Further shoot elongation was achieved on MS medium containing 0.5 mg/l GA3. The elongated micro shoots were rooted on MS medium fortified with 2.0 mg/l IAA and 0.8 mg/l IBA on highest percentage of rooting response (80±2.16), the maximum number of root per explants (18.4±1.50) and root length (4.2±1.34 cm) was recorded. The well developed shoot and rooted plantlets were successfully transferred into paper cups containing vermiculite, sand and soil in the ratio of 1:2:1 and subsequently they were established in the greenhouse. The results of this study provide the first successful report on in vitro direct plant regeneration from shoot tip explants of A. baccifera L.
... The preparation process for squash explants is similar to that for cucumbers: immature cotyledons are excised from 1-day-old seedlings. However, especially in the case of squashes, care should be taken not to remove the cotyledonary nodes around the shoot apex by breaking off the hypocotyl ( Figure 2C), because only this part retains regeneration ability, as reported previously (Ananthakrishnan et al. 2003;Kathiravan et al. 2006;Lee et al. 2003;Zhang et al. 2008). For the purpose of generating more transgenic shoots, we use the proximal parts of cotyledons, cut longitudinally into two pieces, as explants ( Figure 2B, D). ...
Cucumber (Cucumis sativus L.) and Cucurbita species (squashes, pumpkins, and gourds), belonging to the Cucurbitaceae family, are among the major vegetable crops in the world. Transgenic approaches could contribute to the accumulation of new knowledge of these species and to the development of elite cultivars. Despite this, research reports using transformants of these species are very limited so far. One of the reasons for this may be that although there are effective transformation methods, these methods are not well known among researchers. In the present review, we describe efficient protocols for the transformation of cucumber and squash plants and mention possible pitfalls in and advice for following these protocols. In addition, we discuss the current progress of genetic transformation research using cucumbers and squash, including genome editing.
... Members of the genus Cucurbita are used by different human cultures across the globe and are a notable source for nutrition and medicinal treatments. Cucurbita maxima produces the largest number of different fruit shapes of any species in this economically important clad, and misidentification of the species can lead to confusion when using this cucurbit for botanical research [12]. The species epithet of Cucurbita maxima suggests vigorous growth of fruit, which is the most massive of any cucurbit. ...
Cucurbita maxima is an incredibly diverse species, and it is suggested to have more cultivated forms than any other crop. It has many medicinal uses including anti-diabetic, anti-oxidant, anticancer, and anti-inflammatory properties. It is also a major food source for wide variety of world cultures, due to its fiber content, carbohydrates, β-carotene, vitamins, alkaloids, minerals, fatty acids, flavo-noids, and polysaccharides. Detection of root-associated microbes in C. maxima has not been well documented in the scientific lit-erature. A multi-phase approach was implemented to first verify fungal associations in C. maxima, and then second, to document the colonization of fungi in C. maxima grown in a conventional agricultural system. Cucurbita maxima grown in southwest Ohio showed relationships with members of the Zygomycota and Plasmodiophorid Slime Molds. This study provides a first report of a relationship between Olpidium spp. and Plasmodiophorid Slime Molds in C. maxima in the United States. The images provided in this manuscript are the first photographic documentation of these organisms in C. maxima to date. This is also a first report of Plasmodiophorid Slime Molds in any member of the family Cucurbitaceae
A method of Agrobacterium mediated plant regeneration using petiole explants and transformation of pBI121 CaMV 35S:GUS construct and MDC123 CaMV 2 × 35S:Cas9:MCAGL6-sgRNA construct having Bar gene as plant selection marker was optimized for the first time in bitter melon. Different combination of cytokinins (BAP, Kinetin, TDZ) and auxins (2,4-D, IAA, NAA) were used in callus induction media (CIM) and shoot induction media (SIM) to regenerate plants from petiole (P) and leaf (L) explants taken from three varieties (BSS578, KSP1522, and KSP1277) and abbreviated as LCIM/PCIM and LSIM/PSIM. Leaf calli failed to induce shoot regeneration despite of 44.66 to 46.33% callus induction. However, significantly higher percentage (90.9%) of multiple shoot proliferation found in PSIM2 using petiole as explant with the callus induction of 57% in KSP1522. Based on the varietal response for callus and shoot induction, petiole explant mediated transformation protocol was selected and optimized for pre-culture period, acetosyringone concentration and phosphinothricin concentration (Bar gene as a selectable marker). Agrobacterium mediated transformation of GUS with plant selection marker kanamycin (80 mg L−1) showed 10–11% transformation efficiency using 3 days old pre-cultured explants and 250 µM acetosyringone. Similar transformation efficiency percentage was observed for plant transformation of gene with phosphinothricin (0.5 mg L−1) as plant selection. Thus, protocol efficiently works for both GUS and Bar insertion into bitter melon plants. The in vitro regeneration of bitter melon plant using petiole explant was more effective than leaf explant and variation in the percentage of plant regeneration capacity was highly varietal responsive.
Phytohormones are one of the predominant players in regulating
out and out plant growth as well as development. Majority of the
physiological and biochemical processes in the sessile plant are regulated
and coordinated by the activity of phytohormones such as abscisic acid
(ABA), gibberellins (GAs), ethylene (ETH), auxin (indole-3-acetic acid
(IAA)) and cytokinins (CKs). These five phytohormones (auxin, CK, GA,
ABA, and ETH) are sometimes referred to as the “classical”
phytohormones. There are some more recent additions to the family of
phytohormones viz- brassinosteroids (BR), salicylates
(SA), jasmonates (JA) and single peptides. The role of phytohormones is
to regulate genetically programmed developmental changes or reactions
to environmental stimuli by acting near or far from their sites of synthesis.
External application of phytohormones can be done for improvement of
plant growth, yield, and photosynthetic pigments by modulation of
physiological and biochemical processes. Therefore, it is crucial to study
and understand the physiological role played by these phytohormones in
plants so as to explore their all-out utility in boosting vegetable
production.
Pumpkin (Cucurbita maxima), belonging to the family Cucurbitaceae, is a well-known edible plant, cultivated and abundantly used as herbal medicine and functional food. Pumpkins have long been used in traditional medicine to treat a wide range of conditions, including intestinal parasites, tumors, hypertension, arthritis, hyperglycemia, inflammation, dyslipidemia, and bacterial or fungal infections.
Virus diseases are a major production constraint for pumpkin. Recessive resistance to squash leaf curl China virus and tomato leaf curl New Delhi virus has been mapped in Cucurbita moschata (Duchesne ex Poir.) breeding line AVPU1426 to chromosomes 7 and 8, respectively. Molecular markers tightly associated with the resistance loci have been developed and were able to correctly predict resistance and susceptibility with an accuracy of 99% for squash leaf curl China virus resistance and 94.34% for tomato leaf curl New Delhi virus in F2 and back cross populations derived from the original resistance source AVPU1426. The markers associated with resistance are recommended for use in marker-assisted breeding.
Genetic engineering is commonly used to improve the agronomic traits of crops. However, genetic transformation in pumpkin remains a challenge. Conducting transformation trials, we accidentally created transgenic L1 periclinal chimeras in pumpkins.
Using our modified Agrobacterium ‐mediated transformation, we generated transgenic L1 periclinal chimeras which have high value in research on development of the meristem.
Fluorescence observations of transformed L1 cells enabled us to reveal cell fates. These L1 cells can develop into stomata, epidermal hairs, seed coat, and epidermis of the root, stem, leaf, flower, and fruit. These periclinal chimeras can be propagated vegetatively with minimal risk of transgene flow.
This study offers new perspectives on development of the meristem and a promising technique for creating transgenic periclinal chimeras in plants.
Nowadays, agricultural waste byproducts are exploited in the food industry rather than discarded. Pumpkin is one of the most significant vegetable crops that is widely consumed in farmland and certain urban regions. The current study was designed to measure the phytochemical constituents, food application, health benefits, and toxicity of pumpkin and pumpkin byproducts. Pumpkins and pumpkin byproducts (seeds, leaf, and skin/peel) can be utilized as functional ingredients. Different parts of the pumpkin contain bioactive compounds including carotenoids, lutein, zeaxanthin, vitamin E, ascorbic acid, phytosterols, selenium, and linoleic acid. Pumpkin is used in various food sectors as a functional food, including baking, beverages, meat, and dairy industries. Furthermore, the leaves and pulp of the pumpkin are used to produce soups, purees, jams, and pies. Different parts of pumpkins have several health benefits such as antidiabetic, antioxidant, anticancer, and anti-inflammatory effects. Therefore, this review paper elaborates on the pumpkins and pumpkin byproducts that can be used to develop food products and may be valuable against various diseases.
A study was conducted to examine the physical and chemical properties of pumpkin. The analysis focused on fresh and ripe pumpkin to assess their physical and chemical composition. The findings revealed that pumpkins exhibited a yellow to golden yellow color, indicating a significant presence of β-carotene at 11.23 mg/100g, making them a rich source of antioxidants. Additionally, the study analyzed the chemical composition, revealing a moisture content of 6.10%, carbohydrate content of 5.46%, protein content of 0.95%, and fat content of 0.21%. Furthermore, the study measured the acidity of the pumpkin at 0.008% and the pH value at 4.57. These results provide valuable insights into the chemical properties of pumpkin, highlighting its nutritional composition.
Colour is one of the important attributes which influences the acceptance of a food product. Carotenoids are a class of compounds which impart colour into food products. The micronutrient in the fresh pumpkin such as carotene was found to be in good amounts (8 mg /100 g), while minerals such as P (27.8 mg/100 g) and Fe (1.5 mg/100 g) were present in considerable amounts. In the present study, attempts were made to store the carotene rich pumpkin flour (PF) at both ambient and refrigerated temperature for 6 months and its application in the preparation of vermicelli. The PF and PF fortified vermicelli samples were evaluated for the stability of β-carotene during storage. It was observed that at 4°C, the retention of β-carotene in PF was 25.5% at 6 months, while at 30°C, the retention was 11.45% for the same duration. The retention of β-carotene in vermicelli was higher at higher levels of PF incorporation (85.19%) as compared to lower levels of application (6.35%). However, a significant decrease in carotene during storage was observed in both PF incorporated vermicelli and PF stored under both refrigerated and ambient conditions.
Pumpkin (Cucurbita maxima), belonging to family Cucurbitaceae, is a well-known edible plant, cultivated and abundantly used as herbal medicine and functional food. This review aimed to investigate functional and nutraceutical constituents present in all three parts (peel, flesh and seeds) of pumpkin, and health benefits of these nutrients. Pumpkin fractions are rich source of nutritional components like oils, proteins, carbohydrates and minerals. Certain classes of phytochemical compounds including, phenolics, flavonoids, tocopherols, carotenoids, terpenoids, cucurbitacin, moschatin and phytosterols have been discovered from pumpkin fractions. Considerable evidences from several epidemiological studies upon animals and human trials have been present about pharmacological actions of pumpkin bio actives. Therapeutically, pumpkin peel, flesh and seeds contain active ingredients, acting as effective antioxidants and antimicrobial agents. No part of pumpkin fruit should be discarded during processing as these parts possess excellent phytochemistry, capable of impacting positive health impacts. All these parts of pumpkin can be used in the form of powders or extracts to isolate and characterize bio-actives and to utilize them in food and pharmaceutical industries as therapeutic agents. Instead of consumption in raw form, technologies should be implemented to develop novel and innovative nutraceuticals and pharma foods from health beneficial constituents of pumpkin.
The objective of this study was to investigate the effects of ultrasonic treatment on the aroma profile of pumpkin juice. Samples were treated according to the experimental design, with a numerically controlled ultrasonic instrument at an ultrasound frequency of 45 kHz under various conditions (treatment times of 10, 20, and 30 min, sample temperature of 25(± 2)∘C, and powers of 180, 240, and 300 W). The aromatic profiles of pumpkin juice showed that a total of 52 volatile components were detected in the 10 groups of pumpkin juices, including 16 alcohols, 11 alkane olefins, 6 aldehydes, 4 ketones, 4 esters, and 11 others. Alcohol compounds had the highest content among all volatile compounds; the most abundant were 1-Octen-3-ol, (E, Z)-3, 6-Nonadien-1-ol, 1-hexanol, and (Z) -3-Nonen-1-ol. Compared to the untreated samples of juices, ultrasonic treatment leads to the formation of new compounds or the disappearance of compounds found in the untreated samples. The findings presented in this study provide a deeper understanding of the application of ultrasound to enhance the volatile components of pumpkin juice.
In poly cystic ovary syndrome multiple cysts formation occurs in female ovaries. It is highly prevalent in developing countries like Pakistan and India. Its symptoms include irregular menstrual cycle, abnormal hair growth pattern, acne and mood swings. This disease is also associated with imbalance in androgen levels (which include testosterone and estrogen) and high insulin levels also called hyperinsulinemia. It also contributes in the development of type 2 diabetes, heart disease and cancers. Seed cycling is a modern method used in alternate medicine of treating PCOS by using different seeds in the different stages of menstrual cycle. In this review article we will find out how seed cycling is effective in the treatment of PCOS. Flax, sesame, pumpkin and sunflower seed combinations are being used at different stages of female menstrual cycle. Flaxseeds contain lignans, omega 3 fatty acids and fibre which help in relieving the symptoms of polycystic ovaries. Sesame and sunflower seeds contain high number of fatty acids and lignans which helps in balancing hormonal disturbances in female. Losing weight by having diet rich in fibre improve the level of cholesterol and it is also beneficial decreasing the level of insulin resistance, which if not treated can progressed into type 2 diabetes mellitus and its complications. Herbal medicine in Ayurveda has shown promising effects in relieving pre- menstrual symptoms, hormonal fluctuations and the co-morbidities.
The Cucurbita genus, often called as cucurbits, include several economically important fruits and vegetable crops like cucumber, gourds, squash, watermelon, pumpkin, and melon. Several of the species including Cucurbita pepo and C. maxima have nutritional value and are utilized in folk medicine for treating gastrointestinal diseases and intestinal parasites. Such an activity is attributed to the presence of fatty acids, glycosides, resins, sterols, carotenoids, phenols, tocopherols, saponins, steroids, and terpenoids such as cucurbitacins. Squash and pumpkin are the major members of Cucurbitaceae family used as food and animal feed. The two major pumpkin varieties C. pepo and C. maxima are considered as highly polymorphic with manifold nutritional, including food preservative abilities and medicinal activities. Development of transgenic pepo for tolerance to various biotic and abiotic stresses coupled with nutritional value is the need of the hour. Very few reports are available on transgenic production of pepo. The present review summarizes the regeneration and transformation protocols for developing transgenic C. pepo and C. maxima and future prospects.
Cucurbita maxima is a species of cucurbit native to Central America, that has been shown to have many medicinal uses includ-ing anti-diabetic, anti-cancer, and anti-inflammatory properties. This economically important member of the Cucurbitaceae is not thoroughly documented in the scientific literature, and this manuscript fills in a major gap that is missing by providing the first high resolution Scanning Electron Microscopy images of Cucurbita maxima pollen to date. The individual pollen grain is a monad with apolar spherical morphology, echinate projections, and is 100μm in diameter. Researchers conducting palynological or paleopalynol-ogy studies will be able to use these images as a literature reference for Cucurbita maxima.
Nearly 80% of herbaceous plants form relationships with Arbuscular Mycorrhizal Fungi (AMF), which are often beneficial to the host, by acquiring limited resources, such as phosphorus and water. The cucurbit, Cucurbita maxima is an incredibly diverse species, and suggested to have more cultivated forms than any other crop species. C. maxima has many medicinal uses, including anti-diabetic, anti-oxidant, anticancer, and anti-inflammatory properties, and is also a major food source due to its fiber content, carbohydrates, β-carotene, vitamins, alkaloids, minerals, fatty acids, flavonoid, and diverse polysaccharide content. Currently, the scientific literature is missing definitive documentation of an AMF association with Cucurbita maxima in the United States of America. This study utilizes light microscopy as supportive evidence to show an arbuscular mycorrhizal relationship with different varieties of Cucurbita maxima.
Transformation of cucumber cv. Endeavor was attempted using three Agrobacterium tumefaciens strains (a supervirulent leucinopine type, an octopine type and a nopaline type), each harbouring one of three binary vectors which contained an acidic chitinase gene from petunia, and basic chitinase genes from tobacco and bean, respectively, driven by the CaMV 35S promoter. Petiole explants were inoculated with a bacterial suspension (10(8) cells·ml(-1)), cocultivated for 48-96 h and placed on Murashige and Skoog (MS) medium with 5.0 μM each of 2,4-D and BA, 50 mg·l(-1) kanamycin and 500 mg·l(-1) carbenicillin. The frequency of embryogenic callus formation ranged from 0 to 12%, depending on strains/vectors used and length of cocultivation, with the highest being obtained using the leucinopine strain with petunia acidic chitinase gene. The kanamycin-resistant embryogenic calli were used to initiate suspension cultures (in liquid MS medium with 1.0/1.0 μM 2,4-D/BA, 50 mg·l(-1) kanamycin) for multiplication of embryogenic cell aggregates. Upon plating of cell aggregates onto solid MS medium with 1.0/1.0 μM NAA/BA and 50 mg·l(-1) kanamycin, calli continued to grow and later differentiated into plantlets. Transformation by the leucinopine strain and all three vectors was confirmed by PCR amplification of the NPT II gene in transgenic calli and plants, in addition to Southern analysis. Expression of the acidic chitinase gene (from petunia) and both basic chitinase genes (from tobacco and bean) in different transgenic cucumber lines was confirmed by Western analyses.
[Link at : http://link.springer.com/article/10.1007/BF00232459]
A cDNA expression library in lambda MOSEIox derived from poly(A)+ RNA from pumpkin endosperm was screened immunologically with a polyclonal antibody raised against partially purified gibberellin (GA) 2 beta,3 beta-hydroxylase from pumpkin endosperm. A recombinant fusion protein encoded by a selected positive clone catalyzed 3 beta-hydroxylation of GA15, GA24, GA25, and GA17 and of GA12-aldehyde, GA12, GA9, and GA20, albeit less efficiently. The fusion protein also catalyzed 2 beta-hydroxylation of the C20 GAS GA25, GA13, and, as identified putatively, GA28. The full-length clone contains an open reading frame of 1041 nucleotides encoding 346 amino acid residues with a predicted molecular weight of 38,992 and pI of 7.2. Transcript levels of this gene and of the previously cloned GA 7-oxidase and 20-oxidase genes from pumpkin endosperm rose until day 2 after the start of imbibition of the mature seeds, but only at one-two hundredth to one-six thousandth of the level found in the endosperm, as determined by quantitative reverse transcriptase-polymerase chain reaction. In contrast, GA 7-oxidase, 20-oxidase, and 3 beta-hydroxylase enzyme activities were present in cell-free systems prepared from embryos of mature seeds and decreased after imbibition.
The first step in gibberellin biosynthesis is catalyzed by copalyl diphosphate synthase (CPS) and ent-kaurene synthase. We have cloned from pumpkin (Cucurbita maxima L.) two cDNAs, CmCPS1 and CmCPS2, that each encode a CPS. Both recombinant fusion CmCPS proteins were active in vitro. CPS are translocated into plastids and processed by cleavage of transit peptides. For CmCPS1 and CmCPS2, the putative transit peptides cannot exceed the first 99 and 107 amino acids, respectively, because longer N-terminal deletions abolished activity. Levels of both CmCPS transcripts were strictly regulated in an organ-specific and developmental manner. Both transcripts were almost undetectable in leaves and were abundant in petioles. CmCPS1 transcript levels were high in young cotyledons and low in roots. In contrast, CmCPS2 transcripts were undetectable in cotyledons but present at significant levels in roots. In hypocotyls, apices, and petioles, CmCPS1 transcript levels decreased with age much more rapidly than those of CmCPS2. We speculate that CmCPS1 expression is correlated with the early stages of organ development, whereas CmCPS2 expression is correlated with subsequent growth. In contrast, C. maxima ent-kaurene synthase transcripts were detected in every organ at almost constant levels. Thus, ent-kaurene biosynthesis may be regulated through control of CPS expression.
Six summer squash ( Cucurbita pepo L.) cultivars were regenerated via somatic embryogenesis using cotyledons excised from germinated or nongerminated seeds. Genotypes included were zucchini, commercial F 1 hybrids, `President', `Seneca Zucchini', `Jade'; the noncommercial inbred line `Caserta Inbred 557311'; and two yellow squash hybrids `Dixie' and `Seneca Butterbar'. Somatic embryogenesis was initiated in induction medium containing 22.62 μ m 2, 4-D, and embryos were germinated in maturation medium containing 0.27 μ m NAA and 0.23 μ m kinetin. Plants were elongated and rooted on basal medium without hormones. All media contained carbenicillin at 500 mg·liter –1 . Sixty-one percent of the `Seneca Butterbar' cotyledons produced somatic embryos when kept on induction medium for 10 weeks. Overall, 7% of the initial explants produced plantlets, and regeneration efficiency was calculated as 0.3 plantlets per initial explant. The relative production of plants from cotyledons that were kept on induction medium for different time periods were determined for `Caserta Inbred 557311' and `Seneca Zucchini'. All cotyledons produced somatic embryos after 11 to 17 weeks on induction medium. However, plantlet production was optimal with explants kept on induction medium for 13 weeks for `Seneca Zucchini' and for 15 weeks for `Caserta Inbred 557311', producing an average of 4.5 and 9.3 plants per explant, respectively, from 90% to 70% of the explants. We recovered plants from all six cultivars; thus, our regeneration protocol may be applicable to other genotypes. The high percentage of regenerants obtained indicates that the regeneration method is efficient enough to be adapted successfully to squash transformation experiments. Chemical names used: α-carboxybenzylpenicillin (carbenicillin); 2,4-dichlorophenoxyacetic acid (2,4-D); 6-furfurylaminopurine (kinetin); α-napthaleneacetic acid (NAA).
The first step in gibberellin biosynthesis is catalyzed by copalyl diphosphate synthase (CPS) and ent-kaurene synthase. We have cloned from pumpkin (Cucurbita maxima L.) two cDNAs, CmCPS1 and CmCPS2, that each encode a CPS. Both recombinant fusion CmCPS proteins were active in vitro. CPS are translocated into plastids and processed by cleavage of transit peptides. For CmCPS1 andCmCPS2, the putative transit peptides cannot exceed the first 99 and 107 amino acids, respectively, because longer N-terminal deletions abolished activity. Levels of both CmCPStranscripts were strictly regulated in an organ-specific and developmental manner. Both transcripts were almost undetectable in leaves and were abundant in petioles. CmCPS1 transcript levels were high in young cotyledons and low in roots. In contrast,CmCPS2 transcripts were undetectable in cotyledons but present at significant levels in roots. In hypocotyls, apices, and petioles, CmCPS1 transcript levels decreased with age much more rapidly than those of CmCPS2. We speculate that CmCPS1 expression is correlated with the early stages of organ development, whereasCmCPS2 expression is correlated with subsequent growth. In contrast, C. maxima ent-kaurene synthase transcripts were detected in every organ at almost constant levels. Thus,ent-kaurene biosynthesis may be regulated through control of CPS expression.
Plant regeneration from tissue cultures of summer squash (Cucurbita pepo L. ev. YC60) has been observed. Embryogenic callus tissues were initiated when cotyledons of mature seeds were excised and cultured on Murashige and Skoog (MS) medium supplemented with either 22.7 μ m 2,4-D or a combination of 4.7 μ m 2,4,5-T, 4 μ m BA, and 0.5 μ m kinetin. Clusters of somatic embryos were found in callus tissue. Maturation of these somatic embryos was effected by transfer of embryogenic callus tissues to MS supplemented with 0.5 μ m NAA and 0.25 μ m kinetin. Regenerated mature plants were morphologically normal and set fruits containing seeds that germinated normally. Chemical names used: 6-benzylaminopurine (BA); 2,4-dichlorophenoxyacetic acid (2,4-D); α - napthaleneacetic acid (NAA); 2,4,5-trichlorophenoxyacetic acid (2,4,5-T).
Five different genotypes from in vitro as well as greenhouse grown melon plants were shown to be highly responsive for in vitro shoot formation from leaf explants when placed on basic MS medium supplemented with 1 mg/l 6-benzylaminopurine. In addition, a very suitable regeneration system was obtained when cotyledon pieces of mature seeds were incubated on the same culture medium. In this case, the first shoots already appeared after 10 days of incubation, and hundreds of shoots were formed on the cut surface 3 to 4 weeks later. Explants from mature cotyledons derived from seedlings did not lead to any shoot formation.
Embryogenic callus derived from pumpkin hypocotyl segments was induced and maintained for 15 years on MS medium supplemented with the auxins IBA (4.9 μM), 2, 4-D (4.5 μM) or IAA (5.7 μM). On induction media continued embryo maturation and development of adult plants typically failed. Therefore, small embryogenic clumps and individually isolated embryos were subcultured two to four times on one of the conversion media: MS supplemented with 1.5% sucrose and (a) no hormone, (b) 2.9 μM IAA, (c) 5.7 μM IAA, (d) 11.4 μM IAA, (e) 12 μM IEt, (f) 3.8 μM ABA or (g) 2% activated charcoal. The cell line and the kind of auxin used in the induction and maintenance medium, both had a marked influence on the development of plantlets. The best result was achieved with a line that has been induced and maintained for 15 years on MS with IBA. In the IBA line, out of 100 embryos, 77 developed into plantlets on MS medium supplemented with 11.4 μM IAA.
Plant regeneration from tissue cultures of summer squash, Cucurbita pepo L., cv. YC60, has been observed. Somatic embryos organized from shoot apex derived callus cultured on Murashige and Skoog (MS) medium supplemented with 1.2 mg/l 2,4,5-trichlorophenoxyacetic acid, 0.8 mg/l benzylaminopurine, and 0.1 mg/l kinetin. Embryos developed into plantlets by transfer of immature somatic embryos to MS medium with 0.05 mg/l NAA and 0.05 mg/l kinetin. Regenerated plants appeared morphologically normal and set fruits with seeds which could germinate normally.
Adventitious shoots formed on the proximal cut edges of different cotyledonary explants of watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai; cvs. Sweet Gem and Gold Medal] cultured on Murashige and Skoog's (MS) medium with 1 mgl(-1) 6-benzyladenine (BA). Light (16-h photoperiod, about 7 Wm(-2) cool-white fluorescent lamps) was essential for shoot formation. To obtain transformed plants, cotyledonary explants of 'Sweet Gem' were cocultured with Agrobacterium tumefaciens LBA4404, a disarmed strain harboring a binary vector pBI121 carrying the CaMV 35S promoter-β-glucuronidase (GUS) gene fusion used as a reporter gene and NOS promoter-neomycin phosphotransferase gene as a positive selection marker, for 48 h on MS medium with 1 mgl(-1) BA and 200 μM β-hydroxyacetosyringone. After 48 h of culture, explants were transferred to medium with 1 mgl(-1) BA 250 mgl(-1) carbenicillin, and 100 mgl(-1) kanamycin and cultured in the light. Adventitious shoots formed on the explants after 4 weeks of culture. When subjected to GUS histochemical assay, young leaves obtained from the shoots showed a positive response at a frequency of up to 16%. Preculturing cotyledonary explants on MS medium with 1 mgl(-1) BA for 5 d enhanced the competence of the cells to be transformed by Agrobacterium. Southern blot analysis confirmed that the GUS gene was incorporated into the genomic DNA of the GUS-positive regenerants. The transformed plants were grown to maturity.
Cotyledon explants of muskmelon (Cucumis melo L., cv. Amarillo Oro) seedlings were co-cultivated with disarmed Agrobacterium tumefaciens strain LBA4404 that contained the binary vector plasmid pBI121.1. The T-DNA region of this binary vector contains the Nopaline synthase/neomycin phosphotransferase II (NPTII) chimeric gene for kanamycin resistance and the Cauliflower Mosaic Virus 35S/β-glucuronidase (GUS) chimeric gene. After infection, the cotyledon pieces were placed in induction medium containing 100 mg/l kanamycin. Putative transformed shoots were obtained, followed by the development of morphologically normal plantlets. The transgenic nature of regenerants was demonstrated by polymerase chain reaction, Southern blot analysis, plant growth on medium selective for the transgene (NPTII) and expression of the co-transformed GUS gene. Factors affecting the transformation procedure are discussed.
A method of high frequency shoot induction was established using excised hypocotyl segments bearing cotyledons (referred to as cotyledonary hypocoryl explants) of cucumber (Cucumis sativus L.) seedlings. Adventitious shoots were obtained from the upper excised hypocotyls of cucumber seedlings incubated on Murashige and Skoog (MS) medium containing 2.0 mg/L zeatin. The effects of cotyledon size and hypocotyl length of seedlings on shoot organogenesis were also examined. A higher frequency (80–90%) of adventitious shoots was obtained from explants of 3- and 5-day-old seedlings with either 1 cotyledon or 2 half-cotyledons, and a short hypocotyl length (2 mm). However, explants with long hypocotyls and 2 cotyledons produced fewer shoots. Adventitious shoots from cotyledonary hypocotyl explants were removed and rooted in MS medium containing 1.0 mg/L IAA. The rooted plantlets developed into normal plants after acclimatization to ambient humidity levels.
The frequency of tetraploidy in melon plants (Cucumis melo L.) regenerated from somatic embryos, adventitious shoots, shoot primordia and in plants propagated from axillary buds was compared. Tetraploids were found in plants regenerated from somatic embryos, adventitious shoots and shoot primordia, and the frequency of tetraploids in the three culture methods was 31%,30% and 4%, respectively. No tetraploidy was observed in plants propagated from axillary buds. The highly frequent appearance of tetraploidy in regenerated plants appears to be a universal phenomenon in tissue culture of melon.
A single regeneration procedure using cotyledon examples effectively regenerated five commercially grown muskmelon cultivars. This regeneration scheme was used to facilitate gene transfers using either Agrobacterium tumefaciens or microprojectile bombardment methods. In both cases, the transferred genes were from the T-DNA region of the binary vector plasmid pGA482GG/cp cucumber mosaic virus-white leaf strain (CMV-WL), which contains genes that encode neomycin phosphotransferase II (NPT II), [beta]-glucuronidase (GUS), and the CMV-WL coat protein (CP). Explants treated with pGA482GG/cpCMV-WL regenerated shoots on Murashige and Skoog medium containing 4.4 [mu]m 6-benzylaminopurine (BA), kanamycin (Km) at 150 mg[center dot]liter[sup [minus]1] and carbenicillin (Cb) at 500 mg[center dot]liter[sup [minus]1]. The authors' comparison of A. tumefaciens- and microprojectile-mediated gene transfer procedures shows that both methods effectively produce nearly the same percentage of transgenic plants. R[sub 0] plants were first tested for GUS or NPT II expression, then the polymerase chain reaction (PCR) and other tests were used to verify the transfer of the NPT II, GUS, and CMV-WL CP genes.
A rice chitinase cDNA (RCC2) driven by the CaMV 35S promoter was introduced into cucumber (Cucumis sativus L.) through Agrobacterium mediation. More than 200 putative transgenic shoots were regenerated and grown on MS medium supplemented with 100 mg/l kanamycin.
Sixty elongated shoots were examined for the presence of the integrated RCC2 gene and subsequently confirmed to have it. Of
these, 20 were tested for resistance against gray mold (Botrytis cinerea) by infection with the conidia: 15 strains out of the 20 independent shoots exhibited a higher resistance than the control
(non-transgenic plants). Three transgenic cucumber strains (designated CR29, CR32 and CR33) showed the highest resistance
against B. cinerea: the spread of disease was inhibited completely in these strains. Chitinase gene expression in highly resistant transgenic
strains (CR32 and CR33) was compared to that of a susceptible transgenic strain (CR20) and a control. Different responses
for disease resistance were observed among the highly resistant strains. CR33 inhibited appressoria formation and penetration
of hyphae. Although CR32 permitted penetration of hyphae, invasion of the infection hyphae was restricted. Furthermore, progenies
of CR32 showed a segregation ratio of 3:1 (resistant:susceptible). As the disease resistance against gray mold was confirmed
to be inheritable, these highly resistant transgenic cucumber strains would serve as good breeding materials for disease resistance.
Two-week-old in vitro grown Cucurbita pepo L. intact plants and cotyledons (detached and undetached from the mother-plant)
were transformed by Agrobacterium rhizogenes strain NCPPB 1855, grown for 48 h at 25 °C on YMB medium. All infected material
formed vigorous hairy roots in about seven days. The transformed roots were successfully grown in liquid MS medium without
plant growth regulators for an indefinite number of transfers. Genetic transformation of root DNA was proven by Southern analysis
performed with a rolABC probe and a vir probe. Our results demonstrate that, in contrast with previous literature, A. rhizogenes
could represent an efficient and reproducible system to transform C. pepo plants. Furthermore, we verified that plant age
and incubation times/temperatures of bacterial strain influence transformation efficiency.
Embryogenic callus cultures were established from immature cucumber(Cucumis sativus L.) embryos on E20A (Dumas de Vaulxet al. 1981) or MS (Murashige and Skoog 1962) media supplemented with 6-benzylaminopurine (BAP), α-naphthylacetic acid (NAA) and/or
2,4-dichlorophenoxyacetic acid (2,4-D). Regeneration of plants was observed after a transfer to culture media either without
growth regulators or supplemented with kinetin and NAA. Flow cytometry was employed to estimate DNA ploidy levels. Most of
cell nuclei in young leaf tissues were found in G1 phase with 2C DNA content. Callus cultures were mixoploid with DNA content ranging from 2C to 32C. The frequency of polyploid
cells was increasing with the age of culture and the polyploidization was accompanied by a gradual loss of regeneration ability.
Plants regenerated from callus cultures were classified as diploid (57 %), tetraploid (18 %), octoploid (4 %) and mixoploid
(2n/4n, 4 %) and (4n/8n, 17 %). The results of this study confirmed a close link between the polyploidization and the loss
of totipotencyin vitro. Tetraploid plants obtained in this study have a potential to be used in interspecific crosses where their tetraploid status
could help in overcoming existing breeding barriers due to differences in chromosome number.
Transgenic cucumber (Cucumis sativus L.) plants were successfully obtained from hypocotyl explants inoculated with Agrobacterium tumefaciens, which harbored a binary vector plasmid with NOS-nptII, CaMV 35S-I-gus and CaMV 35S-hph genes. Acetosyringone enhanced the efficiency of transformation at the cut surface cells of hypocotyl explants during five days of co-cultivation. Transformed cells were more effectively selected using 20–30 mg/l hygromycin B than using 50–100 mg/l kanamycin. Shoot regeneration occurred within 4–6 wks, and 12 of 21 regenerated plantlets displayed strong GUS expression in the very young leaves. All of 8 GUS-positive R0 plants examined showed single or a few positive bands by Southern blot analysis. The expression of the CaMV 35S-I-gus gene was observed in various tissues and organs of R0 and R1 transgenic cucumber plants.
The generation of transgenic cucumber (Cucumis sativus L.) plants was achieved by biolistic transformation of a highly embryogenic cell suspension culture using the nptII and uidA gene. Functional expression of the genes in transgenic plants was determined by neomycin phosphotransferase and β-glucuronidase enzyme assays. Southern analysis of DNA isolated from kanamycin-resistant plants confirmed stable integration of the genes as well as multicopy integration and rearrangements. A study of gene expression showed activity of the uidA gene in plants regenerated from kanamycin-resistant calli about one year after bombardment, indicating a high stability of the nonselectable gene.
An efficient system of transformation leading to a majority of transformed diploid plants from leaf explants of Cucumis melo L. var. Cantalupensis (cv. Védrantais) was developed. Several regeneration protocols using cotyledon or leaf explants were analysed with particular emphasis on the regeneration efficiency and the ploidy level of the regenerated melon plants. The use of leaf explants excised from 10 day-old seedlings, cultured in Murashige and Skoog’s medium supplemented with 1 μM 6-benzylaminopurine (BAP) and 1 μM 6-(γ,γ-dimethylallylamino)-purine (2iP), resulted in a high regeneration frequency (73%). In these conditions, more than 84% of the regenerated plants were found to be diploid. Addition of an Agrobacterium-mediated transformation step did not significantly change the percentage (81.8%) of diploid plants regenerated. This protocol was successfully used to produce diploid transgenic melon plants expressing the antisense ACC oxidase gene, encoding ACC oxidase which catalyses the last step of ethylene biosynthesis. Ethylene production and ACC oxidase activity of the leaf explants from transgenic plants was reduced by more than 80% as compared to the control untransformed tissues. This transformation/regeneration method could be routinely used for the introduction of other genes of interest in melon.
A cDNA for thylakoid-bound ascorbate peroxidase of pumpkin was cloned and characterized. Thylakoid-bound ascorbate peroxidase had a high similarity to cytosolic ascorbate peroxidases, and the precursor contained a transit peptide to chloroplasts at its amino-terminus and a putative membrane-spanning region at its carboxy-terminus.
Three cDNA clones (cat1, cat2, cat3) for catalase (EC 1.11.1.6) were isolated from a cDNA library of pumpkin (Cucurbita sp.) cotyledons. In northern blotting using the cDNA-specific probe, the cat1 mRNA levels were high in seeds and early seedlings of pumpkin. The expression pattern of cat1 was similar to that of malate synthase, a characteristic enzyme of glyoxysomes. These data suggest that cat1 might encode a catalase associated with glyoxysomal functions. Furthermore, immunocytochemical analysis using cat1-specific anti-peptide antibody directly showed that cat1 encoding catalase is located in glyoxysomes. The cat2 mRNA was present at high levels in green cotyledons, mature leaf, stem and green hypocotyl of light-grown pumpkin plant, and correlated with chlorophyll content in the tissues. The tissue-specific expression of cat2 had a strong resemblance to that of glycolate oxidase, a characteristic enzyme of leaf peroxisomes. During germination of pumpkin seeds, cat2 mRNA levels increased in response to light, although the increase in cat2 mRNA by light was less than that of glycolate oxidase. cat3 mRNA was abundant in green cotyledons, etiolated cotyledons, green hypocotyl and root, but not in young leaf. cat3 mRNA expression was not dependent on light, but was constitutive in mature tissues. Interestingly, cat1 mRNA levels increased during senescence of pumpkin cotyledons, whereas cat2 and cat3 mRNAs disappeared during senescence, suggesting that cat1 encoding catalase may be involved in the senescence process. Thus, in pumpkin, three catalase genes are differentially regulated and may exhibit different functions.
A cDNA encoding stromal ascorbate peroxidase (sAPX) was isolated using poly(A)+ RNA of pumpkin cotyledons by RT-PCR. The cDNA encodes a polypeptide with 372 amino acids and shares complete sequence identity with pumpkin thylakoid-bound ascorbate peroxidase (tAPX), except for the deletion of a putative membrane spanning region located in the carboxy domain of tAPX. Southern blot hybridization and analysis of intron structure indicated that mRNAs for sAPX and tAPX, whose suborganellar localizations in chloroplasts are different, are produced by alternative splicing. Immunoblot analysis showed that the accumulation of sAPX and tAPX was differently regulated during germination and subsequent greening of pumpkin cotyledons.
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Cloning and expression of a gibberellin 2[beta],3[beta]-hydroxylase cDNA from pumpkin endosperm
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