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Cultivar identification of ‘Yuzu’ ( Citrus junos Sieb. ex Tanaka) and related acid citrus by leaf isozymes
Abstract
Leaf extracts of 27 ‘Yuzu’ and related acid citrus cultivars were analyzed using polyacrylamide gel electrophoresis for isozyme variation of glutamate oxaloacetate transaminase (GOT) and shikimate dehydrogenase (SDH). SDH yielded 12 different isozyme phenotypes and six cultivars were discriminated by this enzyme alone. GOT produced 10 different isozyme phenotypes and four cultivars were separated. When both enzyme systems were taken together, 16 cultivars (59%) were uniquely discriminated and the rest could be classified into four groups of 2–4 cultivars each. Mutation originated cultivars could not be discriminated. Differences between cultivars suggested that isozymes may provide useful markers for cultivar identification.
... These techniques provide a better estimation of the genetic diversity and structure of populations ( WEISING et al. 1995;ANDRÉS et al. 1999). Isozymes are closely related to gene products and their electrophoretic mobility, resulting from differences in size and shape of enzyme molecules, are good indicators for genetic diversity ( RAHMAN et al. 2000). Many researches have used electrophoresis of isoenzymes as an efficient tool for classification and discrimination between species of higher plants ( ELLSTARND & LEE 1987;APAVATJRUT et al. 1999). ...
... Many researches have used electrophoresis of isoenzymes as an efficient tool for classification and discrimination between species of higher plants ( ELLSTARND & LEE 1987;APAVATJRUT et al. 1999). Studies on isoenzymes have been accomplished on a great number of pine species from North America and also some Mediterranean pines, Pinus sylvestris populations and P. halepensis-brutia complex in Europe ( SCHILLER et al. 1986;SCALTSOYIANNES et al. 1994;ANDRÉS et al. 1999;RAHMAN et al. 2000). The present study was conducted to compare and discriminate wild type with conical and ball shaped types, using the epicuticular wax layer and its micromorphological variability on stem, needles and seeds, peroxidase activity and variation of its isozymes as well as lignin and protein content. ...
Mondell pine (Pinus eldarica Medw.) is a rare and drought resistant pine, found naturally only in semi desert environment southeast of Tbilisi, Georgia, and has been probably introduced to Iran more than 800 years ago. This species has gradually altered both in shape and growth rate in Nashtifan-Khaf, and generated two species nova which so called as ball shaped and conical, according to characteristics of their shape and smaller stature. This study was conducted to discriminate these species nova using epicuticular wax features and certain biochemical characteristics i.e., peroxidase electrophoretic pattern (PAGE), the activity of soluble (SPO), ionically (IPO) and covalently (CPO) wall-bound fractions of peroxidase, lignin and total protein content. Observation via scanning electron microspopy showed differences that in adaxial and abaxial surface of needles, and also seed surface, in ball shaped type with the others. But none of the biochemical characteristics had significant difference in these three types. The electrophoretic analysis showed that the peroxidase isozyme is not suitable marker for discriminating the species nova from wild type, in spite of obvious micro- and macromorphological differences. Results of current study, propose the use of molecular markers and cytogenetic studies in order to specify cause of species nova occurrence in the study area. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)
... Therefore, the knowledge of genetic variation and relationship among different genotypes is an important consideration for strengthening the citrus industry in India. Morphological (Jaskani et al., 2006;Altaf and Khan, 2008 ;Sharma et al., 2004) and isozyme markers (Ashari et al., 1989;Elisirio et al., 1999;Rahman et al., 2001 ;Fang et al., 1997) have been employed in assessing the underlying genetic variation of citrus genotypes, however, these do not portray the reliable genetic relationships among the genotypes. This is due to the limited number of morphological and biochemical markers, low level of polymorphism, unknown genetic control of these traits, environmental interactions, stage specific identification and inadequate sampling of the genome . ...
Genetic variability and fingerprint profiles of 19 indigenous and exotic mandarin genotypes introduced from different parts of India and USA were determined using 60 SSR markers. Of the 57 SSR markers amplified, a total of 96 alleles were detected by 39 polymorphic SSR loci and maximum 5 alleles were amplified with an average of 2.46 alleles per primer pair. The CAT01 was the highly informative marker as it revealed maximum number of alleles (5), PIC value (0.75) and genetic diversity (0.79). Twenty six SSRs revealed specific/unique alleles and identified nine genotypes including all the hybrids. Across the genotypes, maximum number of alleles (83) was detected in Daisy hybrid and the percentage of polymorphic marker was maximum (80.32) in Nova hybrid. The markers with low number of alleles were able to differentiate the varieties with specific alleles. The higher average expected heterozygosity (35.6%) with in a mandarin group as compared to the average observed heterozygosity (27.2%) may be explained by selfing, which reduced the proportion of heterozygotes. The genotypes were classified in three clusters i.e. cluster-I, cluster-II and cluster-III. All the indigenous genotypes (selections) were grouped in cluster-I and it had maximum genetic similarity coefficient. However, the exotic genotypes (hybrids) were grouped in cluster-II and cluster-II. Clustering was according to the breeding history of genotypes but independent to their geographic origin. The low observed heterozygosity frequency, PIC value, and number of alleles explained the narrow genetic base in the present set of mandarin genotypes.
... The diversity and genetic relatedness in Citrus spp. have been evaluated by isozymes technique since 1980 [9]. The most influential carcinogen is benzo(a)pyrene (BaP), which is embryo toxic and teratogenic in animals [10]. ...
... Therefore, the knowledge of genetic variation and relationship among different genotypes is an important consideration for strengthening the citrus industry in India. Morphological (Jaskani et al., 2006;Altaf and Khan, 2008 ;Sharma et al., 2004) and isozyme markers (Ashari et al., 1989;Elisirio et al., 1999;Rahman et al., 2001 ;Fang et al., 1997) have been employed in assessing the underlying genetic variation of citrus genotypes, however, these do not portray the reliable genetic relationships among the genotypes. This is due to the limited number of morphological and biochemical markers, low level of polymorphism, unknown genetic control of these traits, environmental interactions, stage specific identification and inadequate sampling of the genome . ...
Genetic variability and fingerprint profiles of 19 indigenous and exotic mandarin genotypes introduced from different parts of India and USA were determined using 60 SSR markers. Of the 57 SSR markers amplified, a total of 96 alleles were detected by 39 polymorphic SSR loci and maximum 5 alleles were amplified with an average of 2.46 alleles per primer pair. The CAT01 was the highly informative marker as it revealed maximum number of alleles (5), PIC value (0.75) and genetic diversity (0.79). Twenty six SSRs revealed specific/unique alleles and identified nine genotypes including all the hybrids. Across the genotypes, maximum number of alleles (83) was detected in Daisy hybrid and the percentage of polymorphic marker was maximum (80.32) in Nova hybrid. The markers with low number of alleles were able to differentiate the varieties with specific alleles. The higher average expected heterozygosity (35.6%) with in a mandarin group as compared to the average observed heterozygosity (27.2%) may be explained by selfing, which reduced the proportion of heterozygotes. The genotypes were classified in three clusters i.e. cluster-I, cluster-II and cluster-III. All the indigenous genotypes (selections) were grouped in cluster-I and it had maximum genetic similarity coefficient. However, the exotic genotypes (hybrids) were grouped in cluster-II and cluster-II. Clustering was according to the breeding history of genotypes but independent to their geographic origin. The low observed heterozygosity frequency, PIC value, and number of alleles explained the narrow genetic base in the present set of mandarin genotypes.
... The literature available on C. junos as a host for citrus bacterial canker is very limited (Table 2). Often, the data provided are given for a single uncharacterised C. junos accession (Table 2), although different accessions are well known and characterised (Rahman et al., 2001). A single study (Deng et al., 2010), investigating the susceptibility level of five Chinese C. junos accessions, suggested that levels of partial resistance are cultivar-dependent. ...
Following a request from the European Commission, the European Food Safety Authority (EFSA) Plant Health (PLH) Panel analysed a dossier submitted by the Japanese authorities in order to clarify the host status of Citrus junos with regard to Xanthomonas citri pv. citri and Xanthomonas citri pv. aurantifolii, causal agents of citrus bacterial canker, and to indicate whether C. junos fruit could represent a pathway for the introduction of citrus bacterial canker into the European Union. In a previous opinion in the year 2014, the EFSA PLH Panel concluded that commercial fresh citrus fruit is generally pathway and that no commercially important Citrus species or variety can be considered as immune to citrus bacterial canker. In the current assessment, the EFSA PLH Panel analysed the two scientific papers provided by the Japanese authorities, as well as 16 additional papers identified through a systematic literature review. The PLH Panel considered that the conclusions of its previous opinion remain valid and that convergent lines of evidence provide sufficient demonstration that C. junos is a host of X. citri pv. citri and X. citri pv. aurantifolii. Therefore, there is no reason to consider the C. junos fruit differently from other citrus species. Consequently, the assessment of the general citrus fruit pathway from the 2014 opinion still applies. Uncertainties on these conclusions are a result of the scarce scientific evidence published on this subject in addition to the methodological and reporting limitations of the published papers.
... Isozymes have been used as biochemical markers to study taxonomic relationships between Citrus species (Protopapadakis and Papanikolaus 1999). This method is useful in distinguishing cultivars derived by sexual reproduction but does not generally distinguish between cultivars differentiated by mutations (Roose 1988;Herrero et al. 1996;Rahman et al. 2001). In the recent two decades, a wealth of molecular phylogenetic investigations have been carried out to examine genetic relationships among Citrus and its relatives. ...
Application of molecular markers, have now been increasingly adopted to address the problems in Citrus taxonomy. Compared to morphological data, molecular tools provide abundant information, highly efficient and are insensitive to environmental factors. Molecular markers has provided an ideal means for identifying genotypes, estimation of relatedness between different accessions and following inheritance of economically important characters. In Citrus, a wide variety of DNA based markers has been used in order to study their genetic variation as well as phylogenic and taxonomic relationship among different genera. RAPD markers provide a fast and easy approach for taxonomic classification and cultivar-typing of Citrus fruits. SSR have proven to be the marker of choice in Citrus breeding research, because of their variability, ease of use, accessibility of detection and reproducibility. ISSR, SRAP, CAPS-SNP, AFLP are also used to study the genetic diversity of Citrus throughout the world. In addition, cpDNA is especially useful in phylogenetic analyses due to its evolutionary conservatism, relative abundance in plant tissue, small size and pre dominant uniparental inheritance.
... Seedling morphology, such as leaf shape and thorniness, was carefully compared with nucellar seedlings obtained from polyembryonic seeds. Only those suspected to be nucellar were analyzed for hybridity with three glutamate oxaloacetate transaminase isozyme genes, Got-1, Got-2, and Got-3, according to the procedure described by Rahman et al. (2001) and Ngo et al. (2011). Zygotic seedlings with the hybrid characteristics were grafted onto 20-year-old 'Miyagawa-wase' satsuma mandarin trees to enhance the growth of the seedlings. ...
... Hal ini merupakan indikator yang baik unruk keragaman genetik. sehingga isoenzim dapat digunakan untuk mengidentifikasi kultivar pamelo (Phan at al. 2006), Citrus junos dan kerabat jeruk asam (Rahman at al. 2001). membedakan bibit jeruk triploid dan diploid (King at al. 1996). ...
... Since 1980, the isozymes technique has been used to evaluate diversity and genetic relatedness in Citrus spp. (Ashari et al., 1989;Elisirio et al., 1999;Rahman et al., 2001). More recently, new molecular techniques, such as RAPD, RFLP (Abkenar et al., 2004;Jena et al., 2009), AFLP (Campos et al., 2005;JinPing et al., 2009) and microsatellite markers (Koehler et al., 2003;Ghanbari et al., 2009) have been used to identify Citrus spp. with high accuracy. ...
This study aimed to identify Bhutanese mandarin accessions using morphological characteristics. A total of 30 accessions were selected from six districts in the major mandarin-growing regions in Bhutan. Samples of 15 leaves, 5 flowers and 15 fruit were collected randomly from each accession. A total of 23 characters were evaluated from the trees, leaves, flowers and fruit from each plant. A dichotomous key constructed from the morphological qualitative characters was able to classify the accessions to the district level. There were statistically highly significant differences (P < 0.01) for leaf length, leaf width, fruit weight, fruit diameter, fruit length, epicarp width, number of seeds and total soluble solids (TSS). The floral morphology showed little variation among trees and districts. The accessions from Dagana had the highest fruit weight with 107.2 g followed by Samtse, Zhemgang, Sarpang, Trongsa and Tsirang with fruit weights of 96.1, 93.3, 87.6, 78.9 and 58.3 g, respectively. The highest TSS were found in the accessions from Trongsa (12.9 °Brix) followed by Zhemgang (12.1 °Brix), Dagana (11.2 °Brix), Sarpang (10.9 °Brix), Tsirang (10.6 °Brix) and Samtse (10.3 °Brix). Accessions from Samtse were early maturing (mid October) while accessions from Tsirang were late maturing (February–March).
... Isozymes have been used as biochemical markers to study taxonomic relationships between citrus species (Rahman and Nito, 1994;Protopapadakis and Papanikolaus, 1999). This method is useful in distinguishing cultivars derived by sexual reproduction but does not generally distinguish between cultivars differentiated by mutations (Roose, 1988;Herrero et al., 1996;Rahman et al., 2001). Recently, various DNA markers have been used extensively to study phylogenetic relationships among plants (Whitkus et al., 1994). ...
Inter-simple sequence repeat (ISSR) markers were used to study phylogenetic relationships among 33 citrus genotypes including several undefined local or native varieties as well as some known varieties in the Fars Province of Iran.Based on 234 total ISSR fragments and 209 polymorphic fragments, a genotype similarity dendrogram was constructed using UPGMA cluster analysis. The 33 genotypes were classified into six major groups with, within-group similarity values of ≥0.65. Group 1 included Citrus sinensis and Citrus reticulata, group 2 common sour orange and ‘Peach’ sour orange, Group 3 ‘Bakraee’ and Volkameriana and three selections of sweet lime. ‘Bakraee’ is an undefined variety with morphological similarities to both sweet lime and mandarin. It is probably a hybrid between these two varieties. Group 4 included Lisbon lemon (Citrus limon) and two undefined selections ‘Rock lemon’ and ‘Pear-shaped lemon’. They might be mutant or hybrid lemons. Group 5 included a single genotype (D3) with little molecular similarity to other genotypes. Group 6 could be divided into two subgroups. One including citron (Citrus medica) and three local varieties of ‘Otroj’, ‘Bidkhoni’ from Darab and ‘Bidkhoni’ from Fassa. Based on their morphological and molecular similarities with citron, they are regarded as citron variants. The other subgroups of group 6 included Citrus aurantifolia, Citrus latifolia and 11 undefined genotypes with close morphological and molecular similarities to lime. The latter were considered as lime variants.
... Swingle (1967) has hypothesized that 'Yuzu' is a hybrid between Ichang papeda (Citrus ichangensis) and mandarin, whereas Tanaka (1954) described 'Yuzu' as a high-quality species and claimed that it was not a hybrid. Hirai et al. (1986) have reported that 'Yuzu' is not a hybrid between Ichang papeda and mandarin (Rahman et al., 2001). 'Alibert' and 'Rangpur' were clustered in the same branch at a similarity level of ~0.78. ...
Citrus production with its many varieties is of importance since it provides economically important products for Turkish exports. Sour orange is a rootstock commonly used for propagating the different scion varieties. Knowledge of the genetic diversity of the rootstock accessions would be useful in order to improve citrus breeding programs. We studied genetic relationships and diversity of 51 accessions of sour orange (Citrus aurantium) and their relatives using SSR (simple sequence repeat) and SRAP (sequence-related amplified polymorphism) molecular markers. Twenty-one SRAP primer combinations were tested on these accessions and relatives, producing 167 polymorphic fragments, with a mean of 8.0 and a mean polymorphism information content value of 0.47. Seventeen SSR primers also produced 30 polymorphic fragments, with a mean of 1.4 per primer and a mean polymorphism information content value of 0.39. The unweighted pair-group method with arithmetic average analysis using combined SSR and SRAP data showed a similarity range from 0.71 to 1.00 among the accessions. In the cluster analysis, sour orange relatives were indicated as a separate group from sour orange. 'Macrophylla' and 'Mexican lime' were the accessions most distinct (0.71) from the others. We conclude that genetic diversity in these sour orange accessions is lower and some of them were identical.
... Each cultivar though closely linked to each other by their appearance, they had somewhat difference in the insect-resistance and/or aroma. Taxonomists identified yuzu cultivars by its morphology (leaf shape, flower colour, fruit size and seed) or using isozyme analysis (Rahman, Nito, & Isshiki, 2001). It is the fact that yuzu is available in a diversity of cultivars and its flavour has been extensively studied. ...
The essential oils of six different yuzu cultivars, Kumon (KUM), Nagano (NAG), Yasu (YAS), Jimoto (JIM), Komatsu Sadao (KOS) and Komatsu Koichi (KOK), were extracted by cold-pressing method. A total of 69 compounds of the six samples were identified. Application of GC-olfactometry and aroma extraction dilution analysis technique in three-fold stepwise dilution of the neat oil for all samples indicated eight odourants with the highest flavour dilution (FD) values. Those were limonene, α-pinene, α- and β-phellandrene, myrcene, γ-terpinene, (E)-β-farnesene and linalool. ‘KOS’ was differentiated from the other oil samples by showing the highest number of components having yuzu-like odour notes and also from the PCA analysis of the FD-factor values. This is the first time the aroma characteristics of yuzu essential oils of specified cultivars were investigated.
In order to consideration and distinguishing of Mondell pine species (Pinus eldarica) and its two natural generated morphotypes, needles samples were collected from three years old seedlings in similar conditions and then extracted. Protein band patterns of the samples were studied and compared using electrophoresis method on polyacrylamide gel in the SDS-PAGE system. 12 morphological traits related to seedlings and mature individuals of the three pines, compared with each other. The amount of total protein also obtained using Bradford procedure and content of chlorophyll a and b measured by spectrophotometer with Harborne method. One dimensional electrophoresis of proteins showed perfectly similar isozyme patterns for each type (i.e. Mondell pine, Ball-shaped and Conical-shaped pines). Furthermore the amount of total protein and chlorophyll a and b contents of samples did not showed significant difference using ANOVA test. But 11 features of morphological measured traits were more in Mondell pine than the new generated pines. These results indicate the nonexistence of biochemical differences among these pines in spite of obvious structural and morphological differences.
In order to distinguish the Mondell pine species (Pinus eldarica) from its two natural generated
morphotypes (i.e. Ball-shaped and Conical-shaped pines), qualitative alteration of peroxidase enzyme and
the assessment of the enzyme activity were compared by use of polyacrylamide electrophoretic method
on PAGE system. Furthermore, lignin contents of the needle and stem samples along with 13 macro
morphological characteristics of individuals were compared with each other. Results showed that the
isozyme pattern of three types of the studied pines, the activity of this enzyme and lignin contents were
similar. Whereas, the morphological traits comparison, indicate clear differences in the pines. These, state
that despite of obvious morphological differences and application of peroxidase enzyme in physiological
changes, plant evolution studies, genetic consideration and identifying genus, species and subspecies but
at least soluble portion is not suitable marker for distinction of these pines. The results of this study also,
propose to use of molecular markers and cytogenetically studies in order to specify the cause of this
occurrence.
طبق آمارنامه محصولات باغبانی سال 1395 وزارت جهاد کشاورزی، سطح بارور باغ¬هاي¬ كشور 4/2 ميليون هكتار می-باشد که دراینبین، ميوه¬هاي نیمه گرمسیری 5/28 درصد از سطح زیر کشت محصولات باغبانی را به خود اختصاص داده است. مرکبات یکی از مهمترین انواع میوههای گرمسیری و نیمه گرمسیری محسوب میشوند که نقش عمدهای را در تولید محصول و تأمین سلامت جامعه به جهت ارزش غذایی و طعم مطبوع دارا میباشند. امروزه ارقام هیبرید مرکبات سبب ایجاد پتانسیلهای جدید در تولید و مصرف این میوهها شده است. شکل، طعم و ارزش غذایی خاص آنها سبب افزایش بازارپسندی گردیده است. در این مقاله مروری به معرفی چند میوه هیبرید به نام¬های تجاری بکرایی،Papeda ,Chironja, Yuzu, Kabbad, Blood lime, Orangelo, Tangelo که خصوصیات مطلوبی دارند، پرداخته می¬شود. برخی از این خصوصیات عبارتاند از: گسترش فصل برداشت، بهبود صفات کیفی (طعم، عطر، رنگ گوشت و پوست، شکل) و صفات کمی (اندازه میوه و اندازه بذر، عمر انبارداری ، بافت، اسیدیته، قند، ترکیبات باارزش غذایی و دارویی).
Randomly Amplified Polymorphic DNA (RAPD) markers were used to evaluate genetic similarity and interrelationship among 31 acid citrus species and cultivars, including sour oranges (six accessions); 'Yuzu' (four accessions) and its relatives (21 accessions). Out of the 60 decamer primers screened, 27 were selected which produced 108 markers; 76 of which were polymorphic. Species or cultivar-specific RAPD markers were also found. A dendrogram based on genetic distance implied that sour oranges were very distinct from 'Yuzu' and its relatives. 'Yuzu' accessions were very closely linked to each other, however; for the other specimens genetic polymorphism could easily be detected by RAPDs and the genetic variation between accessions was quite high and revealed their different origins. In this study some RAPDs allowed the distinction of very close cultivars, for instance 'Kabosu' from 'Aka kabosu'.
Yuzu (Citrus junos) is the most popular commercial acid citrus grown in Japan; however, the botanical origin of Yuzu as hybrid or non-hybrid Citrus species is confusing and controversial. In order to clarify the hybrid origin of Yuzu and its relationship with other acid citrus cultivated in Japan, two experiments were carried out in this study, in the first; PCR-RFLP analyses of three regions for each of chloroplast DNA (cpDNA; rbcL-ORF106, trnD-trnT, trnH-trnK) and mitochondrial DNA (mtDNA; nad7/exon2-exon3, nad7/exon3-exon4, 18s-5s) were performed in 26 cultivars of acid citrus, three true Citrus species, i.e., mandarin, pummelo and citron; and one papeda, ichang papeda (C. ichangensis). In the second experiment those two unique cpDNA restriction patterns of Yuzu found in one region of chloroplast DNA (trnH-trnK) were assessed in 22 mandarins to find their maternal relationship. Based on the results of this study only four cultivars of acid citrus, 'Matsuda Sudachi', 'Zuishoyu', 'Hanayu' and 'Yuko' showed both cpDNA and mtDNA restriction patterns as same as Yuzu. Also, the two unique cpDNA restriction patterns of Yuzu were not observed in the mandarin accessions, therefore the hybrid origin of Yuzu was not confirmed in this study. The data revealed that Yuzu acted as female progenitor in the origin of four acid citrus grown in Japan.
In acid citrus, genetic control of male sterility, an important characteristic for breeding seedless fruit cultivars, is not precisely known because of the presence of barriers such as polyembryony and a long juvenile phase. In this study, 22 crosses with 16 male-fertile acid citrus cultivars were carried out and the zygotic seedlings were grafted onto adult satsuma mandarin trees to enhance flowering. Four crosses with two monoembryonic and male-sterile citrus plants (HY16 and 'Kiyomi') were also carried out and zygotic seedlings showing precocious flowering were used to examine the inheritance of male sterility. Of the 26 crosses, 21 with six cultivars as seed parents generated male-sterile and male-fertile zygotic seedlings with various segregation ratios, whereas five crosses with 'Sudachi' as a seed parent generated only male-fertile zygotic seedlings. The sterile anthers were categorized into undeveloped and underdeveloped by their size in most progenies. The result of these crosses showed that eight cultivars with male-sterile cytoplasmic factors of 'Yuzu', lemon, or satsuma mandarin generated male-sterile zygotic seedlings, while 'Sudachi' with male-fertile cytoplasmic factors of pummelo generated only male fertile zygotic seedlings. The result also suggested that a dominant nuclear fertility-restorer gene system comprising one epistatic gene R1 and two complementary genes R2 and R3 controls the restoration of male fertility and male-sterile anther size in acid citrus plants with sterile cytoplasm (S). The complementary gene R3 is located downstream from R2, and gene R1 is epistatic to R2 and R3 genes. Genotypes (S)r1r1r2r2-- are responsible for undeveloped anthers while genotypes (S)r1r1R2-r3r3 are responsible for underdeveloped anthers. The identified genotypes for male sterility in major acid citrus cultivars will contribute to breeding seedless acid cultivars.
Yuzu (Citrus ichangensis × C. reticulate) fruit is an important functional food that possesses several health-promoting properties. It has been widely used in commercial medical products, healthy foods, and cosmetics in many countries. Yuzu is a rich source of wide variety of non-nutritive compounds, such as flavonoids, anthocyanins, phenolic acids, carotenoids, and tannins; as well as nutritive compounds such as sugars, proteins, vitamins, fibers, and minerals. Yuzu fruit (juice, peel, and seeds) and its bioactive compounds have been demonstrated to have numerous functional properties, such as antioxidant, anti-inflammatory, anticancer, antiplatelet, angiogenesis, and antimicrobial properties, both in vitro and in vivo. These diverse applications provided by the yuzu fruit (juice, peel, and seeds) and its bioactive compounds are of great industrial importance. This review summarizes the composition, nutritional values, and functional properties of yuzu fruit, and their biological activity in relation to their potential impact on human health.
The phylogenetic relationships within the genus Citrus and related species found in Korea were analyzed by comparing the trnL-trnF intergenic spacer sequences of the chloroplast DNA (cpDNA). All 21 accessions had trnL-trnF sequences of identical length, 372 base pairs (bp). Of the 377 aligned positions in the trnL-trnF regions of these species, 67 sites were variable (17.8%, 67/377). Pairwise sequence divergences in the trnL-trnF region ranged from 0 to 3.2% in the ingroup accessions and from 8.8 to 10.9% between the in- and the outgroup species Phellodendron amurense. The greatest divergence (3.2%) was between C. aurantium and C. natsudaidai. A phylogenetic tree was generated based on maximum parsimony (MP) analysis of the trnL-trnF sequences data from 22 accessions, including the outgroup taxon, producing 17 equally most parsimonious trees. The consensus tree contained eight independent clusters. Cluster I contained C. platymamma, which has a collared fruit shape, unique among Korean Citrus. Cluster II contained mostly mandarin and pummelo accessions. Cluster V contained C. sunki and six accessions of C. unshiu. Four of six C. unshiu accessions had identical sequences. In addition, Nagami kumquat (Fortunella margarita) was phylogenetically close to Citrus, whereas the trifoliate orange (Poncirus trifoliate) was distant from Korean Citrus species, both local species and cultivars. These results confirmed that analyses of cpDNA trnL-trnF sequences can detect enough polymorphisms to differentiate Korean Citrus species and to determine their phylogenetic relationships.
PCR-RFLP analyses of three regions for each of chloroplast DNA (cpDNA; rbcL-ORF106, trnD-trnT, trnH-trnK) and mitochondrial DNA (mtDNA; nad7/exon2-exon3, nad7/exon3-exon4, 18S-5S) were performed in 26 cultivars of acid citrus grown in Japan to identify polymorphisms and classify them. The polymorphisms
were compared with those of three true Citrus species, i.e., mandarin, pummelo and citron. Ichang papeda (C. ichangensis) was also included in this study to find its relationship with Yuzu. Inter-species cpDNA variation was recognized and the
acid citrus were divided into three groups, namely; I (‘Yuzukichi’ and ‘Kinkoyu’), II [sour oranges (‘Kaiseito’, ‘Daidai’
and ‘China daidai’), ‘Nansho daidai’, ‘Kiku daidai’, C. sudachi (‘Mushi yukaku’, ‘Yushi yukaku’ and ‘Yushi mukaku’), C. sphaerocarpa (‘Kabosu’ and ‘Aka kabosu’), C. kizu (‘Taninaka kizu’, ‘Kinosu’ and ‘Kizu’), ‘Zanbo’, ‘Mochiyu’, ‘Jabara’ and ‘Naoshichi’], and III [Yuzu (‘Tetraploid’, ‘Tochikei
yuzu’ and ‘Yamanekei yuzu’), ‘Matsuda sudachi’, ‘Zuishoyu’, ‘Hanayu’ and ‘Yuko’]. CpDNA restriction patterns of the three
true Citrus species differed from each other as well as from those of ichang papeda. CpDNA restriction patterns of group I of the acid
citrus were identical to those of mandarins. Group II showed the same as pummelos. CpDNA restriction patterns of group III
were differed from those of the three true Citrus species in the three regions. This group was differed from ichang papeda after digestion of trnH-trnK PCR products with TaqI, HinfI and AluI, while they showed identical restriction patterns in two regions, rbcL-ORF106 and trnD-trnT. Citrons and ichang papeda were placed in groups IV and V, respectively. Based on mtDNA restriction patterns, the acid citrus
were divided into three groups; i, ii and iii. In groups i and ii accessions of groups I and II of cpDNA were placed with
mandarins and pummelos, respectively. In group iii accessions of group III of cpDNA were placed with ichang papeda. Citrons
were placed in a distinct group, iv.
Allozyme variation at 15 loci resolved from 12 enzyme systems was studied in 205 accessions of cultivated Camellia japonica . All loci were polymorphic and a total of 64 alleles were detected. The sensitivity of electrophoresis in distinguishing clones was high, with 95.4% of the clones uniquely characterized by their 15-locus genotypes. Replicates of cultivars collected in different localities were allozymically identical. The hypothesis that some cultivars originated as sports was tested in a number of cultivar groups. The allozyme data failed to support this hypothesis for some groups, suggesting that in these cases cultivars of independent origin have become incorrectly incorporated into bonafide clone groups during their long history of cultivation.
Starch gel electrophoresis of leaf tissue samples was used to distinguish pineapple [ Ananas comosus (L.) Merr.] cultivars. Isozyme genotypes at two peroxidase and three phosphoglucomutase loci allowed the unique identification of 15 of the 27 cultivars examined. The hypotheses that some cultivars originated as either sports or hybrids were confirmed by the allozyme data.
Leaf extracts of 163 Japanese persimmon cultivars ( Diospyros kaki L.) and six other Diospyros species were analyzed for isozyme variation of glucose phosphate isomerase (GPI, E C 5.3.1.9) and malate dehydrogenase (MDH, E C 1.1.1.37). With both systems, the bands were sharp and well-resolved, and intracultivar polymorphism was absent. Isozyme phenotypes were more varied with GPI than with MDH. With Japanese persimmons, GPI yielded 24 different banding patterns, and six cultivars were uniquely discriminated by this enzyme alone. MDH produced only three different banding patterns, with none of the cultivars discriminated. When both enzyme systems were taken together, 18 cultivars were uniquely discriminated and the rest could be classified into 22 groups of 2 to 18 cultivars each.
Seedling populations of three cultivars of trifoliate orange [ Poncirus trifoliata (L.) Raf.] were grown in a greenhouse from seed collected in three different years and four different locations. Starch gel electrophoresis of four heterozygous loci was used to distinguish nucellar (apomictic) seedlings from zygotic seedlings. Minimum estimates of the frequency of zygotics among 23 seed samples ranged from 0% to 76%, with an overall mean of 18%. All zygotics apparently originated from self-pollination or pollination by other trees of trifoliate orange. Samples collected from the same tree in different years had significantly different frequencies of zygotics. Differences among cultivars in frequency of zygotics were obscured by the large environmental effects. Samples stored for 8 months before planting generally did not differ in frequency of zygotics from those stored for 5 months. Mean height of zygotics was less than that of nucellars in all samples measured, significantly less in most samples. However, many zygotics were so similar to nucellars in height, leaf size, thorn length, petiole length, and stem diameter that they would be difficult to identify using these traits.
The distinction between apomictic nucellar and zygotic 5-month-old seedlings of Citrus was made using two genetically defined isozyme markers: phosphoglucose isomerase and phosphoglucose mutase. Of 123 seedlings of attempted crosses of ‘King’ (seed parent) × ‘Parsons Special’, 18 had the ‘King’ genotype and were therefore nucellars. Segregation ratios for both markers were as expected among the zygotics.
The chrysanthemum (Dendranthema grandiflora Tzvelev.) cultivars 'Dark Charm', 'Salmon Charm', 'Coral Charm' and 'Dark Bronze Charm' are either radiation-induced mutants or spontaneous sports of 'Charm' and constitute a family or series of plants that primarily differ in flower color. These cultivars, which were difficult to differentiate genetically by DNA amplification fingerprinting (DAF), were easily identified by using arbitrary signatures from amplification profiles (ASAP). Genomic DNA was first amplified with three standard octamer arbitrary primers, all of which produced monomorphic profiles. Products from each of these DNA fingerprints were subsequently reamplified using four minihairpin detainer primers. The 12 primer combinations produced signatures containing ≃37% polymorphic character loci, which were used to estimate genetic relationships between cultivars. Forty-six (32%) unique amplification products were associated with individual cultivars. The number of ASAP polymorphisms detected provided an estimate of the mutation rate in the mutant cultivars, ranging from 0.03% to 1.6% of nucleotide changes within an average of 18 kb of arbitrary amplified DAF sequence. The ASAP technique permits the clear genetic identification of somatic mutants and radiation-induced sports that are genetically highly homogeneous and should facilitate marker assisted breeding and protection of plant breeders rights of varieties or cultivars.
Isoenzyme staining was used to characterize 55 of 78 raspberry cultivars (Rubus idaeus L., R. × neglectus Peck, and R. occidentalis L.). Six enzymes were needed to achieve this characterization: isocitrate dehydrogenase, malate dehydrogenase, phosphoglucoisomerase, phosphoglucomutase, shikimic acid dehydrogenase, arid triose phosphate isomerase. The 23 cultivars that were not uniquely characterized were grouped into eight groups of two and two groups of three and four. Two of these groups comprised black raspberry cultivars, all of which were similar isozymically. Isoenzymes could not distinguish between the cultivar Willamette and a spine-free mutant of the cultivar. Analysis of cultivars obtained from several sources revealed that raspberry cultivar mislabeling exists but is not very prevalent. Regular isoenzyme analysis of raspberry cultivars held by germplasm repositories, certified and other propagators, and breeders is both feasible and advisable for early detection of cultivar mislabeling.
it>Citrus rootstocks are commercially propagated from open-pollinated seed, which usually produces uniform populations of nucellar seedlings. Morphological traits are used to identify any off-type seedlings. In this study, seedling populations of 15 rootstock cultivars were evaluated morphologically and for their isozyme genotypes at 7 loci. The populations varied in degree of polyembryony and in percentages of plants that were morphologically or isozymically variable. Two cultivar populations contained only typical nucellar seedlings. Four populations contained a few morphologically variant plants that had isozyme genotypes typical for the cultivar. The remaining rootstock populations contained plants that varied in morphology, isozyme profile, or both. Isozymes were useful for the detection of zygotic seedlings that were not detected by their morphology. The zygotic seedlings, with one exception, appeared to arise from self-pollination.
Leaf tissue samples of seven cultivars of Citrus limon, one of Citrus aurantifolia, one cultivar of Citrus latifolia, one cultivar of Citrus meyeri and two cultivars of local selections were analyzed for isozyme variation for malate dehydrogenase (MDH), glutamate oxaloacetate transaminase (GOT), tetrazolium oxidase (T.O.) and esterase (EST) to examine the biodiversity of farmers' selections and commercial cultivars. The GOT zymograms consisted of bands in two zones. The GOT isozymes are characterized by co-dominant expression. The enzyme differences were found to be adequate for distinction at the species level. MDH isozymes showed one or three bands. Two major areas of T.O. activity were identified. Zymograms for EST should prove useful as a diagnostic tool for cultivar identification in view of the extensive polymorphism of this enzyme.
Fifty-four apple ( Malus domestica Borkh.) cultivars were characterized electrophoretically using 6 isozyme systems. Intracultivar variation in isozyme phenotype was not observed, whereas intercultivar polymorphism was sufficient to permit reliable and unambiguous identification of nearly every cultivar. The most useful isozyme systems for distinguishing among the cultivars were 6-phosphogluconate dehydrogenase and aspartate aminotransferase. Sports could not be distinguished from the original cultivar. The genetic basic of several polymorphisms was known, enabling the comparison of the isozyme genotype observed in a hybrid with that predicted on the basis of parental genotypes. The 6-phosphogluconate dehydrogenase genotype of ‘Spartan’ indicated that ‘Yellow Newtown’ may not have been the paternal parent.
The genetic control of isozymes from Citrus and its near relatives was determined for three gene/enzyme systems: glutamate oxaloacetate transaminase, phosphoglucose isomerase and phosphoglucose mutase. These enzymes are controlled by four genes having 19 codominant alleles, 12 of which occur in Citrus subg Citrus. Formal genetic studies were carried out with F, biotypes and F1 populations of known origin. When biotypes were grouped into traditionally recognized species to examine genetic affinities within and between species, a remarkable pattern of uniformity of genotype combinations was found within a species, and every species had an unique combination. Because many economically important cultivars produce asexual(nucellar) as well as sexual(zygotic) embryos, a central problem of the breeder is to distinguish these when plants are young, long before fruiting. Isozyme markers can be used with varying degrees of efficiency, depending on the genotypes of the particular parents, to distinguish nucellar seedlings from those of zygotic origin.
Leaf isozymes of four enzymes in citrus were analyzed using polyacrylamide gel electrophoresis to demonstrate the phylogenic relationship of the cultivars. Two loci for the superoxide dismutase(SOD) (EC1.15.1.1), one locus for the peroxidase (EC 1.11.1.7) and polyphenol oxidase (EC1.10.3.2), respectively were examined. In addition to the two loci of glutamate oxal-acetate transaminase(GOT) (EC2.6.1.1), reported in the former study, a new locus was detected in the present report. These experiments suggest that the mandarin cultivars originated from three genetic sources ; Indian, Chinese, and Japanese ones. Most of the Chinese mandarins including 'Kinokuni' showed identical genotypes, while the 'Ponkan' and 'Dancy tangerine' differed from the Chinese cultivars in their genotypes at the Px locus, and they were assumed to have originated from India as suggested by T. Tanaka. On the other hand, the Japanese mandarins were considered to be genetically affected by the'Chinese mandarins. But some of the Japanese mandarins including 'Yatsushiro' and 'Koji' exhibited genetic effects from Tachibana, the wild type of the mandarin in J.apan. The genotypes of the sweet orange cultivars were homogeneous and similar to those 0L the Chinese mandarins. Sour orange was heterozygous at several loci, and was assumed to be a hybrid between mandarin and pummelo. Among the pummelo cultivars, some heterogeneity was detected at some loci.
Leaf isozymes of glutamate oxaloaceate transaminase (EC 2.6.1.1; GOT) in the "true citrus fruit trees" group of Citrinae were analyzed using polyacrylamide gel electrophoresis to elucidate phylogenetic relationships among them. Segregation at each of three loci, Got-1, Got-2 and Got-3 of GOT isozymes in the progenies obtained from intergeneric crosses between Citrus and Microcitrus species followed a simple Mendelian inheritance, suggesting that Microcitrus and Citrus species have similar genetic systems and the species of these two genera are closely related to each other. Some common alleles at the three loci controlling GOT isozymes were recognized between the genera of "true citrus fruit trees". These findings suggest that a genetic identity is conserved among the "true citrus fruit trees" group; even some of them climatically, geographically and reproductively isolated.
PCR-RFLP of 14 accessions of six Annona species namely A. cherimola Mill., A. squamosa L., A. reticulata L., A. glabra L., A. muricata L. and A. montana Macfad. and A. cherimola x A. squamosa were studied. One set of Sequence Tagged Site (STS) primers was used in order to amplify the position of cpDNA fragment: 'rbc L-ORF106'. The amplified fragment was digested with six kinds of restriction enzymes (four base cutter). Enzymes average generated two to eight bands in each sample. Seven different restriction patterns were observed when digested with Rsa I, but other enzymes showed four to six. The pair-wise distance values ranged from 0.017 to 0.607 with the minimum between A. squamosa and A. cherimola x A. squamosa and the maximum between A. glabra and A. cherimola x A. squamosa. Eight most parsimonious trees were obtained using Swofford parsimony method, and 50 % majority-rule consensus tree was obtained from them. These results indicated that the cultivated and wild species grouped separately and the A. cherimola x A. squamosa was placed between its parents in the cultivated species. A. glabra clustered singly in the dendrogram demonstrating that this species is different from the others. We believe that these results would give basic information in the phylogenetic studies of Annona species using a large number of species.
Isozymes of GOT and MDH in leaves of 105 citrus biotypes were analyzed using acrylamide gel electrophoresis. At least 3 loci were detected on these two enzymes. From the analysis, Natsumikan (Citrus natsudaidai), Hassaku (C. hassaku) and some other citrus cultivars are considered to be hybrids of pummelo (C.grandis) and mandarin as suggested. Moreover, Iyo (C.iyo), which has been assumed to be a hybrid of sweet orange (C.sinensis) and mandarin, was found to have a genetic effect from pummelo. Sour oranges (C.aurantium) had unique allele A at Got-1. In addition to the sour orange, the same allele was found only in lchang lemon (C.wilsoni), Bergamot (C. bergamia) and 'Hirado' pummelo. While, Tachibana (C.tachibana), Shiikuwasha (C. depressa), and Korai-tachibana (C. nippokoreana), which are reported to be native to Japan and its neighboring areas, had an unique allele A at Got-2. This allele was also found in Hyuganatsu (C.tamurana) and some biotypes originated in Japan, but not found in mandarins originated in mainland China, India, Philippines and other areas. While, Yuzu (C.junos) and its relatives had B at Got-2 and B at Mdh-1.
A set of 31 Cucurbita pepo L. commercial cultivars with different fruit shapes (morphotypes) was evaluated for resistance to powdery mildew of cucurbits (Erysiphe cichoracearum and Sphaerotheca fuliginea) under field conditions of a natural infection. During vegetative growth, the infection degree (ID) was assessed four times on leaves (L) and twice on stems and petioles (S). The `Area below the curve' values of the disease infection progress were calculated separately for leaves (ABC-L) and stems and petioles (ABC-S). Data were subjected to one-way analyses of variance and mean separation was performed using LSD multiple range tests. Significant differences in field resistance were found between morphotypes. Cultivars with scallop and acorn fruit shapes were the most resistant. The straightneck, crookneck and ornamental gourd groups showed medium levels of field resistance. The zucchini, cocozelle, pumpkin and vegetable marrow groups of morphotypes were the most susceptible. Isozyme spectra of peroxidases for 11 selected C. pepo cultivars representing different groups of morphotypes with various levels of field resistance were analysed as well. They could be ranged into three basic groups which correspond with the cultivar's level of field resistance. This phenomenon is discussed as a potential biochemical marker in C. pepo selection.
The genetic control of isozymes from Citrus and its near relatives was determined for three gene/ enzyme systems: glutamate oxaloacetate transaminase, phosphoglucose isomerase and phosphoglucose mutase. These enzymes are controlled by four genes having 19 codominant alleles, 12 of which occur in Citrus subg Citrus. Formal genetic studies were carried out with F1 biotypes and F1 populations of known origin. When biotypes were grouped into traditionally recognized species to examine genetic affinities within and between species, a remarkable pattern of uniformity of genotype combinations was found within a species, and every species had an unique combination. Because many economically important cultivars produce asexual(nucellar) as well as sexual(zygotic) embryos, a central problem of the breeder is to distinguish these when plants are young, long before fruiting. Isozyme markers can be used with varying degrees of efficiency, depending on the genotypes of the particular parents, to distinguish nucellar seedlings from those of zygotic origin. THERE have been several attempts to render a taxonomy reflecting species relationships in Citrus, a diverse and difficult group comprised of two subgenera, Citrus and Papeda (Swingle, 1943; Swingle and Reece, 1967). The former includes all the edible cultivars. Papeda includes six species with non-edible fruits. The number of species to be recognized in Citrus subg Citrus, and which biotypes to include in which species, constitute the central problems in Citrus taxonomy. The best known taxonomies, those of Swingle (1943) and Tanaka (1969), differ widely in the number of admitted species. Swingle recognized 16 in Citrus, Tanaka, 159. At another extreme, Scora (1975) and independently, Barrett and Rhodes (1976), recognized just three basic species exclusive of those of the subgenus
The taxonomic classification of the wild Lima bean complex needs to be assessed to select species for use in breeding programs and to identify genetic resources for conservation. The objectives of this study were to determine the genetic relationships among, and the phylogeny of, wild Lima bean (Phaseolus lunatus L.) and related species (P. augusti Harms, P. bolivianus Piper, P. pachyrrhizoides Harms, and P. rosei Piper) from South America and to identify specific genetic reserves for conservation. These relationships were investigated by means of amplified restriction fragment length polymorphism (AFLPs) on total genomic DNA. The 122 accessions formed a cluster that was distant from common bean (P. vulgaris L.), confirming earlier morphology and hybridology data. Two gene pools of wild Lima beans were confirmed. One was widely distributed in neotropical lowlands, while the other was restricted to the western Andes, in Ecuador and northern Peru. The study also revealed the existence of a third group of wild Lima bean distributed in the Departments of Boyaca and Cundinamarca, Columbia. The three species P. augusti, P. bolivianus, and P. pachyrrhizoides differed very little, certainly not sufficiently to merit a separate taxonomic ranking at the species level. The accessions could be grouped instead according to four geographic origins: Ecuador and northern Peru; Department of Junin, Peru; Departments of Cuzco and Apurimac, Peru; and Bolivia and northwestern Argentina. Results from this study should result in a better selection of parental materials in breeding programs and point to areas where germplasm collections and conservation are needed.
Nine isozyme systems were investigated by polyacrylamide gel electrophoresis as a means of identifying the six species of Fortunella. By examining the characteristic isoenzymatic patterns, it was found that there was considerable variability between the different species. The relationships among the six species of Fortunella were inferred from the similarity matrix cata. The results indicate that no pairs of species are identical and the degree of relationship between the species was low, suggesting that the six species of Fortunella are independent.
A cross between the monoembryonic Clementine MA3 SRA 649 used as seed parent and the polyembryonic Carvalhais mandarin used as pollen progenitor was carried out in the SRA INRA-CIRAD, S. Giuliano, Corsica. Cuttings of 61 plants putatively resulting from this cross were sent to the Centro de Citricultura, Faro, Portugal for further agronomic evaluation under field trials. Isozyme analysis allowed the identification of 59 plants as having sexual origin, however, two plants that showed isozyme patterns identical to their seed parent were impossible to identify as being either of zygotic or of somatic origin. Isozymes confirmed Carvalhais mandarin as the pollen parent of 42 progeny plants, while RAPD analysis confirmed Carvalhais paternity over the remaining 19 plants. A fast and low pollutant method for extraction of Citrus total genomic DNA suitable for RAPD analysis is reported.
Isozyme variability in peach (Prunus persica L. Batsch) cultivars was quantified and its distribution among freestone, clingstone and nectarine groups was studied using the Shannon-Weaver diversity index. Esterases, peroxidases and malate dehydrogenases from foliar extracts of 26 cultivars were analyzed by starch gel electrophoresis. Little variability was found, with two, three and four different electrophoretic patterns for each of the enzymatic systems. Twenty percent of the total variability was found between groups. The lowest variability within groups was found among nectarines. These results agree with those obtained in Spanish cultivars. The low isozymic variation reflects a reduced genetic variability, possibly as a result of the founder effect of a few genotypes from which the current cultivars were bred.
A genetic diversity analysis involving 49 lychee (Litchi chinensis Sonn.) accessions using eight enzyme systems encoding 12 loci (Idh-1, Idh-2, Mdh-2, Per-1, Pgi-2, Pgm-1, Pgm-2, Skdh, Tpi-1, Tpi-2, Ugpp-1, and Ugpp-2) revealed moderate to high levels of genetic variability. Cluster analysis of the isozyme data from 40 genetically different accessions of the total 49 identified three groups at the 50% level of genetic similarity, the largest of which contained 32 of the 40 accessions distributed in three sub-groups. The groups including the three sub-groups differed markedly both in frequency and composition of alleles at different loci. On average, 77% of the loci were polymorphic with an overall mean of 2.2 alleles per locus and an observed heterozygosity of 0.387. The unbiased genetic identities (I) between groups ranged from 0.809 between Groups 1b and 2 to 0.937 between Groups 1b and 1c. Group 2 has diverged from the other groups with an average genetic identity of 0.821. Summing over all 11 polymorphic loci, 16% of gene diversity was due to differentiation between groups and 84% to that within groups. Comparison of isozyme fingerprints revealed that some accessions identically named, such as ‘No mai tsz’, ‘Kwai mi’, and ‘Hak ip’, possessed different isozyme genotypes, whereas some others with different names displayed identical isozyme genotypes.
Citrus leaf extracts were analyzed by starch gel electrophoresis for isozymes of hexokinase (HK), isocitrate dehydrogenase (IDH), leuclne aminopeptldase (LAP), malate dehydrogenase (MDH), and malic enzyme (ME). The first three showed a single region of activity suggesting each is specified by a single gene. Banding patterns indicated HK and LAP are monomeric, and IDH is dimeric. Evidence of four alleles of Hk and ldh and three of Lap was found in the nine species of cultivated Citrus and Poncirus trifoliata . Isozymes of MDH occurred in three gel regions, are apparently specified by four genes two of which, Mdh-1 and Mdh-2 , are polymorphic. Zymograms indicated that MDH is dimeric but the subunits of Mdh-1 do not dimerize with those of Mdh-2 . ME is specified by two genes but only one was polymorphic. ME is apparently monomeric. Isozymes of citrus provide molecular tags to determine the genetic origin of citrus seedlings that can be either asexual from the nucellus or sexual from a zygote. A very large proportion of all possible seedlings either from selfing or from crossing can now be distinguished as to their genetic origin with isozyrnes.
Acid Citrus in Shikoku Island
- K Kimura
- T Taninaka
Kimura, K., Taninaka, T., 1988. Acid Citrus in Shikoku Island. Harada Co., Tokushima, Japan (in Japanese).
Acid Citrus in Shikoku and Kyushu Island
- K Kimura
- T Taninaka
Kimura, K., Taninaka, T., 1990. Acid Citrus in Shikoku and Kyushu Island. Harada Co., Tokushima, Japan (in Japanese).
Genetic relationships in Citrus and related genera
- M J Asins
- R Herreo
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- L Navarro
- B Manicom
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Asins, M.J., Herreo, R., Pina, J.A., Carbonell, E.A., Navarro, L., 1996. Genetic relationships in Citrus and related genera. In: Manicom, B., Robinson, J., du Plessis, S.F., Joubert, P., van Zyl, J.L., du Preez, S. (Eds.), Proceedings of the International Society of Citriculture, Vol. 1. Dynamic Ad, Nelspruit, South Africa, pp. 248±253.
Leaf isozymes as genetic markers in Citrus Identi®cation of nucellar and zygotic seedlings of citrus with leaf isozymes The botany of Citrus and its wild relatives of the orange subfamily (family Rutaceae, subfamily Aurantioideae) The Citrus Industry
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Soost, R.K., Torres, A.M., 1981. Leaf isozymes as genetic markers in Citrus. In: Matsumoto, K., Oogaki, C., Kozaki, I. (Eds.), Proceedings of the International Society of Citriculture, Vol. 1. Aiko Printing Co., Tokyo, Japan, pp. 7±10. Soost, T., Williams, E., Torres, A.M., 1980. Identi®cation of nucellar and zygotic seedlings of citrus with leaf isozymes. HortScience 15 (6), 728±729. Swingle, W.T., 1967. The botany of Citrus and its wild relatives of the orange subfamily (family Rutaceae, subfamily Aurantioideae). In: Webber, H.J., Batchelor, L.D. (Eds.), The Citrus Industry, Vol. 1. University of California, Riverside, pp. 129±474. Tanaka, T., 1954. Species problem in citrus. Japanese Society for the Promotion of Science, Tokyo, 152 pp.
Study of the Acid Citrus in Japan with Special Reference to its Species, Morphology and Histology
- K Kimura
Kimura, K., 1996. Study of the Acid Citrus in Japan with Special Reference to its Species, Morphology and Histology. Harada Co., Tokushima, Japan (in Japanese with English summary).
Acid Citrus in Japan
- K Kimura
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Kimura, K., Taninaka, T., 1995. Acid Citrus in Japan. Harada Co., Tokushima, Japan (in Japanese).
Acid citrus cultivars related to the yuzu (Citrus junos Sieb. ex Tanaka) in Japan
- T Taninaka
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Taninaka, T., Otoi, N., Moromoto, J., 1981. Acid citrus cultivars related to the yuzu (Citrus junos Sieb. ex Tanaka) in Japan. In: Matsumoto, K., Oogaki, C., Kozaki, I. (Eds.), Proceedings of the International Society of Citriculture, Vol. 1. Aiko Printing Co., Tokyo, Japan, pp. 73±76.
Japanese Society for the Promotion of Science
- T Tanaka