Regeneration in Jatropha curcas: factors affecting the efficiency of in vitro regeneration. Ind Crops Prod
Factors influencing in vitro regeneration through direct shoot bud induction from hypocotyl explants of Jatropha curcas were studied in the present investigation. Regeneration in J. curcas was found to be genotype dependent and out of four toxic and one non-toxic genotype studied, non-toxic was least responsive. The best results irrespective of genotype were obtained on the medium containing 0.5 mg L−1 TDZ (Thidiazuron) and in vitro hypocotyl explants were observed to have higher regeneration efficiency as compared to ex vitro explant in both toxic and non-toxic genotypes. Adventitious shoot buds could be induced from the distal end of explants in all the genotypes. The number of shoot buds formed and not the number of explants responding to TDZ treatment were significantly affected by the position of the explant on the seedling axis. Explants from younger seedlings (≤15 days) were still juvenile and formed callus easily, whereas the regeneration response declined with increase in age of seedlings after 30 days. Transient reduction of Ca2+ concentrations to 0.22 g L−1 in the germination medium increased the number of responding explants.
Available from: Zeng-Fu Xu
- "The efficiency of the in vitro plant regeneration system, the efficiency of Agrobacterium tumefaciens transformation, and the specific antibiotic selection procedure are key factors in plant genetic transformation (Kajikawa et al. 2012; Li et al. 2008; Mao et al. 2009). To date, shoot regeneration systems for J. curcas have been successfully established using various explants such as cotyledons, epicotyls, hypocotyls , leaves, petioles, nodes, and stems (Khurana-Kaul et al. 2010; Kumar and Reddy 2010; Sharma et al. 2011; Singh et al. 2010; Sujatha and Mukta 1996; Toppo et al. 2012). Some regeneration systems have been utilized in genetic transformation protocols employing A. tumefaciens (Khemkladngoen et al. 2011; Kumar et al. 2010; Li et al. 2008; Mao et al. 2009; Misra et al. 2012; Pan et al. 2010) or particle bombardment (Joshi et al. 2011; Purkayastha et al. 2010). "
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ABSTRACT: Jatropha curcas is considered a potential biodiesel feedstock crop. Currently, the value of J. curcas is limited because its seed yield is generally low. Transgenic modification is a promising approach to improve the seed yield of J. curcas. Although Agrobacterium-mediated genetic transformation of J. curcas has been pursued for several years, the transformation efficiency remains unsatisfying. Therefore, a highly efficient and simple Agrobacterium-mediated genetic transformation method for J. curcas should be developed. We examined and optimized several key factors that affect genetic transformation of J. curcas in this study. The results showed that the EHA105 strain was superior to the other three Agrobacterium tumefaciens strains for infecting J. curcas cotyledons, and the supplementation of 100 mM acetosyringone slightly increased the transient transformation frequency. Use of the appropriate inoculation method, optimal kanamycin concentration and appropriate duration of delayed selection also improved the efficiency of stable genetic transformation of J. curcas. The percentage of β-glucuronidase positive J. curcas shoots reached as high as 56.0 %, and 1.70 transformants per explant were obtained with this protocol. Furthermore, we optimized the root-inducing medium to achieve a rooting rate of 84.9 %. Stable integration of the T-DNA into the genomes of putative transgenic lines was confirmed by PCR and Southern blot analysis. Using this improved protocol, a large number of transgenic J. curcas plantlets can be routinely obtained within approximately 4 months. The detailed information provided here for each step of J. curcas transformation should enable successful implementation of this transgenic technology in other laboratories.
Plant Biotechnology Reports 11/2015; DOI:10.1007/s11816-015-0377-0 · 1.19 Impact Factor
Available from: Ivan Luciano Pequeño-Granado
- "eron la induc - ción de brotes mediante el uso de BAP en combinación con AIB . Existen diversos trabajos relacionados con la regeneración de Jatropha curcas L . desarrollados a tra - vés de la organogénesis directa , utilizando diferentes tipos de explantes tal es el caso de ( Sujatha y Mukta , 1996 ; Kumar y Reddy , 2010 ; Kumar et al . , 2010a ; Sharma et al . , 2011 , Kumar y Reddy , 2012 ) que a través de peciolos , cotiledones e hipocotilos lograron la regeneracion de la especie . Así mismo , Khurana - Kaul et al . ( 2010 ) ; Kumar et al . ( 2010c ) ; Misra et al . ( 2010a ) ; Kumar et al . ( 2011 ) , lograron la regeneración directa a partir de hojar ; mientras que , Wei et al . ( 2004 ) logró b"
Available from: Amit Sagervanshi
- "The majority of the reports for J. curcas have used plant growth regulators (PGRs) such as 6-benzylaminopurine (BAP), indoleacetic acid (IAA), indole-3-butyric acid (IBA), gibberellic acid (GA 3 ), kinetin (KN), and thidiazuron (TDZ) for culture generation using different explants, including nodal segments, shoot apices, hypocotyls, epicotyls, juvenile cotyledons, leaves, petioles, and stems with various regeneration efficiency (Sujatha et al. 2005; Jha et al. 2007; Shrivastava and Banerjee 2008; Khurana-Kaul et al. 2010; Singh et al. 2010; Varshney and Johnson 2010; Khemkladngoen et al. 2011; Sahoo et al. 2012; Zhang et al. 2013). The significant role of CuSO 4 and Ca 2+ in the shoot bud induction enhancement has been reported (Khurana-Kaul et al. 2010; Sharma et al. 2011). In our study, "
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ABSTRACT: Jatropha curcas L. is attaining worldwide interest as an important biofuel crop. Experiments were conducted to improve the prevailing micropropagation technique as well as to develop a new ex vitro rooting method for J. curcas plant regeneration. Regeneration and ex vitro rooting efficiency was enhanced by augmenting the culture medium with abscisic acid (ABA). Different concentrations of 6-benzylaminopurine (BAP) and indole-3-butyric acid (IBA) were tested for callus generation from both in vitro and in vivo explants (leaf and petiole) on Murashige and Skoog (MS) medium. The best regenerative callus was achieved on MS medium supplement-ed with BAP (4.44 μM) and IBA (2.45 μM) from in vitro-cultured petioles. Highest regeneration (91%) was achieved by culturing petiole callus on MS medium supplemented with BAP (8.88 μM), IBA (0.49 μM), and ABA (1.9 μM), whereas 61% regeneration was obtained from in vitro leaf callus. Shoot proliferation and elongation was achieved on BAP (2.22 μM) and IAA (8.56 μM) with 10–13 shoots per explants. Highest rooting (65%) was achieved from M1 shoots (BAP, IAA, and ABA) on MS medium supplemented with IBA (2.45 μM), naphthaleneacetic acid NAA (0.54 μM), and 0.02% activated charcoal. Ex vitro rooting of 1-mo-old M1 shoots obtained from the charcoal-containing medium resulted optimum rooting (>72%) when transferred to polybags containing ster-ile sand. The plantlets were successfully acclimatized in soil with more than 98% survival rate in the greenhouse.
In Vitro Cellular & Developmental Biology - Plant 06/2014; 50(5). DOI:10.1007/s11627-014-9628-y/fulltext.html · 0.98 Impact Factor
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