A grobacterium-mediated genetic transformation in hot chilli (Capsicum annuum L. var. Pusa jwala)

Manoharan, Muthusamy; Vidya, Chinnasamy; Sita, G. Lakshmi

doi:10.1016/s0168-9452(97)00231-8

Cited by 63 publications

(32 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The unbound fractions were collected in 0.1 M potassium phosphate buffer, and the bound fractions were eluted in 1 M NaOH and subjected to CS assay after dialysis against 0.1 M potassium phosphate buffer. reports on genetic transformation in sweet pepper (31,32) and also hot chili (33). As an extension of the present study, regulating the pungency levels in Capsicum cultivars by using csy1 gene would have wide applications in food, pharmaceutical, and allied industries.…”

Section: Discussionmentioning

confidence: 55%

Characterization of capsaicin synthase and identification of its gene (csy1) for pungency factor capsaicin in pepper (Capsicumsp.)

Prasad

Kumar

Gururaj

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

Section: Discussionmentioning

confidence: 55%

Characterization of capsaicin synthase and identification of its gene (csy1) for pungency factor capsaicin in pepper (Capsicumsp.)

Prasad

Kumar

Gururaj

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…cotyledonary leaves, hypocotyls, shoot tips, leaf discs and nodal segments have been studied to test their regeneration potential but the response for different explants varied from one genotype to another. However, in red pepper, hypocotyls and cotyledonary leaves had responded well than other explants reported so far (Gunay and Rao 1978;Ramirez-Malagon and Ochoa-Alejo 1996;Manoharan et al 1998). In preliminary stage, we have started with all possible explants such as cotyledonary leaves, leaf discs and hypocotyls.…”

Section: Discussionmentioning

confidence: 99%

“…has been reported for diverse explants such as cotyledons (Gunay and Rao 1978;Ebida and Hu 1993;Manoharan et al 1998;Golegaonkar and Kantharajah 2006), hypocotyls (Gunay and Rao 1978;Ramirez-Malagon and Ochoa-Alejo 1996), leaf discs (Sanatombi and Sharma 2008) and nodal segments (Ahmad et al 2006). However, these reports are genotype-specific and consequently, the regeneration protocol as well as transformation procedure need to be established for each commercial cultivar for exploiting the potential of genetic engineering.…”

Section: Introductionmentioning

confidence: 99%

“…Agrobacterium-mediated transformation is widely applied for red pepper transformation but its efficiency relies on the effective shoot regeneration and the genotype. The protocols available for red pepper transformation either could not be reproduced successfully or with very low efficiency in different genotypes (Manoharan et al 1998;Zhu et al 1996;Lee et al 2004;Li et al 2003).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

An improved plant regeneration and Agrobacterium - mediated transformation of red pepper (Capsicum annuum L.)

Kumar

Sharma

Chattopadhyay

et al. 2012

Physiol Mol Biol Plants

View full text Add to dashboard Cite

Capsicum annuum (red pepper) is an important spice cum vegetable crop in tropical and subtropical countries. Here, we report an effective and reproducible auxin free regeneration method for six different red pepper cultivars (ACA-10, Kashi Anmol, LCA-235, PBC-535, Pusa Jwala and Supper) using hypocotyl explants and an efficient Agrobacterium-mediated transformation protocol. The explants (hypocotyls, cotyledonary leaves and leaf discs) collected from axenic seedlings of six red pepper cultivars were cultured on either hormone free MS medium or MS medium supplemented with BAP alone or in combination with IAA. Inclusion of IAA in the regeneration medium resulted in callus formation at the cut ends of explants, formation of rosette leaves and ill defined shoot buds. Regeneration of shoot buds could be achieved from hypocotyls grown in MS medium supplemented with different concentrations of BAP unlike other explants which failed to respond. Incorporation of GA 3 in shoot elongation medium at 0.5 mg/l concentration enhanced the elongation in two cultivars, LCA-235 and Supper, while other cultivars showed no significant response. Chilli cultivar, Pusa Jwala was transformed with βC1 ORF of satellite DNA β molecule associated with Chilli leaf curl Joydebpur virus through Agrobacterium tumefaciens. Transgene integration in putative transformants was confirmed by PCR and Southern hybridization analysis.

show abstract

“…Genetic transformation is extremely difficult in pepper plants, and the efficiency of transformation is very low. Although a number of transformation protocols have been published (Manoharan et al, 1998;Mihalka et al, 2000;Kim et al, 2001), investigations into the functions of pepper genes remain problematic due to the lack of stable and effective genetic transformation procedures. Moreover, transformation itself is a complex, labor-intensive procedure, with low efficiency (Baulcombe, 2004;Benedito et al, 2004;Burch-Smith et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Optimization of virus-induced gene silencing in pepper (Capsicum annuum L.)

Wang¹,

Li²,

Gong³

et al. 2013

Genet. Mol. Res.

View full text Add to dashboard Cite

ABSTRACT. Virus-induced gene silencing is currently a powerful tool for the study of gene function in plants. Here, we optimized the protocol for virus-induced gene silencing, and investigated factors that affect the efficiency of tobacco rattle virus-induced gene silencing in pepper plants. Consequently, an optimal protocol was obtained by the syringe-infiltration method in the leaves of pepper plants. The protocol involves 2-leaf stage plants, preparing the Agrobacterium inoculum at a final OD 600 of 1.0 and then growing the inoculated plants at 22°C. Using this protocol, we achieved high efficiency in silencing CaPDS in different cultivars of pepper plants. We further used the CaPOD gene to illustrate the general reliability of this optimized protocol. Viral symptoms were observed on the leaves of inoculated plants of the Early Calwonder cultivar 25 days post-inoculation, indicating that this protocol can also be used to silence other genes in pepper plants. Real-time polymerase chain reaction analyses revealed that the expression levels of CaPDS and CaPOD were dramatically reduced in inoculated leaves compared to 2493 ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 12 (3): 2492-2506 (2013 Virus-induced gene silencing in pepper control plants. These results demonstrate that the optimized protocol can be applied to functional genomic studies in pepper to investigate genes involved in a wide range of biological processes.

show abstract

A grobacterium-mediated genetic transformation in hot chilli (Capsicum annuum L. var. Pusa jwala)

Cited by 63 publications

References 26 publications

Characterization of capsaicin synthase and identification of its gene (csy1) for pungency factor capsaicin in pepper (Capsicumsp.)

Characterization of capsaicin synthase and identification of its gene (csy1) for pungency factor capsaicin in pepper (Capsicumsp.)

An improved plant regeneration and Agrobacterium - mediated transformation of red pepper (Capsicum annuum L.)

Optimization of virus-induced gene silencing in pepper (Capsicum annuum L.)

Contact Info

Product

Resources

About