Optimal Media for In-Vitro Regeneration of Two Local Genotypes of Chili Pepper (Capsicum annuum L.) from West Sumatera

Abstract

Genetic engineering strategy in chilli peppers (Capsicum annuum L.) has been applied so far to improve its genetic capacity for instance against some important diseases. This strategy needs an established system for calli preparation as well as transformant regeneration. In most cases, this necessity is genotype dependent especially for some local genotypes. Based on this, we optimized media for calli induction, shoot and root regeneration of our three local cultivars namely Kampung, Kopay, and Trisula. Media combinations basically were composed of MS and combined with some growth factors, for instance, IAA, BAP, NAA and TDZ with different combinations for each calli induction, shoot and root regeneration. All media compositions were supplemented with 30 g/L sucroses. The best medium for calli induction was MS media supplemented with 4 mg/L BAP, 0.5 mg/L IAA and L2 vitamin (Thiamin-HCl and Pyrodoxine-HCl). While the best medium for shoot induction and elongation was MS supplemented with 1mg/L BAP, 5 mg/L AgNO 3 , 2 mg/L GA 3 , 2 mg/L Calcium Pantothenate and L2 vitamin. Furthermore, the best medium for root induction and elongation was WPM (Woody Plant Medium) supplemented with 0.4 mg/L NAA. This finding should provide optimal media for calli induction and shoot-root regeneration for the three local cultivars Kampung, Kopay, and Trisula.

Optimization of Plant Regeneration in Different Pepper (Capsicum annuum L.) Lines

“…The shoot structures were generally obtained from nutrient media containing high cytokinin (3-5 mg L -1 BA) and low auxin (0.5 mg L -1 IAA). When the literature is examined, BA and IAA are the most commonly used plant growth regulators and combinations in organogenesis studies in Capsicum (Gunay and Rao, 1978;Agrawal et al, 1989;Arroyo and Revilla, 1991;Christopher and Rajam, 1996;Ramírez-Malagón and Ochoa-Alejo, 1996;Hyde and Phillips, 1996;Sanatombi and Sharma, 2006;Sanatombi and Sharma, 2008;Renfiyeni et al, 2017;Haque and Ghosh, 2018;Gammoudi et al, 2017). We observed that the shoot structures were generally obtained from nutrient media containing high cytokinin (3-5 mg L -1 BA) and low auxin (0.5 mg L -1 IAA).…”

“…This may be due to the different Capsicum species used. Several studies have reported that IAA is an effective auxin for organogenesis studies in C. annuum L. However, the genotype effect is still an important factor in Capsicum genus (Venkataiah and Subhash, 2001;Renfiyeni et al, 2017).…”

“…There are many studies using different explants to improve the organogenesis protocol in the Capsicum genus. Different explants such as cotyledon and hypocotyl (Gunay and Rao, 1978;Agrawal et al, 1989;Arroyo and Revilla, 1991;Hyde and Phillips, 1996;Christopher and Rajam, 1996;Franck-Duchenne et al, 1998;Husain et al, 1999;Venkataiah et al, 2003;Joshi and Kothari, 2007;Sanatombi and Sharma, 2008;Orlinska and Nowaczyk, 2015;Renfiyeni et al, 2017;Hegde et al, 2017a, b;Gammoudi et al, 2017;Bhutia et al, 2018), zygotic embryo (Agrawal and Chandra, 1983;Arous et al, 2001), young seedlings (Phillips and Hubstenberger, 1985;Ebida and Hu, 1993), stem, leaf, root, shoot tip, embryo (Agrawal et al, 1989;Madhuri and Rajam, 1993;Christopher and Rajam, 1996;Venkataiah et al, 2003;Sharma, 2007a, 2007b;Renfiyeni et al, 2017;Bhutia et al, 2018), mature seeds (Ezura et al, 1993), shoot tip (Madhuri and Rajam, 1993), injured hypocotyl (Ramírez-Malagón and Ochoa-Alejo, 1996), seedling and embryonal explants (Dabauza and Pena, 2001), epicotyl (Renfiyeni et al, 2017), shoot buds (Haque and Ghosh, 2018) were used for in vitro organogenesis of Capsicum.…”