Establishment of efficient and rapid regeneration system for some diploid wild species of Arachis

Abstract

Though peanut tissue culture has advanced to a considerable extent using a number of explants with the subsequent production of transgenic plants, wild Arachis species appeared to be recalcitrant to using similar explants. In this study, the use of cotyledonary nodes as explants prepared from 7-day old seedlings resulted in the development of a simple and rapid regeneration protocol for five diploid wild species including A. diogoi, A. stenosperma, A. duranensis, A. cardenasii and A. correntina belonging to the genus Arachis for producing multiple shoots. Shoot bud initiation was observed 10 to 15 days after culture initiation. Responding cotyledonary nodes with shoot buds were subcultured to lower levels of cytokinin and finally to MS basal medium for further shoot development and elongation. Production of multiple shoots was observed in all the five diploid species with a maximum of 9 to 16 shoots were obtained per explant in the primary cultures. The number of shoot buds increased significantly with repeated explant subculturing with recovery up to 45 shoots from responding explants. These shoots were rooted efficiently on MS medium supplemented with 1 mg l -1 naphthalene acetic acid and the time taken from explanting to the transfer of shoots to potting mixture was about 12 weeks. All rooted shoots were successfully established in soil in glass house and further transferred to field. These plants survived to maturity and set seed.

Thidiazuron‐Induced Tissue Culture Regeneration from Quartered‐Seed Explants of Arachis Paraguariensis

“…Even though several studies involving diff erent species have shown that genotype plays a critical role during in vitro regeneration (Bailey et al, 1993;Srinivasan et al, 2010), it is desired that a regeneration protocol be applicable to a wide range of genotypes or even species. The protocols developed in this study were suitable across six diff erent genotypes of A. paraguariensis.…”

“…Likewise, the induction of tetraploidy is another vital technique that could broaden the available gene pool as well as eliminate hybridization barriers that are due to diff erences in ploidy levels. Interestingly, tissue culture of cultivated peanut has witnessed a remarkable improvement in the past few decades; however, many of the wild Arachis species appear recalcitrant to similar methods (Srinivasan et al, 2010). Specifi cally for A. paraguariensis, limited success has been achieved in the regeneration of plants from mature leafl ets (Dunbar and Pittman, 1992), protoplasts (Li et al,…”

RNAi-mediated Control of Aflatoxins in Peanut: Method to Analyze Mycotoxin Production and Transgene Expression in the Peanut/<em>Aspergillus</em> Pathosystem

“…Recombine p2+4ENTR into pCAPD 29 (NCBI Accession: KC176455.1) using LR clonase II enzyme mix according to manufacturer's instructions to make plasmid p5XCAPD, and transform it into Escherichia coli DH5α using standard techniques followed by partial sequencing. Note: The complete RNAi insert is shown in 33 . Move tissues to SIM (500 μM cefotaxime and 100 μM kanamycin) for shoot formation, with bi-weekly transfers for 2 months.…”

High-efficiency regeneration of peanut (Arachis hypogaea L.) plants from leaf discs

“…Leaflets are the most widely used explants in peanut tissue culture. Several other types of explants, such as cotyledonary nodes (Srinivasan et al, 2010), epicotyls, hypocotyls (Marion et al, 2008), axillary meristems (Singh and Hazra, 2009) and cotyledons (Bhatnagar et al, 2010; *Corresponding author. E-mail: jzhang@ippcaas.cn.…”