Cefotaxime, a cephalosporin antibiotic, and different ethylene inhibitors, such as silver nitrate, cobalt chloride, nickel chloride and O-acetyl salicylic acid, significantly delayed the loss of regeneration potential in embryogenic cultures of Pennisetum americanum. In the presence of these chemicals, ethylene content in the atmosphere of the culture vessel was less than that of the control. Cefotaxime, silver nitrate and O-acetyl salicyclic acid did not have any effect on callus growth based on fresh weight, while growth based on dry weight was enhanced by O-acetyl salicyclic acid.
The influence of indoleacetic acid (IAA) and some IAA-amino acid conjugates in combination with benzyladenine (BA) on in vitro shoot regeneration from leaf discs of peanut and pigeonpea was investigated. The frequency of shoot regeneration and average number of shoot buds produced was dependent on the type of auxin present in the medium. Highest frequency of plant regeneration in peanut was induced by BA in combination with IAA or IAA-l-alanine, while in pigeonpea BA in combination with IAA or IAA-l-aspartic acid produced best results. Four hundred plants of peanut and 25 plants of pigeonpea were transplanted to soil.Abbreviations: BA-benzyladenine, IAA-indoleacetic acid, IAA-l-alanine-indole acetyl-l-alanine, IAA-aspartic acid -indole acetyl-l-aspartic acid, IAA-glycine -indole acetyl-l-glycine, IAAphenylalanine -indole-aeetyl-l-phenyl-alanine
Seed and seedling expiants of pigeonpea were evaluated for organogenesis and somatic embryogenesis. De novo plant regeneration through organogenesis was obtained from mature cotyledons, primary leaves and roots of seedlings. Production of multiple shoots from the cotyledonary node was observed in cultures of whole seeds on 6-benzylaminopurine enriched medium. Somatic embryos were induced from immature cotyledons and embryonal axes, however, well-developed plants could not be derived from these embryos. The regenerants obtained through organogenesis were transferred to the field and grown to maturity.
Transgenic peanut plants were produced using Agrobacterium mediated gene transfer. Primary leaf explants of peanut were co-cultivated with Agrobacterium tumefaciens LBA 4404 harbouring the binary plasmid pBI 121 (conferring β-glucuronidase activity and resistance to kanamycin) and cultured on regeneration medium supplemented with kanamycin to select putatively transformed shoots. They were rooted and plants were transferred to soil. Stable integration and expression of the transgenes were confirmed by NPT II assay, Southern blot hybridization and GUS assay.
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