SummaryPeanut (Arachis hypogaea L.) is a major species of the family, Leguminosae, and economically important not only for vegetable oil but as a source of proteins, minerals and vitamins. It is widely grown in the semi-arid tropics and plays a role in the world agricultural economy. Peanut production and productivity is constrained by several biotic (insect pests and diseases) and abiotic (drought, salinity, water logging and temperature aberrations) stresses, as a result of which crop experiences serious economic losses. Genetic engineering techniques such as Agrobacterium tumefaciens and DNA-bombardment-mediated transformation are used as powerful tools to complement conventional breeding and expedite peanut improvement by the introduction of agronomically useful traits in high-yield background. Resistance to several fungal, virus and insect pest have been achieved through variety of approaches ranging from gene coding for cell wall component, pathogenesis-related proteins, oxalate oxidase, bacterial chloroperoxidase, coat proteins, RNA interference, crystal proteins etc. To develop transgenic plants withstanding major abiotic stresses, genes coding transcription factors for drought and salinity, cytokinin biosynthesis, nucleic acid processing, ion antiporter and human antiapoptotic have been used. Moreover, peanut has also been used in vaccine production for the control of several animal diseases. In addition to above, this study also presents a comprehensive account on the influence of some important factors on peanut genetic engineering. Future research thrusts not only suggest the use of different approaches for higher expression of transgene(s) but also provide a way forward for the improvement of crops.
Pigeon pea [Cajanus cajan (L.) Millsp.] (Family: Fabaceae) is an important legume crop cultivated across 50 countries in Asia, Africa, and the Americas; and ranks fifth in area among pulses after soybean, common bean, peanut, and chickpea. It is consumed as a major source of protein (21%) to the human population in many developing countries. In India, it is the second important food legume contributing to 80% of the global production. Several biotic and abiotic stresses are posing a big threat to its production and productivity. Attempts to address these problems through conventional breeding methods have met with partial success. This paper reviews the chronological progress made in tissue culture through organogenesis and somatic embryogenesis, including the influence of factors such as genotypes, explant sources, and culture media including the supplementation of plant growth regulators. Comprehensive lists of morphogenetic pathways involved in in vitro regeneration through organogenesis and somatic embryogenesis using different explant tissues of diverse pigeon pea genotypes are presented. Similarly, the establishment of protocols for the production of transgenics via particle bombardment and Agrobacterium-mediated transformation using different explant tissues, Agrobacterium strains, Ti plasmids, and plant selectable markers, as well as their interactions on transformation efficiency have been discussed. Future research thrusts on the use of different promoters and stacking of genes for various biotic and abiotic stresses in pigeon pea are suggested.
Agrobacterium-mediated genetic transformation was performed using embryonic axes explants of pigeon pea. Both legume pod borer resistant gene (cry1Ac) and plant selectable marker neomycine phosphor transferase (nptII) genes under the constitutive expression of the cauliflower mosaic virus 35S promoter (CaMV35S) assembled in pPZP211 binary vector were used for the experiments. An optimum average of 44.61% successfully hardened dot blot Southern hybridization positive plants were obtained on co-cultivation media supplemented with 200 µM acetosyringone without L-cysteine. The increased transformation efficiency from a baseline of 11.53% without acetosyringone to 44.61% with acetosyringone was further declined with the addition of different concentrations of L-cysteine to co-cultivation media. Transgenic shoots were selected on 50 and 75 mg L -1 kanamycin. Rooting efficiency was 100% on half-strength Murashige and Skoog medium with 20 g L -1 sucrose and 0.5 mg L -1 indole butyric acid in the absence of kanamycin. Furthermore, 100% seed setting was found among all the transgenic events. The plants obtained were subjected to multi-and nochoice tests to determine the behavioral responses and mortality through Helicoverpa armigera bioassays on the leaf and relate their relationship with the expression of cry1Ac protein which was found to be less in leaf as compared to the floral buds, anther, pod, and seed.
In vitro regeneration of pigeon pea through organogenesis and somatic embryogenesis was demonstrated with pigeon pea cv. JKR105. Embryonic axes explants of pigeon pea showed greater regeneration of shoot buds on 2.5 mg L −1 6-benzylaminopurine (BAP) in the medium, followed by further elongation at lower concentrations. Rooting of shoots was observed on half-strength Murashige and Skoog (MS) medium with 2 % sucrose and 0.5 mg L −1 3-indolebutyric acid (IBA). On the other hand, the regeneration of globular embryos from cotyledon explant was faster and greater with thidiazuron (TDZ) than BAP with sucrose as carbohydrate source. These globular embryos were maturated on MS medium with abscisic acid (ABA) and finally germinated on half-strength MS medium at lower concentrations of BAP. Comparison of regeneration pathways in pigeon pea cv. JKR105 showed that the turnover of successful establishment of plants achieved through organogenesis was more compared to somatic embryogenesis, despite the production of more embryos than shoot buds.
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