Abstract
Transfer of foreign genes into higher plants mediated by Agrobacterium tumefaciens is a standard technique in plant genetic engineering. We have produced of morphologically normal and fertile transgenic plants from leaf explants inoculated with Agrobacterium tumefaciens strain LBA 4404 carrying binary vector pCAMBIA 1319Z, the latter of which contains Cry1Ac gene for making insect tolerant blackgram plant, a neomycin phosphotransferase (nptII) gene and a β-glucuronidase (GUS) gene (uidA) for histochemical assay of the transgene. The transformed green shoots, selected and rooted on medium containing kanamycin, tested positive for nptII and uidA genes by histochemical assay and PCR analysis. These plantlets were established in soil and grown to maturity to collect the seeds. Analysis of T0 plants showed the expression and integration of uidA into the plant genome. GUS activity in leaves, roots, flowers, anthers and pollen grains was detected by histochemical assay. For insect bioassay, the transgenic plants were toxic to bollworm larvae and lessened the damage caused by their feeding.
Highlights
Black gram [Vigna mungo (L.) Hepper] is an important leguminous source of protein for a large segment of the vegetarian population in the developing countries of Asia
For the first time, the conditions for establishing an A. tumefaciens-based transformation pro
The expression of Cry1Ac gene through Agrobacterium-mediated transformation and tolerant to bollworm (Helicoverpa armigera) insect was first reported in cotton [24], Transgenic potato was formed by the entry of Cry 1la1(Cry V) gene into potato genome and become tolerant to potato tuber moth [Phthorimaea operculella and Symmetris chematangolias (Lepidoptera: Gelechiidae)] [25], transgenic brinjal was made by the expression of Cry 1Ab gene into brinjal through Agrobacterium mediated transformation and tolerant to fruit
Summary
Black gram [Vigna mungo (L.) Hepper] is an important leguminous source of protein for a large segment of the vegetarian population in the developing countries of Asia. India is the largest producer and consumer of black gram in the world, but production is limited due to its high susceptibility to lepidopteron pest [3]. Flowers and developing pods are eaten by Heliothis virescence & Maruca caterpillar which cause a large reduction in both yield and quality. These borer insets-pests can be controlled to some extent by the spray of different chemicals and pesticides before the harvest of black gram fruits but these chemicals have harmful effect on human health. Genetic transformation of large-seeded legumes in general and V. mungo in particular has been difficult and challenging [4]-[10], considerable progress has been achieved in some grains and legumes as reported in Pea by [11], in mungbean by [12] and in chickpea by [13]-[15]
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