Green gram (Vigna radiata L. Wilczek) is one of the important pulse crops in India belongs to the family Fabaceae. The production and productivity is severely affected by abiotic stresses like drought and salinity. Both control and transformed plants of green gram variety 'IPM-02-03' were evaluated for moisture stress. Soil moisture stress was imposed at the reproductive stage by withholding irrigation completely for 7 days. Morphological, physiological and biochemical parameters were evaluated for drought tolerance at 0 th , 3 rd and 7 th days with 100%, 50% and 25% water holding capacity, respectively during drought stress. In almost all the cases under moisture stress, transgenic plants showed higher yield and 100 seed weight. Pigments like chlorophyll and carotenoids accumulation was drastically decreases with increase in moisture stress. Higher accumulation of proline and antioxidant enzymes in transgenic plants than control showed better survival in stress condition. The present study suggests that, over expression of TIP1 gene in transformed plant can enhance better survival under drought stress condition.
Background: Green gram is grown in many parts of India as a source of dietary protein (21-25%). It is an important nitrogen fixing crop which fixes atmospheric nitrogen (119-140 kg/ha) to soil and enhance the soil productivity. In the present investigation, efficient Agrobacterium-mediated genetic transformation of Vigna radiata L. (Wilczek) has been achieved with VrTIP1 gene for abiotic stress resistance i.e. moisture and salinity stress.
Methods: Four days old shoot tip and cotyledonary node were used for in vitro regeneration with MS medium supplemented with BAP 2.0 mg/l, kinetin 0.5 mg/l and 50 mg/l kanamycin for co-cultivation with Agrobacterium tumefaciens strains, LBA 4404. The modified binary vector pCXSN, EHA105 containing hygromycin phosphotransferase II (hpt II) marker genes and a synthetic TIP1 gene under a constitutive CaMV35S promoter were used for transformation of Vigna radiata L. cotyledonary node explants. Putative transformants selected from hygromycin resistant shoots were subsequently rooted on MS medium supplemented with 1.0 mg/l NAA and later transferred to sterile vermiculite followed by transfer to the transgenic green house.
Result: The T1 plants were produced from PCR positive T0 plants and analysed for presence and integration of transgenes in putative T1 plants were confirmed by polymerase chain reaction (PCR) amplification of 752 bp of hpt II fragment. This protocol can be effectively used for transferring new traits in greengram and other legumes for their quantitative and qualitative improvements.
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