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Putative Transgenic Plants through in Planta Transformation against Phytophthora Foot Rot in Black Pepper (Piper nigrum L.)
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Black pepper (Piper nigrum L.), popular as ‘king of spices’ is one of the important export earning spice crop of India. One of the serious reasons causing the low productivity of black pepper during the last decades was the incidence of foot rot disease caused by the Phytophthora capsici in the pepper plantations of Kerala, the major pepper producer state in India. Since all the available cultivated varieties are susceptible to this disease, an attempt was done for utilising the disease resistance available in the related pepper species by adopting a different plant transformation approach. The in planta transformation via pollen tube pathway was done in the black pepper variety Panniyur-2, using the total exogenous DNA of Piper colubrinum , a wild relative species of Piper resistant to the dreaded foot rot disease caused by Phytophthora capsici. The resulting putative transformant seeds were germinated in vitro by embryo rescue technique. These germinated putative transformants were later subjected to in vitro multiplication, elongation and ischolar_maining. These cultures were screened in vitro in the ischolar_maining phase by incorporating the toxic culture filtrate of the pathogen P. capsici in the ischolar_maining media. The ischolar_mained putative transformant plantlets were hardened and screened artificially for disease tolerance under ex vitro conditions. The survived seedlings were planted out. The RAPD analysis of the plantlets with the decamer primers OPA 08 and OPG 08 have shown variation in banding pattern compared to the DNA recipient parent P. nigrum variety Panniyur-2.
Keywords
Black Pepper, Piper nigrum, Piper colubrinum, In planta Transformation, Pollen Tube Pathway Transformation, In vitro Germination, Embryo Rescue, Disease Screening, RAPD Analysis.
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