Transformation Of Pepper Using Binary Expression Vector With Bt And CpTI

Pepper (Capsicum annuum L.) is a common vegetable, has high nutritional and medicinal value and high market demand. While in the production, the insect-pest serious influences its development of yield and quality seriously, genetic engineering breeding technology provides a new way to creat insect-resistant pepper varieties. In this experiment,’S1’ and ’405’was selected as the peppers material to study the differentiation rate of different geno-type, explent type, BR concentration and Mannose concentration, which optimized the system of Pepper regeneration. This study used Agrobacterium-mediated to conducted the transformation of exogenous genes and received new insect-resistance Pepper material. The test results were as follows:l.The result of differentiation rate of petiole cotyledons, class Flamingo-bill and hyp-ocotyls showed that class Flamingo-bill was the strongest one with the highest regen-eration and differentiation, which reached47.0%, the hypocotyls followed by it, the lowest one was petiole cotyledons, almost no differentiation.2. The compare result of two pepper materials’differentiation rate class Flamingo-bill showed that the differentiation rate of pepper material’S1’was higher than pepper materia1’405’. The differentiation rate of pepper material’S1’ is47%and pepper materia1’405’is just30%. We can conclude that the regenerative differentiation capacity of pepper material’S1’ is more stronger than pepper material’405’.3. When treated the class Flamingo-bill of pepper material with different BR concentrations range from0.001mg/L to0.010mg/L, the differentiation rate of class Flamingo-bill improved obviously. When the BR=0.01mg/L, this change reached the top.4The result of treating the class Flamingo-bill of pepper material with different Mannose concentrations showed that12g/L Sucrose+18g/L Mannose is the best ratio to select Resistant sho-ots.5. Using this study optimized genetic regeneration system obtained transgenic plants of two pepper materials. PCR analysis and Southern blotting confirmed two insect-resistant genes have been integrated into the genome of the pepper successfully. PCR positive rate was16.40%, the conversion rate of0.55%; Southern blotting results show that a total of five pepper plants were positive pepper, the positive rate was45.45%.6. The results of the Chlorophenol red detection, RT-PCR detection and feeding insect experiments found that Bt genen and CpTI gene can express in transgenic pepper material plants. These To pepper provide new varieties to cultivate double insect-resistant genes pepper.