Construction Of Rnai Vectors Of 3 Bzip Candidate Genes From Capsicum Annuum And Its Agrobacterium Mediated Pepper Transformation

Capsicum annuum L. is one of the most important vegetables all over china, and also an important exporting crops in China. A vast of reduction in its yield is usually caused by continuous exposure to various stresses including abiotic such as drought, abnormal temperature, and biotic stresses such as diseases. Molecular mechanism of pepper stresses stress resistance is important for its genetic improvement. Previous studies suggested that transcription factors play great roles in the adaptation of plants to various stresses, and basic leucine zipper is one of the most conservative transcription factors and exist widely in eukaryote. Members of the bZIP family have been identified in the genome of Arabidopsis, rice and soybean and play important roles in the regulation of plant growth and development, transduction of light signal, defense reaction, response of biotic and abiotic stresses as well as ABA mediated signal transduction. So far little is known about the roles of members of bZIP family in pepper and it is presumed that bZIPs may also play important roles in environmental adaptation of pepper, and the functional study of bZIPs is important for molecular mechanism elucidation and efficient genetic improvement of pepper stresses resistance.The RNAi vectors of 3 bZIP candidate genes isolated from Capsicum annuum cDNA library of leaves challenged with pathogen infection in the previous studies were constructed in this study, by using the 5'or 3'untranslated regions of the full length cDNAs of the 3 candidate genes and the gateway compatible pHELLSGATE 12 vector. The 3 RNAi vectors were further transformed into the genome of a local pepper variety p120 by Agrobacterium mediated method and the 8,9,12 clones of T0 regenerated Kan resistant plants of the 3 RNAi vectors was acquired, and 4,5,8 corresponding transgenic plants were verified by PCR amplification primed by the specific primers of the corresponding 3 bZIP regions, and the corresponding T1 lines were also acquired by self- pollination. The results of this study laid an important base for the further functional analysis of the 3 candidate genes by RNAi analysis.