The Isolation And Genetic Transformation Of TaCBL3 In Wheat (Triticum Aestium L.)

CBL proteins (calcineurin B-like protein) were identified as a group of calcium sensor unique to plants and some protozoa in recent years. CBLs activate the downstream calcium signaling transduction pathways through interacting with CIPKs ( CBL Interacting Protein Kinase). The Ca2+ -CBL-CIPK signaling pathway plays important roles in response to various abiotic stresses such as high salt, drought, low temperature in planta.In the present research, a full-length cDNA sequence encoding TaCBL3 was cloned. TaCBL3 over expression vector was constructed and delivered into developing wheat scutulum by particle bombardment.The main results are as follows:(1) We first searched the ESTs of TaCBL genes in database and then assembled them into TaCBL genes. Specific primers were designed and a TaCBL cDNA sequence was cloned by RT-PCR.(2) Sequence alignment and phylogenetic tree analysis showed that this gene was highly homologous to OsCBL3, therefore it was designated as TaCBL3 gene. Arabidopsis and rice CBL proteins generally have four EF-hand structures, while only three EF-hand structures were found in TaCBL3.(3) It is reported that the OsCBL3 gene plays a important role in multiple abiotic stresses, such as salt, drought and cold. Therefore, in order to further study the role of TaCBL3 in abiotic stress, and to breed new wheat varieties which are resistant to abiotic stresses, TaCBL3 gene over-expression vector was constructed by inserting it into pAHC25, and the over-expression vector was introduced into developing wheat scutulum in Zhengmai 9023 by particle bombardment, and regenerated transgenic wheat was obtained.(4) The vector of pAHC25 contains the bar gene, so we identified the transgenic plants of wheat through the PCR amplification of bar gene, and gained seven positive transgenic wheat.The results thus obtained in this research laid the base for further investigating the role of TaCBL3 in response to abiotic stresses in wheat.