Cloning and genetic transformation of melon CmHSP83

• 1、Huazhong Agricultural University，College of Horticulture and Forestry，Key Laboratory of Horticultural Plant Biology (HAU), MOE, Wuhan 430000, China

Abstract：Melon (Cucumis melon L.) seedlings often suffer from waterlogging stress in rain season. Screening genes associated with waterlogging tolerance in melon is useful for breeding tolerant cultivars.A gene that encoding a heat shock protein 83(CmHSP83) were selected as candidates being associated with waterlogging tolerance in melon by RNA-seq analysis.In this project,the CDS of CmHSP83 were cloned and the sequence were analyzed by bioinformatics methods. The results revealed that CmHSP83 protein belongs to HSP90 family and may locate in endoplasmic reticulum.An anaerobic response element(ARE) and several other cis-regulatory elements associated with stress response exist in the up-stream regulatory sequence of CmHSP83. QRT-PCR analysis indicated that the expression level of CmHSP83 gene can be induced by waterlogging,drought and salt stress. Subsequently, the over-expression vector, PTCK303-CmHSP83, was successfully constructed. Ultimately, the gene was transformed into the melon 'M63' by Agrobacterium mediated transformation and six trangenetic plants were obtained. PCR analysis of T1 lines indicated that foreign gene was inherited from T0 plant. All the above work has laid a good foundation for the further analysis of gene function.

Keywords： melon waterlogging tolerance heat shock protein transformation