关键词: 葡萄, SnRK2家族, 基因克隆, 荧光定量, 基因表达

Abstract:

Full length cDNA sequence of 8 grape SnRK2 genes（VvSnRK2）were cloned from grapevine Ruby Seedless using RT-PCR，sequence analysis showed that the N-terminal domain of VvSnRK2 was highly conserved，while divergent in the C-terminal. The phylogenyetic analysis showed that VvSnRK2s could be divided into three groups. The VvSnRK2s protein carry all acidic amino acids and they all hydrophilic proteins. All VvSnRK2 but VvSnRK2.2，VvSnRK2.8 genes carry 9 exons. The predicted secondary structure suggested that the main structure of eight proteins are alpha helix，beta turn and random coil. The subcellular localization were predicted in the cytoplasmic. The analysis of cis-regulatory elements showed that VvSnRK2s contained one or more of ABRE，DRE/CRT，LRTE elements except VvSnRK2.1，VvSnRK2.2 and VvSnRK2.6. Quantitative real-time PCR data indicate that VvSnRK2.7 and VvSnRK2.8 were expressed at the highest-level in the roots and stems，which were as 3.8 and 5.0 times as leaves，respectively. Under the 0 ℃ and–4 ℃ treatments，VvSnRK2.2 was down-regulated，while the expression was 0 of VvSnRK2.8. Under the 30 ℃ treatment，VvSnRK2.2 and VvSnRK2.5 were up-regulated 3.8 and 3.6 folds，respectively. VvSnRK2.1 and VvSnRK2.2 were response to salt stress，while VvSnRK2.5 was highly response to drought stress.

Key words: Vitis vinifera, SnRK2 family gene, cloning, quantitative PCR, gene expression