马铃薯DGK基因家族克隆及表达分析  

Cloning and Expression Analysis of DGK Gene Family in Potato(Solanum tuberosum)

作　　者：王迪 何双双 单雅成 黄彦 曾相琴 田周娥 胡新喜[1] 秦玉芝[1] 林原 WANG Di;HE Shuang-Shuang;SHAN Ya-Cheng;HUANG Yan;ZENG Xiang-Qin;TIAN Zhou-E;HU Xin-Xi;QIN Yu-Zhi;LIN Yuan(College of Horticulture,Hunan Agricultural University/Hunan Provincial Potato Engineering and Technology Research Centre/Key Laboratory for Vegetable Biology of Hunan Province,Changsha 410128,China)

机构地区：[1]湖南农业大学园艺学院/湖南省马铃薯工程技术研究中心/蔬菜生物学湖南省重点实验室,长沙410128

出　　处：《农业生物技术学报》2024年第12期2715-2730,共16页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(3190150807)。

摘　　要：二酰基甘油激酶(diacylglycerol kinase,DGK)是脂质信号通路中的关键酶,在植物激素信号传导、生物和非生物胁迫中起着重要作用。本研究参考模式植物拟南芥(Arabidopsis thaliana)中AtDGKs蛋白序列以及DGK保守结构域信息,分析马铃薯(Solanum tuberosum)中StDGKs基因的序列特点,对此多基因家族进行序列克隆和分析,并对其在干旱和盐胁迫响应以及马铃薯块茎发育中的表达模式进行分析。结果表明,从马铃薯基因组数据库(S.tuberosum group Phureja DM1-3 v6.1)中检索出11个StDGKs,并成功克隆出其中6个基因StDGK1、StDGK2、StDGK5、StDGK6、StDGK7和StDGK8。序列分析结果显示,6个StDGKs基因全长为1460~2220 bp,序列比对相似性超过95%。实时荧光定量PCR结果显示,6个StDGKs的表达均受到干旱和盐胁迫诱导,干旱处理下,与对照组相比,StDGK2、StDGK8在12 h、StDGK6在4 h的表达量达到峰值;盐胁迫处理下,StDGK1、StDGK5、StDGK7的表达量在4 h达到峰值,StDGK2、StDGK6在15 min达到峰值。此外,本研究发现,StDGKs可能在马铃薯块茎发育初期发挥重要作用,StDGK1、StDGK8在块茎发育早期至中期的表达量逐渐升高,在块茎膨大后期又缓慢下降。本研究为深入解析StDGK基因家族功能提供基础资料。Diacylglycerol kinase(DGK)is a key enzyme in lipid signalling pathways and plays an important role in phytohormone signalling,biotic and abiotic stresses.This study analysed the sequence characteristics of the StDGKs genes in potato(Solanum tuberosum)with reference to the protein sequence of the model plant Arabidopsis thaliana AtDGKs as well as information on the conserved structural domains of DGK,and sequence cloning and analysis of this multigene family were performed,followed by analysis of its expression pattern in response to drought,salt stress and potato tuber development.The results showed that 11 StDGKs were screened from the potato genome database(S.tuberosum group Phureja DM1-3 v6.1)and 6 of them(StDGK1,StDGK2,StDGK5,StDGK6,StDGK7,StDGK8)were successfully cloned.Sequence analysis showed that the full length of the 6 StDGKs were 1460~2220 bp,and the similarity of sequence comparison was more than 95%.Real-time fluorescence quantification PCR showed that the expression of 6 StDGKs were induced by drought and salt stress.Compared with the control,the expression of StDGK2 and StDGK8 were significantly up-regulated and peaked at 12 h of drought treatment and StDGK6 at 4 h.The expression of StDGK1,StDGK5,StDGK7 were significantly higher than that of the control under salt stress treatment and peaked at 4 h,and that of StDGK2 and StDGK6 peaked at 15 min of salt treatment.In addition,this study found that StDGKs played an important role in the early stage of potato tuber development.For example,the expression of StDGK1 and StDGK8 gradually increased from the early to the middle stage of tuber development,and then slowly decreased at the late stage of tuber expansion.This study provides basic materials for a deeper understanding of StDGKs gene function.

关 键 词：马铃薯 二酰基甘油激酶(DGK) 非生物胁迫 块茎发育 表达模式 

分 类 号：S532[农业科学—作物学]