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ARF基因干扰表达对不同发育阶段和贮藏条件马铃薯酶活性的影响
引用本文:周香艳,张宁,刘柏林,裴瑞芳,司怀军,王蒂.ARF基因干扰表达对不同发育阶段和贮藏条件马铃薯酶活性的影响[J].草业学报,2016,25(4):133-139.
作者姓名:周香艳  张宁  刘柏林  裴瑞芳  司怀军  王蒂
作者单位:1.甘肃农业大学生命科学技术学院,甘肃 兰州 730070; 2.甘肃农业大学农学院,甘肃 兰州 730070; 3.甘肃省作物遗传改良与种质创新重点实验室,甘肃省干旱生境作物学省部共建国家重点实验室培育基地,甘肃 兰州 730070
基金项目:甘肃农业大学盛彤笙科技创新基金项目(GSAU-STS-1333),国家自然科学基金项目(31160298),甘肃省杰出青年基金项目(1308RJDA011),甘肃省农业科技创新项目(GNCX-2012-49)资助。
摘    要:以前期研究获得的马铃薯栽培品种“甘农薯2号”ADP核糖基化因子(ARF)基因干扰表达转化植株为材料,用qRT-PCR法对转基因植株的ARF基因表达量进行了测定,结果表明,不同发育阶段转基因叶片中ARF基因的相对表达量先降低,生长后期略有增加,表明ARF基因干扰表达量随马铃薯生长发育发生改变。ARF基因干扰表达影响马铃薯叶片中酶活性,不同发育阶段转基因叶片与对照相比,多酚氧化酶(polyphenol oxidase, PPO)活性升高11.61%~27.84%,硝酸还原酶(nitrate reductase, NR)活性提高21.10%~41.32%,磷脂酶D(phospholipase D, PLD)活性降低2.88%~57.64%,蔗糖磷酸合成酶(sucrose phosphate synthase, SPS)活性提高29.00%~39.57%;不同温度(4℃和室温)贮藏的块茎中ARF基因的相对表达量变化趋势一致:均先降低,再升高,但前者较后者ARF基因相对表达量显著降低。室温较4℃贮藏的转基因块茎PPO活性升高30.44%~56.28%,NR活性提高17.41%~40.92%,PLD活性降低24.39%~85.11%,SPS活性提高30.89%~45.78%。室温较4℃贮藏的非转基因块茎PPO活性升高25.11%~70.66%,NR活性提高36.07%~89.62%,PLD活性降低11.35%~72.64%,SPS活性提高27.31%~61.33%。本研究通过探讨ARF基因干扰表达对马铃薯生理生化特性的影响,为进一步研究ARF基因在马铃薯生长发育调控中的作用提供一定的理论基础。

关 键 词:马铃薯  ARF基因  干扰表达  转基因植株  酶活性
收稿时间:2015-04-14

Effects of RNAi expression of the ARF gene on enzyme activity at different developmental stages and storage temperatures in potato
ZHOU Xiang-Yan,ZHANG Ning,LIU Bai-Lin,PEI Rui-Fang,SI Huai-Jun,WANG Di.Effects of RNAi expression of the ARF gene on enzyme activity at different developmental stages and storage temperatures in potato[J].Acta Prataculturae Sinica,2016,25(4):133-139.
Authors:ZHOU Xiang-Yan  ZHANG Ning  LIU Bai-Lin  PEI Rui-Fang  SI Huai-Jun  WANG Di
Institution:1.College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China; 2.College of Agronomy, Gansu Agricultural University, Lanzhou 730070, China; 3.Gansu Key Laboratory of Crop Genetic and Germplasm Enhancement, Gansu Provincial Key Laboratory of Aridland Crop Science, Lanzhou 730070, China
Abstract:ADP ribosylation factor (ARF )gene expression interference in a transformed potato cultivar “Gan-nongshu 2”was evaluated.Real time fluorescence quantitative polymerase chain reaction (qRT-PCR)showed that the relative expression of the ARF gene varied in the transgenic plants at different developmental stages and increased slightly at latter growth stages,indicating that ARF gene interference expression varied with ma-turity.Enzyme activity in the leaves was effected by ARF gene interference expression.For leaves of transgenic plants at different development stages,PPO activity increased by 11.61%-27.84%,NR activity increased by 21.10%-41.32%,PLD activity decreased 2.88% -57.64% SPS activity increased by 29.00% - 39.57%when compared with the control.Relative expression of the ARF gene in stored tubers under different storage temperatures (room temperature and 4℃)showed the same trend;initially decreasing and then increasing.For transgenic tubers stored at room temperature,the activity of PPO increased by 30.44%-56.28%,NR activity increased by 17.41% - 40.92%,PLD activity decreased 24.39% - 85.11%,SPS activity increased by 30.89%-45.78% when compared with transgenic tubers stored at 4℃.For non-transgenic tubers stored at room temperature,PPO activity increased by 25.11%-70.66%,NR activity increased by 36.07%-89.62%, PLD activity decreased 11.35%-72.64% and SPS activity increased by 27.31%-61.33%.This study investi-gated the effect of RNAi expression of ARF gene on physiological and biochemical characteristics in potato to provide a theoretical basis for further study of the regulation role of the ARF gene in potato of different growth and stage of development.
Keywords:potato  ARF gene  interference expression  transgenic plants  enzyme activity
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