转匍匐翦股颖AsHSP26.8a拟南芥株系的光合作用研究

doi:10.16420/j.issn.0513-353x.2020-0967

摘要/Abstract

摘要：

通过测定转匍匐翦股颖小热激蛋白基因AsHSP26.8a拟南芥株系的根长、植株质量、蒸腾速率、气孔导度、净光合速率以及PSⅡ光能转化效率,探讨AsHSP26.8a对拟南芥光合作用的影响,并进一步通过qRT-PCR分析了拟南芥内源光合相关基因（rbcL、psaA、psbA、psaE、clpP、rpoB、rpoA、accD）的表达情况来揭示AsHSP26.8a参与光合作用的调控途径。结果表明：与野生型相比,转基因拟南芥根长显著缩短,鲜质量和干质量下降;光合速率、蒸腾速率、气孔导度和PSⅡ光能转化效率均有所下降,光合相关基因的表达也有所下调。由此认为AsHSP26.8a抑制了转基因拟南芥光合相关基因的表达,使转基因拟南芥的光合作用减弱,从而进一步抑制了转基因拟南芥根系生长和干物质的积累。

关键词: 匍匐翦股颖, 叶绿体小热激蛋白, AsHSP26.8a, qRT-PCR, 光合作用, 转基因, 拟南芥

Abstract:

In this paper,effects of a creeping bentgrass small heat shock protein,AsHSP26.8a,on photosynthesis in transgenic Arabidopsis thaliana were studied by measuring root length,plant weight,transpiration rate,stomatal conductance,net photosynthetic rate and PSⅡlight energy conversion efficiency of AsHSP26.8 transgenic A. thaliana. Furthermore,the expression of endogenous photosynthesis-related genes（rbcL,psaA,psbA,psaE,clpP,rpoB,rpoA,accD）in A. thaliana were analyzed by qRT-PCR to reveal the regulation pathway of AsHSP26.8a in plant photosynthesis. Results showed that compared with the wild type,the root length of transgenic A. thaliana was significantly shortened,the fresh weight and dry weight decreased,and the photosynthetic rate,transpiration rate,stomatal conductance and photochemical efficiency of transgenic A. thaliana decreased,and the expression of photosynthesis-related genes in transgenic A. thaliana was also down-regulated. It is suggested that AsHSP26.8a inhibits the expression of photosynthesis-related genes in transgenic A. thaliana,which weakened the photosynthesis of transgenic A. thaliana,and subsequently leaded to the inhibition of root growth and dry matter accumulation of transgenic A. thaliana.

Key words: Agrostis stolonifera, chloroplast small heat shock protein, AsHSP26.8a, qRT-PCR, photosynthesis, transgenic, Arabidopsis thaliana

中图分类号:

朱俊飞, 李鑫, 董康挺, 唐志菲, 边秀举, 王丽宏, 李会彬, 孙鑫博. 转匍匐翦股颖AsHSP26.8a拟南芥株系的光合作用研究[J]. 园艺学报, 2021, 48(8): 1619-1625.

ZHU Junfei, LI Xin, DONG Kangting, TANG Zhifei, BIAN Xiuju, WANG Lihong, LI Huibin, SUN Xinbo. Effect of Creeping Bentgrass Small Heat Shock Protein AsHSP26.8a on Transgenic Photosynthesis in Arabidopsis thaliana[J]. Acta Horticulturae Sinica, 2021, 48(8): 1619-1625.

图/表 4

表1 qRT-PCR引物信息表

Table 1 List of primers used in qRT-PCR

基因 Gene	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse primer
rbcL（ATCG00490）	F：TTGGCAGCATTCCGAGTAACTCCT;	R：CTGGTAAGTCCATCGGTCC
psaA（ATCG00350）	F：CTACTTTGCCACCCACTGC;	R：TGAGTGCTTTAGGGCGTCC
psbA（ATCG00020）	F：GCATAGCACTGAATAGGGAGCCG;	R：GCGACCTTGGATTGCTGTTGC
psaE（AT2G20260）	F：CGTGTCTTTCTTGCCGATGA;	R：TGGGTTTGGTGGCAGTAGC
clpP（ATCG00670）	F：GGAGGAGCAATTACCAAACG;	R：GCTTGGGCTTCTGTTGCTGAC
rpoB（ATCG00190）	F：GCGAAAGAATCCTCCTATGC;	R：CCACCTCACATCAATAACTC
rpoA（ATCG00740）	F：TGCGATGCGAAGAGCTTTAC;	R：CAATGATTTCCACAGCGGG
accD（ATCG00500）	F：GAAGGTTCACAAGCGGCTG;	R：GAAATAACTCGCTCAGAACAC
AtActin（AT2G37620）	F：CCGACAGAATGAGCAAAGAGA;	R：CCTCCAATCCAGACAGAGTATTT

图1 转AsHSP26.8a拟南芥株系（TG1、TG3）与野生型（WT）生长情况比较 “*”,“**”表示在0.05和0.01水平上的显著差异。下同。

Fig. 1 Comparison of the growth of ASHSP26.8a transgenic Arabidopsis thaliana lines（TG1,TG3）and wild-type（WT） “*”and“**”indicate significant differences between WT and each TG at P < 0.05 or 0.01 respectively. The same below.

图2 转AsHSP26.8a拟南芥株系（TG1、TG3）与野生型（WT）光合作用指标比较

Fig. 2 Comparison of photosynthetic indexes between AsHSP26.8a transgenic Arabidopsis thaliana lines（TG1,TG3）and WT

图3 拟南芥野生型（WT）和转AsHSP26.8a基因株系（TG1、TG3）的光合基因相对表达量

Fig. 3 The relative expression levels of photosynthetic genes in Arabidopsis thaliana wild-type（WT）and transgenic AsHSP26.8a gene lines（TG1,TG3）

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