青杄PwRbcS 基因克隆及对逆境响应分析

doi:10.13560/j.cnki.biotech.bull.1985.2016.10.019

摘要/Abstract

摘要： RbcS基因编码了植物光合作用中核酮糖1,5二磷酸羧化酶/加氧酶的亚基,这种酶能催化二氧化碳的固定和碳氧化两者之间竞争反应。本实验中通过RACE-PCR方法成功获得了青杄RbcS的cDNA全长,对PwRbcS的cDNA序列进行了生物信息学分析预测,同时利用实时定量PCR技术测定青杄各个组织及非生物胁迫下PwRbcS的表达水平。结果表明：PwRbcS基因cDNA全长936 bp,编码区共552 bp,共编码186个氨基酸。蛋白质分子量为20.712 6 kD,等电点为9.07。组织特异性表达结果发现PwRbcS该基因主要在叶片中表达,且成熟叶表达量最高。非生物胁迫实验结果表明,ABA 和NaCl以及低温处理处理下响应十分明显。在NaCl处理下,表达量先上升后下降,处理6 h时达到对照的14倍,施加ABA时,在处理8 h时上升至对照的24倍,低温胁迫下,表达量先上升后下降,6 h达到对照的12倍,而高温胁迫及PEG（polyethylene glycol）下表达量变化不明显。因此PwRbcS作为一个在木本植物青杄中发现的新的RbcS基因,可能在青杄逆境响应中行使一定的功能。

关键词: Picea wilsonii, RbcS, 生物信息学分析, 组织表达, 胁迫响应

Abstract: RbcS gene encodes a subunit of nuclear plant photosynthesis 1,5 two ribulose bisphosphate carboxylase / oxygenase. In this study,PwRbcS cDNA was obtained using RACE -PCR method. Bioinformatics analysis and real-time PCR technique were used to determine the relative expression of PwRbcS. The results showed that the full-length of PwRbcS cDNA was 936 bp,including the ORF of 552 bp encoding 186 amino acids. The theoretical molecular weight of PwRbcS is 20.712 6 kD and the pI value was 9.07. RT-qPCR showed that PwRbcS is mainly expressed in the needles,especially in mature needles. The expression of PwRbcS changed obviously under the treatment of NaCl,ABA and low temperature,respectively. After NaCl treatment,the expression level of PwRbcS changed significantly,reaching to 14 times than the control at 6 h. The expression level of PwRbcS reached to 24 times than the control after 8 h-ABA-treatment. And under low temperature stress,the expression increased gradually to 12 times than the control and followed by a drop,while under high temperature and PEG stress expression level did not change significantly. Therefore,PwRbcS may play an important role during abiotic stresses in planta.

Key words: Picea wilsonii, RbcS, bioinformatics, tissue expression, stress response

鞠丹, 胡安妮, 张杰, 张凌云. 青杄PwRbcS 基因克隆及对逆境响应分析[J]. 生物技术通报, 2016, 32(10): 154-162.

JU Dan, HU An-ni, ZHANG Jie, ZHANG Ling-yun. Cloning and Stress Response Analysis of PwRbcS in Picea wilsonii[J]. Biotechnology Bulletin, 2016, 32(10): 154-162.

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