甘蔗ASR基因克隆及水分胁迫下的表达分析

【目的】克隆甘蔗ASR（ABA-stress-ripening）基因的序列并检测其在干旱胁迫下的表达量,为甘蔗抗逆胁迫机制的研究提供实验依据。【方法】以甘蔗叶片总RNA为模板,通过RT-PCR扩增ASR基因的cDNA,采用生物信息学软件分析所克隆基因的编码蛋白特性,并用荧光定量PCR分析该基因的表达情况。【结果】克隆得到的cDNA片段长度为402 bp,编码133个氨基酸,命名为SoASR1（GenBank登录号：JQ712581）。同源性分析表明,SoASR1基因在12种植物中的一致性为56%~99%。实时荧光定量分析结果表明,随着甘蔗干旱胁迫时间的延长,SoASR1基因表达量先升高后下降,...

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Published in西南农业学报 Vol. 30; no. 10; pp. 2196 - 2201
Main Author 梁潘霞;邢颖;廖青;黄杏;李长宁;黄太庆;江泽普;李杨瑞
Format Journal Article
LanguageChinese
Published 广西农业科学院资源与环境研究所,广西南宁,530007%中国农业科学院甘蔗研究中心/农业部广西甘蔗生物技术与遗传改良重点实验室/广西甘蔗遗传改良重点实验室,广西南宁,530007 2017
Subjects
ASR基因
干旱胁迫
甘蔗
甘蔗
ASR基因
干旱胁迫
Drought
Sugarcane
ABA-stress-ripening
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ISSN1001-4829
DOI10.16213/j.cnki.scjas.2017.10.007

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Summary:【目的】克隆甘蔗ASR（ABA-stress-ripening）基因的序列并检测其在干旱胁迫下的表达量,为甘蔗抗逆胁迫机制的研究提供实验依据。【方法】以甘蔗叶片总RNA为模板,通过RT-PCR扩增ASR基因的cDNA,采用生物信息学软件分析所克隆基因的编码蛋白特性,并用荧光定量PCR分析该基因的表达情况。【结果】克隆得到的cDNA片段长度为402 bp,编码133个氨基酸,命名为SoASR1（GenBank登录号：JQ712581）。同源性分析表明,SoASR1基因在12种植物中的一致性为56%~99%。实时荧光定量分析结果表明,随着甘蔗干旱胁迫时间的延长,SoASR1基因表达量先升高后下降,干旱胁迫加硅处理的甘蔗叶片SoASR1基因表达量峰值比干旱处理提前34 h出现。【结论】克隆获得甘蔗SoASR1基因,SoASR1基因受到干旱胁迫的诱导表达,在转录水平上参与了甘蔗抗干旱胁迫反应,可作为候选基因用于甘蔗抗旱机制研究。施硅能进一步提高甘蔗抗旱性。
Bibliography:Sugarcane ; Drought ; ABA-stress-ripening
Objective】To provide experimental basis for further research on regulation mechanism of ASR in response to drought,ASR（ ABAstress-ripening） gene from sugarcane was cloned and its expression level under drought stress was detected.【Method】Based on the total RNA of sugarcane leaves,the cDNA of ASR gene was amplified by RT-PCR,the characteristics of the deduced protein were analyzed by using bioinformatics software and its expression was analyzed by quantitative real-time PCR. 【Result】The results showed that the cDNA of ASR gene which obtained by RT-PCR,was 402 bp in full length,and it encoded a putative ASR protein with 133 amino acids,named SoASR1,its GenBank accession number was JQ712581. Comparison of the amino acids sequences homology in SoASR1 protein from 12 different species indicated that the ASR protein had 56 % to 99 % identity in amino acids sequence with other plants. The results of quantitative realtime PCR analysis showed that the mRNA of SoASR1 was increas
ISSN:1001-4829
DOI:10.16213/j.cnki.scjas.2017.10.007