| 杉木天冬酰胺合成酶基因的克隆与表达分析 |
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| 引用本文： | 饶丽莎,官清娜,林思祖,叶义全,许珊珊.杉木天冬酰胺合成酶基因的克隆与表达分析[J].热带作物学报,2018,39(5):931-939. |
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| 作者姓名： | 饶丽莎 官清娜 林思祖 叶义全 许珊珊 |
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| 作者单位： | 1. 福建农林大学林学院
2. 国家林业局杉木工程技术研究中心 |
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| 摘 要： | 为了解天冬酰胺合成酶(asparagine synthetase,AS)基因在杉木生长发育及逆境中的作用机制,以杉木种子及其萌发的幼苗为材料,利用RT-PCR和RACE技术获得杉木ASN基因的c DNA序列(Cl ASN1)和基因组序列,并对该基因在不同生长发育阶段和逆境胁迫下的表达模式和Asn含量变化进行分析。结果表明:Cl ASN1基因c DNA全长1 609 bp,编码443个氨基酸;生物信息学分析结果表明,该基因为含跨膜结构的不稳定亲水蛋白,无信号肽,定位于过氧化物酶体,具有多样的磷酸化位点;进化树分析结果显示,该基因与北美云杉和樟子松亲缘关系最近;Cl ASN1基因组序列全长3 210 bp,含10个外显子和9个内含子;q PCR结果表明,该基因从浸种24 h到幼苗期表达量呈上升趋势,且在干旱胁迫、铝胁迫和光暗处理下均可诱导表达;除铝胁迫24 h和持续光照24 h外,Cl ASN1表达模式与Asn含量的变化趋势相同,此结果表明Cl ASN1基因通过催化Asn的合成参与杉木幼苗的生长发育及胁迫响应。
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| 关 键 词： | 杉木 ASN基因 克隆 表达分析 |
Cloning and Expression Analysis of Asparagine Synthetase Gene from Cunninghamia lanceolata |
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| RAO Lisha,GUAN Qingna,LIN Sizu,YE Yiquan,XU Shanshan.Cloning and Expression Analysis of Asparagine Synthetase Gene from Cunninghamia lanceolata[J].Chinese Journal of Tropical Crops,2018,39(5):931-939. |
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| Authors: | RAO Lisha GUAN Qingna LIN Sizu YE Yiquan XU Shanshan |
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| Institution: | 1 College of Forestry, Fujian Agriculture and Forestry University;
2 Engineering & Research Center of Chinese Fir, State Forestry Administration; |
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| Abstract: | In order to investigate the underlying mechanism of ASN gene of Cunninghamia lanceolata in the growth stages and under abiotic stresses, RT-PCR combined with RACE was employed to clone the complete cDNA sequence and DNA sequence from Cunninghamia lanceolata seedling, and the expression of ClASN1 gene and the Asn content under various growth stages and abiotic stresses were analyzed. The results showed that the complete cDNA sequence of ClASN1 was 1 609 bp, encoding 443 amino acids. The bioinformatics software predicted that ClASN1 protein was unstable and hydrophilic with transmembrane structure without signal peptide, located in peroxidase and with various phosphorylation sites. Anglicizing phylogenetic tree of ASN genes in different plants indicated that ClASN1 had the closest relative with Picea sitchensis and Pinus sylvestris. The full length of genomic ClASN1 was 3 210 bp, which contained 10 exons and 9 introns. qPCR results showed that the expression of ClASN1 increased with the time elapsed from seed germination to seedling stage, and was also induced under drought stress, aluminum stress and light dark stress, moreover, the general expression tendencies of ClASN1gene accorded with the content of Asn under various stages from seed germination to seedling growth and different stresses expect for aluminum stress 24 h and light stress 24 h. In conclusion, the above results suggested that ClASN1 may play a role in the various stages from seed germination to seedling growth and stress responses. |
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| Keywords: | Cunninghamia lanceolata ASN gene cloning expression analysis |
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