| 摘要: |
| 酸性转化酶作为蔗糖代谢中的关键酶,在还原糖积累型荔枝中表达及酶活性显著高于蔗糖积累型荔枝。为了全面了解酸性转化酶的生物学特征,该文通过运用生物信息学方法对荔枝果实酸性转化酶LcSAI基因的基本理化性质、蛋白二级结构、亲水性/疏水性、跨膜结构域、信号肽、磷酸化位点、保守结构域、系统进化进行系统的分析; 利用qRT-PCR技术对LcSAI在‘妃子笑'不同组织和果实不同发育阶段进行表达分析。结果表明:(1)荔枝果实酸性转化酶为定位于液泡的亲水性不稳定蛋白,无信号肽,其蛋白的二级结构主要有无规则卷曲和延伸链构件,并散布于整个蛋白;(2)在N-端含有一个跨膜区,包含两个保守结构域,位于N-端的Pfam DUF3357结构域和Glyco_32结构域,属于糖基水解酶基因家族32超家族;(3)系统进化分析结果显示LcSAI与龙眼酸性转化酶基因同源,不同组织间LcSAI表达水平为雄花>根>嫩茎>种子>嫩叶>雌花>果皮>老叶,果实不同发育阶段LcSAI表达具有特异性。该研究为深入研究LcSAI果实酸性转化酶基因调控蔗糖代谢途径机理提供数据依据。 |
| 关键词: 荔枝, 酸性转化酶, 生物信息学, 表达分析 |
| DOI：10.11931/guihaia.gxzw201910017 |
| 分类号:Q943 |
| 文章编号:1000-3142(2021)06-0989-08 |
| Fund project:国家重点研发计划(2019YFD1000900); 中央级公益性科研院所基本科研业务专项(1630062021010); 国家荔枝龙眼产业技术体系(CARS-32-20); 广东省自然科学基金(2018A030307012)[Supported by the National Key R & D Program of China(2019YFD1000900); Special Program of Central Public Research Institutes for Basic Research(1630062021010); China Litchi and Longan Industry Technology Research System(CARS-32-20); Natural Science Foundation of Guangdong Province(2018A030307012]。 |
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| Bioinformatics and expression analysis of acid invertase LcSAI of Litchi chinesis |
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DONG Chen, WEI Yongzan, WANG Yi, ZHENG Xuewen, LI Weicai*
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Institute of South Subtropical Corp Research, Chinese Academy of Tropical Agricultural Science, Key Laboratory
of Tropical Fruit Biology, Ministry of Agriculture, Zhanjiang 524091, Guangdong, China
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| Abstract: |
| As a key enzyme in sucrose metabolism, acid invertase expression and enzyme activity was significantly higher in reducing sugar accumulation litchi than in sucrose accumulation litchi. In order to fully understand the biological characteristics of acid invertase, we systematically analyzed the basic physical and chemical properties, protein secondary structure, hydrophilic/hydrophobic, transmememal domain, signal peptide, phosphorylation site, conserved domain and phylogenetic evolution of LcSAI by bioinformatics, and analyzed the expression of LcSAI in different tissues and fruits at different developmental stages of Litchi chinesis ‘Feizixiao' by using qRT-PCR. The results were as follows:(1)The acid invertase of litchi fruit was a hydrophilic unstable protein localized to vacuole, and there was no signal peptide. The secondary structure of the protein mainly consisted of irregularly coiled and extended chain members, which were scattered throughout the protein.(2)LcSAI contained a transmembrane region at the N-terminus, including two conserved domains, the N-terminal Pfam DUF3357 domain and the Glyco_32 domain, belonging to the glycosyl hydrolase gene family 32 superfamily. Phylogenetic evolution indicated LcSAI homology with the longan acid invertase gene, the LcSAI expression levels were male flower > root > young stem > seed > young leaf > female flower > peel >mature leaf; the expression of LcSAI in different stages of fruit development was specific, LcSAI expression reached a peak in the second week, then LcSAI expression dropped sharply in the third week, gradually increased in the fourth and fifth weeks, and decreased to a minimum in the eighth week of fruit development. This study provides a theoretical reference for further study of the mechanism of LcSAI fruit acid invertase gene regulation of sucrose metabolism pathway. |
| Key words: Litchi chinensis, acid invertase, bioinformatics, expression analysis |
