黑皮果蔗中碱性蔗糖转化酶基因ShINV6的克隆与表达分析  

Molecular Cloning and Expression Analysis of a Neutral/Alkaline Invertase Gene(ShINV6)from‘Badila’

作　　者：李和平 张树河 李瑞美 潘世明 LI Heping;ZHANG Shuhe;LI Ruimei;PAN Shiming(Institute of Subtropical Agriculture,Fujian Academy of Agricultural Sciences,Zhangzhou,Fujian 363005,China)

机构地区：[1]福建省农业科学院亚热带农业研究所,福建漳州363005

出　　处：《热带作物学报》2022年第3期484-490,共7页Chinese Journal of Tropical Crops

基　　金：福建省公益类科研院所基本科研专项(No.2019R1030-1);福建农林大学国家甘蔗工程技术研究中心开放课题(No.NERO2018.1.5);国家糖料产业技术体系建设项目(No.CARS-170707)。

摘　　要：在高等植物中,蔗糖转化酶广泛参与调节植物的生长发育、生物和非生物胁迫应答等过程,在控制光合产物在不同库组织的分配上起重要作用,是决定作物经济产量的关键酶。本研究以黑皮果蔗‘Badila’为实验材料,克隆了一个果蔗蔗茎中碱性转化酶基因ShINV6,并对其序列特征、表达特性及其编码蛋白的酶学特征进行了分析。生物信息学分析发现ShINV6基因cDNA全长共2859 bp,其中编码区为1254 bp,共编码417个氨基酸,其中精氨酸(Arg)含量最高,为12.7%,分子质量为46.29 kD,理论等电点为10.89,进化关系分析发现ShINV6蛋白与甘蔗、玉米、高粱等单子叶作物的中碱性蔗糖转化酶高度同源,具有典型的Glyco_hydro_100结构域,多重序列比对发现ShINV6蛋白的C端比其他转化酶少约130个氨基酸,细胞定位和亲水性分析预测ShINV6蛋白可能为细胞质中的亲水性蛋白。在果蔗生长发育过程中,观测蔗茎中的蔗糖含量变化,发现随着种植时间的延长蔗糖含量逐渐增加,其中10月至11月是糖分积累最多的月份。荧光定量PCR结果显示9月份ShINV6表达量达到最大值,结合果蔗生长期生物产量增长规律推测ShINV6很可能参与了蔗茎生长旺盛期光合作用产物蔗糖在蔗茎(库)中的卸载过程,把蔗糖转化成己糖以供蔗茎快速生长。In higher plants,invertase is widely involved in regulating plant growth,development,biological and abiotic stress response,and plays an important role in controlling the distribution of photosynthates in different bank tissues.It is a key enzyme that determines crop economic yield.In this study,the neutral/alkaline invertase ShINV6 was cloned from the stem of‘Badila’.The sequence characteristics,expression characteristics and enzymatic characteristics of its encoding protein were analyzed.Bioinformatics analysis found that the full length of ShINV6 cDNA was 2859 bp,in-cluding 1254 bp of ORF that encoding 417 amino acids,and the content of arginine(Arg)was 12.7%.The molecular weight of ShINV6 was 46.29 kD and the PI value was 10.89.Evolutionary relationship analysis showed that ShINV6 protein was highly homologous with the neutral/alkaline invertase of monocotyledon crops such as sugarcane,maize and sorghum.It has typical Glyco_hydro_100 domain,multiple sequence alignments revealed that the C-terminal of ShINV6 was about 130 amino acids less than other invertases.Cell localization and hydrophilic analysis predict that ShINV6 may be hydrophilic protein in cytoplasm.During the growth and development of sugarcane,the change of su-crose content in sugarcane stems was observed.It was found that the sucrose content increased gradually with the ex-tension of planting time,and the most sugar accumulation month was from October to November.QRT-PCR results showed that ShINV6 expression reached its maximum in September,and combined with regulation of biological yield growth in sugarcane growing period,it was speculated that ShINV6 might be involved in the unloading process of su-crose which the product of photosynthesis in the peak growth stage of sugarcane stems(library),and sucrose was trans-formed into hexaccharide for the rapid growth of sugarcane stems.

关 键 词：BADILA 中碱性转化酶 ShINV6 基因克隆 

分 类 号：S566.1[农业科学—作物学]