锥栗种仁转录组及淀粉和蔗糖代谢相关酶基因的表达分析  

Transcriptome Analysis for Developing Kernel and Expression Analysis of Starch and Sucrose Metabolism-related Genes in Castanea henryi

作　　者：张琳[1] 范晓明[1] 林青[1] 向晖[1] 汪武[1] 袁德义[1] 

机构地区：[1]中南林业科技大学经济林培育与保护省部共建教育部重点实验室/经济林培育与利用湖南省2011协同创新中心,长沙410004

出　　处：《植物遗传资源学报》2015年第3期603-611,共9页Journal of Plant Genetic Resources

基　　金：国家"十二五"科技支撑计划课题(2013BAD14B04)

摘　　要：本研究采用高通量测序技术对淀粉积累高峰期的锥栗种仁进行转录组测序分析,对得到的Unigene进行功能注释、分类及代谢通路分析,并进一步通过实时定量PCR方法分析了7个与淀粉和蔗糖代谢相关酶基因在锥栗种仁发育过程中的表达特征。结果显示:de novo组装后共获得53629条Unigene序列,序列平均长度为746 bp;通过与其他核酸、蛋白质数据库的Blast搜索比对,共26739条Unigene序列获得基因注释,占All-Unigene的49.86%;与COG数据库比对后将其注释的14413条Unigene序列划分成25类;GO功能注释的33926条Unigene基因共分成细胞组分、分子功能和生物过程3大类58个分支;与KEGG数据库比对结果注释的5277条Unigene序列划分为116条代谢通路;7个与淀粉和蔗糖代谢相关酶基因表达量变化趋势中,CH.29636(淀粉合成酶,SS),CH.11971(淀粉分支酶,SBE)两个基因随着种仁的发育呈现逐渐上升的趋势,而其余5个基因(CH.31302、CH.33690、CH.19238、CH.30128、CH.13088)表达量随着种仁的发育呈先上升后下降的趋势,该结果与锥栗种仁发育期淀粉和蔗糖的变化规律一致。这些信息为锥栗果实品质形成相关功能基因的研究提供了重要依据。Castanea henryi is an important woody grain plants in south of China. The current research on the fruit development of C. henryi mainly focuses on fruit component analysis, fruit quality analysis, and fruit processing,whereas the molecular mechanisms involved in fruit development remain to be uncovered. Transcriptome sequencing and analysis were conducted on kernel in the beginning of starch synthesis peak phase of C. henryi using Illumina platform-based strategy. Expression patterns of seven starch and sucrose metabolism-related Unigenes were assayed with real-time quantitative PCR （qPCR） method. The results showed that de novo assembly genera- ted a total of 53629 Unigenes with average length of 746 bp. Among them,26739 unigenes were annotated against NR, NCBI and Swiss-Port protein databases, 14413 Unigenes were assigned 25 categories by comparing with COG （Clusters of Orthologous Groups） database, and 33926 Unigenes were assigned 3 categories including cellular component, molecular function and biological process and 58 branches with GO （ Gene Ontology） database. Functional annotation against KEGG （Kyoto Encyclopedia of Genes and Genomes） database identified 5277 Unigenes which were mapped to 116 metabolic pathways. Moreover, the qPCR analysis showed that two Unigenes CH. 29636 and CH. 11971 had a gradual increase,while the other 5 genes（ CH. 31302, CH. 33690, CH. 19238, CH. 30128,and CH. 13088）increased at the early stage and then decreased. The expression profiles of the 7 genes were in accordance with the accumulation of starch and sucrose in the developing fruits of C. henryi. This study provided a foundation for further characterizing the functional genes involved in the formation of fruit quality in C. henryi.

关 键 词：锥栗 RNA-SEQ 种仁 实时荧光定量PCR 淀粉 蔗糖 

分 类 号：S664.2[农业科学—果树学]