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期 刊 ：植物遗传资源学报 2015年 16卷 3期

关键词：锥栗；RNA-Seq；种仁；实时荧光定量PCR；淀粉；蔗糖；

Keywords:Castanea henryi, RNA-seq, kernel, real-time RT-PCR, starch, saccharose,

摘 要 ：锥栗是我国南方重要的木本粮食树种，当前对锥栗果实发育的研究主要集中在果实品质分析及果品加工等方面，而对锥栗果实发育过程的分子机理研究尚未见报道。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53 629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26 739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14 413条Unigene序列划分成25类；GO功能注释的33 926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5 277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。

Abstract:Castanea henryi is an important woody grain plants in south of China. The current research on the fruit development of C. henryihenryi 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 generated a total of 53 629 Unigenes with average length of 746 bp. Among them, 26 739 unigenes were annotated against NR, NCBI and Swiss-Port protein databases, 14 413 Unigenes were assigned 25 categories by comparing with COG (Clusters of Orthologous Groups) database, and 33 926 Unigenes were assigned 3 categories (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 5 277 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 are in accordance with the accumulation of starch and sucrose in the developing fruits of C. henryi. This study provides a foundation for further characterizing the functional genes involved in the formation of fruit quality in C. henryi.

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