红肉火龙果成熟茎的全长转录组测序分析及蔗糖代谢相关基因分离  

Full-length transcriptome sequencing snalysis and sucrose metabolism-related gene isolation of mature stems from red pitaya

作　　者：郑乾明 王小柯 马玉华[1] ZHENG Qian-ming;WANG Xiao-ke;MA Yu-hua(Institute of Pomology Science,Guizhou Academy of Agricultural Science,Guiyang 550006,China)

机构地区：[1]贵州省农业科学院果树科学研究所,贵阳550006

出　　处：《西南农业学报》2022年第3期581-587,共7页Southwest China Journal of Agricultural Sciences

基　　金：贵州省科学技术基金项目[黔科合基础(2017)1178]。

摘　　要：【目的】分离红肉火龙果成熟茎中表达的基因全长序列,为研究参与蔗糖合成和降解等重要代谢途径的候选基因生理功能提供基础。【方法】提取红肉火龙果成熟茎总RNA,采用基于Pacbio Sequel平台的转录组测序进行全长转录组分析。对获得的序列进行功能注释和分类,并与此前基于Illumina平台的转录组测序结果进行比较分析。【结果】全长转录组测序共获得30313条高质量转录本,平均长度为1829 bp。预测获得编码序列(Coding sequence,CDS)数量为40255个,长度为297~5202 bp,平均长度为1179 bp;共计13939条转录本在公共数据库被注释,占所有转录本数量的45.98%;在NR数据库中注释的转录本数量占比为45.49%,注释转录本数量较多的物种为甜菜和菠菜,占比分别为50.95%和24.90%。与此前基于Illumina平台的转录组测序联合分析,获得若干在成熟茎中表达的蔗糖代谢相关基因的全长序列。【结论】基于全长转录组测序获得的转录本长度显著长于Illumina平台的转录组测序,有利于后续分离和研究红肉火龙果成熟茎中重要代谢途径的功能基因全长。【Objective】The full-length sequences of genes expressed in the mature stems of Hylocereus polyrhizus were isolated,which provided a basis for studying the physiological functions of candidate genes involved in important metabolic pathways such as sucrose synthesis and degradation.【Method】Total RNA was extracted from mature stems of H.polyrhizus,and transcriptome sequencing basing on the Pacbio Sequel platform was used for full-length transcriptome analysis.The obtained sequences were annotated and classified,and compared with the transcriptome sequencing basing on the Illumina platform.【Result】A total of 30313 high-quality transcripts with an average length of 1829 bp were obtained by full-length transcriptome sequencing.The estimated number of Coding sequences(CDS)was 40255,with a length of 297-5202 bp and an average length of 1179 bp.A total of 13939 transcripts,accounting for 45.98%of all transcripts,were annotated in the public databases.The number of annotated transcripts in NR database was 45.49%,and the number of annotated transcripts in sugar beet and spinach were 50.95%and 24.90%,respectively.In combination with Illumina-based transcriptome sequencing,several full-length sucrose metabolism-related genes that were highly expressed in mature stems were obtained.【Conclusion】The transcript length from full-length transcriptome sequencing was significantly longer than that obtained by Illumina transcriptome sequencing,which was useful for the subsequent isolation of full-length functional genes involved in important metabolic pathways in the mature stem of H.polyrhizus.

关 键 词：成熟茎 基因全长序列 序列注释 蔗糖代谢 基因分离 

分 类 号：S667.9[农业科学—果树学]