黄独微型块茎低温离体保存的转录组分析

doi:10.7525/j.issn.1673-5102.2018.01.012

摘要/Abstract

摘要： 以黄独微型块茎为材料，进行了4℃低温离体保存，并对其进行了转录组分析。结果表明：原始数据进行质量预处理后，对照组（Con25）和处理组（Con4）的reads有效比分别为98.67%和98.69%；对拼接序列去重复后最终得到了219792个长度大于200bp的transcripts（177Mb）和161066个Unigenes（99Mb）；Unigenes的NR注释数目最多的10个物种分别为出芽短梗霉EXF-150、葡萄、可可、水稻粳稻组、无油樟、谷子、出芽短梗霉变种namibiae CBS 147.97、麻疯树、短至生黑醋杆菌CBS 110374和无花果拟盘多毛孢w106-1；样本注释基因最多的5个KOG是一般的功能预测、信号转导机制、翻译后修饰和蛋白质周转以及伴侣、翻译和核糖体结构和生物合成、能源生产和转换；样本KEGG注释Unigenes最多的5个pathway分别为碳代谢、氨基酸生物合成、糖酵解途径、淀粉蔗糖代谢和丙酮酸代谢。注释到的Unigenes个数为26006，注释到的不同酶数为1177，映射到的不同pathway数为327；样本基因的功能在Biological Process分类中主要聚集于cellular process和metabiolic process，在Cellular Component分类中主要聚集于cell和cell part，在Molecular Function分类中主要聚集于binding和catalytic activity；在黄独微型块茎低温离体保存中，共获得164145个差异表达基因，其中有63305个基因表达上调，有100840个基因表达下调；差异表达基因部分极其显著的GO term有液泡继承、单链断裂修复、纺锤体伸长、麦芽糖分解代谢过程、甘露糖基转移酶的活性、甘露糖磷酸转移酶活性、细胞壁甘露糖蛋白的生物合成过程、G1期细胞有丝分裂周期早期细胞芽和有丝分裂纺锤体定位的建立；差异表达基因部分极其显著的pathway有原发性胆汁酸的生物合成、类固醇激素的合成、氯代环己烷和氯苯降解、双酚A降解、氟苯甲酸降解、糖胺聚糖合成硫酸软骨素、糖胺聚糖合成硫酸乙酰肝素、甲苯降解、萘的降解和核黄素代谢。本实验结果可为黄独微型块茎的种质保存和后续萌发提供理论依据。

关键词: 黄独, 微型块茎, 低温离体保存, 转录组分析

Abstract: With Dioscorea bulbifera L. microtubers, we studied the conservation in vitro at 4℃ low temperature and its transcriptome analysis. After the original data were pretreated, the effective ratio of reads in the control group(Con25) and the treatment group(Con4) was 98.67% and 98.69%, respectively. The 219792 transcripts of 200 bp(177Mb) and 161066 Unigenes(99Mb) were obtained after the removal of the splicing sequence; 10 species of the most NR annotations in the Unigene was Aureobasidium pullulans EXF-150, Vitis vinifera, Theobroma cacao, Oryza sativa Japonica Group, Amborella trichopoda, Setaria italica, Aureobasidium pullulans var. namibiae CBS 147.97, Jatropha curcas, Aureobasidium melanogenum CBS 110374 and Pestalotiopsis fici W106-1, respectively. The 5 KOG of the annotated most unigenes were general function prediction only, signal transduction mechanisms, posttranslational modification, protein turnover, chaperones, translation, ribosomal structure and biogenesis and energy production and conversion. The 5 pathway of the KEGG annotated most unigenes were carbon metabolism, biosynthesis of amino acids, glycolysis/gluconeogenesis, starch and sucrose metabolism and pyruvate metabolism. The number of annotated unigenes was 26 006, the number of different annotated enzymes was 1 177, and the number of different pathway was mapped to 327. Gene function of samples in Biological Process classification mainly gathered at cellular process and metabolic process, in Cellular Component classification it gathered at cell and cell part, in Molecular Function classification mainly concentrated in binding and catalytic activity. In conservation in vitro at low temperature of D.bulbifera L. microtubers, a total of 164 145 differentially expressed genes was obtained, of which 63305 genes were up-regulated and 100 840 genes were down regulated. Extremely significant GO terms of the differentially expressed genes were vacuole inheritance, single strand break repair, mitotic spindle elongation, maltose catabolic process, mannosyltransferase activity, mannosylphosphate transferase activity, cell wall mannoprotein biosynthetic process, G1 phase of mitotic cell cycle, incipient cellular bud site and establishment of mitotic spindle orientation. Extremely significant pathways of the differentially expressed genes were primary bile acid biosynthesis, steroid hormone biosynthesis, chlorocyclohexane and chlorobenzene degradation, bisphenol degradation, fluorobenzoate degradation, glycosaminoglycan biosynthesis-chondroitin sulfate, glycosaminoglycan biosynthesis-heparan sulfate, toluene degradation, naphthalene degradation and riboflavin metabolism. The experimental results provides a theoretical basis for germplasm conservation of D.bulbifera L. microtuber and its subsequent germination.

Key words: Dioscorea bulbifera L., microtuber, conservation in vitro at low temperature, transcriptome analysis

中图分类号:

S632.1

洪森荣, 吴辉, 钟露, 熊思敏, 罗霞, 刘行. 黄独微型块茎低温离体保存的转录组分析[J]. 植物研究, 2018, 38(1): 100-109.

HONG Sen-Rong, WU Hui, ZHONG Lu, XIONG Si-Min, LUO Xia, LIU Xing. Transcriptome Analysis of Dioscorea bulbifera L. Microtubers Conserved in vitro at Low Temperature[J]. Bulletin of Botanical Research, 2018, 38(1): 100-109.

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