熱脅迫下水仙花葉片的轉錄組分析

宋姣敏 徐小萍 陳裕坤 陳桂信 賴鐘雄

摘 ?要??為研究水仙響應高溫脅迫過程中相關基因的表達及響應熱應激的分子機制，本研究以漳州水仙花為材料，使用BGISEQ-500測序技術，對高溫（30、35?℃）處理及正常生長條件（15?℃）的水仙葉片進行轉錄組學的測序分析。使用Trinity軟件對15（對照）、30、35?℃（高溫脅迫）處理24 h的水仙葉片測序數據進行從頭組裝，利用BLAST軟件進行基因比對注釋，通過DEGseq和PossionDis檢測差異表達基因，并對差異表達基因進行GO和KEGG富集分析。結果表明：本次水仙花葉片測序共獲得112 160條總長度為128 733 815 bp、平均長度為1147 bp的Unigene，其N50以及GC含量分別為1770?bp和42.33%。Nr比對注釋匹配度最高的物種為石刁柏，利用KOG數據庫進行比對，5560條Unigene序列分布于25個功能區域。差異表達基因GO富集顯示與光合系統及膜系統相關的GO term被顯著富集，KEGG分析顯示次生代謝物的生物合成、苯丙烷類生物合成、代謝途徑、光合作用、糖胺聚糖降解、乙醛酸和二羧酸代謝、光合作用-天線蛋白、單萜類生物合成等8條代謝途徑在3個比較組中均顯著性富集，硫胺素代謝途徑是唯一在高溫脅迫條件下均顯著富集的代謝途徑。本研究注釋了大量相關基因序列，通過本研究為改善水仙花植物的耐熱性及耐熱性品種的培育提供了豐富的數據資源和分子基礎。

關鍵詞 ?水仙;高溫脅迫;轉錄組;差異表達基因中圖分類號??S682.2+1??????文獻標識碼??A

Analyses of Leaf Transcriptome under Heat Stress in Narcissus

SONG Jiaomin， XU Xiaoping， CHEN Yukun， CHEN Guixin^*， LAI Zhongxiong^*

College of Horticluture / Horticultural Plant Bioengineering Institute， Fujian Agriculture and Forestry University， Fuzhou， Fujian 350002， China

Abstract ?To explore the pathways of response to heat stress and mechanism of heat-tolerance ofNarcissus tazetta var.?chinensis，?we used the BGISEQ-500 sequencing technology andde novotranscriptome assembly to gain a comprehensive overview of the transcriptome?of the leaves under heat temperature?（30?℃， 35?℃） and normal growth conditions （15?℃）. The data of?RNA-Seq treated 24 h at 15， 30， 35?℃?werede novoassembled using Trinity software.?Gene annotation was performed using BLAST software， and differentially expressed genes were detected by DEGseq and PossionDis. A total of 112 160 unigenes were generated with a toal length of 128 733 815 bp， average length of 1147 bp and N50 of 1770 bp. For annotation functional，Narcissuswas highly similar to Asparagus officinalis species. On the basis of clusters of euKaryotic Orithologous Groups?（KOG） analysis， 5560 unigene sequences were distributed over 25 functional regions. Gene Ontology （GO） enrichment of DEGs showed significant enrichment of GO term associated with photosynthetic systems and membrane systems. KEGG analysis showed?significant enrichment of the pathways?about the biosynthesis of secondary metabolites， phenylpropanoid biosynthesis， metabolic pathways， photosynthesis， glycosaminoglycan degradation， glyoxylic acid and dicarboxylic acid metabolism， photosynthesis-antenna proteins， monoterpenoid biosynthesis， etc. And the thiamine metabolc pathway was the only one that was significantly enriched under high temperature stress conditions. The study annotated a large number of related gene sequences， and would provide abundant data resources and molecular basis for improving the heat tolerance and heat tolerance ofNarcissus.

Keywords ?Narcissus; high temperature stress; transcriptome; differentially expressed gene

DOI10.3969/j.issn.1000-2561.2019.10.011

中國水仙別名雅蒜、天蔥、麗蘭等，是中國的十大傳統名花之一，也是漳州市的市花，屬石蒜科（Amaryllidaceae）水仙屬（Narcissus）植物。不僅具有極高的觀賞價值，還具有較高的經濟價值^[1]。……