基于转录组测序挖掘甜荞自交不亲和相关功能基因  

Mining of Common Buckwheat Self-Incompatibility Related Gene Based on RNA-Seq

作　　者：张建[1] 黄科慧 刘洋 唐恬 吴银环 王于栋 陶建波 王艺钢 姜澳华 刘佳琦 方小梅 易泽林[1] ZHANG Jian;HUANG Kehui;LIU Yang;TANG Tian;WU Yinhuan;WANG Yudong;TAO Jianbo;WANG Yigang;JIANG Aohua;LIU Jiaqi;FANG Xiaomei;YI Zelin(College of Agronomy and Biotechnology,Southwest University,Chongqing 400715,China;Gulang County Sand Prevention and Control Technology Extension Center of Gansu,Gulang Gansu 733100,China)

机构地区：[1]西南大学农学与生物科技学院/重庆市荞麦产业创新团队,重庆400715 [2]甘肃省古浪县防沙治沙技术推广中心,甘肃古浪733100

出　　处：《西南大学学报（自然科学版）》2022年第11期99-113,共15页Journal of Southwest University(Natural Science Edition)

基　　金：重庆市自然科学基金项目(CSTB2022NSCQ-MSX1034);中央高校基本业务费专项(SWU119070,XDJK2019C038,XDJK2019D022);重庆市农委荞麦产业技术体系项目(CQCJT2021001).

摘　　要：普通甜荞属于花柱异长型自交不亲和1年生草本作物,营养价值丰富.为了探明甜荞育性和花型发育的重要功能途径和关键基因,以自交不亲和普通甜荞乌克兰大粒荞和自交亲和甜荞品种甜自21为材料,分别对盛花期的甜自21等柱花,乌克兰大粒荞长柱花和短柱花雌雄蕊进行转录组测序,获得283170条平均长度为1351 bp,N50为2040 bp的Unigenes,其中有68813条差异表达基因(DEGs).GO功能富集分析显示,自交不亲和甜荞中长、短柱花雌雄蕊中的差异基因主要富集在导管形成和导管发育过程中;KEGG代谢通路分析显示,自交不亲和甜荞(S-型和L-型)与自交亲和甜荞(H-型)雌雄蕊中差异基因主要富集在倍半萜和三萜化合物合成途径中;自交不亲和甜荞长、短柱花雌雄蕊中差异基因(L vs S)主要富集在植物激素信号转导途径中.倍半萜和三萜化合物合成途径共富集到77个差异基因,其中三萜化合物合成途径基因在自交不亲和甜荞中下调表达,倍半萜化合物合成途径基因以上调表达为主.植物激素信号转导途径中,与自交亲和型等柱花雌雄蕊比较,生长素、茉莉酸和赤霉素信号通路中的差异表达基因在自交不亲和型长、短柱花雌雄蕊中均高表达.通过qRT-PCR对15个自交亲和性相关差异表达基因进行分析,结果表明:其表达趋势与转录组结果一致,AGL80和PG在短柱花雌雄蕊中高表达,长柱花雌雄蕊中低表达,可能与自交不亲和甜荞长、短柱花发育有关.锌指蛋白基因CCCH,转录因子ARF,MYC,SP11,SUI1与甜荞育性发育相关.该文比较分析了不同花型甜荞雌雄蕊转录组测序结果,获得大量Unigenes数据及差异表达基因的相关信息,发现了导管形成与发育对自交不亲和甜荞花柱发育具有重要影响,明确了萜类化合物、植物激素和转录因子对甜荞育性发育具有重要作用.Common buckwheat(Fagopyrum esculentum),a heteromorphic self-incompatible(SI)species,is the annual herbaceous crops with rich nutritional value.The objective of this study is to explore the important functional pathways and key genes regulating fertility and style development of self-incompatible common buckwheat,which has great significance to further study the molecular mechanism of self-incompatibilities in common buckwheat.Ukraine daliqiao(UD)(heterostyly and self-incompatible)and Tianzi 21(TZ)(homostyled and self-compatible)as materials,RNA-seq were carried out with the samples of pistils and stamens from S-morph flower(S)and L-morph flower(L)of UD,and H-morph flower of TZ(H).A total of 283170 unigenes with an average length of 1351 bp and N50 of 2040 bp were obtained,among which 68813 differentially expressed genes(DEGs)were identified.GO function enrichment analysis showed that the DEGs in L vs S were significantly enriched in“angiogenesis”and“blood vessel development”.KEGG metabolic pathway analysis showed that DEGs of L vs H and S vs H were significantly enriched in the pathways of sesquiterpenoid and triterpenoid biosynthesis.The most of DEGs of L vs S were enriched in the pathway of plant hormone signal transduction.There were 77 DEGs in“sesquiterpenoid and triterpenoid biosynthesis”,among which the gene expression of triterpenoid synthesis pathway was down-regulated and the gene expression of sesquiterpenoid synthesis pathway was mainly up-regulated in self-incompatible common buckwheat.In the plant hormone signal transduction pathway,compared with the self-compatible common buckwheat,the DEGs in auxin,JA and GA had high expression in the self-incompatible common buckwheat.Fifteen DEGs related to self-incompatibility were analyzed by qRT-PCR,and the expression trend was consistent with the results of RNA-Seq analysis.The expression levels of AGL80 and PG were high in the pistils and stamens of S-morph flower and low in the pistils and stamens of L-morph flower in UD,which may be related to

关 键 词：甜荞 雌雄蕊 RNA-SEQ 自交亲和性 花柱发育 

分 类 号：S517[农业科学—作物学]