多组学综合分析白木通自花授粉后幼果脱落机制  

Multi-omics comprehensive analysis of fruitlet abscission mecha⁃nism after self-pollination of Akebia trifoliata ssp.australis

作　　者：李贤航 龙凤 熊毅 陈松树 刘红昌 LI Xian-hang;LONG Feng;XIONG Yi;CHEN Song-shu;LIU Hong-chang(College of Agriculture,Guizhou University/Guizhou Key Laboratory of Propagation and Cultivation on Medicinal Plants,Guiyang,Guizhou 550025,China)

机构地区：[1]贵州大学农学院/贵州省药用植物繁育与种植重点实验室,贵州贵阳550025

出　　处：《南方农业学报》2024年第1期137-150,共14页Journal of Southern Agriculture

基　　金：国家重点研发计划项目(2021YFD1601002);贵州省科技计划项目(黔科合平台人才[2020]2101号)。

摘　　要：【目的】通过多组学综合分析白木通幼果在自花授粉后的脱落机制,为白木通良种繁育提供理论依据。【方法】通过人工授粉技术对白木通栽培种BMTGKAN1进行自花授粉和栽培种间授粉,授粉后第13和18 d取样,利用苯胺蓝染色法分析授粉后花粉管的萌发情况,石蜡切片技术观察授粉后胚珠发育情况,统计不同授粉处理的幼果留存率,并结合转录组和代谢组分析授粉后幼果脱落机制。【结果】自花授粉和栽培种授粉的花粉在BMTGKAN1雌蕊内萌发生长情况相似,但在受精过程中,栽培种间授粉胚珠在授粉后6 h发生核融合,自花授粉组的胚珠面积较小,未发生核融合,授粉后18 d栽培种间授粉的胚珠发育出胚乳和胚,而自花授粉胚珠未发育出胚乳和胚,出现内外珠被分离,且此时幼果留存率存在明显差异,即自花授粉后第18 d幼果留存率仅为5.92%,栽培种间授粉后第18 d幼果留存率高达86.24%。在转录和代谢水平上,栽培种间授粉与自花授粉相比,授粉后第13 d和第18 d差异表达基因(DEGs)分别为444和3646条,差异代谢物(DEMs)分别为427和412个。在相同授粉方式的比较中,自花授粉后第13 d与第18 d相比,有3305个DEGs和457个DEMs,栽培种间授粉第13 d与第18 d相比,有1123个DEGs和420个DEMs。实时荧光定量PCR检测结果与转录组数据变化趋势基本相同,说明转录组数据分析结果相对准确。在各对比组DEGs和DEMs联合分析中,至少在一个对比组中被富集到的KEGG信号通路共有162条,其中有48条在各对比组中均被富集到。植物激素信号转导通路、亚油酸代谢通路、ABC转运蛋白通路和苯丙烷生物合成通路极有可能参与调控白木通自花授粉后的幼果脱落。【结论】BMTGKAN1属于晚期自交不亲和植物,其自花授粉后13~18 d大量脱落幼果,受其体内植物激素信号转导通路、亚油酸代谢通路、ABC转运蛋白通路、苯丙烷生物合成通路调�【Objective】In this study,multi-omics comprehensive analysis methods were used to explore the mecha-nism of fruitlet abscission after self-pollination of Akebia trifoliata ssp.australis,and to provide a theoretical basis for the breeding of improved varieties of A.trifoliata ssp.australis.【Method】Self-pollination and interspecific pollination of A.trifoliata ssp.australis cultivated variety BMTGKAN1 were carried out by artificial pollination techniques.Samples were taken on the 13rd and 18th d after pollination.The pollen tube germination was analyzed by aniline blue staining method,and ovule development was observed by paraffin section technique,and the retention rate of fruitlet under different polli-nation treatments was calculated.The mechanism of fruitlet shedding after pollination was analyzed by transcriptome and metabolome.The germination and growth of pollen in BMTGKAN1 pisticles were similar between self-pollinated and cul-tivated varieties.However,during fertilization,the ovule of cultivated interspecific pollination occurred nuclear fusion at 6 h after pollination.The ovule area of self-pollination group was small,and nuclear fusion did not occur.【Result】At 18 d after pollination,the ovules of inter-specific pollination developed endosperm and embryo,while the ovules of self-pollination developed no endosperm and embryo,and the inner and outer integuments were separated,and the fruitlet re-tention rate was quite different at this time.On the 18th d after pollination,the fruitlet retention rate of self-pollinated was only 5.92%,while that of cultivated species was as high as 86.24%.At the level of transcription and metabolism,com-pared with self-pollination,there were 444 and 3646 differential expressed genes(DEGs)and 427 and 412 differential me-tabolites(DEMs)on the 13rd and 18th d after pollination.In the comparison of the same pollination methods,there were 3305 DEGs and 457 DEMs on the 13rd d after self-pollination compared with the 18th d after self-pollination.There were 1123 DEGs and

关 键 词：白木通 自花授粉 幼果脱落 转录组 代谢组 

分 类 号：S567.190.353[农业科学—中草药栽培]