摘要
为探究藜麦(Chenopodium quinoa,Willd.)花粉发育的分子机制,基于基因组和转录组数据,经同源比对及RT-qPCR表达分析等,预测藜麦中AMS候选基因并克隆其编码序列,随后经病毒介导的基因沉默(VIGS)及双分子荧光互补(BiFC)法进行候选基因的功能验证。结果显示,藜麦中有CqAMSlike1和CqAMSlike2两个基因,其中CqAMSlike1的CDS长1929 bp,而CqAMSlike2的CDS长1920 bp,两基因均在不同发育时期的圆锥花序中表达。与CqAMSlike2相比,CqAMSlike1的表达更与AMS的表达模式相一致,而被进一步用于基因的功能验证。与对照相比,基于烟草脆裂病毒(TRV)沉默CqAMSlike1的藜麦两性花出现部分花丝缩短、花药不裂,花粉败育,单株种子少,种子空瘪率增加的表型。BIFC结果显示CqAMSlike1自身,与CqTDF1和CqMYB80均有互作,预示CqAMSlike1可形成同源或异源聚体调控藜麦绒毡层发育基因的表达。
To investigate the molecular mechanism of pollen development of quinoa(Chenopodium quinoa,Willd.),CqAMS candidate genes were predicted and their coding sequences were cloned based on genomic and transcriptomic data,which included homologous alignment and RT-qPCR confirmation analysis of expression patter genes,followed by virus-indiated gene silencing(VIGS)and bimolecular fluorescence complementation(BiFC)methods to perform functional validation of the candidate genes.The results showed that quinoa had two copies of AMS homologs,named as CqAMSlike1 and CqAMSlike2.The CDS of CqAMSlike1 has 1929 bp,while that of CqAMSlike2 had 1920 bp.Both genes were expressed in flowers at different developing stages of panicles,however,compared with the expression pattern of CqAMSlike2,the expression pattern of CqAMSlike1 was more consistent with the expression pattern of AMS gene in A.thaliana,thus the CqAMSlike1 was chosen for further function verification.Compared with the wild type,CqAMSlike1 is silenced in quinoa plants by VIGS based on tobacco rattle virus(TRV),showed phenotypes such as partial shortened filaments,shriveled,indehiscent stamens,abortive pollens,fewer seeds and increased seed shriveling rate.Further BiFC results showed that CqAMSlike1 could interact with itself,CqTDF1 and CqMYB80,indicating that CqAMS-like1 could form homo or heteropolymers to regulate the genes expression for controlling the development of quinoa anther tapetum.The study laid a basis for elucidating the molecular mechanism of pollen tapetum development,and creating male sterile quinoa germplasm.
作者
张玉敏
林春
刘正杰
林彤
董玉梅
毛自朝
ZHANG Yumin;LIN Chun;LIU Zhengjie;LIN Tong;DONG Yumin;MAO Zichao(College of Agriculture and Biotechnology,Yunnan Agricultural University,Kunming 650201,China;Research Center of Featured Minor Crops,Yunnan Agricultural University,Kunming 650201,China)
出处
《西北农业学报》
CAS
CSCD
北大核心
2024年第4期683-694,共12页
Acta Agriculturae Boreali-occidentalis Sinica
基金
国家自然科学基金(32060421,32260482)
云南省科技厅引智项目(202105AQ130002)。
关键词
藜麦
AMS
病毒介导的基因沉默
花粉发育
蛋白互作
Chenopodiun quinoa
AMS
Virus-indiated gene silence(VIGS)
Pollen devolpment
Protein interation
