流式细胞术的优化及其在水稻无融合生殖材料鉴定中的应用  

Optimization of Flow Cytometry and Its Application in Identification of Apomictic Materials in Rice

作　　者：田钧友 淡俊豪 夏玉梅[2] 詹祎捷 唐宁[2] 王耀 曹孟良 TIAN Junyou;DAN Junhao;XIA Yumei;ZHAN Yijie;TANG Ning;WANG Yao;CAO Mengliang(Longping Branch,Graduate School,Hunan University,Changsha,Hunan 410125,China;State Key Laboratory of Hybrid Rice,Hunan Hybrid Rice Research Center,Changsha,Hunan 410125,China)

机构地区：[1]湖南大学研究生院隆平分院,湖南长沙410125 [2]湖南杂交水稻研究中心杂交水稻全国重点实验室,湖南长沙410125

出　　处：《杂交水稻》2024年第6期9-17,共9页Hybrid Rice

基　　金：湖南省自然科学基金项目(2023JJ40471);湖南省农业科技创新基金项目(2023CX13,2023CX79)。

摘　　要：人工创制水稻无融合生殖体系是杂种优势固定的有效途径,其后代中包含二倍体无性种子和四倍体有性种子,因此准确鉴定两种不同倍性的后代对水稻无融合生殖研究至关重要。利用流式细胞术(flow cytometry,FCM)比较研究了水稻不同发育阶段、不同取材部位对水稻无融合生殖材料倍性检测结果的影响,旨在明确适用于其倍性鉴定的最佳发育阶段和取材部位,并通过表型和根尖染色体制片验证水稻无融合生殖材料后代的倍性。结果表明,幼苗期水稻不同单株的同一部位和同一单株的不同部位间的相对DNA含量波动较大,其中叶鞘的相对DNA含量较稳定;分蘖期水稻节间的相对DNA含量最稳定,可作为FCM检测水稻倍性的最佳材料。研究结果为无融合生殖研究中利用FCM鉴定水稻染色体倍性的材料选择提供科学参考。Artificial creation of rice apomixis system is an effective way to fix heterosis.The offspring of rice apomixis contain diploid asexual seeds and tetraploid sexual seeds.Therefore,accurate identification of these two ploidy types in the offspring is crucial for apomixis research in rice.A comparative study was conducted using flow cytometry(FCM)to investigate the effects of different developmental stages and sampling parts of rice on ploidy detection results.The aim was to determine the appropriate developmental stage and optimal sampling part for ploidy identification of apomictic rice materials,and to verify the ploidy of their offspring through phenotypic observation and root tip chromosome preparation.The results showed that during the seedling stage,the relative DNA contents were fluctuated greatly among the same part of different rice plants and different parts of the same rice plant,whereas the relative DNA contents were stable in the leaf sheath.The relative DNA contents were most stable in the internodes of rice plants at tillering stage,which could be the optimal material for FCM detection of rice ploidy.These findings provide a scientific reference for the selection of materials for the identification of rice chromosome ploidy by FCM in apomixis research.

关 键 词：水稻 流式细胞术 倍性鉴定 无融合生殖 

分 类 号：S511.01[农业科学—作物学] S311