基于流式细胞仪鉴定菊芋倍性方法的建立及应用  

Establishment and application of aploidy method for identification of Helianthus tuberosus L.by flow cytometry

作　　者：赵孟良[1,2,3] 任延靖 田闵玉 钟启文 ZHAO Mengliang;REN Yanjing;TIAN Minyu;ZHONG Qiwen(Academy of Agriculture and Forestry,Key Laboratory of Vegetable Genetics and Physiology,Qinghai University,Xining,Qinghai 810016,China;State Key Laboratory of Plateau Ecology and Agriculture,Qinghai University,Xining,Qinghai 810016,China;College of Horticulture,Northwest A&F University,Yangling,Shaanxi 712100,China)

机构地区：[1]青海大学农林科学院/青海省蔬菜遗传与生理重点实验室,青海西宁810016 [2]青海大学三江源生态和高原农牧业国家重点实验室,青海西宁810016 [3]西北农林科技大学园艺学院,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2021年第3期138-146,共9页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家自然科学基金项目(31660569,31960602);青海省科技厅重点实验室项目(2020-ZJ-Y02);青海省农林科学院基金项目(2018-NKY-006)。

摘　　要：【目的】利用流式细胞仪检测菊芋种质的染色体倍性,为菊芋的种质鉴定及遗传育种研究提供基础数据。【方法】建立基于流式细胞仪检测菊芋倍性的检测体系,并针对不同电压((340,365.3和422 V))、聚乙烯吡咯烷酮(PVP)添加与否、取样部位(老叶、嫩叶、块茎和根)对体系进行优化。利用建立的稳定优化体系对302份菊芋种质资源的倍性进行检测。【结果】针对菊芋材料,选取幼嫩叶片,在样品处理时添加PVP溶液消除多酚及多糖物质的干扰,检测电压选用365.3 V,以流式细胞仪进行倍性检测的效果较好,准确性较高。302份菊芋种质资源倍性检测结果表明,整体变异系数在2.17~9.74,二倍体共3份,仅占资源总数的0.99%;四倍体共25份,占资源总数的8.28%;六倍体共274份,占资源总数的90.73%。【结论】基于流式细胞仪建立的菊芋种质资源倍性鉴定方法检测效果较好,准确性较高。【Objective】In order to provide the basic data for Helianthus tuberosus L.germplasm identification and genetic breeding,this study determined the chromosome ploidy by using flow cytometry in this study.【Method】This study established a testing system based on the flow cytometry(FCM)system,and the system was optimized by using different voltages(340,365.3 and 422 V),adding PVP or not,and using different samples(old leaves,young leaves,tuber,and root).Then,we tested 302 H.tuberosus L.germplasm resources were tested by using above the optimized system.【Result】Young leaves with PVP(eliminating the interference of polyphenols and polysaccharides)under 365.3 V can reveal the had better the best testing effect.Ploidy testing results of 302 H.tuberosus L.germplasm resources showed that the total variation coefficient of variation ranged from 2.17 to 9.74,three(accounted for 0.99%)resources were diploid,25(accounted for 8.28%)resources were tetraploid,and 274(accounted for 90.73%)resources were hexaploid.【Conclusion】The method based on flow cytometry was effective and accurate for testing the ploidy of H.tuberosus L.germplasm resources.

关 键 词：菊芋 流式细胞仪 染色体倍性 倍性检测 种质资源 

分 类 号：S632.9[农业科学—蔬菜学]