Identification of Chromosomal Ploidy and DNAC-Value in Sugar Beet  by Flow Cytometry

doi:10.6048/j.issn.1001-4330.2021.05.005

Abstract

Abstract: 【Objective】 A rapid, accurate and convenient method for determination of chromosome ploidy and DNAC- value in sugar beet by flow cytometry was used to provide reference for the study of chromosome ploidy and molecular biology of the beet. 【Methods】 Fresh leaves of M39-8-4 were taken as samples. The known genome of 'Luopuhongdaike' was set as external standard. Ploidy and genomic content value detection by flow cytometry(FCM) was established for sugar beet. 【Results】 After screening 7 kinds of commonly used cell dissociation solution, Marie's dissociation liquid was the optimal one for sugar beet. The detected variable coefficient (CV) of four sugar beet species and 'Luopuhongdaike' control were smaller than 5.0%,and the loading rate exceeded 200 events/μL with good reproducibility and clear and reliable results. The DNA contents of M39-8-4 Chaoyuan, 02343 Liuqifu,7208 Yikanghe were 821.88,891.94 and 1,759.59 Mb. Lsr88-5-1 was diploid and the DNA content was 822.48 Mb. 【Conclusion】 Flow cytometry could accurately distinguish 4 beet materials with different ploidy and it can be used as a method to identify the ploidy of sugar beet. The ploidy and DNA content detection method established based on FCM for sugar beet can provide reference for the related research on other plants.

Key words: sugar beet; chromosomal; flow cytometry (FCM)

摘要： 【目的】研究流式细胞技术快速、准确、方便测定甜菜体内染色体倍性和DNAC-值,为甜菜染色体倍性、分子生物学领域的相关研究提供参考依据。【方法】以二倍体M39-8-4嫩叶为材料,以已知基因组的新疆野杏洛浦洪待克为外标,建立适合于甜菜染色体倍性和基因组的流式细胞术测定方法。【结果】经7种常用细胞解离液筛选, Marie's为甜菜作物的最佳选择。用Marie's解离液制备的甜菜细胞核悬浮液,测定4份甜菜材料和新疆野杏洛浦洪待克值的变异系数(CV)均小于5.0%,且上样速率超过 200 events/μL,可重复性好,结果清晰、可靠。M39-8-4超原、02343 刘福齐和7208 伊抗褐的DNA含量依次是821.88、891.9和1 759.59 Mb;待测材料LSR88-5-1为二倍体,DNA含量822.48 Mb。【结论】流式细胞术能准确区分不同倍性的甜菜材料;可作为鉴定甜菜倍性的方法。

关键词: 甜菜, 染色体倍性, 流式细胞技术

CLC Number:

S566.5

SHA Hong, GAO Yan,DONG Xinjiu, GAO Weishi, Mayila Yusuyin, YANG Hongze. Identification of Chromosomal Ploidy and DNAC-Value in Sugar Beet by Flow Cytometry[J]. Xinjiang Agricultural Sciences, 2021, 58(5): 822-828.

沙红, 高燕, 董心久, 高卫时, 玛依拉·玉素音, 杨洪泽. 利用流式细胞术检测甜菜染色体倍性和DNAC-值[J]. 新疆农业科学, 2021, 58(5): 822-828.

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Identification of Chromosomal Ploidy and DNAC-Value in Sugar Beet by Flow Cytometry

利用流式细胞术检测甜菜染色体倍性和DNAC-值

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