利用流式细胞术快速鉴定169份香蕉种质资源的染色体倍性  

Ploidy identification of 169 Musa germplasms by flow cytometry

作　　者：吕顺 任毅 王芳 胡桂兵[2] 黄秉智[3] 刘文清 何建齐 刘建平 曾莉莎 周建坤 麦景郁 张珂恒 LU Shun;REN Yi;WANG Fang;HU Guibing;HUANG Bingzhi;LIU Wenqing;HE Jianqi;LIU Jianping;ZENG Lisha;ZHOU Jiankun;MAI Jingyu;ZHANG Keheng(Dongguan Banana and Vegetable Institute, Dongguan 523061, Guangdong, China;College of Horticulture, South China Agricultural University, Guangzhou 510642, Guangdong, China;Institute of Fruit Tree Research, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, Guangdong, China;Agriculture Technology Service Center of Machong Town, Dongguan 523135, Guangdong, China)

机构地区：[1]东莞市香蕉蔬菜研究所,广东东莞523061 [2]华南农业大学园艺学院,广州510642 [3]广东省农业科学院果树研究所,广州510640 [4]东莞市麻涌镇农业技术服务中心,广东东莞523135

出　　处：《果树学报》2018年第6期668-684,共17页Journal of Fruit Science

基　　金：东莞市社会科技发展项目(2013108101043);广东省现代农业科技创新联盟建设项目(2016LM1027)

摘　　要：【目的】利用流式细胞术快速测定169份香蕉种质资源的倍性,以期为香蕉遗传进化以及杂交育种研究提供理论依据。【方法】研究以‘小果野蕉’(AA)为内参,分析香牙蕉(Musa AAA Cavendish)、大蕉(Musa Dajiao)、粉蕉(Musa ABB Pisang Awak)、棱指蕉(Musa ABB Bluggoe)、龙牙蕉(AAB)、贡蕉(AA)、野生蕉等不同类型香蕉种质资源的染色体倍性。【结果】野生蕉、贡蕉均为二倍体,香牙蕉、棱指蕉、龙牙蕉均为三倍体,大蕉和粉蕉多数为三倍体,‘畦头大蕉’‘景洪野大蕉’‘粉杂1号’为四倍体。本研究中的香蕉杂交组合‘高州中把大蕉’ב那邦野蕉’、‘高州中把大蕉’ב广宁野生蕉’、‘高州中把大蕉’ב阿宽蕉’、‘华农中把大蕉’ב阿宽蕉’的后代均为四倍体,而‘广粉1号’ב那邦野蕉’为三倍体。‘海南红蕉’和‘飞亚-25’为三倍体。【结论】香蕉基因组和倍性十分复杂,根据形态特征鉴定香蕉染色体倍性,其结果可靠性很低。以嫩叶为材料,采用FCM技术可以快速、准确鉴定香蕉资源的倍性。【Objective】Banana（Musa spp.） is one of the most important fruits in the world and one of the important economic crops in southern China. Banana germplasms are rich and diverse. Its chromosomal ploidy is quite complicated due to the long evolution. The identification of chromosomal ploidy is the basis for studying the genetic evolution, classification and breeding of banana. The identification of chromosomal ploidy by chromosome squashing is difficult because the banana chromosome witch are small, and the satellite DNA is easy to mix with the small chromosome. Flow cytometry（FCM） is now the prevailing method for identification of chromosomal ploidy in plants, because of the ease of sample preparation and the high sample throughput. In this study, the chromosomal ploidy levels of 169 Musa germplasms were identificated by flow cytometry in order to provide theoretical basis for the study of genetic evolution and hybridization of banana.【Methods】The leaves unopened of 169 Musa germplasms were selected with ice boxes from Banana Germplasm Resource Garden of Dongguan Banana and Vegetable Institute and Banana and Litchi Germplasm Resource Garden in Guangzhou of Na-tional Fruit tree Germplasm. 20-30 mg fresh leaves contained an equal number of sample and standard sample were chopped with a sharp razor blade in a Petri dish containing 1.5 m L OTTO buffer Ⅰ which was precooled. The mixture was placed on ice 5-10 min and filtered with a 22 μm strainer to remove large cellular materials and the remaining 50 μL sample was mixed again after discarding the upper liquid. Then, 200 μL OTTO buffer Ⅱ was added to the mixture. The cell suspension was stained by Propidium Iodide and RNase 5-10 min. In the end, 200 μL cell suspension was used for the analysis of cell flow analyzer. Xiaoguo Yejiao, a known diploid wild banana（M. acuminata） was used as the internal standard, and we analyzed the relative genome size of musa germplasms containing cavendish, pisang awak, pisang mas, bluggoe, wild banana etc. by

关 键 词：香蕉 流式细胞术 染色体倍性 快速鉴定 

分 类 号：S668.1[农业科学—果树学]