宜昌橙(Citrus ichangensis)体细胞中期染色体CMA荧光显带分析  

CMA fluorescence banding of somatic metaphase chromosomes in Citrus ichangensis

作　　者：李春艳[1,2] 李晔[2] 向素琼[2] 李晓林[2] 梁国鲁[2] 汪卫星[2] 

机构地区：[1]西南大学图书馆,重庆北碚400716 [2]西南大学园艺园林学院.重庆市级高校果树学重点实验室,重庆北碚400716

出　　处：《果树学报》2016年第12期1469-1476,共8页Journal of Fruit Science

基　　金：中央高校基本科研业务费(XDJK2012C071)

摘　　要：【目的】宜昌橙是柑橘属抗逆性较强的一类重要野生种质资源,且具有特殊的体细胞中期染色体联会现象,染色体研究对其起源与进化、重要基因的发掘与应用以及染色体结构与组成等研究具有重要意义。宜昌橙体细胞中期染色体CMA荧光显带分析可为丰富其染色体特征、遗传差异与亲缘关系以及体细胞染色体联会现象发生机制探讨等提供理论与技术支持。【方法】选取8个不同类型的宜昌橙为试验材料,采集幼嫩茎尖制备体细胞中期染色体标本,利用CMA荧光显带技术对其进行荧光带型分析。【结果】8个类型宜昌橙均显示出了不同的CMA(+)带型,根据带型特征并结合前人研究将其分为4个不同类型:C型:两臂的端部具有CMA(+)荧光带;D型:长臂或短臂的端部具有CMA(+)荧光带;E型:没有荧光带或较弱;Bst型:宜昌橙随体染色体荧光带型,呈现为一个端部、近着丝粒区域及整个随体具有CMA(+)荧光带。【结论】各供试宜昌橙CMA带型呈现明显的多态性与差异性,且部分同源染色体表现出CMA(+)条带杂合性;CMA显带分析可初步对宜昌橙不同种质进行遗传分类;宜昌橙遗传差异与CMA荧光显带技术在宜昌橙与柑橘属植物染色体特征分析以及系统分类方面具有一定的应用前景。【Objective】Citrus ichangensis Swingle is a very important breeding germplasm of Citrus with high resistances and somatic chromosome association which is a specific phenomenon comparing with other plants. Chromosome research is an important method for studying plant origin and development, gene function and chromosome structure. Here we studied the somatic chromosome fluorescence banding stained by chromomycin A3 in eight different types of C. ichangensis Swingle which would contribute to the research of relationship between structure and composition of chromosomes, origin and evolution of Citrus. This paper explored the chromosome characteristics and the somatic chromosome association in C.ichangensis Swingle as well.【Methods】Fluorescence banding of metaphase chromosomes was analyzed using CMA staining. First, the preparation of somatic metaphase chromosome samples was conducted by wall degradation hypotonic method according to Chen Ruiyang. The procedures were as follows： the tender leaves and stem tips were put into 0.002 mol·L- 18-hydroxyquinoline solution for 2.5 h, and then immersed into mix solution of methylalcohol and acetic acid（3∶1 in volume） for over 2 h, then the samples were digested with 3% cellulase and pectinase solution mix for 2.5 h and washed with deionized water andinserting mix solution of methylalcohol and acetic acid（3：1 in volume） in the tubes carefully. The samples were spread on the glass sides and stained with 5% Giemsa solution. The second, CMA staining followed the methods of M. Guerra and YANG Xiaolin.【Results】Eight accessions had different CMA（＋） bands. According to pattern characteristics and the results of previous studies, CMA banding pattern were divided into four different types： C-type： the ends of the arms possessed CMA（＋） fluorescence bands; D-type： the end of the long arm or short arm possessed CMA（＋） fluorescence band; E-type： no fluorescence band; Bsttype： the end of the long arm, centromeric region and satellite chro

关 键 词：宜昌橙 体细胞 染色体 荧光显带 多态性 杂合性 

分 类 号：S666.4[农业科学—果树学]