玉米B染色体显微分离技术  

Microdissection Technique of B Chromosome in Maize

作　　者：冉佐 姚启伦[1,2] 陈发波 高健[1,2] 李文博 向明芳 李玉洁 Ran Zuo;Yao Qilun;Chen Fabo;Gao Jian;Li Wenbo;Xiang Mingfang;Li Yujie(College of Life Science and Technology,Yangtze Normal University,Fuling,408100;Centre for Green Development and Collaborative Innovation in Wuling Mountain Region,Yangtze Normal University,Fuling,408100;Fuling Jiangbei District Agricultural Service Center,Subdistrict Office of Jiangbei District,Fuling District,Chongqing,Fuling,408000)

机构地区：[1]长江师范学院生命科学与技术学院,涪陵408100 [2]长江师范学院武陵山片区绿色发展协同创新中心,涪陵408100 [3]涪陵区江北街道农业服务中心重庆市涪陵区江北街道办事处,涪陵408000

出　　处：《分子植物育种》2018年第20期6785-6792,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31371633);重庆市科委社会事业与民生保障科技创新专项(2016shmszx80030);重庆市教委优秀成果转化项目(KJZH17133)共同资助

摘　　要：染色体分离是连接细胞遗传学和分子生物学的桥梁技术。本研究以含B染色体的玉米地方品种‘DP11’和自交系‘B73’为材料,通过滴片技术制作染色体制片,采用微玻璃针法显微分离制片中的玉米B染色体。结果表明,采用笑气处理时间2 h、冰乙酸固定时间2 h、酶解时间6 h和滴片距离35 cm的处理技术可制备染色体重叠率低、分散性好且适于染色体分离的玉米B染色体制片。基于单细胞染色体数目、目标染色体的形态大小,在光学显微镜下可对玉米B染色体正确地识别。借助显微操作系统,使用针尖直径为1～5μm的微玻璃针,载物台与微玻璃针针尖呈15°和45°的角度,均可实现玉米B染色体的成功分离。分离玉米B染色体为深化其分子生物学及起源进化研究提供了物质技术依据。Chromosome isolation is a bridge between cytogenetics and molecular biology. In this study, the maize landrace ＇DP11＇ and selfing line ＇B73＇ containing B chromosome were used as materials. The maize B chromosome was isolated by the glass microneedle from a slide prepared with the chromosomal dropping-slide technique. The results indicated that a slide of maize B chromosomes with low chromosome overlap rate, good dispersity, and suitable for isolating a single chromosome could be obtained by the techniques of N20 treatment for 2 h, glacial acetic acid fixation for 2 h, enzymatic hydrolysis for 6 h, and dropping-slide distance of 35 cm. Maize B chromo- somes could be properly identified under the optical microscope based on its morphology and size as well as the number of chromosomes in a cell. With a glass microneedle of 1-5 μm tip diameter, the maize B chromosome might be separated in aid of a micromanipulator set when the angles between the needle tip and bench were 15° and 45°, respectively. The isolation of maize B chromosome would provide material and technical basis for further study on the molecular biology as well as origin and evolution of maize B chromosome.

关 键 词：玉米 B染色体 显微分离 

分 类 号：S513[农业科学—作物学]