摘要
为使烟草染色体标本制备过程中取材更方便,本研究以异源五倍体烟草(2n=5x=58)为材料,观察了不同大小子房、不同预处理方法对烟草染色体制片效果的影响。首先,子房大小按花冠与花萼长度比例分为6类;其次,预处理方法分为2类,即:(1)室温下于0.002mol/L的8-羟基喹啉中预处理2、4和6h;(2)7℃下于0.002mol/L的8-羟基喹啉中预处理12、24和36h。结果显示,当花冠长度小于或等于花萼长度的1/2时,子房中分裂期细胞的比例最大,为97.00±17.82个/1000个。该时期子房于7℃经24h预处理后,染色体数目与形态清晰可辨。利用上述方法制作云烟87(2n=4x=48)和N.plumbaginifolia(2n=2x=20)的染色体标本,均获得较好的结果。此外,将上述方法制作的五倍体烟草染色体经5SrDNA-FISH,获得了清晰可辨的5个不同信号,与预期一致。可见,幼嫩子房可用于烟草染色体标本的制作,且用改良的预处理方法制备的染色体标本可用于染色体计数和核型分析,也可用于原位杂交分析。
In order to obtain materials of Nicotiana plants easily when preparing chromosome specimens,in this study,allopentaploid tobacco (2n=5x=58) was used as materials to observe the effects of ovary sizes and pretreatment methods on Nicotiana plants chromosome specimen preparation.First,ovaries were divided into 6 types based on corolla length/calyx length;and then,two pretreatments were compared,one was pretreatment in 0.002 mol/L 8-hydroxy-quinoline solution at normal atmospheric temperature for 2,4 and 6 h,the other one was pretreatment in 0.002 mol/L 8-hydroxy-quinoline solution at 7 ℃ for 12,24 and 36 h.When the length of the corolla was less than or equal to 1/2 of the calyx,the ratio of mitotic cells reached the highest,being 97.00±17.82 in 1000 cells.When ovaries with the most mitotic cells were pretreated at 7 ℃ for 24 h,the number and configuration of chromosomes were clear and distinguishable.The above mentioned methods were applied in chromosome specimen preparation of Yunyan87 (2n=4x=48) and N.plumbaginifolia (2n=2x=20),and nice results were obtained,respectively.In addition,5S rDNA-FISH was carried out on chromosomes of N.plumbaginifolia,and expected recognizable signals were found on 5 different chromosomes.These results indicated that tender ovary could be used to prepare chromosome specimens of Nicotiana plants.Chromosome specimens prepared with modified method were not only beneficial to chromosomes counting and karyotype analysis,but also can be applied to in situ hybridization analysis.
作者
杨垚
张艳
党江波
梁国鲁
郭启高
庞静
YANG Yao;ZHANG Yan;DANG Jiangbo;LIANG Guolu;GUO Qigao;PANG Jing(College of Horticulture and Landscape,Southwest University,Chongqing 400715,China;Chongqing Tobacco Research Institute,Chongqing Municipal Tobacco Company,Chongqing 400715,China;Zunyi Tobacco Company of Guizhou Province,Meitan Branch,Zunyi,Guizhou 564100,China)
出处
《中国烟草科学》
CSCD
北大核心
2019年第4期56-61,共6页
Chinese Tobacco Science
基金
中国烟草总公司重庆市公司科技项目“重庆自育烤烟新品系深化培育与示范”（NY20170401070001）
关键词
烟草
幼嫩子房
染色体制片
原位杂交
Nicotiana
tender ovary
chromosome specimen preparation
in situ hybridization
