| 浒苔染色体制备及核型初步分析 |
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| 引用本文： | 赵晓惠,施锦婷,张建恒,康新宇,丁晓玮,何培民,文钦琳,杨晓倩.浒苔染色体制备及核型初步分析[J].海洋渔业,2019,41(4):444-452. |
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| 作者姓名： | 赵晓惠 施锦婷 张建恒 康新宇 丁晓玮 何培民 文钦琳 杨晓倩 |
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| 作者单位： | 上海海洋大学海洋生态与环境学院,上海,201306;上海海洋大学海洋生态与环境学院,上海201306;高知大学海洋植物学研究室,日本高知 783-8502;上海海洋大学海洋生态与环境学院,上海201306;江苏省海洋生物产业技术协同创新中心,连云港222005 |
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| 基金项目： | 国家重点研发计划;国家自然科学基金;上海市科委科技英才杨帆计划项目;公益性行业科研专项 |
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| 摘 要： | 由于浒苔染色体制备难度较大,至今还没有准确染色体数目及核型分析报道。以引发我国黄海绿潮优势种浒苔(Ulva prolifera)为研究对象,采用酶解-荧光法制备浒苔染色体,并比较分析染色体制备过程中秋水仙素预处理时间、酶解时间和滴片高度等3个关键因素对染色体制片效果的影响,以建立浒苔染色体制备方法。结果表明:用质量浓度为0.2%的秋水仙素处理12 h,染色体浓缩程度适宜,分散均匀,形态清晰;用质量浓度为2%的纤维素酶、果胶酶、离析酶,混合酶解20 h效果最好,能去除细胞壁、细胞膜和多糖物质;30 cm高度下滴片,能使细胞分散均匀,可以获得质量较高的浒苔染色体。核型分析显示,浒苔雌、雄配子体染色体数目为9条,孢子体染色体数目为18条。研究结果可为未来浒苔遗传特征分析以及浒苔功能基因定位研究奠定基础。
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| 关 键 词： | 浒苔 配子体 孢子体 染色体 核型分析 |
Karyological observation on Ulva prolifera chromosomes |
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| ZHAO Xiao-hui,SHI Jin-ting,ZHANG Jian-heng,KANG Xin-yu,DING Xiao-wei,HE Pei-min,WEN Qin-lin,YANG Xiao-qian.Karyological observation on Ulva prolifera chromosomes[J].Marine Fisheries,2019,41(4):444-452. |
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| Authors: | ZHAO Xiao-hui SHI Jin-ting ZHANG Jian-heng KANG Xin-yu DING Xiao-wei HE Pei-min WEN Qin-lin YANG Xiao-qian |
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| Institution: | (College of Marine Ecology and Environment, Shanghai Ocean University, Shanghai201306, China;Graduate School of Kuroshio Science, Kochi University, Kochi783-8502, Japan;Jiangsu Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resource, Lianyungang Jiangsu222005, China) |
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| Abstract: | Ulva prolifera is one of the causing species of the world’s largest green tides in the Yellow Sea, China. Due to difficulties in chromosomal preparation, large numbers of chromosomes and their relatively small size of Ulva, few researches have been reported. To establish the optimal approach for analyzing chromosomes of U. prolifera, the karyotypes and chromosomes of U. prolifera were observed after a series of treatments, including pretreatment with 0.2% colchicines for about 12 h, and mixture of enzymes prior to dropping from 30 cm height overhead glass slides for spreading the surface coat, which are the optimal conditions for chromosomes preparation. Pretreatment with colchicine can destroy and inhibit the formation of microtubules in the spindle, which increases the number of cells at the metaphase stage. The degree of enzymatic hydrolysis directly affects the quality of the chromosomes. Dropping suspended chromosomes onto a slide breaks down the enzyme lysed extracellular envelope and the nuclear membrane, leading to well dispersed chromosomes. The prepared chromosomes were stained with 4′,6-diamidino-2-phenylindole(DAPI), a fluorescent probe sensitive and specific to DNA. The results indicated that the chromosomes were well distributed, and the morphology was clear under the treatment of colchicine for 12 h. Enzymolysis for about 20 h can effectively remove the cell wall, cell membrane and polysaccharides, the cells were evenly dispersed, and the chromosomes were clearly punctured or rod-like. When the drop height was about 30 cm, the cells were well dispersed and the chromosomes could be easily observed. The results showed that the chromosome numbers of the haploid male and female gametophytes were both 9, and there were 18 in diploid sporophytes. Based upon the relative size of chromosome, the karyotypes of the female or male gametophyte chromosomes were primarily analyzed. All the results laid a solid foundation for the basic technique for localization of molecular markers on Ulva chromosomes. |
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| Keywords: | Ulva prolifera gametophytes sporophytes chromosome karyotype |
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