Fundamental study for citrus chromosome mapping

柑橘染色体定位的基础研究

• 批准号: 08660034
• 负责人: KITAJIMA Akira
• 金额: $ 1.15万
• 依托单位: Kochi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08660034/
• 关键词: citrus; chromosome; karyotype snslysis; fluorescent stain

Major purpose of this study is to develop new chromosome preparation method for citrus karyotype analysis that is important of gene chromosome mapping, and the second is to try citrus karyotype analysis using this new method and to develop useful chromosome banding method for identification of citrus chromosome.1.Optimum enzymatic treatment was 120 min.incubation of 2% cellulase + 1% pectolyase or 90 min.incubation of 3% cellurase + 2% pectolyase for root tip of in vitro cultured plantlet, and 90min.incubation of 3% cellurase + 1% pectolyase for young leaf tip from orchard tree. The chromosome length of the longest one in each chromosome setreached 4-10mum that was over 2-5 times of the longest chromosome length which had been reported.2.The chromosome numbers of 2n=18 in Poncirus and orange, 2n=20-21 in 'Tosa-Buntan' and 'Suisho-Buntan', 2n=19 in satsuma mandarin, 2n=20 in Fortunella and 2n=19-20 in yuzu were major in this investigation. The karyotyape analysis of Poncirus and orange indicated that both had 2 pairs of metacentric and 7 pairs of submetacentric chromosome, and 6 paires of Poncirus chromosome and 5 pairs of orange chromosome were possible to be identified.3.G banding and N banding method was not available for citrus chromosome. C banding method showed unique banding pattern but repeatability was very low in the present study. CMA/DAPI stain showed stable unique band in 'Tosa-Buntan' chromosome. There were 3 types of chromosome that having 2 bands, 1band and no band.

本研究的主要目的是建立柑橘染色体核型分析的新方法，为柑橘染色体定位提供新的方法；其次是尝试利用该方法进行柑橘染色体核型分析，并建立适合柑橘染色体鉴定的染色体显带方法。1.以2%纤维素酶+1%果胶水解酶或3%纤维素酶+2%果胶水解酶处理试管苗根尖120分钟，3%纤维素酶+1%果胶水解酶处理90分钟，果园幼叶尖处理90分钟。每条染色体中最长的一条染色体长度可达4-10微米，是已报道的最长染色体长度的2-5倍以上。2.本研究的主要染色体数目为2n=18，‘Tosa-Buntan’和‘Suisho-Buntan’为2n=20-21，温州蜜柑为2n=19，Fortunella为2n=20，榆树为2n=19-20。对柑橘和柑橘的核型分析表明，它们都有2对中着丝粒染色体和7对亚中着丝粒染色体，有可能鉴定出6对中着丝粒染色体和5对橙色染色体。3.柑橘染色体不适用于G带和N带。C显带方法显示出独特的带型，但重复性很低。CMA/DAPI染色在‘Tosa-Buntan’染色体上显示稳定的独特条带。染色体有3种类型，有2条带，1条带和无带。