[1]姚启伦,李玉洁,陈发波,等.玉米染色体滴片关键技术及其优化[J].江苏农业学报,2019,(02):255-261.[doi:doi:10.3969/j.issn.1000-4440.2019.02.003]
　YAO Qi-lun,LI Yu-jie,CHEN Fa-bo,et al.Key techniques and their selective preference of maize chromosomal dropping-slides[J].,2019,(02):255-261.[doi:doi:10.3969/j.issn.1000-4440.2019.02.003]
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玉米染色体滴片关键技术及其优化()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2019年02期
页码:
255-261
栏目:
遗传育种·生理生化
出版日期:
2019-04-30

文章信息/Info

Title:
Key techniques and their selective preference of maize chromosomal dropping-slides
作者:
姚启伦1; 2; 李玉洁1; 陈发波1; 2; 高健1; 2; 李文博1; 2; 方平1; 2; 李红艳1
(1.长江师范学院生命科学与技术学院，重庆408100；2.长江师范学院武陵山片区绿色发展协同创新中心，重庆408100)
Author(s):
YAO Qi-lun1; 2; LI Yu-jie1; CHEN Fa-bo1; 2; GAO Jian1; 2; LI Wen-bo1; 2; FANG Ping1; 2; LI Hong-yan1
(1.Department of Life Sciences and Technologies, Yangtze Normal University, Chongqing 408100, China;2.Correlative Innovation Centre for Green Development in the Wulingshan Region, Yangtze Normal University, Chongqing 408100, China)
关键词:
玉米; 染色体; 滴片; 优化
Keywords:
maize; chromosome; dropping-slides; optimization
分类号:
S513
DOI:
doi:10.3969/j.issn.1000-4440.2019.02.003
文献标志码:
A
摘要:
探讨优化玉米染色体滴片关键因子处理效果，寻求一种操作简单、染色体分散度和完整性好，并且适于玉米染色体分离操作的制片方法。本研究以玉米自交系B73根尖为材料，在单因素试验的基础上采用四因素二次回归正交组合设计，分析玉米滴片技术中4个关键因子的处理效果。结果表明，N2O处理时间、冰乙酸固定时间、酶解时间和滴片距离对染色体滴片效果均存在最佳处理效应。基于二次回归方程的统计方法得到了符合实际的优化的染色体滴片技术方案，即采用N2O处理时间2.0 h、冰乙酸固定时间2.0 h、酶解时间6.0 h和滴片距离35.0 cm。由此方案制作了高质量的玉米染色体制片，保证了细胞中染色体的分散性和完整性。
Abstract:
To explore the effect of optimizing the key factors on maize chromosome, a method of moking dropping-slides with simple operation, good chromosome dispersion and integrity, and good suitability for the operation of chromosome separation was sought. In this study, the root tips of maize inbred line B73 were used as the material. Based on the single factor experiment, four factors quadratic regression right cross design was used to analyze the effects of four factors on chromosomal dropping-slides of maize. The results showed that the treatment time of nitrous oxide, the fixation time of glacial acetic acid, enzymolysis time and the dropping-slide distance all had the best effect on the chromosomal dropping-slide of maize. Based on the statistical method of quadratic regression equation, the optimized chromosome dropping-slides technique was obtained, which consisted of two hours of nitrous oxide treatment, two hours of glacial acetic acid fixation, six hours of enzymatic hydrolysis and 35 cm of dropping-slide dislance. High-quality maize chromosome dropping-slides were mode by the scheme, and the dispersibility and integrity of chromosomes in cells were ensured.

参考文献/References:

[1]李懋学，张赞平. 作物染色体研究技术[M]. 北京：中国农业出版社，1996：58-92.
[2]马婷玉，李明，吴燕燕，等．穿心莲染色体制片优化及核型分析［J］．江苏农业科学，2017,45(7):33-36.
[3]MCFEE A F，SAYER A M，SAOMAA S I，et al. Methods for improving the yield and quality of metaphase preparations for FISH probing of human lymphocyte chromosomes[J]. Environ Mol Mutagen，1997，29：98-104.
[4]PENG R H，LIU F，HU X，et al. Microdissection and microcloning of chromosome 5 in Gossypium arboreum[J]. Plant Mol Biol Rep，2012，30(5)：1218-1228.
[5]JONES N，HOUBEN A. B chromosomes in plants：escapees from the A chromosome genome[J]. Trends in Plant Science，2003，8：417-423.
[6]HU Z M，CUI L H，WANG L L，et al. Isolation of single chromosomes and chromosomal fragments of Lilium regale[J]. Acta Genetica Sinica，1997，24(3)：278-281.
[7]杨大翔. 遗传学实验 [M]. 3版.北京：科学出版社，2016：3-98.
[8]潘颀，王芳，郝爱花，等. 新疆紫草不同材料染色体制片技术研究[J]. 新疆农业科学，2013，50(9)：1642-1650.
[9]劳世辉，魏岳荣，盛鸥，等. 菠萝蜜的染色体制片优化及核型分析[J]. 中国农学通报，2011，27(16)：215-219.
[10]陈雪，商超，张晓莹，等. 射干染色体制片技术的探索[J]. 中国园艺文摘，2016(1)：9-12.
[11]刘丹，夏雪，吴益梅，等. 植物染色体制片效果影响因素的解析[J]. 浙江农业科学，2015，56 (10)：1654-1657.
[12]BARCH M J，KNUTSEN T，SPERBUCK T L. AGT cytogenetics laboratory manual[M]. 3rd. Philadelphia：Lippincott Raven Publishers，1991：39-46.
[13]JONES R N，GONZALEZ-SANCHEZ M，GONZALEZ-GARCIA M，et al. Chromosomes with a life of their own[J]. Cytogenet Genome Res，2008，120(3/4)：265-280.
[14]DENG W T S，LUCAS J N，LEUNG C S，et al. A new method for improving metaphase chromosome spreading[J]. Cytometry Part A the Journal of the International Society for Analytical Cytology，2003，51(1)：46-51.
[15]贾建. 沙果果醋醋酸发酵工艺条件的优化[J]. 中国调味品，2015，40(4)：42-45.
[16]叶双峰，叶书有，张影. 回归最优设计进行无花果施肥的研究[J]. 江苏林业科技，2001，28(2)：25-27.
[17]HENEGARIU O，HEEREMA N A，WRIGHT L L，et al. Improvements in cytogenetic slide preparation：controlled chromosome spreading，chemical aging and gradual denaturing[J]. Cytometry，2001，43(2)：101-109.
[18]MASONBRINK R E，BIRCHLER J A. Sporophytic nondisjunction of the maize B chromosome at high copy numbers[J]. J Genet Genomics，2010，37(1)：79-84.
[19]LAMB J C，MEYER J M，CORCORAN B，et al. Distinct chromosomal distributions of highly repetitive sequences in maize[J]. Chromosome Research，2007，15(1)：33-49.
[20]WANG H，WAN Y. A modified method for improving the quality of metaphase preparations[J]. Chinese Journal of Birth Health & Heredity，2013，21(7)：64-65.
[21]徐道娜，薛志强，李世栋，等. 西瓜染色体压片技术改进研究[J]. 西北农业学报，2007，16(6)：301-304.
[22]姚启伦. 玉米染色体制片技术中关键因子处理效应及优化[J]. 湖南农业大学学报，2007，33(4)：419-422.
[23]KATO A，LAMB J C，BIRCHLER J A. Chromosome painting in maize using repetitive DNA sequences as probes for somatic chromosome identification[J]. Proc Natl Acad Sci USA，2004，101(37)：13554-13559.
[24]KATO A. Air drying method using nitrous oxide for chromosome counting in maize[J]. Biotech Histochem，1999，74(3)：160-166.
[25]AHLADA M R，RAO I，VICTOR R，et al. Slide making and prototype of a dropper device for chromosomal preparations[J]. Int J Hum Genet，2005，5(3)：213-216.

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备注/Memo

备注/Memo:
收稿日期：2018-01-12 基金项目：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） 作者简介：姚启伦（1964-），男，重庆万州人，博士，教授，主要从事玉米遗传育种研究，（Tel）13908253365;（E-mail）yql641@aliyun.com 通讯作者：陈发波，（Tel）13983320701;（E-mail）chenfabo963@126.com
更新日期/Last Update: 2019-05-05