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作物学报 ›› 2011, Vol. 37 ›› Issue (06): 1116-1123.doi: 10.3724/SP.J.1006.2011.01116

• 研究简报 • 上一篇    

应用ISSR分子标记绘制红麻种质资源DNA指纹图谱

汪斌1,祁伟1,**,兰涛1,陈惠端2,徐建堂1,粟建光3,李爱青4,祁建民1,*   

  1. 1福建农林大学/作物遗传育种与综合利用教育部重点实验室, 福建福州 350002; 2 福建农林大学机电工程学院, 福建福州 350002; 3 中国农业科学院麻类研究所, 湖南长沙 410205; 4 安徽省种子管理总站, 安徽合肥 230001
  • 收稿日期:2010-09-16 修回日期:2011-03-06 出版日期:2011-06-12 网络出版日期:2011-03-24
  • 通讯作者: 祁建民, E-mail: qijm863@163.com, Tel: 0591-83644898
  • 基金资助:

    本研究由福建省科技重点项目(2008N0005), 福建省科技重点项目(2007N0011), 国家麻类产业技术体系建设(nycytx-19-s12), 国家公益性行业计划(nyhyzx07-018-3)项目资助。

Establishment of DNA Fingerprintings of Kenaf (Hibiscus Cannabinus L.) Germplasm Resources with ISSR Molecular Markers

WANG Bin1,**,QI Wei1,**,LAN Tao1,CHEN Hui-Duan2,XU Jian-Tang1,SU Jian-Guang3,LI Ai-Qing4,QI Jian-Ming1,*   

  1. 1 Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2 College of Mechanical and Electrical Engineering, Fujian Agriculture and Forestry University, Fuzhou 350002, China; 3 Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, China; 4 Anhui Province Seeds Management Station, Hefei 230001, China
  • Received:2010-09-16 Revised:2011-03-06 Published:2011-06-12 Published online:2011-03-24
  • Contact: 祁建民, E-mail: qijm863@163.com, Tel: 0591-83644898

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被引次数

72

摘要： 以6份红麻种质资源为材料, 对UBC807~UBC80等80个ISSR引物进行筛选, 筛选出多态性好的ISSR引物20个。利用这20个ISSR引物扩增来自国内外84份红麻种质资源, 共获得230条谱带, 平均每个引物扩增出11.5条谱带, 其中多态性谱带185条, 多态性条带比率为80.43%, 表明供试的红麻种质资源遗传多样性较丰富。以供试84份红麻种质资源的ISSR扩增谱带为基础, 建立了供试材料扩增条带指纹数据库的Excel文件。根据指纹图谱唯一性原则, 采用自行开发的DNA指纹数据分析软件, 再从20个多态性好的ISSR引物中遴选出UBC 813、UBC 825、UBC 836、UBC 888和UBC 889引物, 绘制出82个红麻种质资源的DNA指纹图谱, 为红麻种质资源分子身份证的构建奠定了基础。

关键词: 红麻, 种质资源, DNA, ISSR, 指纹图谱

Abstract: Kenaf (Hibiscus cannabinus L.) is an important economic crop in China. To identify germplasm resources and establish the data base of DNA fingerprintings, we adopted six kenaf germplasm resources to screen 80 ISSR primers, 20 of which were polymorphic. Then 20 ISSR primers were used to amplify 84 kenaf germplasm resources introduced and preserved from home and abroad. Totally 230 bands were produced,the average number of DNA bands amplified by each primer was 11.5, and the number of polymorphic DNA bands was 185. The polymorphic proportion of DNA bands was 80.43%, which indicated the abundant genetic diversity of kenaf germplasm resources preserved. Based on the DNA bands amplified from 84 kenaf germplasm resources, the Excel data base for PCR amplified bands of the kenaf germplasm resources was established. According to the principle of uniqueness of the fingerprintings, the DNA fingerprints of 82 kenaf germplasmwere constructed with five ISSR primers (UBC813, UBC825, UBC836, UBC888 and UBC889) selected from 20 polymorphic primers, using the analyzing software designed by our laboratory programmed and based on image manipulation. The fingerprintings provide a basis of the molecular identification of kenaf germplasm resources. In this research, the DNA fingerprintings system was proved to be feasible and reliable.

Key words: Kenaf (Hibiscus cannabinus L.), Germplasm resource, DNA, ISSR, Fingerprinting

[1]Cheng Z(程舟), Sameshima K(鲛岛一彦), Chen J-K(陈家宽). The study, processing and utilization of kenaf in Japan. Plant Fiber Products (中国麻业), 2001, 23(3): 16-24 (in Chinese)
[2]Dempsey J M. Fiber Crops. Gainesville: University Presses of Florida, 1975. pp 300-325
[3]Huang P-K(黄培坤), Deng L-Q(邓丽卿), Su J-G(粟建光), Li A-Q(李爱青). Identification and utilization study of kenaf germplasms from foreign countries. China’s Fiber crops (中国麻作), 1989, 11(4): 5-9 (in Chinese)
[4]Li A-Q(李爱青). The visiting report of jute and kenaf germplasms in Kenya. China’s Fiber crops (中国麻作), 1990, 12(1): 16-21 (in Chinese)
[5]Howard A, Howard G L C. Studies in Indian fibre plants 2 on some new varieties of Hibiscus cannabinus L. and Hibiscus sabdariffa L. Indian Dep Agric, Mem Bot Serv, 1911, 4: 9-36
[6]Bhardwaj H L, Rangappa, M, Webber III C L. Effects of row spacing, fertilizers, and varieties on kenaf yield in Virginia. In: Proceedings of the International Kenaf Association Conference, Albuquerque, NM 8, 1996. pp 21-26
[7]Menzel M Y, Wiloson F D. Chromosomes and crossing behaviour of Hibiscus cannabinus. Am J Bot, 1961, 48: 651-657
[8]Deng L-Q(邓丽卿), Su J-G(粟建光), Huang P-K(黄培坤), Li A-Q(李爱青). The study of morphology and tasonomy of kenaf germplasms. China’s Fiber crops (中国麻作), 1991, 13(4): 16-20 (in Chinese)
[9]Li A-Q(李爱青), Dai Z-G(戴志刚), Chen J-Q(陈基权), Su J-G(粟建光). Studies on identification and classification of kenaf (Hibiscus cannabinus L.) germplasm. J Plant Genet Resour (植物遗传资源学报), 2006, 7(3): 352-355 (in Chinese with English abstract)
[10]Tao A-F(陶爱芬), Qi J-M(祁建民), Li A-Q(李爱青), Fang P-P(方平平), Lin L-H(林荔辉), Wu J-M(吴建梅), Wu W-R(吴为人). The analysis of genetic diversity and relationship of elite kenaf germplasm based on inter-simple sequence repeats. Acta Acron Sin (作物学报), 2005, 31(12): 1668-1671 (in Chinese with English abstract)
[11]Lin L-H(林荔辉), Wang B(汪斌), Tao A-F(陶爱芬), Wu J-M(吴建梅), Sun Z-Q(孙志强), Fang P-P(方平平), Qi J-M(祁建民). The analysis of genetic diversity of kenaf germplasm based on inter-simple sequence repeats. J Wuhan Bot Res (武汉植物学研究), 2008, 26(3): 240-244 (in Chinese with English abstract)
[12]Li H(李辉), Li D-F(李德芳), Chen A-G(陈安国), Tang H-J(唐慧娟), Li J-J(李建军), Huo G(霍光). The breeding of kenaf male sterility and ISSR marker of male sterile gene. Chin Agric Sci Bull (农业生物技术科学), 2008, 24(8): 80-83 (in Chinese with English abstract)
[13]Huo G(霍光), Li D-F(李德芳), Chen A-G(陈安国), Li J-J(李建军), Tang H-J(唐惠娟). Genetic diversity analysis of 44 shares of Hibiscus cannabinus L. germplasm resources using ISSR molecular marker. J Anhui Agric Sci (安徽农业科学), 2009, 37(21): 9890-9892 (in Chinese with English abstract)
[14]Wang Z-H(王忠华). DNA fingerprinting technology and its application in crop germplasm resources. Mol Plant Breed (分子植物育种), 2006, 4(3): 425-430 (in Chinese with English abstract)
[15]Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T, Hornes M, Frijters A, Pot J, Peleman J, Kuiper M, Zabeau M. AFLP: A new technique for DNA fingerprinting. Nucl Acids Res, 1995, 23: 4407-4414
[16]Yuan L-X(袁力行), Fu J-H(傅骏骅), Warburtor M, Li X-H(李新海), Zhang S-H(张世煌), Khairallah M, Liu X-Z(刘新芝), Peng Z-B(彭泽斌), Li L-C(李连城). Comparison of genetic diversity among maize inbred lines based on RFLPs, SSRs, AFLPs and RAPDs. Acta Genet Sin (遗传学报), 2000, 27(8): 725-733 (in Chinese with English abstract)
[17]Li L(李丽), Zheng X-Y(郑晓鹰). Utilization of AFLP molecular markers in identification of Chinese cabbage varieties. Mol Plant Breed (分子植物育种), 2006, 4(5): 685-689 (in Chinese with English abstract)
[18]Li M(李鸣), Tan Y-M(谭裕模), Li Y-R(李杨瑞), Li R-B(李容柏), Gao G-Q(高国庆). AFLP molecular analysis of genetic diference between cultivars in sugarcane (Saccharum officinarum L.). Acta Acron Sin (作物学报), 2004, 30(10): 1008-1013 (in Chinese with English abstract)
[19]Williams J G, Kubelik A R, Libak K J, Rafalski J A, Tingey S V. DNA polymorohisms amplified by arbitrary primers are useful as genetic markers. Nucl Acids Res, 1990, 18: 6531-6535
[20]Welsh J, McClelland M. Fingerprinting genomes using PCR with arbitrary primers. Nucl Acids Res, 1990, 18(24): 7213-7218
[21]Zietkiewicz E, Rafalski A, Labuda D. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics, 1994, 20: 176-183
[22]Blair M W, Panaud O, Mccouch S R. Inter-simple sequence repeat (ISSR) amplification for analysis of microsatellite motif frequency and fingerprinting in rice (Oryza sativa L). Theor Appl Genet, 1999, 98: 780-792
[23]Prevost A, Wilkinson M J. A new system of comparing PCR primers appliedto ISSR fingerprinting of potato cultivars. Theor Appl Genet, 1999, 98: 107-112
[24]Tang L-H(唐利华), Xiao Y(肖扬), Bian Y-B(边银丙). ISSR fingerprint analysis and SCAR marker of major cultivated strains of Auricularia auricula in China. Mycosystema (菌物学报), 2008, 27(2): 243-251 (in Chinese with English abstract)
[25]Wang X-Y(王心宇), Chen P-D(陈佩度), Qi Z-J(亓增军), Zhang Z-Z(张政值), Ma Z-Q(马正强). Preliminary study of ISSR marker technique on wheat fingerprinting. J Agric Biotechnol (农业生物技术学报), 2001, 9(3): 261-263 (in Chinese with English abstract)
[26]Guo A-P(郭安平), Zhou P(周鹏), Su J-G(粟建光). Random amplified polymorphic DNA (RAPD) analyses among Hibiscus cannabinus and related species. J Trop Subtrop Bot (热带亚热带植物学报), 2002, 10(4): 306-312 (in Chinese with English abstract)
[27]Xu J-T(徐建堂), Qi J-M(祁建民), Fang P-P(方平平), Li A-Q(李爱青), Lin L-H(林荔辉), Wu J-M(吴建梅), Tao A-F(陶爱芬). Optimized CTAB protocol for extracting genomic DNA from kenaf and improved PCR amplifications of ISSR and SRAP. Plant Fiber Sci China (中国麻业科学), 2007, 29(4): 179-183 (in Chinese with English abstract)
[28]Guo X-Q(郭晓强), Feng Z-X(冯志霞). Jeffreys--the creator of DNA fingerprint map. Bull Biol (生物学通报), 2008, 43(1): 60-61 (in Chinese)
[29]Wang L-X(王立新), Li Y-F(李云伏), Chang L-F(常利芳), Huang L(黄岚), Li H-B(李宏博), Ge L-L(葛玲玲), Liu L-H(刘丽华), Yao J(姚骥), ZhaoC-P(赵昌平). Method of ID constitution for wheat cultivar. Acta Acron Sin (作物学报), 2007, 33(10): 1738-1740 (in Chinese with English abstract)
[30]Zhuang J-Y(庄杰云), Shi Y-F(施勇烽), Zhuang J-Z(庄杰政), E Z-G(鄂志国), Zeng R-Z(曾瑞珍), Chen J(陈洁), Zhu Z-W(朱智伟). Construction and testing of primary microsatellite database of major rice varieties in China. Chin J Rice Sci (中国水稻科学), 2006, 20(5): 460-468 (in Chinese with English abstract)
[31]Wang F-G(王凤格), Zhao J-R(赵久然), Wang L(王璐), Yi H-M(易红梅), Guo J-L(郭景伦), Dai J-R(戴景瑞), Yuan Y-P(原亚萍), Lu B-S(卢柏山), Yang G-H(杨国航). Determination of SSR core primers for maize hybrid purity identification. J Agric Biotechnol (农业生物技术学报), 2007, 15(6): 964-969 (in Chinese with English abstract)
[32]Chen M-X(陈美霞), Zhang G-Q(张广庆), Qi J-M(祁建民), Zhang X-C(张晓琛), Lin L-H(林荔辉), Lan T(兰涛), Wu W-R(吴为人), Wu J-M(吴建梅), Tao A-F(陶爱芬), Fang P-P(方平平). Preliminary study on construction of a genetic linkage map of kenaf using SRAP and ISSR markers. Plant Fiber Sci China (中国麻业科学), 2008, 30(3): 121-127 (in Chinese with English abstract)
[33]Han Y-C(韩延闯), Diao Y(刁英), Zhou L(周立), Liu J-Y(刘静宇), Zhou M-Q(周明全), Hu Z-L(胡中立), Song Y-C(宋运淳). Construction of the DNA fingerprinting in Nelumbo. J Wuhan Bot Res (武汉植物学研究), 2004, 22(3): 193-196 (in Chinese with English abstract)
[34]Wei Z-W(魏臻武). DNA fingerprint of Medicago sativa variety genomes using SSR, ISSR and RAPD. Acta Pratac Sin (草业学报), 2004, 13(3): 62-67 (in Chinese with English abstract)
[35]Zhou J-L(周建林), Jie Y-C(揭雨成). Isolation of microsatellite loci in kenaf (Hibiscus cannabinus). J Hunan Norm Univ (Nat Sci) (湖南师范大学自然?科学学报), 2006, 29(1): 78-82 (in Chinese with English abstract)
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