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作物学报 ›› 2010, Vol. 36 ›› Issue (2): 202-210.doi: 10.3724/SP.J.1006.2010.00202

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

 云南甘蔗自育品种DNA指纹身份证构建

刘新龙1,2,马丽1,2,陈学宽1,2,应雄美1,2,蔡青1,3,刘家勇1,2,吴才文1,2,*   

  1. 1 云南省甘蔗遗传改良重点实验室，云南开远661600; 2 云南省农业科学院甘蔗研究所，云南开远 661600; 3 云南省农业科学院生物技术与种质资源研究所，云南昆明 650223
  • 收稿日期:2009-07-03 修回日期:2009-09-08 出版日期:2010-02-10 网络出版日期:2009-12-21
  • 通讯作者: 吴才文, E-mail: gksky_wcw@163.com
  • 基金资助:

    本研究由国家科技支撑计划（2007BAD30B02），现代农业产业技术体系建设专项资金（nycytx-024-01-03），国家科技基础条件平台工作项目子专题（2007DKA21002-11）和农业部农林动植物育种工程（2006BAD01A06-4-1）资助。

     

     

     

Establishment of DNA Fingerprint ID in Sugarcane Cultivars in Yunnan, China

LIU Xin-Long1,2,MA Li1,2,CHEN Xue-Kuan1,2,YING Xiong-Mei1,2,CAI Qing1,3,LIU Jia-Yong1,2,WU Cai-Wen1,2,*   

  1. 1 Yunnan Key Laboratory of Sugarcane Genetic Improvement, Kaiyuan 661600, China; 2 Sugarcane Research Institute, Yunnan Academy of Agricultural Sciences, Kaiyuan 661600, China; 3 Biotechnology & Genetic Resources Institute, Yunnan Academy of Agricultural Sciences, Kunming 650223, China
  • Received:2009-07-03 Revised:2009-09-08 Published:2010-02-10 Published online:2009-12-21
  • Contact: WU Cai-Wen, E-mail: gksky_wcw@163.com

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2276

被引次数

51

摘要：

以云南27份甘蔗自育品种为材料，从国际微卫星协会提供的120对SSR引物中筛选出8对多态性丰富、品种区分率高、易统计的引物组成核心引物。8对SSR引物共产生129条带，123个为多态带，多态条带比例为95.35%，多态信息量平均为0.9445，品种相似性系数在0.269~0.767之间，其中引物SMC1047HA，MSSCIR21不仅多态性丰富，而且单个引物就可区分所有品种，是最有效的核心引物。8对核心引物两两组合的效率分析表明，MSSCIR36/MSSCIR2、MSSCIR16/MSSCIR36和MSSCIR36/SMC336BS是高效引物组合，可以完全有效区分所有品种，且品种相似性系数较低；同时使用蔗区种植面积较大的10个主栽品种验证3个高效引物组合，结果表明, MSSCIR16/MSSCIR36是最佳引物组合，不仅能有效区分所有云南自育品种，而且能将云南自育品种与10个主栽品种最有效地区分开。使用品种的国圃号、国家地区代码、育种单位英文缩写、核心引物名称和分子数据组成云南甘蔗自育品种的DNA指纹身份证，不仅包含了品种的重要信息，而且其中的分子数据可用于品种的真伪鉴定和遗传关系分析，为品种的知识产权保护提供有效的科学依据。

关键词: 云南甘蔗品种, SSR, DNA, 指纹, 身份证

Abstract:

To well evaluate and use the cultivars, we should identify them scientifically and accurately by DNA molecular markers. In this paper, 27 cultivars developed by two breeding institutes in Yunnan province were analyzed with SSR marker. Eight pairs of core SSR primers selected from about 120 pairs of SSR primers offered by the International Sugarcane Microsatellite Consortium made up the core primers for DNA fingerprint. A total of 129 bands were acquired by PAGE with the core primers, 123 of which were polymorphic bands, accounting for percentage of polymorphic band (PPB) was 95.35%, and the mean value of polymorphism information content(PIC) reached 0.9445; the genetic similarity coefficient of the cultivars was 0.269–0.767. SMC1047HA and MSSCIR21 with high PIC value could be used to distinguish all cultivars, which were the most efficient single primers. The result of evaluating different primer combinations from eight core primers indicated that MSSCIR36/MSSCIR21, MSSCIR16/MSSCIR36, and MSSCIR36/SMC336BS were very efficient in identifying these Yunnan cultivars, and their similar coefficients were lower than those of other primer combinations. At the same time, three primer combinations were validated with ten main released cultivars. The result showed MSSCIR16/MSSCIR36 was the optimum primer combination, which can be used in constructing DNA fingerprint ID of cultivars. The DNA fingerprint ID was set up, including serial number of National Nursery of Sugarcane Germplasm Resources (NNSGR), country& region code, breeding institute, core primer name and SSR marker data, which not only consists of the important information of cultivars, but also helps researchers to identify cultivars efficiently. At the same time, it can provide reliable scientific evidence for the protection of intellectual property right for these cultivars.

Key words: Yunnan sugarcane culticar, SSR, DNA, Fingerprint, ID

[1] Deng H-H(邓海华), Zhang Q(张琼). Analysis on the parents of commercial varieties released in Mainland China in recent years. Guangdong Agric Sci (广东农业科学), 2006, (12): 7-10 (in Chinese with English abstract)

[2] Chen R-K(陈如凯). Theory and Practice in Modern Sugarcane Breeding (现代甘蔗育种的理论与实践). Beijing: China Agriculture Press, 2003. pp 2-7 (in Chinese)

[3] Gao Y-L(高运来), Zhu R-S(朱荣胜), Liu C-Y(刘春燕), Li W-F(李文福), Jiang H-W(蒋洪蔚), Li C-D(李灿东), Yao B-C(姚丙晨), Hu G-H(胡国华), Chen Q-S(陈庆山). Establishment of molecular ID in soybean varieties in Heilongjiang, China. Acta Agron Sin (作物学报), 2009, 35(2): 211-218 (in Chinese with English abstract)

[4] 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), Zhao C-P(赵昌平). Method of ID constitution for wheat cultivars. Acta Agron Sin (作物学报), 2007, 33(10): 1738-1740 (in Chinese with English abstract)

[5] She H-D(佘花娣), Chen J-T(陈景堂), Huang Y-Q(黄亚群), Zhu L-Y(祝丽英), Chi S-M(池书敏). Progress on variety identification of crop using DNA fingerprinting. J Agric Univ Hebei (河北农业大学学报), 2003, 26(suppl): 28-32 (in Chinese with English abstract)

[6] Xin J-S(辛景树), Guo J-L(郭景伦), Zhang R-B(张软斌). Compare and analysis of a determination report in the variety authenticity and the purity of seeds by molecular maker. Seeds (种子), 2005, 24(1): 58-60 (in Chinese)

[7] Chen Y-H(陈英华), Hou Y-M(侯昱铭), Li H-Z(李宏字), Li M-B(李茂柏), Yuan Y(袁媛), Xu Z-J(徐正进). Establishment of DNA fingerprinting and analysis of genetic diversity among japonica rice cultivars attended regional trials in northeast region of China. Seeds (种子), 2009, 28(3): 28-34 (in Chinese)

[8] Li l(李莉), Song S-F(宋书锋), Liu Q-S(刘全胜), Fu Y-F(付岳峰), Fu X-Q(符习勤), Dai H(戴恒). Establishment of fingerprint map of hybrid rice with SSR markers. J Yangtze Univ(Nat Sci Edn)(长江大学学报×自然科学版), 2008, 5(4): 67-70 (in Chinese with English abstract)

[9] Wu W(武文), Deng Q-Y(邓启云), Zhou L-J(周丽洁), Zhu X-Q(朱校奇), Chen C-G(陈春光), Zhuang W(庄文), Jin D-M(金德敏), Wang B(王斌). Establishment t of DNA fingerprinting with SSR for Y58S and some other elite parents of two -line hybrid rice. Hybrid Rice (杂交水稻), 2008, 23(3): 52-56 (in Chinese with English abstract)

[10] Zhao J-R(赵久然), Wang F-G(王凤格), Guo J-L(郭景伦), Chen G(陈刚), Liao Q(廖琴), Sun S-X(孙世贤), Chen R-M(陈如明), Liu L-Z(刘龙洲). Series of research on establishing DNA fingerprint pool of Chinese new maize cultivars: II. Confirmation of a set of SSR core primers pairs. J Maize Sci (玉米科学), 2003, 11(2): 3-5 (in Chinese with English abstract)

[11] Wang F-G(王凤格), Zhao J-R(赵久然), Guo J-L(郭景伦), Liu L-Z(刘龙洲). Series of research on establishing DNA fingerprinting poll of Chinese new maize cultivars I. The establishment of a standard SSR system fitting for maize cultivars identification. J Maize Sci (玉米科学), 2003, 11(1): 3-6 (in Chinese with English abstract)

[12] Li X-H(李晓辉), Li X-H(李新海), Gao W-W(高文伟), Tian Q-Z(田清震), Li M-S(李明顺), Ma F-M(马凤鸣), Zhang S-H(张世煌). Establishment of DNA fingerprinting database of maize hybrids and its application in parentage identification. Acta Agron Sin (作物学报), 2005, 31(3): 386-391 (in Chinese with English abstract)

[13] Wu Y-S(吴渝生), Yang W-P(杨文鹏), Zheng Y-L(郑用琏). Establishment of fingerprinting for three hybrids and their parents by SSR markers. Acta Agron Sin (作物学报), 2003, 29(4): 496-500 (in Chinese with English abstract)

[14] Li G-Y(李根英), Dreisigacker S, Warburton M L, Xia X-C(夏先春), He Z-H(何中虎), Sun Q-X(孙其信). Development of a fingerprinting database and assembling an SSR reference kit for genetic diversity analysis of wheat. Acta Agron Sin (作物学报), 2006, 32(12): 1771-1778 (in Chinese with English abstract)

[15] Lin D-Z(林道哲), Dong S-W(董顺文), Liu C-H(刘朝辉), Wang J(王军), Qiu Y-X(邱有荣), Huang X(黄缨), Li S-C(李首成). Fingerprinting analysis of twenty six Brassica napus inbred lines based on RAPD. J Anhui Agric Sci (安徽农业科学), 2008, 36(20): 8493-8496 (in Chinese with English abstract)

[16] Chen B-Y(陈碧云), Wu X-M(伍晓明), Zhang D-X(张冬晓), Liu F-L(刘风兰), Lu G-Y(陆光远), Xu K(许鲲), Gao G-Z(高桂珍). SSR marker fingerprinting of winter rapeseed varieties in national field trails. Mol Plant Breed (分子植物育种), 2008, 6(4): 709-716 (in Chinese with English abstract)

[17] Pan Y B, Miller J, Schnell R, Richard E, Wei Q C. Application of microsatellite and RAPD Fingerprints in the florida sugarcane variety program. San Diego：Plant and Animal Genome XI Conference, 2003. 19-28

[18] Pan Y B, Scheffler B, Richard E P. High-throughput genotyping of commercial sugarcane clones with microsatellite(SSR)DNA markers. Sugar Tech, 2007, 9: 176-181

[19] Piperidis G, Rattey A R, Taylor G O, Cox M C. DNA markers: A tool for identifying sugarcane varieties. Austral Sugarcane, 2004, 8(suppl): 1-8

[20] You J-H(游建华), Wu G-C(吴凯朝), Liang J(梁俊), Fang F-X(方锋学), Mo L-X(莫磊兴). Assessment of the genetic diversity among sugarcane cultivars by use SSR marker. J Anhui Agric Sci (安徽农业科学), 2008, 36(10): 3990-3998 (in Chinese with English abstract)

[21] Lao F-Y(劳方业), Liu R(刘睿), He H-Y(何慧怡), Deng H-H(邓海华), Chen Z-H(陈仲华), Chen J-W(陈健文), Fu C(符成), Zhang C-M(张垂明), Yang Y-H(杨业后). AFLP analysis of genetic diversity in series sugarcane parents developed at HSBS. Mol Plant Breed (分子植物育种), 2008, 6(3): 517-522 (in Chinese with English abstract)

[22] Zhuang N-S(庄南生), Zheng C-M(郑成木), Huang D-Y(黄东益), Tang Y-Q(唐燕琼), Gao H-Q(高和琼). AFLP analysis for sugarcane germplasms. Acta Agron Sin (作物学报), 2005, 31(4): 444-450 (in Chinese with English abstract)

[23] 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 difference between cultivars in sugarcane (Saccharum officinarum L.). Acta Agron Sin (作物学报), 2004, 30(10): 1008-1013 (in Chinese with English abstract)

[24] Cai Q(蔡青), Fan Y-H(范源洪), Aitken K, Piperidis G, McIntyre C L, Jackson P. Assessment of the phylogenetic relationships withi n the “Saccharum Complex” using AFLP markers. Acta Agron Sin (作物学报), 2005, 31(5): 551-559 (in Chinese with English abstract)

[25] Aitken K S, Jackon P A, McIntyre C L. A combination of AFLP and SSR markers provides extensive map coverage and identification of homo(eo)logous linkage groups in a sugarcane cultivar. Theor Appl Genet, 2005, 110: 789-801

[26] Liu X-L(刘新龙), Cai Q(蔡青), Bi Y(毕燕), Lu X(陆鑫), Ma L(马丽), Ying X-M(应雄美), Mao J(毛钧). A rapid silver staining method for PAGE used in sugarcane AFLP and SSR molecular markers. Jiangsu J Agric Sci (江苏农业学报), 2009, 25(2): 433-435 (in Chinese with English abstract)

[27] Pan Y B. Highly polymorphic microsatellite DNA markers for sugarcane germplasm evaluation and variety identity testing. Sugar Tech, 2006, 8: 246-256

[28] Zhang Y(张彦), Guo S-W(郭士伟), He B(何冰), Gao D-Y(高东迎). Construction of molecular fingerprinting database for hybrid rice using SSR markers. Jiangsu J Agric Sci (江苏农业学报), 2006, 22(2): 181-183 (in Chinese with English abstract)

[29] 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)

[30] Liang M-S(梁明山), Zeng Y(曾宇), Zhou X(周翔), Hou L-J(侯留记), Li X(李霞). Genetic markers and their applications in identifying crop cultivars. Chin Bull Bot (植物学通报), 2001, 18(3): 257-265 (in Chinese with English abstract)
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