Study on Identification of Ty- Gene,Ty- Gene,Mi Gene and Cf- Gene by Quadruple PCR in Tomato.[J].Northern Horticulture,2013,37(09):119-123.
四重PCR技术鉴定番茄Ty-1、Ty-2、Mi和Cf-5基因研究
- 文章编号:
- 1001-0009(2013)09-0119-05
- 分类号:
- S 641.203.6
- 文献标志码:
- A
- 摘要:
- 以番茄为试材，只利用1次PCR反应体系，同时对与番茄黄化曲叶病毒病（Tomato yellow leaf curl virus）的Ty-1、Ty-2基因、番茄根结线虫病（Root-knot nematode）的Mi基因和番茄叶霉病（Leaf mold）的Cf-5基因紧密连锁的SCAR标记进行了筛选。结果表明：扩增的特异性片段和单引物扩增的片段吻合，即对与Ty-1基因、Ty-2基因、Mi基因和Cf-5基因紧密连锁的片段特异性扩增，所获得的片段大小分别是395、900、750和960 bp。经TaqI酶切后所获得的特异性片段大小与前人试验所得大小基本相同，初步证明了四重PCR检测的准确性。该体系可对Ty-1、Ty-2、Mi和Cf-5 4个抗病基因进行同时筛选，在苗期辅助选择。降低了生产成本，节约时间、人力、物力，为分子标记辅助选择育种奠定了基础。
- Abstract:
- Taking Lycopersicon esculentum as material，Ty-1，Ty-2，Mi and Cf-5 in tomato tightly linked with four SCAR markers respectively resistant to Tomato Yellow Leaf Curl virus，root-knot nematode and tomato leaf mold were amplified and screened by using a single PCR reaction.The results showed that the PCR products were identical with one amplified by a single SCAR primer.Among them，four SCAR markers tightly linked with Ty-1，Ty-2，Mi and Cf-5 genes produced 395，900，750 and 960 bp bands.Cleavaged with the restriction enzyme Taq I，these specific fragments produced almost unanimously the same size with the others.Preliminary proved the veracity of quadruple PCR.The accurate results proved that Ty-1，Ty-2，Mi and Cf-5 resistant genes could be identified simultaneously by using under adaptable condition.Quadruple PCR could be very important for marker-assisted selection during early stage in tomato.Also could reduce production costs，time saving and less labor.If could reasonable used，it will become an important method of breeding work.
参考文献/References:
[1]李景富.中国番茄育种学[M].1版.北京:中国农业出版社,2011:264-265. [2]彭德良.蔬菜线虫病害的发生和防治[J].中国蔬菜,1998(4):57-58. [3]韩文华,许文奎,刘石磊,等.番茄叶霉病生理小种分化及抗病育种研究进展[J].辽宁农业科学,2005(5):34-37. [4]于拴仓,柴敏,郑晓鹰.番茄叶霉病抗性基因Cf-5的CAPS标记建立[J].分子植物育种,2005,3(1):57-60. [5]Zamir D，Ekstein-Michelson I，Zakay Y，et al.Mapping and introgression of a tomato yellow leaf curlvirus tolerance gene,Ty-1[J].Theor Appl Genet,1994,88:141-146. [6]Hanson P M，Green S K，Kuo G.Ty-2,a gene on chromosome 11 conditioning Gemini virus resistance in tomato[J].Rep Tomato Genet Coop,2006,56:17-18. [7]于力,朱龙英,万延慧,等.多重PCR技术鉴定番茄Ty-1和Mi基因[J].上海农业学报,2009,25(2):6-9. [8]宋燕,陈丽静,李君明,等.利用多重PCR反应同时筛选番茄Tm-22和Mi基因[J].植物遗传资源学报,2006,7(2):165-16. [9]许爽,褚云霞,张辉,等.多重PCR技术鉴定番茄Ty-2和Ty-3基因及田间验证[J].分子植物育种,2009,7(5):954-958. [10]于力,朱龙英,万延慧,等.利用多重PCR反应同时筛选番茄Ty-1和Cf-5基因[J].上海农业学报,2009,25(2):6-9. [11]Williamson V M，Ho J Y，Wu F F，et al.A PCR-based marker tightly linked to the nematode resistance gene，Mi，in tomato[J].Theor Appl Genet,1994,87:757-763. [12]Chamberian J S，Gibbs R A，Ranier J E，et al.Detection screening of the duchenne muscular dystrophy locus via multiplex DNA amplification[J].Nucl Acids Res,1988,16:1141-1156. [13]崔玉娟,胡丹东.多重PCR技术在植物学研究中的应用[J].天水师范学院学报,2008;28(5):43-45. [14]Kirschberg O，Schuttler C，Repp R，et al.A Multiplex-PCR to Identify Hepatitis B Virus-genotypes A-F[J].J Clin Virol,2004,29(1):39-43. [15]李君明,陈丽静,宋燕.利用多重PCR反应同时筛选番茄Tm-22和Mi基因[J].农业生物技术学报,2005,13(6):698-702.
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备注/Memo
第一作者简介：刘超勤（1987-），男，硕士，研究方向为番茄种质资源的研究和利用。E-mail:chaoqin0110@163.com.