| 湖南宽皮柑橘EST-SSR反应体系研究 |
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| 引用本文： | 曾柏全,邓子牛,熊兴耀,杨迎花.湖南宽皮柑橘EST-SSR反应体系研究[J].中国农学通报,2009,25(21):244-247. |
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| 作者姓名： | 曾柏全 邓子牛 熊兴耀 杨迎花 |
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| 作者单位： | 1. 湖南农业大学园艺园林学院,长沙,410128;中南林业科技大学院生命科学与技术学院,长沙,410004
2. 湖南农业大学园艺园林学院,长沙,410128 |
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| 基金项目： | 新型分子标记研发及在湖南宽皮柑橘中应用 |
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| 摘 要： | 首次以EST-SSR 技术应用于湖南宽皮柑橘的研究中，进行了引物的设计与筛选，建立了湖南宽皮柑橘的EST-SSR 的反应体系。共筛选了20对引物，其中12对引物扩增多态性较高，研究确定在25 uL的反应体系中, Mg2+浓度在2.5 mmol?L-1, dNTP 0.2 mmol?L-1,上下引物各为0.2umol?L-1, Taq酶浓度为1.0 U效果好 , 最佳PCR程序为94℃预变性１ min，然后94 ℃，1 min；52 ℃，1min；73 ℃，1min，45个循环，最后73 ℃延伸3 min,利用优化后的反应体系成功构建湖南宽皮柑橘EST-SSR的指纹图谱.
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| 关 键 词： | 水分胁迫 水分胁迫 橡胶树 |
| 收稿时间： | 2009-06-09 |
| 修稿时间： | 2009-07-20 |
Study on the EST-SSR Marker Reaction System of mandarins(Citrus Reticulata Blanco) in Hunan |
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| Zeng Baiquan,Deng Ziniu,Xiong Xingyao,Yang Yinhua.Study on the EST-SSR Marker Reaction System of mandarins(Citrus Reticulata Blanco) in Hunan[J].Chinese Agricultural Science Bulletin,2009,25(21):244-247. |
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| Authors: | Zeng Baiquan Deng Ziniu Xiong Xingyao Yang Yinhua |
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| Institution: | Zeng Baiquan, Deng Ziniu, Xiong Xingyao, Yang Yinhua (1Horticulture and Landscape College, Hunan Agriculture University, Changsha 410128; 2College of Life Science, Central South University of Forestry and Technology, Changsha 410004) |
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| Abstract: | With the purpose to clarify the relationships among the native citrus resources in Hunan, EST-SSR markers were developed. The results showed that the clean and stable bands had been detected by twelve pairs of EST-SSR primer which had been selected out from twenty pairs .Each 25μl PCR reaction mixture consisted of genomic DNA, 2.5 mmol/L of MgCl2, 0.2 mmol/L of dNTP, 0.2 μmol/L of enemprimer and 1.0 unit of Taq polymerase .The results showed that the most suitable protocol was initially denaturing at 94℃ for 1 min; then 94 ℃, 60℃ second; 52 ℃, 60 second; and 73 ℃, lmin for 45 cycles., and finally extended at 73℃ for 3 min. It was established the genetic linking maps of mandarins in Hunan with optimizal reaction system. |
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| Keywords: | Citrus Reticulata Blancozz Molecular markerzz Primerzz EST-SSRzz Reaction systemzz |
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