用SSR技术和混合取样方法估算玉米群体间的遗传距离

作物学报 ›› 2007, Vol. 33 ›› Issue (02): 317-321.

用SSR技术和混合取样方法估算玉米群体间的遗传距离

侯本军¹;王铁固^1,2;陈彦惠^1,*;吴连成¹;库丽霞¹

1 河南农业大学农学院，河南郑州450002；2 郑州大学离子束生物工程重点实验室，河南郑州450052

收稿日期:2006-01-04 修回日期:1900-01-01 出版日期:2007-02-12 网络出版日期:2007-02-12
通讯作者: 陈彦惠

DNA Sampling Strategy and Calculation of Genetic Distance between Maize (Zea mays L.) Populations

HOU Ben-Jun¹,WANG Tie-Gu¹²,CHEN Yan-Hui^1*,WU Lian-Cheng¹,KU Li-Xia¹

1 Agronomy College, Henan Agricultural University, Zhengzhou 450002, Henan; 2Ion Beam Biotechnology Key Laboratory, Zhengzhou University, Zhengzhou 450052, Henan, China

Received:2006-01-04 Revised:1900-01-01 Published:2007-02-12 Published online:2007-02-12
Contact: CHEN Yan-Hui

摘要/Abstract

摘要：

The factors affecting the binding characteristics of GBSS with starch granule were studied using a wheat (Triticum aestivum) cultivar Chinese Spring with different temperature treatments. After sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis, we got GBSS concentration by coomassie brilliant blue G-250(CBB G-250) method. The results showed that the temperature affected the binding of GBSS and starch granule. In 50–80℃, the concentration of GBSS unbound from starch increased with the temperature rising, which the maximal concentration was 11.361 μg mL^-1 at 80℃, while that was reduced when the temperature was 85–95℃. Furthermore, Mg²⁺ could also affect the quantity of GBSS unbound from starch. When Mg2+ concentration was lower than 1.75 mmol L^-1, the concentration of GBSS unbound increased. The lower Mg²⁺ concentration, the higher the concentration of GBSS unbound. However, when Mg²⁺ concentration was higher than 2.5 mmol L^-1, it could restrain GBSS unbinding. These results showed that GBSS bound starch granule by non-covalent bonds. The best GBSS extracting conditions were 55 mmol L^-1 Tris-HCl (pH 6.8), 0.75 mmol L^-1 MgCl₂, 2.3% SDS, 5% 2-ME and 10% glycerol, 15 min in boiling water, or 55 mmol L^-1 Tris-HCl (pH 6.8), 0.75 mmol L^-1 MgCl₂, 2.3% SDS, 5% 2-ME and 10% glycerol, 80℃ 30 min. The results are helpful to investigate 3-D structure of biological activated GBSS and mechanism of GBSS binding with starch granule.

关键词: SSR, 玉米, 遗传距离, 遗传多样性, 取样方法

Abstract:

SSR is a good molecule marker with broad application. Bulk sampling is feasible to analyze genetic diversity of maize population by SSR. It is necessary to study influence of genetic distance as using bulk sampling. In present study, the genetic distances between two populations were compared and analyzed by 50 pairs of SSR primers, checked by inbred lines Huangzao 4 and Mo17, and using 60 individuals DNA samples of Golden Queen and Yuzong 5 maize populations with six treatments (the individual DNA samples, the mixed DNA samples with 5 individuals, 10 individuals, 15 individuals, 20 individuals, and 30 individuals, respectively). The results indicated that using the individual DNA samples can detect the alleles with very low frequencies, and make the detected allele number decreased gradually and the frequency of missing alleles increased with the increase of mixed individuals in the treatmen. The results of the genetic distance between two populations calculated by three models suggested that the genetic distances GD, GD(J) and GD(N) showed a gradually increasing trend with the increasing of the mixed individual number, but the increasing extent of GD(J) and GD(N) was wider. There was an obvious deviation between GD(J), GD(N) from mixed samples and the GD from 60 individual DNA samples based on allele frequency. The results and the comparison of the genetic distances based on different calculation methods suggested that the 10 individuals’ DNA mixture is the best sampling strategy when genetic diversity and polymorphism in maize populations are studied by SSR.
If the Nei’s genetic distance GD(N) and Jaccord’s genetic distance GD(J) are calculated according to 0, 1 data, (GD(N)+GD(J))/2 will be calculated close to the genetic distance by allele frequency of individuals. Gene frequency can be studied by using capillary electrophoresis and SSR fluorescence detection. There will be a broad application prospect about bulk sampling strategy with SSR marker.

Key words: SSR, Maize, Genetic distance, Genetic diversity, Sampling method

侯本军;王铁固;陈彦惠;吴连成;库丽霞. 用SSR技术和混合取样方法估算玉米群体间的遗传距离[J]. 作物学报, 2007, 33(02): 317-321.

HOU Ben-Jun;WANG Tie-Gu;CHEN Yan-Hui;WU Lian-Cheng;KU Li-Xia. DNA Sampling Strategy and Calculation of Genetic Distance between Maize (Zea mays L.) Populations[J]. Acta Agron Sin, 2007, 33(02): 317-321.

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