George M L C, Regalado E, Li W, Cao M, Dahlan M, Pabendon M, Warburton M L, Xianchun X, Hoisington D
CIMMYT-Asian Maize Biotechnology Network, c/o IRRI, DAPO 7777, MetroManila, Philippines.
Theor Appl Genet. 2004 Jun;109(1):80-91. doi: 10.1007/s00122-004-1626-8. Epub 2004 Apr 17.
This study focuses on the standardization of techniques across laboratories to enable multiple datasets to be compared and combined in order to obtain reliable and robust wide-scale patterns of diversity. A set of protocols using a core collection of simple sequence repeat (SSR) markers, reference lines and standard alleles, plus a common system of allele nomenclature, was adopted in the study of maize genetic diversity in a network of laboratories in Asia. Pair-wise allele comparisons of the reference lines, done to assess the general agreement between datasets from four laboratories, showed error rates (raw) ranging from 5.8% to 9.7%, which were reduced to less than 8% after adjustments of correctable errors, and further reduced to less than 6% after the exclusion of all markers with greater than 10% individual error rates. Overall, 45% of the total mismatches were due to frameshift errors, 39% to wrong allele size, 15% to failed amplification and 1% to "extra" alleles. Higher genetic similarity values of the reference lines were achieved using fewer markers with data of higher quality rather than with more markers of questionable quality. Cluster analysis of the merged datasets showed the lines from southern China to be highly diverse, falling into six of the seven clusters observed and all well represented by tester lines. The lines from Indonesia fell into five of six groups, with two main groups represented by tester lines. The CIMMYT lines developed for the Asian region showed a relatively narrow genetic base, falling in two out of seven and in three out of six clusters in China and Indonesia, respectively. In contrast to the case in southern China where 95% of the lines clustered separately from the CIMMYT lines, lines in the Indonesian breeding program show a closer relationship with the CIMMYT lines, reflecting a long history of germplasm exchange.
本研究聚焦于跨实验室技术的标准化,以使多个数据集能够进行比较和整合,从而获得可靠且稳健的大规模多样性模式。在亚洲实验室网络开展的玉米遗传多样性研究中,采用了一套使用简单序列重复(SSR)标记核心集合、参考系和标准等位基因的协议,以及一个通用的等位基因命名系统。对参考系进行成对等位基因比较,以评估来自四个实验室的数据集之间的总体一致性,结果显示错误率(原始)在5.8%至9.7%之间,在纠正可纠正错误后降至8%以下,在排除所有个体错误率大于10%的标记后进一步降至6%以下。总体而言,总错配的45%归因于移码错误,39%归因于等位基因大小错误,15%归因于扩增失败,1%归因于“额外”等位基因。使用较少但质量较高的数据标记而非较多质量存疑的标记,可实现参考系更高的遗传相似性值。合并数据集的聚类分析表明,中国南方的品系具有高度多样性,落入观察到的七个聚类中的六个,且所有聚类均由测试系很好地代表。印度尼西亚的品系落入六个组中的五个,有两个主要组由测试系代表。为亚洲地区培育的国际玉米小麦改良中心(CIMMYT)品系显示出相对较窄的遗传基础,在中国和印度尼西亚分别落入七个聚类中的两个和六个聚类中的三个。与中国南方95%的品系与CIMMYT品系分开聚类的情况相反,印度尼西亚育种计划中的品系与CIMMYT品系关系更密切,这反映了种质交换的悠久历史。