Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (18): 3699-3708.doi: 10.3864/j.issn.0578-1752.2012.18.003

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Construction of Methylation Linkage Groups A and B in Sorghum with MSAP and SSR Markers and Analysis of Methylation Sites and Patterns

 DUAN  Yong-Hong, WANG  Ming, SUN  Yi, YANG  Wu-De   

  1. 1.山西农业大学农学院，山西太谷 030801
    2.山西省农业科学院生物技术研究中心，太原 030031
    3.农业部黄土高原作物基因资源与种质创制重点实验室，太原 030031
  • Received:2012-03-30 Online:2012-09-15 Published:2012-06-29

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Abstract: 【Objective】 The genetic linkage groups of A and B with methylation sensitive markers of Sorghum bicolor L. were constructed, and the methylation sites and methylation patterns were analyzed. 【Method】 F2 segregating population with 150 individuals derived from the sorghum cross B2V4 ×1383-2 was analyzed based on MSAP and SSR markers, and linkage groups were constructed using Map maker/EXP (version 3.0) and Map/Draw 2.1. 【Result】 Methylation linkage groups were constructed. Group A was composed of 30 loci covering 93.7 cM, of which 20 loci were MSAP markers, including 13 loci from EcoRⅠ/MspⅠ enzyme digestion, 7 loci from EcoRⅠ/HpaⅡenzyme digestion, and 10 loci were SSR markers. Group B was composed of 43 loci covering 90.4 cM, of which 39 loci were MSAP markers, including 19 loci from EcoRⅠ/MspⅠenzyme digestion, 20 loci from EcoRⅠ/HpaⅡenzyme digestion and 4 loci were SSR markers. There were only methylation marker products existed from EcoRⅠ/MspⅠ enzyme digestion on linkage group A-a, but on other linkage groups, the methylation markers from both EcoRⅠ/HpaⅡ and EcoRⅠ/MspⅠenzyme digestion were found. Additionally, Group A-b and Group B-b each had a slightly dense methylation resign, nearby Xtxp302 and Xtxp304, while, a highly dense methylation regions, nearby Xtxp 296 in linkage group B-a, with clusters of the EcoRⅠ/Msp I and EcoRⅠ/HpaⅡenzyme digestion MSAP markers were revealed. Based on polymorphism of methylation fragments between parents and their segregation among the F2 population, cytosine methylation patterns between hybrid and their parents were divided into two major groups. 【Conclusion】 MSAP markers can detect a large number of different methylated fragment, with anchor SSR markers, it is an efficient technique in constructing methylation genetic linkage groups or methylation genetic linkage map. High-density methylation regions were revealed near SSR markers Xtxp 302 on Group A-b, Xtxp 96 on Group B-a and Xtxp 304 on Group B-b. Among these loci the number of demethylated loci was higher than the number of methylated loci in hybrid F1 compared to its parents，which might imply a decreased methylation level in the hybrid genome.

Key words: sorghum, SSR, MSAP, DNA methylation

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