玉米SLAF标记的开发及其在玉米果皮纤维素含量BSA分析中的应用

doi:10.3864/j.issn.0578-1752.2018.08.001

Abstract

Abstract: 【Objective】 Reducing cellulose content in pericarp is one of the important goals for quality improvement of sweet maize. However, the research focusing on cellulose content in pericarp of maize is limited and the related gene has not been identified. In order to map the chromosome regions and candidate genes controlling cellulose content in pericarp of sweet maize, specific-locus amplified fragment-sequencing (SLAF-seq) and bulked segregant analysis (BSA) were conducted in this study. 【Method】 A recombinant inbred line (RIL) population was constructed using E327 and G5-1 as parents. In F6 generation of RILs, the cellulose contents in pericarp of each line were detected. According to cellulose content results, the lines with relatively high and low cellulose contents in pericarp were selected for SLAF tags identification and BSA. For BSA, DNA was extracted from two bulked populations as well as two parent lines and digested with HaeⅢ and Hpy166Ⅱ restriction enzymes. After digestion, the 414-464 bp DNA fragments were recovered and used for Illumina library construction which were subsequently sequenced by 2nd generation sequencing technology. Based on the polymorphic SLAF tags developed in this analysis, the SNPs-cellulose content association analysis was performed to map the chromosome regions associated with cellulose content in pericarp of sweet maize. Then, the genes located on the associated regions were found. To further reduce the number of candidate genes, the genes in Arabidopsis homological to these genes were identified and annotated. Based on the published works related on the functional analysis of these Arabidopsis genes, we further identified the candidate genes regulating cellulose content in pericarp of sweet maize. 【Result】The sequencing results of Illumina libraries were consistent with expected. As a result, 73 786 polymorphic SLAF tags were identified, which were uniformly distributed on 10 chromosomes. A total of 523 395 SNPs in those polymorphic tags were identified from the SLAF-sequencing data. Genes responsible for cellulose content in pericarp were mapped onto 6 chromosome regions of maize genome via association analysis and all these 6 chromosome regions were located on the 5th chromosome. In total, 47 gene loci in those regions and 9 genes in these associated regions were identified as candidate genes in further analysis. 【Conclusion】Through SLAF-sequencing based bulking segregated analysis, the cellulose content related genes in pericarp of sweet maize were mapped, suggesting that this method can be applied in gene mapping for other traits.

Key words: maize, pericarp, cellulose, specific-locus amplified fragment-sequencing, bulked segregant analysis

DU LongGang, WANG MeiXing. SLAF-marker Development and Its Application in BSA Analysis of Cellulose Content in Pericarp of Maize Kernel[J].Scientia Agricultura Sinica, 2018, 51(8): 1421-1430.

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