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High Amylose Maize Germplasm Cluster Analysis and Development of Molecular Markers for SbeⅠ and SbeⅡb Genes

Author: NingLiHua
Tutor: ChenHuaBang
School: Shandong Agricultural University
Course: Crop Genetics and Breeding
Keywords: Corn High amylose Cluster analysis Molecular marker-assisted breeding
CLC: S513
Type: Master's thesis
Year: 2011
Downloads: 82
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Abstract

Corn (Zea mays L.), the large scale planting of a crop, corn starch is widely used, the amylose contained in the industrial application is very wide. The lower proportion of ordinary amylose corn, isolated from the common maize grain amylose high cost, which limits the application of amylose. Therefore, to carry out high-amylose maize breeding has an important significance to the development of the production and processing of high amylose. Domestic high-amylose corn severe shortage of resources, the introduction of high-amylose corn germplasm from abroad effectively broaden the high amylose maize germplasm. Control amylose content of major gene the ae gene, as well as many modifier genes common work. According to published NCBI on published sequence cloning the ae and sbeI of genes of high amylose corn, the development of the appropriate molecular markers, and eventually for high amylose maize molecular marker-assisted breeding. The results show that: 1 with 25 pairs of SSR molecular marker amplification of 16 high amylose maize inbred lines and normal corn genome, a total of 65 alleles were detected, 18 were genetic similarity coefficient (0.446 ^{0 .923). UPG MA cluster analysis, 16 high amylose maize inbred lines divided into two categories, four were divided to the SS in germplasm group, and another 12 were divided to the NSS germplasm group. Invaluable GMES-0067, Public amylose content is up to more than 70% of the germplasm of this the germplasm is divided into NSS germplasm group. Genetic diversity analysis of high-starch corn inbreds, the study tested inbred lines heterotic groups, provide the basis for high-amylose corn breeding and germplasm improvement. The primers were designed according to the ae published on the NCBI gene sequence (AF072725), GEMS-0067, LH324, BC300, I115, and Chang 7-2 ae gene cDNA were cloned. Sequence alignment analysis showed that, high amylose starch corn and common corn of the cDNA sequence with a high homology, but the measured high amylose starch corn the ae gene of the 9th outside exon sequences of all the missing high amylose starch corn and common corn the ae gene The cDNA sequence there are several different base difference. Exon 3 of the outer 9 difference at design primers, exon gene 8 and 10 outside the election of a sequence as labeled primers. The materials containing the ae gene in corn could amplify a 750bp specific band, while in the ordinary corn amplified a 1576bp specific bands, called the ae-InDel mark. Use of the mark to detect 16 kinds of high-amylose maize inbred lines and 10 normal maize inbred lines, high amylose maize inbred line genome can amplify the 750bp bands amplified in normal maize inbred lines a 1576bp bands in the F1 generation of high amylose corn and normal corn amplified 750bp and 1576bp bands. Codominant the mark as ae gene function markers. According to the the published NCBI on sbeI gene's sequence (NM ₀ 01.1119 million) designed cited material, respectively, clone 16 high straight chain starch corn genes of sbeI the cDNA, sequence alignment analysis showed that, high amylose starch corn and database the cDNA sequence sbeI in having a high homology, but 15 nt changes the GMES-0067 as measured sbeI gene, wherein 5 nucleotide changes result in amino acid mutation in the locus, and other high amylose corn sbeI The predicted protein sequence with online announced sbeI sequence differences. 5. GMES-0067's sbeI gene in which a change corresponding sites of amino acid nucleotide 1330 (CA) developed successfully able to mark, the primers amplified high amylose starch corn and common corn's genome, were amplified out of a 906bp of specific bands, but after digestion with AluI, only GMES-0067 bar with size has not changed, and other materials are cut into two small fragments (617bp and 289bp). Three with the method detection F1 (GMES-0067x Chang 7-2), so the mark as the GMES-0067 sbeI alleles of codominant molecular markers.}

High Amylose Maize Germplasm Cluster Analysis and Development of Molecular Markers for SbeⅠ and SbeⅡb Genes

Abstract

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