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Genetic Analysis and Gene Mapping of a Floral Organ Development Mutant in Arabidopsis

Author: FuHuiLing
Tutor: ZhouYongMing
School: Huazhong Agricultural University
Course: Crop biotechnology
Keywords: Arabidopsis 3-34 mutant Map-based cloning InDel markers
CLC: Q943
Type: Master's thesis
Year: 2009
Downloads: 65
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Abstract


The model plant Arabidopsis, plays an important role in plant genome research . Arabidopsis mutant identification and gene function analysis of Arabidopsis functional genomics research . This study aimed to developmental mutant Arabidopsis floral organ on recently obtained in this laboratory for detailed genetic analysis and map-based cloning separation control gene mutation traits . This experiment studied mutant 3-34 separation offspring from Arabidopsis T-DNA insertion mutation system NH ( Columbia ecotype ) . The mutant in the process of plant growth and development performance the mutant trait : the main stem is thin and short , and some can not even bolting ; darker leaves , moss stems elongated or distorted ; buds clustered more and most developmental completely ; petals ranging in number two , three , four and five , some flower pistil missing . Preparation T-DNA vector (pBI121) , 35S region of the designed primer four primer 35s-3/YM14 , 35s-3/FsaL , CTF / CTR and GFPR / GFPF to detect vector sequences , but the PCR results showed is negative i.e. does not detect the T-DNA insertion . As the female parent Ler-WT ,3 -34 mutant parent build 1200 per plant F2 population . Phenotypic analysis found that the groups 234 mutant , in line with the pattern of inheritance of single-gene recessive mutation . First, the BSA method for screening the average distribution in Arabidopsis 22 InDel on chromosome 5 markers , found only in the last on the fifth chromosome markers ciw10 appear polymorphism between two Bulked pool , that to illustrate ciw10 with the target gene linkage . Were developed in ciw10 downstream 17 InDel mark the filter F2 population consisting of 42 normal phenotype and 52 mutant phenotype , the target gene is located on between MRI1 and ciw10 . 18 InDel markers also developed using the same method , the group consisting of filter 182 mutant , and finally the target gene is positioned in the mark MUP24-1 and MAEl-3 87.2kb within shared to 57 InDel tag, wherein the newly developed 35 . In this study, the final position of the target gene , cloning and in-depth analysis has laid a good foundation .

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CLC: > Biological Sciences > Botany > Plant Cell Genetics
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