An efficient reverse genetics platform in the model legume Medicago truncatula

New Phytologist - Tập 201 Số 3 - Trang 1065-1076 - 2014
Xiaofei Cheng1, Mingyi Wang1, Hee‐Kyung Lee1, Million Tadege1, Pascal Ratet2, Michael K. Udvardi1, Kirankumar S. Mysore1, Jiangqi Wen1
1Division of Plant Biology The Samuel Roberts Noble Foundation 2510 Sam Noble Parkway Ardmore OK 73401 USA
2Institut des Sciences du Végétal CNRS Avenue de la Terrasse 91198 Gif sur Yvette Cedex France

Tóm tắt

Summary Medicago truncatula is one of the model species for legume studies. In an effort to develop legume genetics resources, > 21 700 Tnt1 retrotransposon insertion lines have been generated. To facilitate fast‐growing needs in functional genomics, two reverse genetics approaches have been established: web‐based database searching and PCR‐based reverse screening. More than 840 genes have been reverse screened using the PCR‐based approach over the past 6 yr to identify mutants in these genes. Overall, c. 84% (705 genes) success rate was achieved in identifying mutants with at least one Tnt1 insertion, of which c. 50% (358 genes) had three or more alleles. To demonstrate the utility of the two reverse genetics platforms, two mutant alleles were isolated for each of the two floral homeotic MADS‐box genes, MtPISTILATA and MtAGAMOUS. Molecular and genetic analyses indicate that Tnt1 insertions in exons of both genes are responsible for the defects in floral organ development. In summary, we have developed two efficient reverse genetics platforms to facilitate functional characterization of M. truncatula genes.

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Thông tin
Thông tin xuất bản

New Phytologist

Tập 201 Số 3

1065-1076

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