Mutation loci and intragenic selection marker of the granule-bound starch synthase gene in waxy maize |
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Authors: | Jian Liu Tingzhao Rong Wanchen Li |
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Institution: | (1) Maize Research Institute, Sichuan Agricultural University, Ya’an, Sichuan, 625014, China;(2) Key Laboratory of Crop Genetic Resources and Improvement, (Sichuan Agricultural University), Ministry of Education, Ya’an, Sichuan, 625014, China |
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Abstract: | Four pairs of specific PCR primers have been designed on the basis of the sequence of the granule-bound starch synthase gene
(GBSS; dominant non-waxy gene Wx) and used to amplify its homologous sequence from thirteen waxy and two non-waxy inbred lines. Results from electrophoresis
indicated that the recessive waxy gene was wx, derived from the dominant non-waxy gene Wx by mutation at its 3′ end. The sequence of the mutated 3′ end was amplified by the TAIL-PCR technique. Sequence alignment
showed that the mutation of the wx gene was caused by transposition of the aldehyde dehydrogenase gene rf2. Two pairs of specific primers were designed on the basis of the sequence difference between the dominant gene Wx and its mutated recessive allele wx and used as intragenic selection markers to identify individual plants of genotypes WxWx, Wxwx, and wxwx by PCR amplification from the segregating population of the F2 generation crossed between waxy and non-waxy inbred lines. Iodine solution staining and starch component assay showed that
all the 35 F2 plants identified as genotype WxWx produced non-waxy kernels of the F3 generation and that all 33 F2 plants identified as genotype wxwx produced waxy kernels of the F3 generation. This result can be used to improve the selection efficiency of waxy maize breeding and for selection of other
single genes and major polygenes. |
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Keywords: | Granule-bound starch synthase gene Intragenic selection marker Mutation locus TAIL-PCR Waxy maize |
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