Characterization of active miniature inverted-repeat transposable elements in the peanut genome |
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Authors: | Kenta Shirasawa Hideki Hirakawa Satoshi Tabata Makoto Hasegawa Hiroyuki Kiyoshima Sigeru Suzuki Sigemi Sasamoto Akiko Watanabe Tsunakazu Fujishiro Sachiko Isobe |
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Institution: | (1) Department of Plant Genome Research, Kazusa DNA Research Institute, 2-6-7 Kazusa-Kamatari, Kisarazu Chiba, 292-0818, Japan;(2) Peanut Plant Breeding Field, Chiba Prefectural Agriculture and Forestry Research Center, 199 He, Yachimata Chiba, 289-1113, Japan |
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Abstract: | Miniature inverted-repeat transposable elements (MITEs), some of which are known as active non-autonomous DNA transposons,
are found in the genomes of plants and animals. In peanut (Arachis hypogaea), AhMITE1 has been identified in a gene for fatty-acid desaturase, and possessed excision activity. However, the AhMITE1 distribution and frequency of excision have not been determined for the peanut genome. In order to characterize AhMITE1s, their genomic diversity and transposition ability was investigated. Southern blot analysis indicated high AhMITE1 copy number in the genomes of A. hypogaea, A. magna and A. monticola, but not in A. duranensis. A total of 504 AhMITE1s were identified from the MITE-enriched genomic libraries of A. hypogaea. The representative AhMITE1s exhibited a mean length of 205.5 bp and a GC content of 30.1%, with AT-rich, 9 bp target site duplications and 25 bp terminal
inverted repeats. PCR analyses were performed using primer pairs designed against both flanking sequences of each AhMITE1. These analyses detected polymorphisms at 169 out of 411 insertional loci in the four peanut lines. In subsequent analyses
of 60 gamma-irradiated mutant lines, four AhMITE1 excisions showed footprint mutations at the 109 loci tested. This study characterizes AhMITE1s in peanut and discusses their use as DNA markers and mutagens for the genetics, genomics and breeding of peanut and its
relatives. |
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