云南地方稻waxy基因序列多样性分析

利用特异引物Wx-F/AG-2对来源于云南省16个州市64个县的252份地方稻种waxy基因中含有微卫星序列(CT)n和第一内含子的序列进行PCR扩增并测序。结果表明在268个碱基的序列内检测到4个变异位点:第一内含子上游56位处(CT)n存在(CT)10、(CT)11、(CT)12、(CT)14、(CT)16、(CT)17、(CT)18、(CT)19、(CT)20、(CT)21等10种变异;(CT)11、(CT)18、(CT)17、(CT)12、(CT)10等5种单倍型是云南优势类型,合计占供试材料的92.85%;籼稻以(CT)10、(CT)11、(CT)12为主,而粳稻以(CT)11、(CT)12、(CT)17和(CT)18为主;第一内含子+1位存在G/T变异,81.75%品种是G,T只出现在(CT)16、(CT)17、(CT)18和(CT)20的品种中,其频率在籼稻中为13.48%,粳稻20.86%,水稻16.17%,陆稻22.35%,粘稻10.47%,糯稻42.62%。+86-88位存在ATA/GTA/A--3种变异;+128位处存在(AATT)5和(AATT)6 2种变异。根据这4个变异位点,可将252个云南地方稻种归为16种单倍型,其中Wx4(32.54%)、Wx9(13.89%)、Wx12(12.7%)、Wx5(12.3%)、Wx1(8.33%)、Wx11(7.94%)是主要类型,合计87.7%,其他类型频率较低。籼/粳亚种、水/陆稻和粘/糯中存在单倍型种类和单倍型频率两方面的差异,籼稻/粳稻、水/陆稻和粘/糯稻各亚种或生态型均有独享的单倍型,共享单倍型频率也存在差异,表明亚种间或生态型间发生了一定的遗传分化。单倍型地理分布分析表明,临沧、普洱单倍型种类最丰富,以之为中心向外扩展,单倍型种类有减少的趋势,第一内含子+1位的T主要分布在临沧、普洱、西双版纳、德宏等南部地区。本研究揭示了云南地方稻种群体waxy基因的变异和分布特点。

The Diversity Analysis of the waxy gene of the Local Rice Variety from Yunnan

The waxy gene sequences including microsatellite sequence (CT)n and the part of first intron were amplificated and sequenced using the PCR pecific primers Wx-F / AG-2 for 252 rice landraces from 16 areas 64 counties of Yunnan. The results showed that Four variable sites were detected in the waxy gene sequences incuding 260 bases. The (CT)n variation duplicate number 10, 11, 12, 14, 16, 17, 18, 19, 20, 21 was tested at site 56 of the first intron upstream; The variation G / T was present in the first intron 1 site and T only was present in the cultivars with (CT)n length greater than 16 breeds; There were three variations include ATA / GTA /A at 86-88 site; There were two (AATT) n variations (5 and 6) duplicate number at 128 site. According to these four polymorphic sites, the 252 Yunnan rice landraces were classified into 16 genotypes, which Wx4 (32.54%), Wx9 (13.89%), Wx12 (12.7%), Wx5 (12.3%), Wx1 (8.33%), Wx11 (7.94%) were the main type, other types of frequency were lower. There were differentiation in genotype types and genotype frequencies between Indica / japonica subspecies, water / upland rice and sticky / glutinous rice. The indica / japonica, water / upland rice and no-glutinous / glutinous rice subspecies or ecotypes had exclusive genotype, shared genotype frequency also had differences. This indicated that genetic differentiation were occurred between subspecies or ecotypes. Geographical distribution of genotypes showed that Lincang, Puer genotype species were the most abundant, and expansion outward as it the center, genotype species were decreasing trend, the T at the first intron 1 site were mainly distributed in Lincang, Pu'er, Banna, Dehong and other southern regions. This study revealed the waxy gene the variability and distribution of Yunnan rice landraces groups.