水稻长护颖基因Oslsl的遗传分析和序列鉴定

Genetic analysis and sequence identification of long sterile lemmaOslsl)gene in rice

  • 摘要: 【目的】对水稻花器官突变材料进行基因定位和克隆,为深入研究水稻颖花发育的分子机理提供新材料。【方法】分别于孕穗期和成熟期观察具有长护颖表型的籼稻品种沙优选2号(lsl098)和正常表型品系9311的花器官和籽粒形态特征。将lsl098与9311杂交构建F2群体,对长护颖基因OslslLong sterile lemma)进行遗传分析、基因定位和测序分析。【结果】形态观察发现,lsl098的长护颖表型在幼穗分化第5期起可明显观察到,成熟期的长护颖形态近似外稃,呈“V”形开口,不影响结实率和发芽率。遗传分析结果显示,正、反交分别获得的155和208个F2单株中长护颖与正常表型植株数目符合1∶3分离比(x21∶3<x20.05,1=3.84),表明该表型受1对隐性基因控制。通过构建长护颖表型池和正常表型池,采用集团分离分析法确定与护颖表型共分离的分子标记,进一步筛选重组单株,并结合重组单株表型,将Oslsl定位在7号染色体短臂分子标记7M063和G01031之间,2个标记对应日本晴参考基因组物理距离为838.8 kb,定位区段包含已克隆的长护颖基因G1LOC_Os07g04670)。测序分析发现,与日本晴和9311的基因组序列相比,Oslsl序列包含了3个SNP突变,2 bp插入和145 bp缺失。【结论】 lsl098携带的长护颖基因Oslsl是已克隆G1基因的新等位基因。

     

    Abstract: 【Objective】Mapping and cloning of rice flower organ genes could provide novel materials for advanced molecular mechanism research of rice spikelet development.【Method】 The flower organs and grain morphological characters of indica rice variety Shayouxuan 2(lsl098)with long sterile lemma phenotype and line 9311 with normal phenotype were observed at the booting and maturity stages. Rice 9311 was crossed with lsl098 to develop F2 population which was applied to genetic analysis,gene mapping and sequencing analysis of long sterile lemma(Oslslgene.【Result】 Morphological observation srevealed that the long sterile lemma phenotype of lsl098 could be observed at the fifth stage of young panicle differentiation. At maturity,the long sterile lemma was similar to the lemma. It showed a"V"form which did not affect seed setting rate and germination rate. Genetic analysis showed that, the numbers of long sterile lemma plants obtained by forward and reverse crossing were 155 and 208 respectively,and they conformed to a separation of 1:3(x21:3 < x2 0.05, 1=3.84) comapred with normal phenotypic plants in F2 populations,indicating that the phenotype was controlled by one pair of recessive genes. Two DNA bulks containing long sterile lemma plants and normal plants respectively were developed which were applied to identify co-segregated molecular markers associated with long sterile lemma phenotypes using a bulked segregant analysis method. Furthermore, the genotype and phenotype of recombinant plants were screened. As a result,Oslsl was located between markers 7M063 and G01031 on the short arm of chromosome 7. The physical distance of two markers was 838.8 kb according to the genome of Nipponbare. One cloned long sterile lemma gene G1 (LOC_Os07g04670)was harbored in the target region. Genomic sequencing analysis showed that lsl098 included 3 SNP mutations,2 bp insertion and 145 bp deletion comparing with that of 9311 or Nipponbare.【Conclusion】Gene Oslsl derived from lsl098 is a new allele of G1 gene which has been cloned al ready.

     

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