中国水稻科学2017，Vol.31Issue(3)：247-256,10.DOI:10.16819/j.1001-7216.2017.6159
水稻无花粉型核雄性不育突变体whf41的鉴定与基因定位
Characterization and Gene Mapping of a No-pollen Genic Sterile Mutant whf41 in Rice
摘要
Abstract
[Objective]The anther cuticle and wax layer act as structural support and protective barriers for the development of pollen sac in rice. Phenotypic analysis and genetic mapping of the related genes can provide genetic resources and lay theoretical basis for further cloning and molecular mechanism analysis. [Method]We identified a no-pollen male sterile mutant whf41 from the mutant progeny of 60Co-γ-treated indica cultivar Zhonghui 8015. Anthers of the wild-type and whf41 mutant at different development stages were fixed for transverse section analysis and mature anthers of which were randomly selected for scanning electron microscopy analysis. BC1F1, F1 lines and BC1F2, F2 populations derived from whf41/ Zhonghui 8015 and whf41/ 02428 were used for genetic analysis and fine-mapping of the mutation site. [Result] Comparing with wild type,the whf41 mutant exhibited thin and transparent milky anther without mature pollen grains. Transverse section analysis showed that the whf41 mutant displayed aborted pollen grains,none detectable exine and swollen tapetal layer without expected PCD (programmed cell death), finally resulting in fragments of tapetal layer and pollen grains in the chamber. Scanning electron microscopy analysis further revealed the mutant had smoother anther wall, fewer lipids both on the inner and outer sides and fragments of degragated pollens. Genetic analysis revealed that whf41 was controlled by a single recessive genic gene, which was fine-mapped to a 45.6-kb region that harbored nine Open Reading Frames(ORFs) on the short arm of chromosome 3 between markers XD-5 and XD-11. Sequence analysis revealed that the whf41 mutant carried a nucleotide substitution and three nucleotide deletion, which led a substitution of one amino acid (D to M)and a deletion of one amino acid (V) in the fourth exon of one Cytochrome P450 family gene, LOC_Os03g0725, which is named as OsWHF41 and probably responsible for the no-pollen male sterility phenotype. Furthermore, Qpcr analysis showed that the expression level of CYP704B2 and seven anther lipid synthesis and transportation related regulators changed significantly in whf41. [Conclusion]Based on all these results, we deduced that the OsWHF41 gene was a new allelic to CYP704B2 gene and this further confirmed the important role of CYP704B2 in anther lipid synthesis and pollen exine formation .关键词
水稻/whf41/隐性核雄性不育/基因定位Key words
rice/whf41/recessive genic male sterility/gene mapping分类
生物科学引用本文复制引用
轩丹丹,程式华,孙廉平,张沛沛,张迎信,吴玮勋,杨正福,占小登,沈希宏,曹立勇..水稻无花粉型核雄性不育突变体whf41的鉴定与基因定位[J].中国水稻科学,2017,31(3):247-256,10.基金项目
国家公益性农业科技研究专项(20140302,20150308) （20140302,20150308）
国家转基因重大专项(2014ZX08001-002) （2014ZX08001-002）
中国农业科学院创新工程超级稻育种创新团队、水稻杂种优势研究机理研究创新团队资助项目(CAAS-ASTIP-2013-CNRRI). （CAAS-ASTIP-2013-CNRRI）
