DONG Bangning,LONG Wuhua,LIU Xi,et al.Phenotypic analysis and gene cloning of a dwarf mutant htr in rice[J].Journal of Nanjing Agricultural University,2017,40(4):573-583.[doi:10.7685/jnau.201612014]





Phenotypic analysis and gene cloning of a dwarf mutant htr in rice
董邦宁 龙武华 刘喜 刘世家 田云录 陈亮明 江玲 王益华 万建民
南京农业大学作物遗传与种质创新国家重点实验室, 江苏 南京 210095
DONG Bangning LONG Wuhua LIU Xi LIU Shijia TIAN Yunlu CHEN Liangming JIANG Ling WANG Yihua WAN Jianmin
State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China
rice(Oryza sativa L.)dwarfgene cloningtranscription factorAP2
[目的] 对水稻矮秆突变体htr进行表型分析与基因克隆,为水稻育种提供新的矮秆基因资源。[方法]对矮秆突变体htr茎秆进行细胞学观察,并将htr与‘N22’和‘9311’杂交构建F2群体,提取隐性极端个体定位基因HTR[结果]htr是一个稳定遗传的矮秆突变体,其株高、穗长、千粒质量、分蘖数等性状均降低,但其叶片变宽,叶色深绿,茎秆增粗。成熟期对htr倒二节间进行切片观察发现:相比‘9311’,htr在茎秆的纵轴方向细胞未能正常伸长,横向细胞数目增多,细胞变小。遗传分析表明突变体htr的矮秆性状受1对隐性单基因控制。利用F2群体中隐性极端个体将HTR定位在第5染色体长臂ttr-1和ttr-2两个分子标记之间,物理距离大约在144 kb范围内。测序结果发现htr中编码AP2转录因子(AP2-like ethylene-responsive transcription factor)基因发生点突变,导致保守氨基酸发生替换,亚细胞定位显示HTR定位于细胞核中。HTR呈现组成型表达,但以在茎秆中表达量最高。[结论]矮秆突变体htr突变性状受一对编码AP2转录因子的隐性单基因控制。HTR基因的克隆对培育厚壁粗秆抗倒伏水稻新品种提供了一定的理论参考。
[Objectives]The aim is to identify rice dwarf mutant htr,localize the target gene and provide new dwarf gene resources for rice breeding. [Methods]Sections of the stem were prepared for cytological observation. The dwarf mutant htr was crossed with ‘N22’ and ‘9311’ to construct F2 populations for genetic analysis and gene mapping,respectively. We selected the recessive extreme individuals to localize the gene HTR. [Results]htr was a stably inherited dwarf mutant. The plant height,panicle length,1 000-grain weight,and tiller number in mature plants all decreased compared with wild type,while the leaf width and culm diameter were increased and the leaf colour of htr was much darker green than wild type. By observing sections of the second internodes at the mature stage,we found in the longitudinal direction of the stem,the cells failed to elongate normally and became small,but the cell number increased. Genetic analysis showed that the dwarf trait of mutant htr was controlled by a pair of recessive gene. The HTR locus was mapped to a region between the markers ttr-1 and ttr-2 on chromosome 5,which was further narrowed down to a 144 kb region. Sequence analysis revealed that the coding region of the AP2-like ethylene-responsive transcription factor had a single nucleotide substitution,resulting in a change in the conserved amino acid sequence in the htr mutant. Subcellular localization assay showed that HTR was located in the nucleus,and the HTR was constitutively expressed,but it had the highest expression in the stem. [Conclusions]The phenotype of htr was controlled by a single recessive gene,which encodes the AP2-like ethylene-responsive transcription factor. The cloning of HTR gene could provide some theoretical reference for cultivating thick-walled and lodging resistant new rice varieties.


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更新日期/Last Update: 2017-07-07