拟南芥雄性不育突变体ms1424的鉴定与基因定位

The identification and gene mapping of a male sterile mutant ms1424 in Arabidopsis thaliana

本研究通过筛选拟南芥Ac-Ds突变体库,获得一种雄性不育突变体male sterile 1424(ms1424)。与野生型相比,ms1424的角果短小、种子数少、茎秆明亮。ms1424花粉的亚历山大红染色正常,而花粉体内、体外萌发结果暗示其花粉与柱头的互作有异常。遗传分析显示ms1424的表型由隐性单基因控制。高通量测序和图位克隆将突变基因定位到Chr1:387247~463498,位于该区间的CER1的突变表型与ms1424相似,而遗传等位杂交实验表明ms1424是cer1的等位突变。进一步的分子实验揭示ms1424缺失了一个6216 bp的DNA片段(包括CER1的3084bp)。与多数cer1突变体的特性相同,ms1424在相对湿度为50%~70%和大于90%环境下分别表现为严重不育和正常可育,这说明ms1424是湿度条件型雄性不育。综上,本研究报道了一个新的拟南芥湿度条件型雄性不育材料。

By screening an Ac-Ds mutant stock of Arabidopsis,we obtained a male sterile mutant named male sterile 1424(ms1424).Compared with wild type,ms1424 had shorter siliques,fewer seeds but brighter stem.Alexander staining of ms1424 pollen was normal,yet results of pollen grain germination in vitro and in vivo tests suggested the pollen-stigma interaction in ms1424 was unusual.The genetic analysis suggested that phenotypes of ms1424 were controlled by a single recessive gene.High-throughput sequencing and map-based cloning mapped the mutated gene into the Chr1:387247–463498 region,and unexpectedly phenotypes of mutated CER1 which is located in this area were similar to that of ms1424.Allelic hybridization test proved that ms1424 was an allele mutation of cer1,and subsequent molecular experiments revealed that ms1424 lost a 6216 bp DNA fragment containing a 3084 bp fragment of CER1.Also,ms1424 had the same properties as most cer1:living in the environment with relative humidity of 50%–70%and over 90%,respectively,ms1424 showed low fertility and normal fertility,suggesting that ms1424 was humidity conditioned male sterile.In a word,this work provides a new humidity-conditional male sterile material of Arabidopsis.