水稻窄叶突变体Narrow leaf 11(nal11)的基因定位
Gene mapping for rice narrow leaf mutant Narrowleaf11(nal11)
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摘要: 目的定位和分析水稻窄叶突变体基因,为水稻叶片发育调控及株型育种提供参考依据.方法用甲基磺酸乙酯(EMS)诱导泸恢17,获得稳定的窄叶突变体(Narrow leaf 11,nal11),调查其与野生型泸恢17抽穗期功能叶的长和宽、分蘖数及成熟期株高.对nal11和绵恢727正反交获得的F2代进行遗传分析及基因定位.结果nal11抽穗期剑叶、倒2叶和倒3叶的宽度与野生型泸恢17存在显著(P<0.05,下同)或极显著(P<0.01,下同)差异,分别为野生型泸恢17的60.7%、57.9%和75.8%,但长度无显著差异(P>0.05);nal11株高为野生型泸恢17的90.3%,存在显著差异;nal11分蘖数极显著增加,为野生型的150.0%.nal11和绵恢727正反交后,F1代均表现正常叶宽,F2代叶宽发生性状分离,正常叶宽与窄叶植株数比例经χ2检验均符合3:1,表明nal11是受核单基因控制的隐性突变.利用SSR标记将nal11定位在水稻第4号染色体RM7290和RM16720标记之间约322 kb范围内,其与2个标记的遗传距离均为0.8 cM,覆盖了32.236 kb的物理区域,在定位区域内有5个注释基因,即Os04g26834、Os04g26850、Os04g26870、Os04g26880和Os04g26841,其序列与前人克隆的窄叶基因无重复.结论获得一个新的水稻窄叶突变体(nal11),其窄叶性状由1个隐性核基因控制,定位于水稻第4号染色体RM7290和RM16720标记之间,对功能叶、株高和分蘖数的表型有明显影响.Abstract: ObjectiveThe present study aimed to map and analyze narrow leaf mutant gene in order to provide the ba-sis for leaf developmental regulation and plant-type breeding of rice. MethodLuhui17 was induced by ethyl methyl sul-fonate(EMS) and then a stable narrow leaf mutant(Narrow leaf 11,nal11) was obtained. Length and width of functional leaves, tiller number and plant height at maturation stage of nal11 were analysed and compared with those of wild type Mi-anhui 727. Orthogonal and reciprocal crosses between nal11 and Mianhui 727 were carried out and F2 population was ob-tained, and genetic analyzis and gene mapping were conducted. ResultWidths of flag leaf and the 2nd and 3rd leaves from the top of nal11 at heading stage were significantly(P<0.05,the same below) or extremely significantly(P<0.01,the same below)different from those of wild type Mianhui 727, and were 60.7%, 57.9%, and 75.8% of the value for wild-type Mianhui 727 respectively. However, there was no difference in length of functional leaves between them(P>0.05). Plant height of nal11 was 90.3% of that of wild type Mianhui 727 and there was significant difference between them. Tiller num-ber of nal11 extremely significantly increased and was 150.0% of that of wild type Mianhui 727. In F1 generation of recip-rocal and orthogonal crosses between nal11 and Mianhui 727, the leaf width was normal in F1, but trait segregation occurred in leaf width of F2 populations. The ratio of rice plants with normal and narrow leaves was conformed as 3:1 by χ2 test. The results indicated that nal11 was recessive mutation controlled by nuclear gene. Using simple sequence repeat (SSR) marker, nal11 gene was located in a range of about 322 kb between RM7290 and RM16720 on chromosome 4 of rice. The genetic distances between nal11 and the two SSR markers were both 0.8 cM, covering 32.236 kb physical do-main. There were five annotation genes(Os04g26834,Os04g26850,Os04g26870,Os04g26880 and Os04g26841) in the region. The sequences were different from those of narrow leaf genes cloned by previous researchers. Conclusion A new narrow leaf mutant (nal11) is obtained. The narrow leaf trait is controled by one recessive nuclear gene. It is located between RM7290 and RM16720 on chromosome 4 of rice, and has obvious influences on functional leaf, plant height and tiller number.