水稻矮化剑叶卷曲突变体dcfl1的鉴定与基因精细定位

发布时间：2018-07-27 09:16

【摘要】：【目的】对一个水稻矮化剑叶卷曲突变体进行鉴定与基因定位,为水稻等禾谷类作物剑叶形态发育及分子改良奠定基础。【方法】在籼型水稻恢复系缙恢10号的甲基磺酸乙酯(EMS)突变库中筛选到一个隐性矮化剑叶卷曲突变体,命名为dcfl1(dwarf and curled flag leaf 1)。田间小区种植,全生育期内观察dcfl1和野生型的株型变化。苗期利用扫描电镜观察叶鞘内表皮细胞大小;孕穗期和抽穗期利用石蜡切片观察剑叶基部形态;开花期测定剑叶、倒2叶和倒3叶的叶绿素含量;成熟期考查株高、有效穗数、穗实粒数、结实率和千粒重等主要农艺性状。配制西农1A/dcfl1杂交组合,利用F1和F2群体进行遗传分析,并利用F2隐性群体进行基因定位。【结果】生育期内,突变体dcfl1都表现出矮化性状。dcfl1叶鞘内表皮细胞长度明显比野生型要短,达到了极显著水平。与野生型相比,穗长、倒1节间和倒2节间均显著变短,倒3节间和倒4节间无显著变化。抽穗期dcfl1剑叶的叶片和叶鞘连接处硬化,剑叶基部展开受阻,半边叶片向内卷曲,剑叶上部和中部正常,其他叶片也正常。农艺性状调查发现,dcfl1的有效穗数为14.24,极显著高于野生型的11.62,穗粒数、实粒数、结实率和千粒重等则无显著变化。此外,dcfl1的叶色略深,剑叶、倒2叶和倒3叶的叶绿素a含量均极显著高于野生型,类胡萝卜素含量也略有升高,但仅剑叶达到极显著差异水平,叶绿素b的含量则无显著变化。西农1A/dcfl1的F1群体中,株高和剑叶表型与野生型一致。F2群体中分离出正常和突变两种表型,突变表型与dcfl1类似,植株株高变矮,剑叶基部特异卷曲,说明矮化和剑叶基部特异卷曲是一对共分离性状。且两种表型分离比符合3㑳1,表明dcfl1突变型受1对隐性核基因控制。利用620株F2隐性单株,最终将DCFL1精细定位在第3染色体短臂In Del标记Ind03-11和Ind03-6之间78 kb的物理范围内,包含15个注释基因,为DCFL1的克隆和水稻剑叶形态发育机理研究奠定了基础。【结论】dcfl1是一个水稻矮化剑叶基部特异卷曲突变体,基因精细定位在第3染色体78 kb的物理范围内。
[Abstract]:[objective] to identify and localize a dwarf flag leaf curl mutant in rice. [methods] A recessive dwarf flag leaf curl mutants were screened from the (EMS) mutant library of Jinhui 10, an indica rice restorer line. Named dcfl1 (dwarf and curled flag leaf 1). The plant types of dcfl1 and wild type were observed during the whole growth period. At seedling stage, the size of epidermal cells in leaf sheath was observed by scanning electron microscope, the basal morphology of flag leaf was observed by paraffin section at booting stage and heading stage, the chlorophyll content of flag leaf, second leaf and third leaf were measured at flowering stage, and the plant height and effective panicle number were examined at maturity stage. The main agronomic characters were the number of grains per ear, seed setting rate and 1000 grain weight. Xinong 1A/dcfl1 hybrids were prepared, genetic analysis was carried out by F1 and F2 populations, and gene mapping was carried out by using F2 recessive populations. [results] during the growth period, The length of epidermal cells in leaf sheath of mutant dcfl1 was significantly shorter than that of wild type, and reached a very significant level. Compared with the wild type, the panicle length, the top 1 Internode and the inverted 2 Internode were significantly shorter, while the inverted 3 Internode and the inverted 4 Internode had no significant change. At heading stage, the leaves and sheaths of dcfl1 hardened at the junction, the base of the blade was blocked, half of the leaf curled inward, the upper and middle of the flag leaf were normal, and the other leaves were normal. It was found that the number of effective panicles was 14.24, which was significantly higher than that of wild type (11.62). There was no significant change in the number of grains per panicle, the number of solid grains, seed setting rate and 1000-grain weight. In addition, the leaf color of Dcfl1 was slightly deeper, the content of chlorophyll a in the flag leaf, the second leaf and the third leaf were significantly higher than those in the wild type, and the carotenoid content was also slightly increased, but the content of chlorophyll b in the flag leaf reached a very significant difference level, but there was no significant change in the content of chlorophyll b. In F1 population of Xinong 1A/dcfl1, normal and mutant phenotypes were isolated from F1 population of Xinong 1A/dcfl1. The phenotypes of plant height and flag leaf were the same as wild type. F2 population. The mutant phenotype was similar to that of dcfl1, the plant height became shorter, and the base of flag leaf was curly. The results showed that dwarfing and special curl of the base of flag leaf were a pair of co-segregation characters. The two phenotypic segregation ratios were in accordance with 3 ~ 1, indicating that the dcfl1 mutation was controlled by a pair of recessive nuclear genes. Using 620F2 recessive single plant, the DCFL1 was located in 78 kb between Ind03-11 and Ind03-6 tagged with in Del on chromosome 3, containing 15 annotated genes. [conclusion] dcfl1 is a special curly mutant at the base of dwarf flag leaf of rice, and the gene is located within the physical range of 78kb on chromosome 3.
【作者单位】：西南大学水稻研究所/转基因植物与安全控制重庆市重点实验室;
【基金】：中央高校基本科研业务费项目(XDJK2013A023) 重庆市研究生科研创新项目(CYS14043)
【分类号】：S511

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