【摘要】：【目的】對一個水稻矮化劍葉卷曲突變體進(jìn)行鑒定與基因定位,為水稻等禾谷類作物劍葉形態(tài)發(fā)育及分子改良奠定基礎(chǔ)�！痉椒ā吭诙i型水稻恢復(fù)系縉恢10號的甲基磺酸乙酯(EMS)突變庫中篩選到一個隱性矮化劍葉卷曲突變體,命名為dcfl1(dwarf and curled flag leaf 1)。田間小區(qū)種植,全生育期內(nèi)觀察dcfl1和野生型的株型變化。苗期利用掃描電鏡觀察葉鞘內(nèi)表皮細(xì)胞大小;孕穗期和抽穗期利用石蠟切片觀察劍葉基部形態(tài);開花期測定劍葉、倒2葉和倒3葉的葉綠素含量;成熟期考查株高、有效穗數(shù)、穗實(shí)粒數(shù)、結(jié)實(shí)率和千粒重等主要農(nóng)藝性狀。配制西農(nóng)1A/dcfl1雜交組合,利用F1和F2群體進(jìn)行遺傳分析,并利用F2隱性群體進(jìn)行基因定位。【結(jié)果】生育期內(nèi),突變體dcfl1都表現(xiàn)出矮化性狀。dcfl1葉鞘內(nèi)表皮細(xì)胞長度明顯比野生型要短,達(dá)到了極顯著水平。與野生型相比,穗長、倒1節(jié)間和倒2節(jié)間均顯著變短,倒3節(jié)間和倒4節(jié)間無顯著變化。抽穗期dcfl1劍葉的葉片和葉鞘連接處硬化,劍葉基部展開受阻,半邊葉片向內(nèi)卷曲,劍葉上部和中部正常,其他葉片也正常。農(nóng)藝性狀調(diào)查發(fā)現(xiàn),dcfl1的有效穗數(shù)為14.24,極顯著高于野生型的11.62,穗粒數(shù)、實(shí)粒數(shù)、結(jié)實(shí)率和千粒重等則無顯著變化。此外,dcfl1的葉色略深,劍葉、倒2葉和倒3葉的葉綠素a含量均極顯著高于野生型,類胡蘿卜素含量也略有升高,但僅劍葉達(dá)到極顯著差異水平,葉綠素b的含量則無顯著變化。西農(nóng)1A/dcfl1的F1群體中,株高和劍葉表型與野生型一致。F2群體中分離出正常和突變兩種表型,突變表型與dcfl1類似,植株株高變矮,劍葉基部特異卷曲,說明矮化和劍葉基部特異卷曲是一對共分離性狀。且兩種表型分離比符合3㑳1,表明dcfl1突變型受1對隱性核基因控制。利用620株F2隱性單株,最終將DCFL1精細(xì)定位在第3染色體短臂In Del標(biāo)記Ind03-11和Ind03-6之間78 kb的物理范圍內(nèi),包含15個注釋基因,為DCFL1的克隆和水稻劍葉形態(tài)發(fā)育機(jī)理研究奠定了基礎(chǔ)。【結(jié)論】dcfl1是一個水稻矮化劍葉基部特異卷曲突變體,基因精細(xì)定位在第3染色體78 kb的物理范圍內(nèi)。
[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.
【作者單位】： 西南大學(xué)水稻研究所/轉(zhuǎn)基因植物與安全控制重慶市重點(diǎn)實(shí)驗(yàn)室;
【基金】：中央高校基本科研業(yè)務(wù)費(fèi)項(xiàng)目(XDJK2013A023) 重慶市研究生科研創(chuàng)新項(xiàng)目(CYS14043)
【分類號】：S511

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