【摘要】：水稻(Oryza sativa L.)是世界上重要的糧食作物之一,世界上有一半的人口以稻米為食。干旱是影響產(chǎn)量的重要非生物脅迫之一。近些年,我國城鎮(zhèn)化進程的加快,更是加劇了我國水資源的短缺,因此,研究節(jié)水抗旱稻至關重要。根系是水稻重要的吸水器官,較深的根系分布層可以增加水稻的吸水量,提高水稻的避旱性。對控制深根比基因的精細定位,能夠幫助我們揭示抗旱機理,加快節(jié)水抗旱稻選育進程,同時對保障我國糧食安全也具有重要意義。本實驗室自2004年以來利用珍汕97B與IRAT109構建重組自交系(Recombinant Inbred Lines,RIL)群體,并利用213個SSR分子標記構建了覆蓋全基因組的圖譜。通過多年多點鑒定,共檢測到四個深根比主效QTL,分別位于第1、2、4、7號染色體。通過回交,結合目標QTL連鎖標記前景選擇和全基因組背景選擇,構建以珍汕97B為背景,含有目標QTL的4個近等基因系(Near Isogenic Line,NIL)。在此基礎上,本文針對位于第2號染色體上深根比主效QTL-q RDR-2進行了精細定位,結果如下:1.BC_4F_1前景與背景檢測:本實驗室于2015年春,完成含有目標QTL-q RDR-2的BC_4F_1構建,結合前景連鎖標記及背景標記基因型檢測,選擇目標區(qū)段連鎖標記基因型雙雜合,背景與珍汕97B盡量相似單株,在63個BC_4F_1中共篩選出32個單株。2.q RDR-2重組交換單株的篩選及重組交換位置的確定:將含有目的片段的近等基因系BC_4F_1發(fā)展為含有8000株的BC_4F_2群體,利用初定位區(qū)間兩側連鎖標記RM6與RM240進行重組交換單株的篩選,共篩選出289株重組交換單株。通過分析重組交換單株目標區(qū)段標記基因型,發(fā)現(xiàn)其中14個標記發(fā)生重組,包括22個重組交換單株。3.q RDR-2精細定位:本課題于2016年6月,將22個重組交換單株的自交后代種植于上海金山廊下基地,每個重組交換單株種植42個BC_4F_3單株。利用“籃子法”對每個單株進行表型鑒定,并結合基因型檢測,將QTL-q RDR-2定位于分子標記Q2-323與Q2-12之間,物理距離為570kb。4.第二次重組交換單株的篩選及重組交換位置的確定:為了進一步縮小區(qū)間距離,在第一次定位的基礎上,利用QTL區(qū)間兩側連鎖標記Q2-323與Q2-12,從4000個F_4單株中,篩選出106個重組交換單株。通過分析重組交換單株目標區(qū)段標記基因型,發(fā)現(xiàn)其中6個標記發(fā)生重組,共包括12個重組交換單株。5.候選基因預測:通過生物信息學分析,并結合本實驗室根尖RNA-seq數(shù)據(jù),在目標區(qū)段預7個候選基因:LOC_Os02g48360、LOC_Os02g48710、LOC_Os02g48770、LOC_Os02g49160、LOC_Os02g49440、LOC_Os02g49460、LOC_Os02g49720,其中LOC_OS02g49160為已知基因OsPIN1。
[Abstract]:Rice (Oryza sativa L.) It is one of the most important food crops in the world. Half of the world's population feeds on rice. Drought is one of the important abiotic stresses affecting yield. In recent years, the quickening process of urbanization in China has aggravated the shortage of water resources in China, so it is very important to study water-saving and drought-resistant rice. Root system is an important water absorption organ of rice. The deeper root distribution layer can increase the water absorption of rice and improve the drought avoidance of rice. The precise mapping of the genes controlling the deep root ratio can help us to reveal the mechanism of drought resistance, accelerate the breeding process of water-saving drought-resistant rice, and also play an important role in ensuring the food security in China. Since 2004, Zhenshan 97B and IRAT109 were used to construct recombinant inbred line (Recombinant Inbred Lines,RIL) population, and 213 SSR molecular markers were used to construct the genome map. After many years of identification, four deep root specific dominant QTL, were found to be located on chromosome 1, chromosome 2 and chromosome 7, respectively. By backcrossing, combining target QTL linkage marker foreground selection and whole genome background selection, four near-isogenic lines (Near Isogenic Line,NIL with Zhenshan 97B background and target QTL were constructed. On this basis, a detailed mapping of deep root specific dominant QTL-q RDR-2 on chromosome 2 was carried out. The results are as follows: 1.BC_4F_1 foreground and background test. The construction of BC_4F_1 containing target QTL-q RDR-2 was completed. Combined with foreground linkage marker and background marker genotype detection, the target segment linkage marker genotype double heterozygosity was selected, and the background was similar to that of Zhenshan 97B single plant. A total of 32 individual plants were screened out of 63 BC_4F_1. 2. Screening of Q RDR-2 recombinant exchange single plants and determination of the recombination exchange site: BC_4F_1, a near-isogenic line containing the target fragment, was developed into a BC_4F_1 containing 8000 strains. The BC_4F_2 community, Two sides linkage marker RM6 and RM240 were used to screen the recombinant exchange single plant, and a total of 289 recombinant exchange single plants were screened. Based on the analysis of target segment marker genotypes of recombinant exchange single plant, it was found that 14 of these markers were recombined, including 22 recombinant exchange single plants. 3. Q RDR-2 fine location: this study was conducted in June, 2016. The inbred progenies of 22 recombinant exchange plants were planted in Jinshan Corridor, Shanghai, and 42 BC_4F_3 plants were planted per recombination exchange single plant. The QTL-q RDR-2 was located between Q2-323 and Q2-12 with the physical distance of 570kb.4. the phenotype of each plant was identified by "basket method" and combined with genotype detection. Screening of the second recombination exchange single plant and determination of the recombination exchange location: in order to further reduce the interval distance, on the basis of the first location, the linkage markers Q2-323 and Q2-12 of the two sides of the QTL interval were used, and the sequence of Q2-323 and Q2-12 were obtained from 4 000 FSCL plants. 106 recombinant exchange single plants were screened. By analyzing the target segment marker genotypes of the recombinant exchange single plant, it was found that 6 of the markers were recombined, including 12 recombinant exchange single plants. Candidate gene prediction: through bioinformatics analysis, combining with RNA-seq data of root tip of our laboratory, 7 candidate genes: LOC_Os02g48360,LOC_Os02g48710,LOC_Os02g48770,LOC_Os02g49160,LOC_Os02g49440,LOC_Os02g49460,LOC_Os02g49720, in target region are predefined. Where LOC_OS02g49160 is a known gene OsPIN1.
【學位授予單位】：華中農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S511

【參考文獻】

相關期刊論文 前10條

1 蘇暉;仲軍;王云智;羊彬;趙田芬;;水稻根系研究的現(xiàn)狀及展望[J];上海農(nóng)業(yè)科技;2016年03期

2 楊潔;赫佳;王丹碧;施恩;楊文宇;耿其芳;王中生;;InDel標記的研究和應用進展[J];生物多樣性;2016年02期

3 來長凱;張文銀;賀奇;孫建昌;王興盛;殷延勃;;寧夏水稻抗旱性鑒定指標的篩選研究[J];種子;2015年08期

4 田又升;謝宗銘;吳向東;王志軍;葉春秀;張國麗;;水稻種質(zhì)資源萌發(fā)期抗旱性綜合鑒定[J];干旱地區(qū)農(nóng)業(yè)研究;2015年04期

5 李香玲;馮躍華;;水稻根系生長特性及其與地上部分關系的研究進展[J];中國農(nóng)學通報;2015年06期

6 鄭愛泉;;DNA分子標記在小麥遺傳育種中的應用[J];陜西農(nóng)業(yè)科學;2014年07期

7 敬禮恒;劉利成;梅坤;陳光輝;;水稻抗旱性能鑒定方法及評價指標研究進展[J];中國農(nóng)學通報;2013年12期

8 邢延豪;周延清;楚素霞;郭靜佩;周鵬;范念斯;;CAPS標記技術及其應用進展[J];江蘇農(nóng)業(yè)科學;2011年05期

9 張文英;智慧;柳斌輝;李明哲;李偉;謝俊雪;宋慶榮;刁現(xiàn)民;;干旱脅迫對谷子孕穗期光合特性的影響[J];河北農(nóng)業(yè)科學;2011年06期

10 徐群;袁筱萍;余漢勇;王一平;湯圣祥;魏興華;;水稻苗期抗旱性的QTL分析[J];中國水稻科學;2010年05期

相關博士學位論文 前1條

1 李一博;水稻粒形、粒重基因GS5及堊白基因Chalk5的克隆與功能研究[D];華中農(nóng)業(yè)大學;2011年

相關碩士學位論文 前10條

1 宋從志;水稻第7號染色體深根比主效QTL的精細定位[D];華中農(nóng)業(yè)大學;2016年

2 姜雪;水稻苗期耐旱性基因位點的發(fā)掘[D];華中農(nóng)業(yè)大學;2015年

3 王嬌;水稻抗旱性相關性狀的QTL定位分析[D];寧夏大學;2015年

4 盛浩聞;水稻湘資3150抗稻瘟病基因Pi48的精細定位與克隆及近等基因系構建[D];湖南農(nóng)業(yè)大學;2014年

5 張卓一;水稻第4染色體抗旱性相關基因的定位與挖掘[D];華中農(nóng)業(yè)大學;2014年

6 王培;水稻深根比遺傳分析[D];上海海洋大學;2014年

7 關成冉;利用近等基因系精細定位一個水稻抽穗期QTL-qHd3-1[D];揚州大學;2011年

8 孔會利;水稻糙米蛋白質(zhì)含量主效QTL的精細定位以及近等基因系的構建[D];華中農(nóng)業(yè)大學;2010年

9 李艷;水稻品種苗期抗旱性及鑒定指標篩選的研究[D];四川農(nóng)業(yè)大學;2006年

10 張玉屏;水稻旱作條件下根系生長特性的研究[D];安徽農(nóng)業(yè)大學;2001年

，

本文編號：2328749