本文關(guān)鍵詞： 水稻 稻瘟病抗性 Pi9基因 分子標(biāo)記輔助選擇育種 抗病純系　出處：《湖南農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：水稻是世界上重要的糧食作物,稻瘟病是水稻最主要的真菌性病害之一,流行年份可使水稻減產(chǎn)10%～30%,是水稻高產(chǎn)穩(wěn)產(chǎn)的主要限制因素。實(shí)踐證明,培育和合理利用抗病品種是控制此病害最經(jīng)濟(jì)有效和對環(huán)境安全的手段,因此需要培育抗譜廣、抗性穩(wěn)定并且持久的抗稻瘟病水稻品種。本研究根據(jù)廣譜持久抗瘟基因Pi9的內(nèi)含子序列設(shè)計(jì)特異功能標(biāo)記Ins2-1,通過分子標(biāo)記輔助選擇和連續(xù)回交育種,定向改良21份水稻受體親本的稻瘟病抗性。主要研究結(jié)果如下：1.通過對對實(shí)驗(yàn)室保存的10個(gè)稻瘟菌菌株進(jìn)行培養(yǎng)、繼代及純化過程了解了在番茄-燕麥培養(yǎng)機(jī)上稻瘟菌菌株的大致生長周期與產(chǎn)孢情況。其中：有3個(gè)小種保存完好,7個(gè)小種已經(jīng)被污染；純化后獲得了對應(yīng)的7個(gè)無污染的小種；對這10個(gè)小種進(jìn)行誘導(dǎo)產(chǎn)孢,觀察產(chǎn)孢情況發(fā)現(xiàn),兩個(gè)小種產(chǎn)孢能力較強(qiáng),5個(gè)小種的產(chǎn)孢能力一般,3個(gè)小種不產(chǎn)孢；觀察各小種菌絲顏色發(fā)現(xiàn),KOH的菌絲為黑色,195-2-2、X2007A-7菌絲為白色,其余7個(gè)菌株菌絲均為深灰黑色；通過對這10個(gè)稻瘟菌菌株的培養(yǎng),可以得知各稻瘟菌菌株在番茄-燕麥培養(yǎng)基上的生長周期約為20 d。2.利用25份稻瘟菌菌株對感病對照材料C039、21份水稻受體材料及抗稻瘟病基因Pi9供體75-1-127進(jìn)行室內(nèi)接種表型鑒定,利用田間病圃發(fā)病檢驗(yàn)所有供試水稻材料的稻瘟病抗性,結(jié)果表明供試受體材料抗譜不廣,抗性不強(qiáng),需要進(jìn)行抗性改良；另外,抗譜表明這些親本中可能含有抗菌譜與Pi9互補(bǔ)的新稻瘟病抗性基因,通過回交改良這些受體親本的稻瘟病抗性,或許可以得到抗譜比Pi9供體親本75-1-127更廣,同時(shí)抗性也更強(qiáng)的新品種。3.分子標(biāo)記Ins2-1在Pi9基因供體親本75-1-127及21份受體親本材料之間存在穩(wěn)定且明顯的多態(tài)性,多態(tài)率為100%。在大圍山天然病圃對所有供試材料進(jìn)行誘導(dǎo)發(fā)病后,利用分子標(biāo)記Ins2-1對豐源B×75-1-127與E32×75-1-127 BC6F1群體中的隨機(jī)取樣單株進(jìn)行基因型鑒定,以檢測分子標(biāo)記的輔助選擇效率,結(jié)果表明Ins2-1標(biāo)記基因型的稻瘟病抗性表型選擇效率為100%,證明此標(biāo)記能夠用于MAS育種實(shí)踐中作為選擇的依據(jù)。4.連續(xù)數(shù)年利用分子標(biāo)記Ins2-1進(jìn)行MAS育種,綜合田間農(nóng)藝性狀考察,目前已經(jīng)獲得5個(gè)組合的數(shù)個(gè)BC6F3株系,5個(gè)組合的BC6F2群體,3個(gè)組合的BC6F1群體,另外還有7個(gè)回交代數(shù)較低的組合,為進(jìn)一步定向改良這20份受體親本的稻瘟病抗性、培育抗病純系打下了良好基礎(chǔ)。5.利用特異顯性分子標(biāo)記Ins2-1輔助選擇將Pi9基因轉(zhuǎn)育到受體親本1701中,并通過連續(xù)回交和自交,獲得了含Pi9基因且遺傳背景接近受體親本的BC6F2群體；通過對BC6F3株系標(biāo)記基因型和室內(nèi)接種表型鑒定,篩選出一個(gè)抗病純系“抗瘟1701”,為培育抗稻瘟病水稻新品種(組合)奠定了基礎(chǔ)。
[Abstract]:Rice is one of the most important food crops in the world. Rice blast is one of the most important fungal diseases in rice. Breeding and rational use of disease-resistant varieties are the most economical, effective and environmentally safe means to control the disease, so it is necessary to develop a broad spectrum of resistance. In this study, we designed a special marker Ins2-1 based on the intron sequence of the broad-spectrum persistent blast resistance gene Pi9, and used molecular marker assisted selection and continuous backcross breeding. The rice blast resistance of 21 rice receptor parents was improved. The main results were as follows: 1. 10 strains of rice blast were cultured in the laboratory. In the course of subculture and purification, the growth cycle and sporulation of rice blast strain on Tomato oat culture machine were studied. Among them, 3 species were well preserved and 7 species had been contaminated. After purification, 7 species without pollution were obtained, and the spore production of the 10 species was induced. The results showed that the sporulation ability of two species was strong, the sporulation ability of 5 species was general, and 3 species were not. The mycelium of Koh was black 195-2-2X 2007A-7, the other 7 strains were dark gray black, and the 10 strains of rice blast were cultured, the results showed that the hypha of Koh was black and the other 7 strains were dark gray black, and the results showed that the mycelium of Koh was white and the other 7 strains were dark gray black. It can be known that the growth cycle of each rice blast strain on Tomato oat medium is about 20 d.2.21 rice receptor materials and Pi9 donor of blast resistance gene 75-1-127 were identified by 25 strains of rice blast strain. The resistance to blast of all rice tested in the field disease nursery was tested. The results showed that the resistance spectrum was not wide and the resistance was not strong, so resistance improvement was needed. The resistance spectrum indicated that these parents may contain new blast resistance genes that complement Pi9 with antibacterial spectrum. By backcrossing, the resistance to blast of these recipient parents may be wider than that of Pi9 donor parent 75-1-127, and the resistance spectrum of these parents may be wider than that of Pi9 donor parent 75-1-127, and the resistance spectrum of these parents may be wider than that of Pi9 donor parent 75-1-127. At the same time, there was a stable and obvious polymorphism of molecular marker Ins2-1 between 75-1-127 and 21 recipient parents of Pi9 gene donor parents, and the polymorphic rate was 1000.After Daweshan natural disease nursery, all the tested materials were induced to develop disease. Genotypes of randomly sampled individual plants in Fengyuan B 脳 75-1-127 and E32 脳 75-1-127 BC6F1 populations were identified by molecular marker Ins2-1. The results showed that the efficiency of phenotypic selection of rice blast resistance with Ins2-1 marker genotype was 100, which proved that the marker could be used as the basis for selection in MAS breeding practice. The molecular marker Ins2-1 was used for MAS breeding for several years, and the field agronomic characters were investigated. At present, five combinations of several BC6F3 strains, five combinations of BC6F2 populations, three combinations of BC6F1 populations, and seven combinations with lower back metasomatism have been obtained to further improve the blast resistance of the 20 recipient parents. The breeding of disease-resistant pure lines laid a good foundation .5.Using specific dominant molecular marker Ins2-1 assisted selection to transfer Pi9 gene into recipient parent 1701, and through continuous backcrossing and self-crossing, the BC6F2 population containing Pi9 gene and genetic background close to receptor parent was obtained. Based on the identification of marker genotypes and indoor inoculation phenotypes of BC6F3 strains, a resistant pure line "resistance to blast 1701" was selected, which laid a foundation for the breeding of new rice varieties (combinations) resistant to rice blast.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S435.111.41

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