本文關(guān)鍵詞：水稻類病斑突變體spl34的遺傳分析與精細(xì)定位　出處：《西南大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

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【摘要】：類病斑突變體(Disease lesion mimic mutants)是指植物在未受逆境脅迫或病原菌侵害的情況下,在其葉片或葉鞘等部位自發(fā)產(chǎn)生類似于過敏反應(yīng)時(shí)所形成的壞死病斑的一類突變體。在類病斑的形成過程中通常會(huì)伴隨著細(xì)胞程序性死亡,活性氧的積累,防衛(wèi)相關(guān)基因的表達(dá)量上調(diào)以及水楊酸、茉莉酸和乙烯等信號分子的合成量增加。在大量已報(bào)道的類病斑突變體中,很多還表現(xiàn)出抗性增強(qiáng)的特性,因此類病斑突變體可作為人類研究植物抗病機(jī)理和細(xì)胞程序性死亡的良好材料。在水稻中,由于很多類病斑突變體對稻瘟病和白葉枯病表現(xiàn)出抗性增強(qiáng)的特性,因此,類病斑突變體能夠?yàn)槿祟惻嘤剐詢?yōu)良的品種提供豐富的種質(zhì)資源。本研究利用化學(xué)誘變劑EMS(ethyl methane sulfonate)處理水稻秈型恢復(fù)系“縉恢10號”從其后代中篩選到一個(gè)遺傳穩(wěn)定的類病斑突變體spl34。以野生型“縉恢10號”和突變體spl34為材料,進(jìn)行了形態(tài)學(xué)鑒定、農(nóng)藝性狀考察、光合色素含量測定、葉綠體的熒光顯微鏡觀察、組織化學(xué)分析、生理生化指標(biāo)測定等方面的研究,并以表型正常的西農(nóng)1A與突變體spl34雜交構(gòu)建群體,F1和F2代用于遺傳分析與基因定位,主要研究結(jié)果如下。1.形態(tài)學(xué)鑒定突變體spl34從分蘗后期開始在下部葉片的葉鞘上出現(xiàn)褐色的類病斑,隨后褐斑沿著中脈擴(kuò)散然后分布于整個(gè)葉片。至成熟期,褐色類病斑擴(kuò)散至整個(gè)植株。遮光實(shí)驗(yàn)表明突變體類病斑的形成受光誘導(dǎo)。2.農(nóng)藝性狀考察成熟期分別對野生型和突變體的株高、穗長、穗粒數(shù)、結(jié)實(shí)率和千粒重等農(nóng)藝性狀進(jìn)行考察。結(jié)果顯示,突變體的株高和穗長均較野生型降低,達(dá)到了顯著差異水平,而千粒重、穗粒數(shù)和結(jié)實(shí)率的降低程度則達(dá)到了極顯著差異水平。3.光合色素含量測定抽穗期分別對野生型和突變體spl34的光合色素含量進(jìn)行測定。結(jié)果顯示,突變體spl34的倒一葉光合色素含量均略高于野生型但未達(dá)到顯著差異水平,而倒二葉和倒三葉光合色素含量則均較野生型低且分別達(dá)到了顯著差異水平和極顯著差異水平。4.葉綠體的熒光顯微鏡觀察抽穗期取野生型和突變體spl34的葉片用于熒光顯微鏡觀察發(fā)現(xiàn),突變體spl34的葉綠體產(chǎn)生的紅色熒光現(xiàn)象較野生型的弱,且突變體中出現(xiàn)類病斑的部位葉綠體產(chǎn)生的紅色熒光也較未出現(xiàn)類病斑的部位弱。5.組織化學(xué)分析臺(tái)盼藍(lán)染色和DAB染色結(jié)果顯示,突變體在類病斑形成部位發(fā)生了細(xì)胞程序性死亡和過氧化氫沉積。6.生理生化指標(biāo)測定抽穗期對野生型和突變體spl34的各項(xiàng)生理指標(biāo)進(jìn)行檢測。結(jié)果顯示,突變體中CAT、POD、SOD等抗氧化酶活性均較野生型的低,而且突變體中產(chǎn)生類病斑的部位H_2O_2和O_2~-的含量均比野生型高,·OH的含量則與野生型無明顯差異。7.稻瘟病抗譜測定與抗性鑒定稻瘟病抗譜測定結(jié)果顯示,突變體spl34對ZA生理群的抗病頻率比野生型高,而對ZB生理群的抗病頻率比野生型低,對ZC、ZD、ZE和ZG生理群的抗病頻率則與野生型相同�？剐澡b定結(jié)果顯示,突變體苗瘟、葉瘟病情指數(shù)和穗頸瘟的病穗率均比野生型略高,但差異并不顯著。8.遺傳分析和基因定位遺傳分析結(jié)果表明,突變體spl34的表型受一對隱性核基因控制。該基因定位于第4染色體的LR49和LR52兩個(gè)分子標(biāo)記之間,物理距離為200Kb。測序分析發(fā)現(xiàn)該區(qū)間內(nèi)的候選基因:LOC_Os04g56480的第3449位堿基發(fā)生突變(G3449T),導(dǎo)致氨基酸替換由色氨酸突變?yōu)榘腚装彼�。該基因與已報(bào)道的HM47為等位突變基因。9.目的基因與防衛(wèi)相關(guān)基因的表達(dá)q RT-PCR結(jié)果表明,與野生型相比,目的基因在突變體中的表達(dá)量下降;而用于進(jìn)行表達(dá)分析的病程相關(guān)基因和稻瘟病抗性基因中,除了PR1a、PR1b和NPR1三個(gè)病程相關(guān)基因在突變體中的相對表達(dá)量顯著升高外,其余的病程相關(guān)基因和稻瘟病抗性基因的相對表達(dá)量均未出現(xiàn)上升。
[Abstract]:A lesion mimic mutant (Disease lesion mimic mutants) is a plant without stress or pathogen attack under the condition of spontaneous generation of a class of mutants with similar necrotic lesion formed in the allergic reaction when the leaf sheath or other parts. In the forming process of the lesions usually accompanied by programmed cell death and the accumulation of active oxygen and defense related gene expression and synthesis of salicylic acid, jasmonic acid and ethylene signaling molecules increased. The lesion has been reported in a large number of mutants, many also exhibit characteristics of enhanced resistance, so the lesion mimic mutants can be used as good materials of human research on the mechanism of plant resistance and cell program death. In rice, because a lot of lesion mimic mutants of rice blast and bacterial blight showed enhanced resistance characteristics, therefore, to cultivate a lesion mimic mutant for man The excellent resistant cultivars provide rich germplasm resources. This study uses chemical mutagen EMS (ethyl methane sulfonate) processing of Indica Rice Restorer Line Jinhui 10 "from the progeny of screening to a stable genetic lesion mimic mutant spl34. with wild type" Jinhui 10 "and mutant spl34 as material for the morphological, agronomic traits, photosynthetic pigment content, chloroplast fluorescence microscopy, histochemical analysis of physiological and biochemical indexes and other aspects, and with normal phenotype of Xinong 1A and mutant spl34 hybrid construction group, F1 and F2 for genetic analysis and gene mapping. The main results are as follows.1. morphological identification of mutant spl34 from late tillering began Brown lesion in leaf sheath of lower leaves, then brown midrib and then spread throughout the diffusion along the blade. To maturity Brown, the lesions spread to the whole plant. Experiments show that the formation of mutant shading lesion induced by light.2. agronomic traits in mature stage of wild type and mutant plant height, panicle length, grain number, seed setting rate and grain weight was investigated and other agronomic traits. The results showed that the mutant plant height and panicle length were compared to the wild type decreased, reached the level of significant difference, and 1000 grain weight, grain number and seed setting rate decreased reached respectively for the wild type and spl34 mutant of photosynthetic pigment content were measured at the heading stage was extremely significant difference in the level of.3. of photosynthetic pigment content. The results showed that the leaf photosynthetic mutant spl34 the pigment content was slightly higher than the wild type, but the difference is not obvious, but the two leaves photosynthetic pigment content were lower than wild type and reached significant level and extremely significant difference in the level of.4. Ye Lv Fluorescence microscope observation body leaves at heading stage from the wild type and mutant spl34 for fluorescence microscopy revealed that the red fluorescence phenomenon of spl34 mutant chloroplasts compared to the wild type weak,.5. histochemical analysis of trypan blue staining and DAB staining showed weak red fluorescence produced lesion and mutant site in the chloroplast also is not part of the lesion, the mutant forms occurs heading determination of programmed cell death and hydrogen peroxide deposition of.6. on physiological and biochemical indexes of wild type and mutant spl34 in various physiological indexes were detected in the lesion. The results showed that the mutant CAT, POD, antioxidant enzyme activity SOD were low compared to the wild type. And, in parts of lesion mimic mutants H_2O_2 and O_2~- were higher than those of the wild type, and the content of OH and.7. had no significant difference between the wild type rice blast disease The spectrum of resistance and resistance identification of rice blast resistance spectrum determination showed that the mutant spl34 of ZA resistance frequency physiological group was higher than in the wild type, while the resistance frequency ZB physiological group lower than wild type of ZC, ZD, ZE and ZG physiological group resistance frequency is the same with the wild type. The resistance identification results showed that the mutant seedling blast disease index, leaf blast and Panicle Blast Disease spike rate were slightly higher than the wild type, but the difference was not significant.8. genetic analysis and gene mapping of genetic analysis showed that the phenotype of the spl34 mutant was controlled by a pair of recessive nuclear genes. The gene is located on chromosome LR49 and fourth LR52 two molecular markers, the physical distance of 200Kb. sequencing analysis identified candidate genes within the interval: LOC_Os04g56480 3449th base mutation (G3449T), resulting in amino acid substitution mutation from tryptophan to cysteine. The gene has been reported with HM47 allele The expression of Q RT-PCR gene mutation results of.9. gene and defense related genes showed that, compared with the wild type gene expression in the mutant decreased; and for the analysis of the expression of pathogenesis related genes and blast resistance gene, in addition to PR1a, the relative expression of course three genes PR1b and NPR1 in the mutant significantly, the relative expression of pathogenesis related genes and rice blast resistance genes of the remaining amount did not rise.

【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S435.11

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 Jiying Zhao;Pengcheng Liu;Chunrong Li;Yanyan Wang;Lequn Guo;Guanghuai Jiang;Wenxue Zhai;;LMM5.1 and LMM5.4,two eukaryotic translation elongation factor 1Alike gene family members,negatively affect cell death and disease resistance in rice[J];Journal of Genetics and Genomics;2017年02期

2 潘璐琪;陸雯;李小白;吳殿星;王雪艷;;秈稻93-11類病斑突變體的特征研究[J];核農(nóng)學(xué)報(bào);2015年03期

3 姜玉英;曾娟;陸明紅;劉杰;黃沖;劉萬才;;2015年全國農(nóng)作物重大病蟲害發(fā)生趨勢預(yù)報(bào)[J];中國植保導(dǎo)刊;2015年02期

4 邱結(jié)華;馬寧;蔣漢偉;圣忠華;邵高能;唐紹清;魏祥進(jìn);胡培松;;水稻類病斑突變體lmm4的鑒定及其基因定位[J];中國水稻科學(xué);2014年04期

5 孫惠敏;張春嬌;李保同;潘曉華;;水稻類病斑突變體的研究進(jìn)展[J];上海農(nóng)業(yè)學(xué)報(bào);2014年03期

6 Bao-Hua Feng;Yang Yang;Yong-Feng Shi;Hai-Chao Shen;Hui-Mei Wang;Qi-Na Huang;Xia Xu;Xiang-Guang Lü;Jian-Li Wu;;Characterization and Genetic Analysis of a Novel Rice Spotted-leaf Mutant HM47 with Broad-spectrum Resistance to Xanthomonas oryzae pv. oryzae[J];Journal of Integrative Plant Biology;2013年05期

7 蘇優(yōu)拉;張逸;李嘉賓;陸軍農(nóng);;卟啉的生物合成途徑與化學(xué)合成方法的比較[J];大學(xué)化學(xué);2012年06期

8 Marietta Baraoidan;Hei Leung;;Characterization and Genetic Analysis of a Light-and Temperature-sensitive Spotted-leaf Mutant in Rice[J];Journal of Integrative Plant Biology;2011年08期

9 陳析豐;金楊;馬伯軍;;水稻類病變突變體及抗病性的研究進(jìn)展[J];植物病理學(xué)報(bào);2011年01期

10 ;RLINl,encoding a putative coproporphyrinogen Ⅲ oxidase,is involved in lesion initiation in rice[J];遺傳學(xué)報(bào);2011年01期

相關(guān)博士學(xué)位論文 前1條

1 奉保華;水稻斑點(diǎn)葉突變體HM47的基因克隆與功能分析[D];中國農(nóng)業(yè)科學(xué)院;2015年

，

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