沙打旺9個(gè)品種對(duì)沙打旺黃矮根腐病的抗性機(jī)理研究及種質(zhì)特性綜合評(píng)價(jià)
發(fā)布時(shí)間：2018-11-21 16:57
【摘要】：沙打旺是我國(guó)特有的一種豆科牧草,兼有防風(fēng)固沙等生態(tài)作用。沙打旺黃矮根腐病是由沙打旺埃里磚格孢(Embellisia astragali)病原所引致的系統(tǒng)性真菌病害,該病是限制沙打旺生產(chǎn)的主要因素之一。種植抗病品種是防治該病的主要途徑,但目前我國(guó)尚未抗病品種可用,篩選出抗病種質(zhì)材料是選育出抗病品種的基礎(chǔ)。本論文的研究目的是通過(guò)離體葉片接種、噴霧接種和自然接種3種接種方法,評(píng)價(jià)9個(gè)沙打旺品種對(duì)沙打旺黃矮根腐病的抗性水平,采用聚丙烯酰胺凝膠電泳(SDS-PAGE)、rDNA-ITS基因測(cè)序和高效液相色譜法(HPLC)分析了供試品種的種子貯藏蛋白、親緣關(guān)系和內(nèi)源激素與抗性相關(guān)酶類,綜合分析對(duì)沙打旺黃矮根腐病的抗性機(jī)制以及抗性水平與生物學(xué)特性之間的關(guān)系,為有效選擇出抗病種質(zhì)奠定基礎(chǔ)。本文取得了以下主要結(jié)果:1.采用分子生物學(xué)方法首次確定了栽培沙打旺的分類地位及品種間的親緣關(guān)系。基于ITS序列構(gòu)建的系統(tǒng)發(fā)育樹表明,栽培的沙打旺與與野生沙打旺為同一物種,即斜莖黃芪(Astragalus adsurgens),但栽培沙打旺各品種間的親緣關(guān)系比與野生沙打旺更近,中沙1號(hào)和陜西榆林與其它7個(gè)品種存在明顯差異;在我國(guó)和美國(guó)的主要黃芪屬(Astragalus)(21種)和棘豆屬植物(Oxytropis)(7種)中,斜莖黃芪與漠北黃耆(A.austrosibiricus)的親緣關(guān)系最近,隸屬于黃芪屬驢豆組。2.雜花品種的植株中有部分花穗為白花,中沙1號(hào)品種的植株均為匍匐型植株,而其它品種均為紫色花穗、直立型植株;種子蛋白測(cè)定結(jié)果表明遼寧阜新與中沙1號(hào)相近。3.確定了自然發(fā)病試驗(yàn)中沙打旺黃矮根腐病的病原來(lái)自帶菌的種子,而無(wú)氣流傳播的外來(lái)菌源,該試驗(yàn)首次證明了病菌從死亡植株上產(chǎn)生的孢子在拔節(jié)期侵染葉片,植株的發(fā)病率逐年增加,在第4年發(fā)病率達(dá)到100%,完善了該病的侵染循環(huán)。4.在3個(gè)接種試驗(yàn)中以發(fā)病率為抗病級(jí)別的評(píng)價(jià)標(biāo)準(zhǔn),在同一抗病級(jí)別的品種中以病情指數(shù)等指標(biāo)為評(píng)價(jià)抗病性強(qiáng)弱的次序,確定了高抗品種為內(nèi)蒙早熟陜西榆林,抗性品種為雜花鄂爾多斯固原河南,中抗品種為寧夏彭陽(yáng),低抗品種為遼寧阜新=中沙1號(hào)。陜西榆林和內(nèi)蒙早熟的抗侵入和抗擴(kuò)展強(qiáng),陜西榆林和固原的抗損害能力強(qiáng),中沙1號(hào)在抗侵入等抗性能力上均最弱。高產(chǎn)草品種為內(nèi)蒙早熟,草地持久性最強(qiáng)品種為遼寧阜新,中沙1號(hào)的種子產(chǎn)量高,且營(yíng)養(yǎng)價(jià)值顯著高于其它品種。5.植株體內(nèi)9種生物化學(xué)物質(zhì)測(cè)定結(jié)果顯示,未接種植株中的5種植物內(nèi)源激素含量和4種生化酶活性在品種間存在顯著(P0.05)差異;接種后,所有品種植株體內(nèi)的吲哚乙酸(IAA)含量均顯著(P0.05)升高,過(guò)氧化物酶(POD)的活性均顯著(P0.05)下降,水楊酸(SA)和脫落酸(ABA)的含量以及超氧化物歧化酶(SOD)和苯丙氨酸解氨酶(PAL)的活性均下降(顯著或不顯著),細(xì)胞分裂素(CTK)和過(guò)氧化氫酶(CAT)均上升(顯著或不顯著)。未染病植株體內(nèi)的吲哚乙酸的含量越高,超氧化物歧化酶的活性越高,該品種抗沙打旺黃矮根腐病越強(qiáng),此可作為沙打旺種質(zhì)對(duì)沙打旺黃矮根腐病的抗性水平的指標(biāo)。6.防風(fēng)、核桃青皮和大蒜乙醇提取液對(duì)沙打旺埃里磚格孢有較強(qiáng)的抑菌作用;沙打旺埃里磚格孢及其發(fā)酵產(chǎn)物對(duì)測(cè)試的植物病原真菌均有拮抗特性,抗菌物質(zhì)值得開發(fā)利用。在選育抗病品種時(shí)除了選擇抗病強(qiáng)、草產(chǎn)量高的內(nèi)蒙早熟和陜西榆林等種質(zhì)之外,也應(yīng)吸納草質(zhì)優(yōu)、早熟但抗病弱的中沙1號(hào)的優(yōu)良特性。
[Abstract]:Shakewang is a kind of leguminous grass which is unique in China, and has the ecological effects of preventing wind and sand. Shawwan yellow dwarf root rot is a systemic fungal disease caused by the pathogeny of Embeliscia stagali, which is one of the main factors to limit the production of sand. The disease resistant variety is the main way to control the disease, but at present, the disease resistant variety is not available, and the screening of the disease-resistant germplasm material is the basis for breeding the disease-resistant variety. The purpose of this study was to evaluate the resistance of 9 Shawwan varieties to the root rot of Shawwan yellow dwarf by inoculation, spraying and natural inoculation from the leaves of the body. rDNA-ITS gene sequencing and high performance liquid chromatography (HPLC) were used to analyze the relationship between the resistance mechanism and the resistance level and the biological characteristics of the seed storage protein, the relationship and the endogenous hormone and the resistance-related enzymes of the test variety. and laying a foundation for the effective selection of the disease-resistant germplasm. The following main results are obtained in this paper: 1. The classification status of the cultivated sand and the relationship between the varieties were first determined by the molecular biology method. The phylogenetic tree constructed on the basis of ITS sequence indicated that the cultivated sandblast was the same as that of the wild sand, that is, Astrialus adsurgens, but the relationship between the variety of the cultivated sand and the wild sand was closer to that of the wild sand. In China and the United States, there were significant differences between the first and the other seven species of the genus Astoralis (21 species) and Oxytropis (7 species) in China and the United States, and the relationship between the Stems of the Stems and the North Yellow River (A. asturosibiricus) was the most recent, belonging to the yellow-yellow donkey-bean group. In the plant of the hybrid-flower variety, some of the plants were of white-white flowers, and the plants of the No. 1 variety were the creeping plants, while the other varieties were the purple-colored and erect-type plants. The results of the determination of the seed protein showed that the Fuxin of Liaoning was close to the No. 1 of Zhongsha 1. It was determined that the pathogen of Shawwan yellow dwarf root rot in the natural disease experiment was from the seed of the bacteria, and the source of the foreign bacteria which was free of gas flow, the first time that the spore produced by the pathogen from the dead plant was infected with the leaf during the jointing stage, and the incidence of the plant increased year by year. In the fourth year, the incidence rate was 100%, and the infection cycle of the disease was improved. In the three inoculation tests, the disease-resistance-grade evaluation criteria were used to evaluate the order of the disease resistance in the same disease-resistant variety, and the high-resistant variety was the early-maturing and early-maturing Shaanxi Yulin, and the resistant variety was Erdos Guyuan, Henan. The anti-varieties of the middle and low-resistant varieties are Pengyang, Ningxia, and the low anti-varieties are the No. 1 of Zhongsha, Fuxin, Liaoning. The anti-invasion and anti-expansion of the early-maturing of Yulin and Inner Mongolia in Shaanxi Province are strong, and the anti-damage ability of Yulin and Guyuan in Shaanxi is strong, and the medium sand 1 is the weakest in the resistance ability of anti-invasion and other resistance. The high-yield grass variety is the early-maturing of the Inner Mongolia, the most persistent species of the grass is Liaoning Fuxin, the seed yield of the Zhongsha 1 is high, and the nutritive value is obviously higher than that of the other varieties. The results showed that the content of endogenous hormones and the activity of 4 kinds of biochemical enzymes in the non-inoculated plants were significant (P0.05). The activity of peroxidase (POD) decreased significantly (P0.05), and the content of salicylic acid (SA) and abscisic acid (ABA) and the activity of superoxide dismutase (SOD) and phenylalanine ammonia-lyase (PAL) were both decreased (significant or insignificant). Both cytokinin (CTK) and catalase (CAT) both increased (significantly or not). The higher the content and the higher the activity of superoxide dismutase (SOD), the higher the activity of superoxide dismutase (SOD). The anti-wind, walnut green skin and the garlic ethanol extract have a strong antibacterial effect on the Shawwang Ericeisporites; and the anti-bacterial substance is worthy of development and utilization of the tested plant pathogenic fungi by the Shakewang Ericeisporites and the fermentation products thereof. In the breeding of the disease-resistant variety, in addition to the selection of such germplasm as the disease resistance, the high yield of the grass, the early maturity of the grass and the Yulin of Shaanxi, the excellent characteristics of the medium and high-quality, early-maturing but disease-resistant medium sand 1 should be absorbed.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：S435.4
，
本文編號(hào)：2347608
[Abstract]:Shakewang is a kind of leguminous grass which is unique in China, and has the ecological effects of preventing wind and sand. Shawwan yellow dwarf root rot is a systemic fungal disease caused by the pathogeny of Embeliscia stagali, which is one of the main factors to limit the production of sand. The disease resistant variety is the main way to control the disease, but at present, the disease resistant variety is not available, and the screening of the disease-resistant germplasm material is the basis for breeding the disease-resistant variety. The purpose of this study was to evaluate the resistance of 9 Shawwan varieties to the root rot of Shawwan yellow dwarf by inoculation, spraying and natural inoculation from the leaves of the body. rDNA-ITS gene sequencing and high performance liquid chromatography (HPLC) were used to analyze the relationship between the resistance mechanism and the resistance level and the biological characteristics of the seed storage protein, the relationship and the endogenous hormone and the resistance-related enzymes of the test variety. and laying a foundation for the effective selection of the disease-resistant germplasm. The following main results are obtained in this paper: 1. The classification status of the cultivated sand and the relationship between the varieties were first determined by the molecular biology method. The phylogenetic tree constructed on the basis of ITS sequence indicated that the cultivated sandblast was the same as that of the wild sand, that is, Astrialus adsurgens, but the relationship between the variety of the cultivated sand and the wild sand was closer to that of the wild sand. In China and the United States, there were significant differences between the first and the other seven species of the genus Astoralis (21 species) and Oxytropis (7 species) in China and the United States, and the relationship between the Stems of the Stems and the North Yellow River (A. asturosibiricus) was the most recent, belonging to the yellow-yellow donkey-bean group. In the plant of the hybrid-flower variety, some of the plants were of white-white flowers, and the plants of the No. 1 variety were the creeping plants, while the other varieties were the purple-colored and erect-type plants. The results of the determination of the seed protein showed that the Fuxin of Liaoning was close to the No. 1 of Zhongsha 1. It was determined that the pathogen of Shawwan yellow dwarf root rot in the natural disease experiment was from the seed of the bacteria, and the source of the foreign bacteria which was free of gas flow, the first time that the spore produced by the pathogen from the dead plant was infected with the leaf during the jointing stage, and the incidence of the plant increased year by year. In the fourth year, the incidence rate was 100%, and the infection cycle of the disease was improved. In the three inoculation tests, the disease-resistance-grade evaluation criteria were used to evaluate the order of the disease resistance in the same disease-resistant variety, and the high-resistant variety was the early-maturing and early-maturing Shaanxi Yulin, and the resistant variety was Erdos Guyuan, Henan. The anti-varieties of the middle and low-resistant varieties are Pengyang, Ningxia, and the low anti-varieties are the No. 1 of Zhongsha, Fuxin, Liaoning. The anti-invasion and anti-expansion of the early-maturing of Yulin and Inner Mongolia in Shaanxi Province are strong, and the anti-damage ability of Yulin and Guyuan in Shaanxi is strong, and the medium sand 1 is the weakest in the resistance ability of anti-invasion and other resistance. The high-yield grass variety is the early-maturing of the Inner Mongolia, the most persistent species of the grass is Liaoning Fuxin, the seed yield of the Zhongsha 1 is high, and the nutritive value is obviously higher than that of the other varieties. The results showed that the content of endogenous hormones and the activity of 4 kinds of biochemical enzymes in the non-inoculated plants were significant (P0.05). The activity of peroxidase (POD) decreased significantly (P0.05), and the content of salicylic acid (SA) and abscisic acid (ABA) and the activity of superoxide dismutase (SOD) and phenylalanine ammonia-lyase (PAL) were both decreased (significant or insignificant). Both cytokinin (CTK) and catalase (CAT) both increased (significantly or not). The higher the content and the higher the activity of superoxide dismutase (SOD), the higher the activity of superoxide dismutase (SOD). The anti-wind, walnut green skin and the garlic ethanol extract have a strong antibacterial effect on the Shawwang Ericeisporites; and the anti-bacterial substance is worthy of development and utilization of the tested plant pathogenic fungi by the Shakewang Ericeisporites and the fermentation products thereof. In the breeding of the disease-resistant variety, in addition to the selection of such germplasm as the disease resistance, the high yield of the grass, the early maturity of the grass and the Yulin of Shaanxi, the excellent characteristics of the medium and high-quality, early-maturing but disease-resistant medium sand 1 should be absorbed.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：S435.4
，
本文編號(hào)：2347608
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