沙打旺9个品种对沙打旺黄矮根腐病的抗性机理研究及种质特性综合评价
发布时间：2018-11-21 16:57
【摘要】：沙打旺是我国特有的一种豆科牧草,兼有防风固沙等生态作用。沙打旺黄矮根腐病是由沙打旺埃里砖格孢(Embellisia astragali)病原所引致的系统性真菌病害,该病是限制沙打旺生产的主要因素之一。种植抗病品种是防治该病的主要途径,但目前我国尚未抗病品种可用,筛选出抗病种质材料是选育出抗病品种的基础。本论文的研究目的是通过离体叶片接种、喷雾接种和自然接种3种接种方法,评价9个沙打旺品种对沙打旺黄矮根腐病的抗性水平,采用聚丙烯酰胺凝胶电泳(SDS-PAGE)、rDNA-ITS基因测序和高效液相色谱法(HPLC)分析了供试品种的种子贮藏蛋白、亲缘关系和内源激素与抗性相关酶类,综合分析对沙打旺黄矮根腐病的抗性机制以及抗性水平与生物学特性之间的关系,为有效选择出抗病种质奠定基础。本文取得了以下主要结果:1.采用分子生物学方法首次确定了栽培沙打旺的分类地位及品种间的亲缘关系。基于ITS序列构建的系统发育树表明,栽培的沙打旺与与野生沙打旺为同一物种,即斜茎黄芪(Astragalus adsurgens),但栽培沙打旺各品种间的亲缘关系比与野生沙打旺更近,中沙1号和陕西榆林与其它7个品种存在明显差异;在我国和美国的主要黄芪属(Astragalus)(21种)和棘豆属植物(Oxytropis)(7种)中,斜茎黄芪与漠北黄耆(A.austrosibiricus)的亲缘关系最近,隶属于黄芪属驴豆组。2.杂花品种的植株中有部分花穗为白花,中沙1号品种的植株均为匍匐型植株,而其它品种均为紫色花穗、直立型植株;种子蛋白测定结果表明辽宁阜新与中沙1号相近。3.确定了自然发病试验中沙打旺黄矮根腐病的病原来自带菌的种子,而无气流传播的外来菌源,该试验首次证明了病菌从死亡植株上产生的孢子在拔节期侵染叶片,植株的发病率逐年增加,在第4年发病率达到100%,完善了该病的侵染循环。4.在3个接种试验中以发病率为抗病级别的评价标准,在同一抗病级别的品种中以病情指数等指标为评价抗病性强弱的次序,确定了高抗品种为内蒙早熟陕西榆林,抗性品种为杂花鄂尔多斯固原河南,中抗品种为宁夏彭阳,低抗品种为辽宁阜新=中沙1号。陕西榆林和内蒙早熟的抗侵入和抗扩展强,陕西榆林和固原的抗损害能力强,中沙1号在抗侵入等抗性能力上均最弱。高产草品种为内蒙早熟,草地持久性最强品种为辽宁阜新,中沙1号的种子产量高,且营养价值显著高于其它品种。5.植株体内9种生物化学物质测定结果显示,未接种植株中的5种植物内源激素含量和4种生化酶活性在品种间存在显著(P0.05)差异;接种后,所有品种植株体内的吲哚乙酸(IAA)含量均显著(P0.05)升高,过氧化物酶(POD)的活性均显著(P0.05)下降,水杨酸(SA)和脱落酸(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(显著或不显著),细胞分裂素(CTK)和过氧化氢酶(CAT)均上升(显著或不显著)。未染病植株体内的吲哚乙酸的含量越高,超氧化物歧化酶的活性越高,该品种抗沙打旺黄矮根腐病越强,此可作为沙打旺种质对沙打旺黄矮根腐病的抗性水平的指标。6.防风、核桃青皮和大蒜乙醇提取液对沙打旺埃里砖格孢有较强的抑菌作用;沙打旺埃里砖格孢及其发酵产物对测试的植物病原真菌均有拮抗特性,抗菌物质值得开发利用。在选育抗病品种时除了选择抗病强、草产量高的内蒙早熟和陕西榆林等种质之外,也应吸纳草质优、早熟但抗病弱的中沙1号的优良特性。
[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.
【学位授予单位】：兰州大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S435.4
，
本文编号：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.
【学位授予单位】：兰州大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S435.4
，
本文编号：2347608
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