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沙打旺(Astragalus adsurgens)品种对黄矮根腐病(Embellisia astragali)的抗性评价
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摘要
沙打旺(Astragalus adsurgens)是我国北方广为栽培的重要豆科牧草,抗旱、耐瘠薄,耐风沙,在改善环境、发展生产中发挥着重要作用。埃里砖格孢(Embellisia astragali Li & Nan nov. sp.)引致的黄矮根腐病是造成沙打旺草地早衰的最主要真菌病害之一。本论文通过实验室试验、温室试验和田问试验,评价了我国当前主要沙打旺品种对黄矮根腐病的抗病性,并初步探讨了其抗病机制,参试品种包括鄂尔多斯、固原野生、河南、辽宁阜新、内蒙古早熟、宁夏彭阳、普通、陕西榆林、杂花和中沙一号沙打旺等10个品种。主要研究结果如下:
     1.实验室条件下,10个品种,接种埃里砖格孢后15日龄幼苗的干重为7.00-15.08mg/皿,与未接种的对照苗相比,降低幅度为19.80%-46.79%,其中降低最多的为内蒙古早熟和宁夏彭阳等两个品种,干重分别降低了46.79%和44.35%,降低最少的为中沙一号,其干重比未接种的对照仅降低了19.80%。各品种的发病率、病情指数和死亡率分别为75.70%-90.60%,16.00-78.00,和10.20%-36.60%,其中内蒙古早熟和宁夏彭阳两品种的上述三项指标最高,分别为90.60%和88.76%、78.00和74.00,36.60%和34.11%;陕西榆林和中沙一号两品种的上述指标最低,分别是77.13%和75.70%,20.00和16.00,11.80%和10.20%。
     2.温室条件下,参试的10个品种,接种或不接种埃里砖格孢,盆栽生长24周的单株干重为0.90g-2.17g,与未接种幼苗相比,干重降低17.74%-63.20%;降低最多的为内蒙古早熟,降低最少的为中沙一号。各品种的发病率、病情指数和死亡率分别为33.70%-76.80%,19.17-70.37,和10.91%-55.56%,其中内蒙古早熟和宁夏彭阳两品种的上述三项指标最高,分别为75.50%和76.80%、70.37和67.62,55.56%和51.11%;陕西榆林和中沙一号两个品种的上述指标最低,分别是35.20%和33.70%,19.17和17.27,12.73%和10.91%。
     3.2010和2011连续两年,在甘肃省环县具有发病史的地块开展的田间试验,对10个品种抗病性评价表明,两年平均田间植株干重为5.15g-6.96g,干重较高的品种为中沙一号和辽宁阜新,分别为6.96g和6.69g,干重较低的品种为固原野生和宁夏彭阳,分别为5.15g和5.52g。发病率最高的为内蒙古早熟和宁夏彭阳,发病率分别为82.33%和80.53%,发病率最低为中沙一号,为33.67%。死亡率最高的为固原野生和内蒙古早熟,分别达59.16%和59.26%,最低的为陕西榆林,仅35.46%。两年间平均病情指数以内蒙古早熟和宁夏彭阳为最高,分别为43.69和43.81,而中沙一号最低,为14.69。
     4.相关性分析表明,实验室试验、温室试验和田间试验获得的各品种的发病率、死亡率和病情指数均呈显著正相关(P<0.05),决定系数为0.82-0.99,可用二元回归方程y=8.28+0.18x1+0.15x2表示,其中Y=田间病情指数,X1=实验室试验的病情指数,X2=温室试验的病情指数。实验室和温室试验结果,可以用来初步筛选和评价沙打旺的抗病性,其与沙打旺品种田间抗病性显著相关,决定系数为0.83。
     5.以抗病品种中沙一号和感病品种内蒙古早熟为材料,比较了两个品种的抗病机理。抗病品种和感病品种中孢子萌发后芽管长度分别为79.9μm和103.7μm,侵入点密度分别为34.6个/mm2和38.8个/mm2,侵入率分别为16.7%和58.6%,说明抗病品种上孢子萌发受到抑制,侵入程度明显低于感病品种。抗病品种与感病品种的叶片表皮细胞密度分别为80.30个/mm2和56.55个/mm2,气孔密度分别为10.95个/mm2和18.80个/mm2,栅栏组织层数分别为3.00层和1.75层,栅栏组织细胞密度分别为59.60个/mm和41.2个/mm,角质层厚度分别为20.25μm和9.54μm。相关分析表明,沙打旺对黄矮根腐病的抗性与叶片表皮细胞密度、栅栏组织层数、栅栏组织细胞密度以及角质层厚度等均呈正相关,与气孔密度呈负相关。抗病品种与感病品种丙二醛(MDA)含量峰值分别出现于5d和7d,峰值为16.4μmol·g-1 FW和21.33μmol·g-1 FW。抗病品种与感病品种过氧化物酶(POD)活性峰值分别出现于1d和9d,峰值分别为663.5 U/g FW·min和511.7U/g FW·min。超氧化物歧化酶(SOD)、多酚氧化酶(PPO)、苯丙氨酸解氨酶(PAL)、β-1,3-葡聚糖酶以及几丁质酶这些保护性酶的活性变化与前两者有相似趋势,抗病品种在受到侵染时保护性酶活性上升快、活性高,而感病品种与之相反,受侵染后保护性酶活性上升缓慢、峰值低。
     通过对与病症表现相关的11个形态学、生理学和病理学指标的综合评价与数学分析,初步认为,参试的10个品种中,陕西榆林和中沙一号为抗病品种,普通和杂花沙打旺为中抗品种,鄂尔多斯、固原野生、河南和辽宁阜新为中感品种,内蒙古早熟和宁夏彭阳为感病品种。
Standing milkvetch(Astragalus adsurgens) is an important forage legume. It has the triats of drought tolerance, high herbage dry matter yield and wide adaptability, which enable the crop plays important roles in fixing sand,dune,controling soil and water erosion,improving soil fertility,and development of livestock production. Fungal diseases are one of the limiting factors for standing milkvetch production and persistence. Yellow dawarf root rot disease caused by Embellisia astragali is a disease reported recently. The disease is responsible for the stand decline and poor persistence,low productivity of the standing milkvetch in northern China where the crop is growing. Use of disease resistant cultivar is the most effective method to control the disease. However, there is no information available on resistance to Embellisia astragali among the cultivars used in practice. Therefore, a series of experiments were carried out in laboratory, greenhouse and field for evaluating the resistance of standing milkvetch cultivars to Embellisia astragali. The main results are as the follows:
     1. In laboratory conditions, germination in 3d of all inoculated varieties were 40%-71%, which were 13.1%-177.8% higher than control. The highest germination variety was Neimeng and the lowest germination variety was Zhongsha No.1. Shoot length of all varieties was reduced by E. astragali inoculation, and six varieties, Erdos, Guyuan, Henan, Liaoning, Neimeng and Ningxia, showed significant (P<0.05) and the decreased rate was more than 20%. Varieties of Neimeng (2.57cm) and Ningxia (2.48cm) had a significantly greater (P<0.05)mycelium distribution range compared with other varieties. In contrast, varieties of Shanxi (1.42cm) and Zhongsha No.1 (1.37cm) had a significantly small-scale (P<0.05) mycelium distribution compared with other varieties. Incidence (90.60%、88.76%), mortality (36.60%,34.11%) and DSI (78.00,74.00) of Neimeng and Ningxia were significantly higher than other varieties. Conversely, Shanxi and Zhongsha No.1 had the lowest Incidence, mortality and DSI. In greenhouse and field conditions, emergence, plant height, root length, plant dry weight, incident, mortality and DSI had the same trend with laboratory results. Meanwhile, inoculation reduced the leaf number, leaf thickness, leaf area, stem diameter and all photosynthetic characters, but increased the stem number.
     2. Results from laboratory, greenhouse and field experiments showed very similar trends. Every disease parameter was significantly (P<0.01) correlated with all other disease parameters even if in different experiments (correlation coefficient ranging from 0.85 to 0.99). Multiple linear regression analysis showed DSI of different varieties in the laboratory and greenhouse experiments was significantly (P<0.05) correlated with that of the field experiment. The dualistic linear regression equation between the DSI of the field experiment (y) and that of the laboratory experiment (x1) and the greenhouse experiment (X2) was: y=8.28+0.18x1+0.15x2 (R2=0.83). Emergence and other 10 parameters were used to evaluated the resistance of all varieties to Embellisia astragali with grey relate degree analysis, principal component analysis and other three mathematical models. According to the research reported here, the most resistant varieties of those tested are Shanxi and Zhongsha No.1, the moderately resistant varieties are Putong and Zahua, the moderately susceptible varieties are Erdos, Guyuan, Henan and Liaoning, and the most susceptible varieties are Neimeng and Ningxia.
     3. Plant defence mechanisms are usually complex and composed of multiple layers of defence that are effective against diverse array of pathogens. There were positive correlations between disease resistance and the epidermis cell density, palisade tissue layers, palisade tissue cell density, corneum thickness. But there were negative correlations between disease resistance and stoma density. There were also positive correlations between disease resistance and the activities of all the enzymes such as POD、SOD、PPO、PAL、β-1,3-glucanse and chitinase. But there were negative correlations between disease resistance and MDA activity. After inoculation, the levels of endogenous phytohormones in both resistant and susceptible varieties were increased, especially in susceptible variety Neimeng. Disorders of endogenous phytohormones after inoculation would lead to a series of symptoms even malformations. In resistant varieties, the length of germ tube, penetration density and penetration were significantly (P<0.01) lower than that in susceptible varieties. As colonization of the cortex, hyphae in resistant varieties were much less than that in susceptible varieties in the same time. More over, hyphae spread much.slower in resistant varieties than in susceptible varieties. Papillae were often found at sites of penetration, but the percentages of papillae in resistant varieties were much higher than that in susceptible varieties. Intercellular hyphae penetrated cortical cell and formed haustoria in it. In resistant varieties haustoria was covered by cell wall appositions, and some haustoria were malformed.
引文
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