Fungal Diversity in Rhizosphere Soil of Soybean Fields under Salt-alkali Stress
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摘要: 为进一步了解大豆根际土壤中真菌对盐碱胁迫的适应性，利用DGGE技术，通过浇灌盐碱液（A）及覆盖盐碱土（B）两种盐碱胁迫方式对野生大豆和栽培大豆根际真菌多样性进行分析。结果表明：不同盐碱胁迫方式对大豆根际真菌相似性影响较大，如野生大豆和栽培大豆在相同的盐碱胁迫处理A1浓度下，野生大豆和栽培大豆根际真菌群落结构的相似性可达0.70；适当的盐碱浓度可增加大豆根际真菌的均匀度和Shannon指数，如野生大豆在B1浓度时均匀度为0.99，在A2浓度时Shannon指数为1.87；其中对DGGE特异条带测序与基因库比对，获得一些相对优势的菌株如：Fusarium oxysporum、Actinomucor elegans、Marcelleina tuberculispora、Marcelleina persoonii、Tofieldiaceae environmental sample clone、Uncultured eukaryote clone等。Abstract: To understand the adaptability of fungi in rhizosphere soil of soybean, two saline-alkali stress ways were set which involved irrigating saline solution (A) and covering saline soil (B), to analyze the diversity of rhizospherical fungi of wild soybean and cultivated soybean, throughthe application of DGGE method. The results showed that the saline alkali stress ways had great influence on the similarity of fungi in rhizosphere of soybean. For example, under the same saline alkali stress way and concentration of A1, the similarity ratio between the rhizospherical fungi of wild soybean and cultivated soybean was 0.70. In addition, appropriate saline concentration can increase the uniformity and Shannon index of soybean rhizospherical fungi, for example, the uniformity of wild soybean at concentration of B1 was 0.99, and the Shannon index at concentration of A2 was 1.87. Moreover, the sequencing of DGGE specific bands and the further comparison with gene bank, some comparative advantageous strain were obtained, such as Fusarium oxysporum, Actinomucor elegans, Marcelleina tuberculispora, Marcelleina persoonii, Tofieldiaceae environmental sample clone and Uncultured eukaryote clone etc.
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图 1 引物GC-Fung和NS1的琼脂糖凝胶电泳结果
注：M为DL2000 DNA Marker；1，3为CK；2，5，7，8为野生大豆A处理；11，13，15，17为野生大豆B处理；4，6，9，10为栽培大豆A处理；12，14，16，18为栽培大豆B处理。
Figure 1. Agarose gel electrophoresis of primers GC-Fung and NS1
图 2 野生大豆和栽培大豆在不同盐碱浓度处理下根际真菌的DGGE图谱
注：1、3为CK；2、5、7、8为野生大豆A处理；11、13、15、17为野生大豆B处理；4、6、9、10为栽培大豆A处理；12、14、16、18为栽培大豆B处理。
Figure 2. DGGE patterns of rhizosphere fungi in G. soja and G. max fields under varied salt-alkali stresses
图 3 根际真菌DGGE图谱UPGMA聚类分析
Figure 3. UPGMA cluster analysis on DGGE bands of rhizosphere fungi
表 1 处理方式及浓度配比
Table 1. Treatments and salt-alkali concentrations applied for experimentation
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表 2 真菌群落结构的均匀度、Shannon指数
Table 2. Uniformities and Shannon indices of fungal communities
类别 项目 CK A处理方式 B处理方式 A1 A2 A3 A4 B1 B2 B3 B4 野生大豆 均匀度 0.96 0.7 0.85 0.98 0.45 0.99 0.97 0.97 0.93 Shannon指数 1.88 1.45 1.87 1.76 0.31 1.59 1.88 2.4 1.93 栽培大豆 均匀度 0.95 0.92 0.81 0.94 0.98 0.92 0.78 0.94 0.96 Shannon指数 2.09 2.03 1.3 1.95 1.91 1.65 1.25 2.34 2.21
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表 3 测序条带与对应的同源性菌株
Table 3. Sequencing strips of sample fungiand corresponding homologous strains
条带编号 同源性菌株 1 Fusarium oxysporum 2 Actinomucor elegans 3 Marcelleina tuberculispora 4 Marcelleina persoonii 5 Tofieldiaceae environmental sample clone 6 Uncultured Cystofilobasidiales(aff.Guehomyces) clone 7 Uncultured microeukaryote clone 8 Marcelleina tuberculispora 9 Uncultured eukaryote clone 10 Pseudonectria rousseliana 11 Uncultured fungus clone 12 Uncultured eukaryote clone 13 Fusarium sp. 14 Stachybotrys chlorohalonata strain 15 Fungal sp. 16 Uncultured Davidiellaceae clone 17 Sordariomycetidae sp.
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