间作桃树对石榴枯萎病土壤真菌多样性的改善研究

Effect of intercropping peach trees on the fungi diversity in pomegranate wilt disease soil

【目的】间作套种对病害控制的研究已得到越来越多的关注,本研究旨在探明间作桃树对石榴枯萎病土壤真菌多样性的影响。【方法】以ITS5-1737F和ITS2-2043R为引物,对云南蒙自石榴枯萎病根际土壤(MZD)、健康石榴根际土壤(MZ)以及间作桃树修复石榴枯萎病根际土壤(MZP)样品的真菌ITS区进行扩增,采用Illumina MiSeq高通量测序的方法系统分析3种土壤样品的根际微生物真菌群落组成及多样性。【结果】MZD,MZ,MZP在门水平上的平均序列数目分别为32865、34698、28714,在属水平上的平均序列数目分别为30901,30514,27002,多年生石榴树根际土壤真菌的平均序列数多于间作桃树修复枯萎病石榴树。MZD、MZ、MZP得到的OTUs数分别为1174、1076、1007,韦恩图分析表明,MZD特有OTUs数为152个,MZ特有OTUs数为47个,MZP特有OTUs数为94个。PCA分析结果表明,真菌群落组成间作桃树修复石榴枯萎病根际土壤与石榴枯萎病根际土壤差异较大,石榴枯萎病根际土壤真菌的群落组成通过间作桃树得到了较大程度改变。在属的分类水平上,共鉴定到481种真菌属,间作桃树修复石榴枯萎病土壤与健康石榴土壤差异显著的真菌属有Un--s-Hypocreales sp.,Un--s-Cordycipitaceae sp.,Un--s-Microascales sp.,腐质霉属(Humicola),葡萄穗霉属(Stachybotrys),透孢黑团壳属(Massarina),Stagonosporopsis等21个;间作桃树修复石榴枯萎病根际土壤与石榴枯萎病根际土壤差异显著的真菌属有被孢霉属(Mortierella),壳针孢属(Septoria),腐质霉属(Humicola),葡萄穗霉属(Stachybotrys),透孢黑团壳属(Massarina)等47个。【结论】多年生石榴树根际真菌的平均序列数多于间作桃树枯萎病石榴树,多年生石榴树单一种植的土壤根际真菌趋向于增多。通过间作桃树可以改变石榴枯萎病根际土壤真菌群落组成及多样性,改善土壤微生态平衡,促进石榴健康生长,对保护石榴产业的可持续发展有重要的经济意义和社会效益。

【Objective】The research of intercropping for disease control recently has become a hot issues for many reasons. This study was conducted to investigate the effects of intercropping peach trees on soil fungal diversity of pomegranate wilt. 【Method】ITS5-1737 F and ITS2-2043 R were used as primers to amplified the ITS region of samples collected from healthy pomegranate rhizosphere soil(MZ), wilt disease pomegranate rhizosphere soil(MZD) and wilt disease pomegranate soil intercropping peach trees(MZP) in Mengzi county, Yunnan province. The fungal community composition and diversity of three kinds of rhizosphere soil were analyzed by high-throughput sequencing.【Result】The sequences average number of MZD,MZ,MZP at phylum level were 32 865,34 698,28 714, respectively, and those of MZD,MZ,MZP at genera level were 30 901,30 514,27 002, respectively. The OTUs numbers obtained from MZD,MZ,MZP were 1174、1076、1007, respectively, Venn diagram analysis showed that 152 OTUs were unique to MZD, 47 OTUs to MZ and 94 OTUs to MZP. PCA analysis showed that the fungal community composition of rhizosphere soil from MZP was significantly different from that of MZD, and intercropping peach trees could significantly change the fungal composition of wilt pomegranate rhizosphere soil. At genus level, a total of 481 fungal genera were noted in this research, 21 fungi genus such as Un--s-Hypocreales sp.,Un--s-Cordycipitaceae sp.,Un--s-Microascales sp.,Humicola,Stachybotrys,Massarina,Stagonosporopsis et al were significantly different between healthy pomegranate soil rhizosphere and pomegranate wilt disease soil rhizosphere. 47 fungi genus such as Mortierella,Mortierella,Septoria,Humicola,Stachybotrys,Massarina,Stachybotrys et al were significantly different between pomegranate wilt disease soil intercropping peach trees and pomegranate wilt disease soil rhizosphere.【Conclusion】The average number of fungal sequences in the rhizosphere of pomegranate wilt disease soil was higher than that in pomegranate wilt disease soil intercropping peach trees, and soil fungi tended to increase in pomegranate monoculture. The results indicated that intercropping peach trees could change the fungi composition and diversity of wilt pomegranate rhizosphere soil as well as improving the soil microecological balance and promoting the healthy growth of pomegranate, which was expected to contribute to protect the sustainable development of pomegranate industry.