CAJ | 학술논문

【目的】采用18S rDNA PCR-DGGE技术研究苦楝－小麦农林复合生态系统根际及非根际土壤真菌群落结构，以期揭示农林复合生态系统中伴生树种对作物根际真菌群落的影响，为农林复合经营实践提供理论参考。【方法】提取根际和非根际土壤真菌 DNA，采用通用引物 GC-FR1和 FF390扩增18S rDNA 基因目的片段，扩增片段用变性梯度凝胶电泳分析；不同处理间微生物群落结构的相似性采用非加权组算术平均法分析比较；以物种丰富度指数、Shannon-Wiener多样性指数和均匀度指数3个指标评价各样品真菌多样性；最后将 DGGE 图谱中的优势条带割胶测序并建立系统发育树。【结果】根际真菌群落结构和多样性远比非根际土壤复杂，且根际土壤样品 DGGE电泳图谱中优势条带数也比非根际土壤样品多。进一步采用非加权组算术平均法对不同处理间真菌群落结构的相似性进行分析，发现每个处理组3次重复间的相似度除苦楝根际土壤外均大于70%；在聚类分析图中所有处理明显分为3个组，相似度为51%；小麦根际真菌菌群与苦楝－小麦农林复合系统中的小麦根际真菌相似性最高，相似度为72%，表明苦楝对小麦根际真菌群落的影响不大；苦楝根际真菌菌群与其他各组间差异较大，相似度仅为51%。主成分分析结果也表明所有处理明显分为3个群组。多样性分析结果表明，各处理组根际真菌之间以及非根际真菌间丰富度差异均不明显，而苦楝－小麦农林复合系统中的小麦及苦楝根际真菌丰富度显著高于非根际真菌；根际真菌的 Shannon-Wiener 多样性指数存在显著差异，非根际真菌的Shannon-Wiener多样性指数差异不显著；根际真菌及非根际真菌各组间均匀度指数差异不大，而小麦－苦楝农林复合系统中的小麦以及苦楝根际真菌与非根际真菌均匀度指数存在明显差异。此外，DGGE图谱中的11条优势条带克隆测序结果表明，3条条带为不能培养的真菌，其余8条条带对应的同源性最高的序列分别属于球囊菌门、担子菌门、子囊菌门和半知菌亚门。【结论】小麦－苦楝农林复合系统中的小麦根际真菌丰富度、多样性指数和均匀度均比单纯小麦根际真菌高，表明此复合系统中伴生的苦楝可增加小麦根际真菌的多样性。【目的】採用18S rDNA PCR-DGGE技術研究苦楝－小麥農林複閤生態繫統根際及非根際土壤真菌群落結構，以期揭示農林複閤生態繫統中伴生樹種對作物根際真菌群落的影響，為農林複閤經營實踐提供理論參攷。【方法】提取根際和非根際土壤真菌 DNA，採用通用引物 GC-FR1和 FF390擴增18S rDNA 基因目的片段，擴增片段用變性梯度凝膠電泳分析；不同處理間微生物群落結構的相似性採用非加權組算術平均法分析比較；以物種豐富度指數、Shannon-Wiener多樣性指數和均勻度指數3箇指標評價各樣品真菌多樣性；最後將 DGGE 圖譜中的優勢條帶割膠測序併建立繫統髮育樹。【結果】根際真菌群落結構和多樣性遠比非根際土壤複雜，且根際土壤樣品 DGGE電泳圖譜中優勢條帶數也比非根際土壤樣品多。進一步採用非加權組算術平均法對不同處理間真菌群落結構的相似性進行分析，髮現每箇處理組3次重複間的相似度除苦楝根際土壤外均大于70%；在聚類分析圖中所有處理明顯分為3箇組，相似度為51%；小麥根際真菌菌群與苦楝－小麥農林複閤繫統中的小麥根際真菌相似性最高，相似度為72%，錶明苦楝對小麥根際真菌群落的影響不大；苦楝根際真菌菌群與其他各組間差異較大，相似度僅為51%。主成分分析結果也錶明所有處理明顯分為3箇群組。多樣性分析結果錶明，各處理組根際真菌之間以及非根際真菌間豐富度差異均不明顯，而苦楝－小麥農林複閤繫統中的小麥及苦楝根際真菌豐富度顯著高于非根際真菌；根際真菌的 Shannon-Wiener 多樣性指數存在顯著差異，非根際真菌的Shannon-Wiener多樣性指數差異不顯著；根際真菌及非根際真菌各組間均勻度指數差異不大，而小麥－苦楝農林複閤繫統中的小麥以及苦楝根際真菌與非根際真菌均勻度指數存在明顯差異。此外，DGGE圖譜中的11條優勢條帶剋隆測序結果錶明，3條條帶為不能培養的真菌，其餘8條條帶對應的同源性最高的序列分彆屬于毬囊菌門、擔子菌門、子囊菌門和半知菌亞門。【結論】小麥－苦楝農林複閤繫統中的小麥根際真菌豐富度、多樣性指數和均勻度均比單純小麥根際真菌高，錶明此複閤繫統中伴生的苦楝可增加小麥根際真菌的多樣性。
【목적】채용18S rDNA PCR-DGGE기술연구고련－소맥농림복합생태계통근제급비근제토양진균군락결구，이기게시농림복합생태계통중반생수충대작물근제진균군락적영향，위농림복합경영실천제공이론삼고。【방법】제취근제화비근제토양진균 DNA，채용통용인물 GC-FR1화 FF390확증18S rDNA 기인목적편단，확증편단용변성제도응효전영분석；불동처리간미생물군락결구적상사성채용비가권조산술평균법분석비교；이물충봉부도지수、Shannon-Wiener다양성지수화균균도지수3개지표평개각양품진균다양성；최후장 DGGE 도보중적우세조대할효측서병건립계통발육수。【결과】근제진균군락결구화다양성원비비근제토양복잡，차근제토양양품 DGGE전영도보중우세조대수야비비근제토양양품다。진일보채용비가권조산술평균법대불동처리간진균군락결구적상사성진행분석，발현매개처리조3차중복간적상사도제고련근제토양외균대우70%；재취류분석도중소유처리명현분위3개조，상사도위51%；소맥근제진균균군여고련－소맥농림복합계통중적소맥근제진균상사성최고，상사도위72%，표명고련대소맥근제진균군락적영향불대；고련근제진균균군여기타각조간차이교대，상사도부위51%。주성분분석결과야표명소유처리명현분위3개군조。다양성분석결과표명，각처리조근제진균지간이급비근제진균간봉부도차이균불명현，이고련－소맥농림복합계통중적소맥급고련근제진균봉부도현저고우비근제진균；근제진균적 Shannon-Wiener 다양성지수존재현저차이，비근제진균적Shannon-Wiener다양성지수차이불현저；근제진균급비근제진균각조간균균도지수차이불대，이소맥－고련농림복합계통중적소맥이급고련근제진균여비근제진균균균도지수존재명현차이。차외，DGGE도보중적11조우세조대극륭측서결과표명，3조조대위불능배양적진균，기여8조조대대응적동원성최고적서렬분별속우구낭균문、담자균문、자낭균문화반지균아문。【결론】소맥－고련농림복합계통중적소맥근제진균봉부도、다양성지수화균균도균비단순소맥근제진균고，표명차복합계통중반생적고련가증가소맥근제진균적다양성。
Objective]To explore the influence of tree species on rhizosphere-associated fungal community in an agroforestry ecosystem, and to provide new insights into agroforestry practice, the fungal community structure in rhizosphere and bulk soil in Melia azedarach-Triticum aestivum agroforestry ecosystem was investigated by 18S rDNA PCR-DGGE.[Method]After the fungal DNA was extracted from extraction of nonrhizosphere and rhizosphere-associated soil, and the targeted fungal 18 S rDNA was amplified using universal primers GC-FR1 and FF390 . And then the amplified DNA fragments were analyzed by density gradient gel electrophoresis ( DGGE) . Similarity among different groups was analyzed using the UPGMA ( unweighted pair group method with arithmetic averages) method,and the fungal diversity was evaluated with species richness ( S ) ,Shannon-Wiener index ( H ) and the equitability index ( E ) . Finally,dominant DGGE bands were excised,sequenced and a phylogenetic tree was constructed. [Result]Results showed that the structure and diversity of rhizosphere-associated fungi community were much more complex than that of the bulk soil,and more dominant bands were observed in DGGE profiles of rhizosphere soil samples. The DGGE profiles were further investigated by cluster analysis using the UPGAMA method,revealing that the similarity among the three replicates of each treatment was more than 70% except for the rhizosphere soil of M. azedarach. In clustering diagram,all the samples were grouped into three clusters with a similarity of 51%. The highest similarity ( 72%) was found between the rhizosphere-associated fungal community from wheat and wheat grown under M. azedarach,which indicated that M. azedarach had minor influence on the rhizosphere-associated fungal community of wheat. In addition,only 51% similarity was noticed between the rhizosphere-associated fungal community of M. azedarach and the other groups. The principal-component analysis ( PCA) also demonstrated that all the treatments was divided into three groups. There were no obvious differences in species richness among rhizosphere-associated fungal communities and nonrhizosphere fungal communities. However, the species richness of rhizosphere-associated fungal community from wheat grown under M. azedarach and M. azedarach were higher than nonrhizosphere fungal community. There were significant differences in Shannon-Wiener indexes among rhizosphere-associated fungal communities, while the difference was not significant among nonrhizosphere fungal communities. Additionally,no significant differences were present in equitability index among rhizosphere-associated fungal communities or nonrhizosphere fungal communities,whereas,the equitability index between rhizosphere-associated and nonrhizosphere fungal community of wheat grown under M. azedarach was significantly different,and the same as M. azedarach. Finally,sequencing of eleven dominant DGGE bands showed that 3 of the 11 sequences were uncultured fungi and the rest 8 sequences belonged to Glomeromycota,Basidiomycota,Ascomycota or Deuteromycota. [Conclusion]The species richness,Shannon-Wiener index and equitability index of rhizosphere-associated fungi from wheat grown under M. azedarach were higher than that of monocultured wheat,which indicated that diversity of rhizosphere-associated fungal community of wheat was enriched by trees grown in this agroforestry ecosystem. Our results may provide new reference for agroforestry practice.