不同喀斯特小生境中土壤丛枝菌根真菌的遗传多样性

doi:10.3724/SP.J.1258.2011.01083

摘要/Abstract

摘要：

为探明西南喀斯特地区小生境土壤中丛枝菌根真菌(AMF)的遗传多样性特征, 利用巢式PCR和变性梯度凝胶电泳相结合的分子生物学方法, 对茂兰3种植被类型下的小生境(石缝、石沟、土面) AMF遗传多样性进行了研究。结果发现: 各类小生境都含有丰富的AMF遗传多样性, 灌木林土面的多样性指数和物种丰富度最高, 为4.06和68; 次生林石缝的最低, 为3.16和29, 所研究的9个小生境多样性指数和物种丰富度的平均值分别高达3.67和48, 高于同类研究在其他地区的结果, 这可能主要与喀斯特生态系统复杂的结构和较高的植物多样性有关; 聚类分析显示各类小生境间的AMF群落结构差异显著, 相似性指数最高仅为0.45, 说明小生境所带来的空间异质性对AMF的遗传多样性产生了显著影响; 基因测序显示球囊霉属(Glomus)极有可能是喀斯特地区AMF的优势菌属, 在以后筛选喀斯特地区的高效生态恢复菌种时可重点考虑球囊霉属的一些菌种。

关键词: 丛枝菌根真菌, 遗传多样性, 喀斯特, 小生境, 丰富度

Abstract:

Aims Our objective was to explore the genetic diversity of arbuscular mycorrhizal fungi (AMF) in karst microhabitat.
Methods We used nested polymerase chain reaction and denaturing gradient gel electrophoresis to study the genetic characteristics of AMF sampled from microhabitats of three different vegetation types in Maolan National Nature Reserve of Guizhou Province.
Important findings The AMF genetic diversities in all three microhabitat types were rich. The highest values of biodiversity index (4.06) and species abundance (68) were found in the shrubland soil surface, and the lowest values (3.16 and 29, respectively) were found in the secondary forest rocky crevice. The average values of biodiversity index and species abundance of the 9 microhabitats studied were 3.67 and 48, respectively. These were much higher than those in some other areas; the possible reasons were the complex karst ecological system structure and high plant species diversity. The AMF genetic diversity of different microhabitat types showed significant differences. The highest similarity index was only 0.45, and this means the spatial heterogeneity of different microhabitat types had significant influence on AMF genetic diversity. Glomus was the dominant genus through the karst area and should be a candidate for screening high ecological restoration strains for karst areas.

Key words: arbuscular mycorrhizal fungi, genetic diversity, karst, microhabitat, richness

魏源, 王世杰, 刘秀明, 黄天志. 不同喀斯特小生境中土壤丛枝菌根真菌的遗传多样性. 植物生态学报, 2011, 35(10): 1083-1090. DOI: 10.3724/SP.J.1258.2011.01083

WEI Yuan, WANG Shi-Jie, LIU Xiu-Ming, HUANG Tian-Zhi. Genetic diversity of arbuscular mycorrhizal fungi in karst microhabitats of Guizhou Province, China. Chinese Journal of Plant Ecology, 2011, 35(10): 1083-1090. DOI: 10.3724/SP.J.1258.2011.01083

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图/表 5

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引物 Primer	序列 Sequence	长度 Length	来源 Sources
GeoA2	5′-CCAGTAGTCATATGCTTGTCTC-3′	1.80 kb	Schwarzott & Schüβler, 2001
Geo11	5′-ACCTTGTTACGACTTTTACTTCC-3′	1.80 kb	Schwarzott & Schüβler, 2001
AM1	5′-GTTTCCCGTAAGGCGCCGAA-3′	0.55 kb	Helgason et al., 1998 Simon et al., 1992
NS31-GC	5′-TTGGAGGGCAAGTCTGGTGCC-3′	0.55 kb	Helgason et al., 1998 Simon et al., 1992
Glol	5′-GCCTGCTTTAAACACTCTA-3′	0.23 kb	Comejo et al., 2004

引物 Primer	序列 Sequence	长度 Length	来源 Sources
GeoA2	5′-CCAGTAGTCATATGCTTGTCTC-3′	1.80 kb	Schwarzott & Schüβler, 2001
Geo11	5′-ACCTTGTTACGACTTTTACTTCC-3′	1.80 kb	Schwarzott & Schüβler, 2001
AM1	5′-GTTTCCCGTAAGGCGCCGAA-3′	0.55 kb	Helgason et al., 1998 Simon et al., 1992
NS31-GC	5′-TTGGAGGGCAAGTCTGGTGCC-3′	0.55 kb	Helgason et al., 1998 Simon et al., 1992
Glol	5′-GCCTGCTTTAAACACTCTA-3′	0.23 kb	Comejo et al., 2004

样品 Sample	Shannon多样性指数 Shannon’s diversity index	丰富度 Richness	均匀度 Evenness
原生林石缝 Original forest rocky crevice	3.89	57	0.96
原生林石沟 Original forest rocky gully	3.74	48	0.97
原生林土面 Original forest soil surface	3.33	36	0.93
次生林石缝 Secondary forest rocky crevice	3.16	29	0.94
次生林石沟 Secondary forest rocky gully	3.65	48	0.94
次生林土面 Secondary forest soil surface	3.71	50	0.95
灌木林石缝 Shrubbery rocky crevice	3.68	50	0.94
灌木林石沟 Shrubbery rocky gully	3.77	48	0.97
灌木林土面 Shrubbery soil surface	4.06	68	0.96

样品 Sample	Shannon多样性指数 Shannon’s diversity index	丰富度 Richness	均匀度 Evenness
原生林石缝 Original forest rocky crevice	3.89	57	0.96
原生林石沟 Original forest rocky gully	3.74	48	0.97
原生林土面 Original forest soil surface	3.33	36	0.93
次生林石缝 Secondary forest rocky crevice	3.16	29	0.94
次生林石沟 Secondary forest rocky gully	3.65	48	0.94
次生林土面 Secondary forest soil surface	3.71	50	0.95
灌木林石缝 Shrubbery rocky crevice	3.68	50	0.94
灌木林石沟 Shrubbery rocky gully	3.77	48	0.97
灌木林土面 Shrubbery soil surface	4.06	68	0.96

条带来源(GenBank登录号) Band source (GenBank accession number)	最近相似种属 Closest genera	相似性 Similarity
原生林石缝 Original forest rocky crevice (JN153048)	用DGGE方法分离的属于未培养的球囊霉属的18S RNA菌种 Uncultured Glomus isolate DGGE gel band 22-1 18S ribosomal RNA gene (HQ874638)	88%
原生林石沟 Original forest rocky gully (JN153041)	用DGGE方法分离的属于未培养的球囊霉属的18S RNA菌种 Uncultured Glomus isolate DGGE gel band 11-2 18S ribosomal RNA gene (HQ874635)	86%
原生林土面 Original forest soil surface (JN153042)	用克隆方法分离的属于未培养的球囊霉属的18S RNA菌种 Uncultured Glomus clone OEF89 18S ribosomal RNA gene (EU340303)	99%
次生林石缝 Secondary forest rocky crevice (JN153043)	用DGGE方法分离的属于未培养的球囊霉属的18S RNA菌种 Uncultured Glomus isolate DGGE gel band 22-2 18S ribosomal RNA gene (HQ874639)	95%
次生林石沟 Secondary forest rocky gully (JN153044)	用克隆方法分离的属于未培养的球囊霉属的18S RNA菌种 Uncultured Glomus clone Ap7 18S ribosomal RNA gene (EU350053)	95%
次生林土面 Secondary forest soil surface (JN153040)	用克隆方法分离的属于未培养的球囊霉属的18S RNA菌种 Uncultured Glomus clone FEA6HBX02GK5NA 18S ribosomal RNA gene (GU198582)	93%
灌木林石缝 Shrubbery rocky crevice (JN153045)	未培养的球囊霉属的18S RNA菌种 Uncultured Glomus gene for 18S ribosomal RNA (AB556933)	97%
灌木林石沟 Shrubbery rocky gully (JN153046)	未培养的球囊霉属的18S RNA菌种, Uncultured Glomus partial 18S rRNA gene (AJ563889)	96%
灌木林土面 Shrubbery soil surface (JN153047)	用DGGE方法分离的属于未培养的球囊霉属的18S RNA菌种 Uncultured Glomus isolate DGGE gel band 22-2 18S ribosomal RNA gene (HQ874639)	91%