Comparative analysis of basidiomycetous laccase genes in forest soils reveals differences at the cDNA and DNA levels

Aims

This study was undertaken to investigate laccase-containing basidiomycete communities at the cDNA and DNA levels and to assess the influences of vegetation and soil types on the basidiomycete communities in forest soils.

Materials and methods

Soil samples were collected from the upper soil layers of two typical subtropical forests (a broad-leaved forest developed in Karst limestone soil and an artificial coniferous forest with Pinus massoniana in red soil) in China. The basidiomycete communities were characterized by cloning and sequencing of the laccase genes at both the cDNA and DNA levels. Compositions of lignin monomers were determined by gas chromatography–mass spectrometry.

Results

Most of the laccase genes obtained in this study were new, highlighting the research gap of this functional group. The trends of the basidiomycetous laccase gene diversity among the upper soil layers of the two forests were consistent between the cDNA and DNA levels. The Agaricales had high activity because they dominated all the tested soils. However, the total basidiomycete communities reflected at the cDNA and DNA levels were significantly different due to the presence of some quiescent basidiomycetous groups. Almost all of the lignin components were decomposed from the O to the A layers in the two forests, and laccases produced by Agaricales were likely responsible for the decomposition of guaiacyl monomers. Both vegetation and soil types had great influences on the active laccase-containing basidiomycete communities, primarily via the pH, C/N, and the contents of lignin monomers.

Conclusions

The cDNA- and DNA-level approaches presented good consistency of diversities but different compositions of laccase-containing basidiomycete communities, thus emphasizing the importance of focusing on laccase genes at the cDNA level in future studies. It is the quality but not the quantity of SOM to determine the diversity and composition of the active laccase-containing basidiomycete communities.     

亚热带两种森林土壤担子菌漆酶基因多样性比较

漆酶是降解森林凋落物中木质素的关键酶之一,直接影响着森林生态系统碳循环过程.运用TA克隆、测序技术,研究了两种亚热带森林(原生常绿落叶阔叶混交林和人工马尾松林)凋落物层(O层)和土壤表层(A层,0～20 cm)降解木质素的担子菌漆酶基因多样性.结果表明:同一土壤层位,原生林土壤中担子菌漆酶基因多样性和种群丰富度高于马尾松林;同一森林生态系统,原生林土壤O层中担子菌漆酶基因多样性和种群丰富度略高于土壤A层,而马尾松林则O层明显低于A层;两森林土壤具有相同含漆酶基因的担子菌优势种群,且大部分优势种群与伞菌目小菇属或侧耳属有较高的氨基酸相似性;与原生林土壤A层和马尾松林土壤O层相比,原生林土壤O层和马尾松林土壤A层中含漆酶基因的担子菌种群分布相对均匀;马尾松林O层与A层之间漆酶基因核苷酸序列的相似性较原生林土壤O层与A层之间的高.表明植被和土壤层位显著影响漆酶基因多样性和群落结构,而植被和土壤层位引起的担子菌可利用底物和土壤pH值的差异可能直接驱动这种影响.     

鼎湖山不同演替阶段森林土壤细菌多样性

[目的]了解鼎湖山早、中、后3个演替阶段的3类森林(针叶林、混交林、阔叶林)土壤细菌群落结构及其多样性,为下一步研究不同演替阶段森林土壤微生物的功能及其与植物的相互作用提供依据.[方法]在代表性林区采集土样,从中提取总DNA,利用细菌通用引物27F和1492R PCR扩增16S rDNA并构建文库.从所构建的3个文库中各挑取150个阳性克隆子并对插入片段进行测序,利用Mothur软件对所得序列进行分析.[结果]从针叶林、混交林和阔叶林文库中分别得到122、118和120条有效16S rDNA序列,各代表70、64、72个OTUs(operational taxonomic units,以97％相似性为划分标准).分析结果显示,共检测到8个细菌门类,其中酸杆菌门(Acidobacteria)在针叶林、混交林和阔叶林土样中分别占53.3％,67.8％和60％ ;变形杆菌门(Proteobacteria)分别占29.5％,20.3％和32.5％ ;其它如厚壁菌,放线菌等均不超过10％.3类森林土壤细菌群落结构差异显著(P＜0.05),3者两两间共有的OTU数量占检测到的OTU总数的比例均低于25％,其中阔叶林土壤细菌有着最高的Chao指数(414.2)和Shannon指数(3.90),及最低的Simpson优势度指数(0.0249).[结论]鼎湖山针叶林、混交林和阔叶林3类林区土壤细菌在种群构成上差异显著,其中阔叶林土壤细菌丰富度及多样性相对较高,但3者在大类组成方面比较相似,均为酸杆菌占绝对优势,变形杆菌次之.     

Genome diversity in microbial eukaryotes

The genomic peculiarities among microbial eukaryotes challenge the conventional wisdom of genome evolution. Currently, many studies and textbooks explore principles of genome evolution from a limited number of eukaryotic lineages, focusing often on only a few representative species of plants, animals and fungi. Increasing emphasis on studies of genomes in microbial eukaryotes has and will continue to uncover features that are either not present in the representative species (e.g. hypervariable karyotypes or highly fragmented mitochondrial genomes) or are exaggerated in microbial groups (e.g. chromosomal processing between germline and somatic nuclei). Data for microbial eukaryotes have emerged from recent genome sequencing projects, enabling comparisons of the genomes from diverse lineages across the eukaryotic phylogenetic tree. Some of these features, including amplified rDNAs, subtelomeric rDNAs and reduced genomes, appear to have evolved multiple times within eukaryotes, whereas other features, such as absolute strand polarity, are found only within single lineages.     

Unraveling the genomic diversity of small eukaryotes

A report of the meeting Comparative Genomics of Eukaryotic Microorganisms, 17-22 October 2009, San Feliu de Guixols, Spain.     

Unraveling the genomic diversity of small eukaryotes

A report of the meeting Comparative Genomics of Eukaryotic Microorganisms, 17-22 October 2009, San Feliu de Guixols, Spain.     

Isolation of fungal cellobiohydrolase I genes from sporocarps and forest soils by PCR

Cellulose is the major component of plant biomass, and microbial cellulose utilization is a key step in the decomposition of plant detritus. Despite this, little is known about the diversity of cellulolytic microbial communities in soil. Fungi are well known for their cellulolytic activity and mediate key functions during the decomposition of plant detritus in terrestrial ecosystems. We developed new oligonucleotide primers for fungal exocellulase genes (cellobiohydrolase, cbhI) and used these to isolate distinct cbhI homologues from four species of litter-decomposing basidiomycete fungi (Clitocybe nuda, Clitocybe gibba, Clitopilus prunulus, and Chlorophyllum molybdites) and two species of ascomycete fungi (Xylaria polymorpha and Sarcoscypha occidentalis). Evidence for cbhI gene families was found in three of the four basidiomycete species. Additionally, we isolated and cloned cbhI genes from the forest floor and mineral soil of two upland forests in northern lower Michigan, one dominated by oak (Quercus velutina, Q. alba) and the other dominated by sugar maple (Acer saccharum) and American basswood (Tilia americana). Phylogenetic analysis demonstrated that cellobiohydrolase genes recovered from the floor of both forests tended to cluster with Xylaria or in one of two unidentified groups, whereas cellobiohydrolase genes recovered from soil tended to cluster with Trichoderma, Alternaria, Eurotiales, and basidiomycete sequences. The ability to amplify a key fungal gene involved in plant litter decomposition has the potential to unlock the identity and dynamics of the cellulolytic fungal community in situ.     

Microbial diversity of topographical gradient profiles in Fushan forest soils of Taiwan

To evaluate the microbial diversity of Fushan forest soils, the variation of soil properties, microbial populations, and soil DNA with soil depth in three sites of different altitude were analyzed. Microbial population, moisture content, total organic carbon (C_org), and total nitrogen (N_tot) decreased with increasing soil depth. The valley site had the lowest microbial populations among the three tested sites due to the low organic matter content. Bacterial population was the highest among the microbial populations. The ratios of cellulolytic microbes to the total bacteria in organic layers were high, implying their roles in the carbon cycle. The microbial biomass carbon (C_mic) and nitrogen (N_mic) contents ranged from 130.5 to 564.1 μg g⁻¹ and from 16.7 to 95.4 μg g⁻¹, respectively. The valley had the lowest C_mic and N_mic. The organic layer had the highest C_mic and N_mic and decreased with soil depth. Analysis using denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) amplicons of 16S rDNA showed that the bacterial diversity of the three sites were very similar to each other in the major bands, and the variation was in the minor bands. However, the patterns in PCR-DGGE profile through gradient horizons were different, indicating the prevalence of specific microbes at different horizons. These results suggest that the microbial diversity in the deeper horizons is not simply the diluted analogs of the surface soils and that some microbes dominate only in the deeper horizons. Topography influenced the quantity and diversity of microbial populations.     

Diversity of tRNA genes in eukaryotes

We compare the diversity of chromosomal-encoded transfer RNA (tRNA) genes from 11 eukaryotes as identified by tRNAScan-SE of their respective genomes. They include the budding and fission yeast, worm, fruit fly, fugu, chicken, dog, rat, mouse, chimp and human. The number of tRNA genes are between 170 and 570 and the number of tRNA isoacceptors range from 41 to 55. Unexpectedly, the number of tRNA genes having the same anticodon but different sequences elsewhere in the tRNA body (defined here as tRNA isodecoder genes) varies significantly (10–246). tRNA isodecoder genes allow up to 274 different tRNA species to be produced from 446 genes in humans, but only up to 51 from 275 genes in the budding yeast. The fraction of tRNA isodecoder genes among all tRNA genes increases across the phylogenetic spectrum. A large number of sequence differences in human tRNA isodecoder genes occurs in the internal promoter regions for RNA polymerase III. We also describe a systematic, ligation-based method to detect and quantify tRNA isodecoder molecules in human samples, and show differential expression of three tRNA isodecoders in six human tissues. The large number of tRNA isodecoder genes in eukaryotes suggests that tRNA function may be more diverse than previously appreciated.     

Carbon flux and diversity of nematodes and termites in Cameroon forest soils

Theoretically, there are three principal ways in which ecosystem processes might respond to reductions in species richness. These theories are reviewed, and then considered in the context of a study of the diversity of soil nematodes and termites in near-primary forest sites at Mbalmayo, Cameroon, and the contribution made by these two taxa to carbon fluxes (CO₂ and CH₄) from the forest floor. Nematode abundances average 2.04 × 10⁶ m^-2, and termites between 2933 and 6957 m^-2. The site is the most species-rich yet investigated for both groups anywhere in the world, so that a very large number of species contribute to carbon fluxes. We speculate about how much redundancy might be built into the functioning of both assemblages, and point out the enormous difficulties of resolving such questions, and of producing such detailed species-inventories.     

Mismatch repair genes of eukaryotes

Mismatches that arise during replication or genetic recombination or owing to damage to DNA by chemical agents are recognized by mismatch repair systems. The pathway has been characterized in detail inEscherichia coll. Several homologues of the genes encoding the proteins of this pathway have been identified in the yeastSaccharomyces cerevisiae and in human cells. Mutations in the human geneshMSH2, hMLH1, hPMS1 andhPMS2 have been linked to hereditary nonpolyposis colon cancer (HNPCC) and to some sporadic tumours. Mismatch repair also plays an antirecombinogenic role and is implicated in speciation.