Survival of bacterial DNA and culturable bacteria in archived soils from the Rothamsted Broadbalk experiment

Dried soil samples from many sources have been stored in archives world-wide over the years, but there has been little research on their value for studying microbial populations. Samples collected since 1843 from the Broadbalk field experiment on crop nutrition at Rothamsted have been used to document changes in the structure and composition of soils as agricultural practices evolve, also offering an invaluable record of environmental changes from the pre- to post-industrial era in the UK. To date, the microbial communities of these soils have not been studied, in part due to the well-documented drop in bacterial culturability in dried soils. However, modern molecular methods based on PCR amplification of DNA extracted directly from soil do not require bacterial cells to be viable or intact and may allow investigations into the legacy of bacteria that were present at the time of sample collection.

In a preliminary study, to establish if dried soils can provide a historical record of bacterial communities, samples from the Broadbalk soil archive dating back to 1868 were investigated and plots treated with either farmyard manure (FYM) or inorganic fertilizer (NPK) were compared. As anticipated, the processes of air-drying and milling greatly reduced bacterial viability whilst DNA yields declined less and may be preserved by desiccation. A higher proportion of culturable bacteria survived the archiving process in the FYM soil, possibly protected by the increased soil organic matter. The majority of surviving bacteria were firmicutes, whether collected in 2003 or in 1914, but a wide range of genera was detected in DNA extracted from the samples using PCR and DGGE of 16S rRNA genes. Analysis of DGGE band profiles indicated that the two plots maintained divergent populations. Sequence analysis of bands excised from DGGE gels, from a sample collected in 1914, revealed DNA from - and β-proteobacteria as well as firmicutes. PCR using primers specific for ammonia oxidizing bacteria showed similar band profiles across the two treatments in recently collected samples, however older samples from the NPK plot showed greater divergence. Primers specific for the genus Pseudomonas were designed and used in real-time quantitative PCR to indicate that archived soil collected in 1868 contained 10-fold less pseudomonad DNA than fresh soil, representing around 10⁵ genomes g⁻¹ soil. Prior to milling, dramatically less pseudomonad DNA was extracted from recently collected air-dried soil from the NPK compared to the FYM plot; otherwise, the two plots followed similar trends. Overall bacterial abundance, diversity and survival during the archiving process differed in the two soils, possibly due to differences in clay and soil organic matter content. Nevertheless, the results demonstrate that air-dried soils can protect microbial DNA for more than 150 years and offer an invaluable resource for future research.     

土壤中多菌灵的降解动态及其对土壤微生物群落多样性的影响

采用实验室模拟方法研究了多菌灵在土壤生态系统中的降解动态及其对土壤微生物群落多样性的影响。结果表明,2.0和4.0 mg kg-1多菌灵在土壤中的降解半衰期分别为8.6和6.8 d。试验初期各个处理的土壤微生物群落的AWCD值显著降低,土壤微生物群落的Shannon指数、Simpson指数和McIntosh指数均低于不加多菌灵的对照,微生物群落的丰富度、均匀度和优势度受到抑制,并且多菌灵对土壤微生物的抑制作用随浓度的增加而增强。随着时间的延长,多菌灵对土壤微生物的毒性逐渐减弱,3 d后土壤微生物多样性逐渐恢复到对照水平。     

中国土壤生物学研究的回顾与展望

土壤生物是土壤发生与发展的重要推动力,是土壤生态系统中最为活跃的组成部分。本文在系统查阅近10年来国内外土壤生物学文献的基础上,分析了中国土壤生物学研究的现状特点,从土壤生态学、土壤生物化学、土壤微生物学、土壤?植物系统、土壤动物学几个方面概括了土壤生物学研究的主要进展。结合国际土壤科学发展的前沿和热点问题,建议重点开展土壤生物与全球环境变化、土壤生物与土壤质量、土壤生物多样性与生态功能、微生物宏基因组学与活性物质开发、土壤污染与生物修复、土壤酶学与元蛋白质组技术等方面的研究。     

Genetic diversity of the African hexaploid species Solanum scabrum Mill. and Solanum nigrum L. (Solanaceae)

Two hexaploid species of Solanum sect. Solanum are present in Africa: Solanum scabrum and S. nigrum. Solanum scabrum is a widely cultivated species and is used as a leafy vegetable, as a source of medicine and as a source of ink dye. In previous studies a wide range of morphological diversity has been reported in this species and in some studies subspecies have been proposed. Subspecies are also recognized in S. nigrum. However, it has not been established whether or not the morphological differences are reflected at the genomic level. The present study applies AFLPs to study the genetic diversity in S. scabrum and its relationship to geographical provenance, morphological differences and the possible existence of subspecies within S. scabrum and S. nigrum. The data obtained were analyzed with cluster analysis (using UPGMA and NJ). The results indicate that the genetic variation within S. scabrum was higher within accessions than between accessions. Accessions did not cluster according to their geographical provenance, indicating that accessions from different geographical areas were not significantly different genetically. The clustering reflected neither morphological differences nor domestication status (cultivated or wild). The morphological differences exhibited by S. scabrum could be due to selection by farmers for different plant types. The AFLP derived clustering pattern did not segregate the subspecies recognized in S. scabrum and S. nigrum into separate subclusters.     

Genetic characterization of a collection of chayote, <Emphasis Type="Italic">Sechium edule</Emphasis> (Jacq.) Swartz,in Costa Rica by using isozyme markers

We established protocols for the analysis of genetic diversity in chayote (Sechium edule) by using isozyme markers, thereby determining the level of genetic diversity present in 42 accessions of chayote from Costa Rica. We obtained clear and reproducible zymograms for eight enzyme staining systems: PGM, 6-PGD, PGI, IDH, MDH, SOD, SKD, and EST, and were able to score 14 putative loci. Eight of the 14 loci examined were polymorphic. We found 35 distinct multilocus genotypes among these accessions. Five of these multilocus genotypes were homozygous for all loci. In addition, our data also revealed that most of the multilocus genotypes (24) were heterozygous for only one of the eight loci, and the rest were heterozygous for two or three loci (9 and 4 accessions, respectively). Seven multilocus genotypes were found in two different accessions. Dice similarity coefficient was used to study the relationship between accessions. This analysis, based on the presence and absence of alleles, revealed that accessions collected in the same location seldom shared the same multilocus genotype. The value of isozyme polymorphisms as tools to continue studies on the characterization of chayote is discussed.     

Morphological characterization of seeds of three Australian wild Citrus species (Rutaceae): Citrus australasica F. Muell., C. inodora F.M. Bailey and C. garrawayi F.M. Bailey

The comparative morphology of the seeds of three Australian Citrus species, C. australasica C. inodora and C. garrawayi, was studied. Their seed characteristics were broadly similar to those of the cultivated species of the genus, when observed under light and scanning electron microscopy. Citrus garrawayi differed in seed shape (rounded to triangular) and seed coat morphology (i.e., thicker with longer epidermal protrusions) from C. australasica and C. inodora (rounded surface with flat underside in shape). The well-developed minute epidermal protrusions on the seed coat of C. garrawayi were more similar to those in the cultivated species, C. × sinensis and C. × aurantium. In contrast, the surface topography of C. australasica and C. inodora seeds was more like that of the cultivated species, C. × aurantifolia and C. × limon. Seed morphology, especially surface topography, was found to be a useful tool for taxonomic identification in Australian wild citrus.     

Diversity, distribution and management of yam landraces (Dioscorea spp.) in Southern Ethiopia

Yam (Dioscorea spp.) is widely grown in many parts of Ethiopia and plays a vital role in local subsistence. Nevertheless, its diversity has not been studied in detail. A survey covering 339 farm households and eight districts was conducted in the major yam growing regions of Southern Ethiopia to investigate the diversity and distribution of yam landraces using structured and semi-structured questionnaires. A total of 37 named landraces were recorded, with a range from one to six (mean 2.9) on individual farms. Farmers’ decisions regarding the number and type of landraces maintained was influenced by tolerance of the landraces to drought, their maturity time and market demand. Most landraces had limited abundance and distribution, and only a few dominant landraces were widely grown. There was also variation amongst districts with respect to diversity, distribution and abundance of the landraces found. In the majority of the localities surveyed, farmers reported a decreasing trend in the number of landraces maintained on individual farms and in the overall yam production. Besides, in those limited areas where yam production is expanding, farmers are increasingly relying on a few selected landraces that mature early. Findings of this study suggest that local farmers in Wolayita and Gamo-Gofa zones maintain considerable yam diversity that remains to be further explored for sustainable utilization and conservation of the available genetic resources.     

Isozyme polymorphism and genetic differentiation in natural populations of an endemic tetraploid species <Emphasis Type="Italic">Avena maroccana</Emphasis> in Morocco

A wild tetraploid oat Avena maroccana Gdgr. was collected from the 11 populations in the periphery of Rommani and Casablanca geographic groups of Morocco. Genetic diversity of the species was investigated using six allozyme systems. Allelic frequencies were scored representing eight polymorphic and five monomorphic loci. Coefficient of gene differentiation (Gst) was 0.3019, which indicated great genetic differentiation. The number of alleles per locus was 2.6154, the percentage of polymorphic loci was 61.54, and the expected heterozygosity was 0.2462 in all populations. Genetic diversity in A. maroccana was high in comparison to self-pollinated species. In total, nine heterozygotes resulting from outcrossing were found in the progeny from M1, M3, M4, M22 and M26. The population of M7 had peculiar alleles Pgd–2SS and Pgd-1SS in high frequency. M9 had the lowest level of diversity out of the 11 populations. Geographic and genetic distances between all the populations were not significantly correlated with each other (r = 0.0996). Cluster analysis showed that two groups, (M1, M22, M2 and M4) and (M3, M23, M8, M5 and M26) were apparently differentiated. Two populations of the Casablanca group, M7 and M9 were independent from each other, and were separated distinctly from the other populations. Genetic diversity of the Rommani and Casablanca groups was almost the same in all the parameters. This was due to the similar man-made habitat such as roadside or rich fertile soil and brown clay soils. The population size of A. maroccana was small and restricted to the narrow central Morocco with great genetic differentiation so that genetic diversity may be reflected from the results of genetic drift and outcrossing heterozygote segregation.