Zhiwei Zhao

Improved tolerance of maize (Zea mays L.) to heavy metals by colonization of a dark septate endophyte (DSE) Exophiala pisciphila

Dark septate endophytes (DSE) are ubiquitous and abundant in stressful environments including heavy metal (HM) stress. However, our knowledge about the roles of DSE in improving HM tolerance of their host plants is poor. In this study, maize (Zea mays L.) was inoculated with a HM tolerant DSE strain Exophiala pisciphila H93 in lead (Pb), zinc (Zn), and cadmium (Cd) contaminated soils. E. pisciphila H93 successfully colonized and formed typical DSE structures in the inoculated maize roots. Colonization of E. pisciphila H93 alleviated the deleterious effects of excessive HM supplements and promoted the growth of maize (roots and shoots) under HM stress conditions, though it significantly decreased the biomass of inoculated maize under no HM stress. Further analysis showed that the colonization of E. pisciphila H93 improved the tolerance of maize to HM by restricting the translocation of HM ions from roots to shoots. This study demonstrated that under higher HM stress, such a mutual symbiosis between E. pisciphila and its host (maize) may be an efficient strategy to survive in the stressful environments.

Recombination and genetic differentiation among natural populations of the ectomycorrhizal mushroom Tricholoma matsutake from southwestern China

Effective conservation and utilization strategies for natural biological resources require a clear understanding of the natural populations of the target organisms. Tricholoma matsutake is an ectomycorrhizal mushroom that forms symbiotic associations with plants and plays an important ecological role in natural forest ecosystems in many parts of the world. It is also an economically very important gourmet mushroom. Because no artificial cultivation is available, natural populations of this species are under increasing threats, primarily from habitat disturbance and destruction. Despite its economical and ecological importance, little is known about its genetics and population biology. Here, using 14 polymerase chain reaction–restriction fragment length polymorphism markers, we analysed 154 strains from 17 geographical locations in southwestern China, a region where over 25% of the global T. matsutake harvest comes from. Our results revealed abundant genetic variation within individual populations. The analyses of gene and genotype frequencies within populations indicated that most loci did not deviate from Hardy–Weinberg equilibrium in most populations and that alleles among loci were in linkage equilibrium in the majority of the local populations. These results are consistent with the hypothesis that sexual reproduction and recombination play an important role in natural populations of this species. Our analyses indicated low but significant genetic differentiation among the geographical populations, with a significant positive correlation between genetic distance and geographical distance. We discuss the implications of our results to the ecology and resource management of this species.

Genetic diversity of the endemic gourmet mushroom Thelephora ganbajun from south-western China

The ectomycorrhizal fungus Thelephora ganbajun is an endemic gourmet mushroom in Yunnan province, south-western China. However, despite its widespread consumer appeal, nutritional value and potential ecological role in natural forests, very little is known about its genetics, diversity and ecology. In this study, we investigated DNA sequence variation at the internal transcribed spacer (ITS) regions among 156 specimens collected from 23 sites of nine regions in Yunnan Province. Our analysis identified a total of 34 ITS haplotypes and these haplotypes were clustered into five distinct phylogenetic groups. The evolutionary divergences among these clades are similar to or greater than many known sister species pairs within the genus Thelephora and the closely related genus Tomentella. Among the 34 ITS haplotypes, 22 were represented by one specimen each and the remaining 12 were each shared by two or more specimens. The most common haplotype contained 68 specimens distributed in 21 of the 23 sites, a result consistent with gene flow among geographical populations. However, analysis of molecular variance (AMOVA) revealed low but significant genetic differentiation among local and regional populations. Interestingly, the Mantel test identified that the extent of genetic differentiation was not significantly correlated with geographical distance. Our study revealed significant genetic divergence within Th. ganbajun and limited but detectable gene flow among geographical populations of this endemic ectomycorrhizal gourmet mushroom.

Lateral gene transfer occurring in haloarchaea: an interpretative imitation study

Lateral gene transfer (LGT) plays an important role in the molecular evolution of haloarchaea. Polyethylene glycol-mediated LGT in haloarchaea has been demonstrated in the laboratory, yet few explanations have been put forward for the apparently common, natural occurrence of plentiful plasmids within haloarchaeal cells. In this study, LGT was induced in two genera of haloarchaea, Haloferax and Halorubrum, by modification of salt concentration of media—a factor that may vary naturally in native haloarchaeal habitat. Minimal growth salt concentrations (MGSCs) of four strains of haloarchaea from these two genera were established, and transformations using two circular double-stranded DNAs (dsDNAs), pSY1 and pWL102, were then produced in media at strain-appropriate MGSCs. The four strains of haloarchaea were transformed successfully by both kinds of dsDNAs with an efficiency of 102–103 transformants per microgram dsDNA. The transformation under reduced salt concentration may be an imitation of natural LGT of dsDNA into haloarchaea when salinity in normally hypersaline environments is altered by sudden introduction of fresh water—for example, by rainfall, snow-melt, or flooding—providing a reasonable interpretation for haloarchaea being naturally richer in plasmids than any other known organisms.

Species clarification of oyster mushrooms in China and their DNA barcoding

Species of the Pleurotus ostreatus complex (oyster mushrooms) are one of the main groups of cultivated edible mushrooms in China, with abundant strains and a complex genetic background. Although the cultivation of oyster mushrooms is a very important industry in China, the inconsistent nomenclature of the complex is controversial and has led to much confusion and economic loss in the mushroom industry. In this study, we performed molecular identification of the species complex based on sequences of the internal transcribed spacer region (ITS), translation elongation factor 1-α (TEF1), and two genes that encode subunits of RNA polymerase II (RPB1 and RPB2). A total of 284 samples with different commercial names gathered from different mushroom spawn preservation centers, companies, and field isolations were investigated via phylogenetic analyses inferred from a single locus or multi-loci. Our analyses indicated that 56% of the strains were labeled with improper scientific names and that all of the strains could be divided into seven lineages, representing one morphological species each, namely P. abieticola, P. eryngii, P. cf. floridanus, P. ostreatus, P. placentodes, P. pulmonarius, and P. tuoliensis. Reference sequences for accurate identification of the taxa were developed or suggested. Meanwhile, we evaluated the suitability of the four candidate segments as DNA barcodes. Our analyses indicated that RPB2 was the most promising candidate segment of DNA barcode for the P. ostreatus species complex, taking into consideration the polymorphisms and other aspects of the four markers.