Novel insights into the kinetics and mechanism of arsenopyrite bio-dissolution enhanced by pyrite.

Arsenopyrite and pyrite often coexist in metal deposits and tailings, thus simultaneous bioleaching of both sulfides has economic (as well as environmental) significance. Important targets in bio-oxidation operations are high solubilization rates and minimized accumulation of Fe(III)/As-bearing secondary products. This study investigated the role of pyrite bioleaching in the enhancement of arsenopyrite dissolution. At a pyrite to arsenopyrite mass ratio of 1:1, 93.6% of As and 93.0% of Fe were solubilized. The results show that pyrite bio-oxidation can promote arsenopyrite dissolution, enhance S0 bio-oxidation, and inhibit the formation of jarosites, tooeleite, and amorphous ferric arsenate. The dry weight of the pyrite & arsenopyrite residue was reduced by 95.1% after bioleaching, compared to the initial load, while only 5% weight loss was observed when pyrite was absent. A biofilm was formed on the arsenopyrite surface in the presence of pyrite, while a dense passivation layer was observed in the absence of pyrite. As(III) (as As2O3) was a dominant As species in the pyrite & arsenopyrite residue. Novel and detailed findings are presented on arsenopyrite bio-dissolution in the presence of pyrite, and the presented approach could contribute to the development of novel cost-effective extractive bioprocesses. ENVIRONMENTAL IMPLICATION: The oxidation of arsenopyrite presents significant environmental hazards, as it can contribute to acid mine drainage generation and arsenic mobilization from sulfidic mine wastes. Bioleaching is a proven cost-effective and environmentally friendly extractive technology, which has been applied for decades in metal recovery from minerals or tailings. In this work, efficient extraction of arsenic from arsenopyrite bioleaching was presented through coupling the process with bio-oxidation of pyrite, resulting in lowered accumulation of hazardous and metastable Fe(III)/As-bearing secondary phases. The results could help improve current biomining operations and/or contribute to the development of novel cost-effective bioprocesses for metal extraction.

Isolation and identification of pathogens of Morchella sextelata bacterial disease.

Morel mushroom (Morchella spp.) is a rare edible and medicinal fungus distributed worldwide. It is highly desired by the majority of consumers. Bacterial diseases have been commonly observed during artificial cultivation of Morchella sextelata. Bacterial pathogens spread rapidly and cause a wide range of infections, severely affecting the yield and quality of M. sextelata. In this study, two strains of bacterial pathogens, named M-B and M-5, were isolated, cultured, and purified from the tissues of the infected M. sextelata. Koch's postulates were used to determine the pathogenicity of bacteria affecting M. sextelata, and the pathogens were identified through morphological observation, physiological and biochemical analyses, and 16S rRNA gene sequence analysis. Subsequently, the effect of temperature on the growth of pathogenic bacteria, the inhibitory effect of the bacteria on M. sextelata on plates, and the changes in mycelial morphology of M. sextelata mycelium were analyzed when M. sextelata mycelium was double-cultured with pathogenic bacteria on plates. The results revealed that M-B was Pseudomonas chlororaphis subsp. aureofaciens and M-5 was Bacillus subtilis. Strain M-B started to multiply at 10-15°C, and strain M-5 started at 15-20°C. On the plates, the pathogenic bacteria also produced significant inhibition of M. sextelata mycelium, and the observation of mycelial morphology under the scanning electron microscopy revealed that the inhibited mycelium underwent obvious drying and crumpling, and the healthy mycelium were more plump. Thus, this study clarified the pathogens, optimal growth environment, and characteristics of M. sextelata bacterial diseases, thereby providing valuable basic data for the disease prevention and control of Morchella production.

Community composition and trophic mode diversity of fungi associated with fruiting body of medicinal Sanghuangporus vaninii.

BACKGROUND: The microbial symbionts of macrofungal fruiting body have been shown to play momentous roles in host growth, development, and secondary metabolism. Nevertheless, there is no report on the fungal diversity of Sanghuangporus, a medicinal and edible homologous macrofungus as "forest gold", which has good effects on antioxidation, boosting immunity and curing stomachache. Here, the diversity and functional group of fungi associated with the fruiting body of the most widely applied S. vaninii were characterized by high-throughput sequencing and FUNGuild tool for the first time. RESULTS: Total 11 phyla, 34 classes, 84 orders, 186 families, and 328 genera were identified in the fruiting body, and our results revealed that the fungal community was dominated by the host fungal taxonomy with absolute superiority (more than 70%), namely, Basidiomycota, Agaricomycetes, Hymenochaetales, Hymenochaetaceae, and genus of Phellinus corrected to Sanghuangporus. Simultaneously, the reads allocated into non-host fungal operational taxonomic units were largely dominated by Ascomycota, Sordariomycetes, Sordariales, Mortierellaceae, and Mortierella. Furthermore, the endophytic fungi were assigned into three trophic modes of "saprotroph" (53.2%), "symbiotroph" (32.2%), and "pathotroph" (14.1%), in which the category of "plant pathogen" was highest enriched with relative abundance of 91.8%, indicating that the endophytic fungi may have the potential to adjust the growth and metabolism of host S. vaninii. CONCLUSION: Altogether, this report firstly provided new findings that can be inspiring for further in-depth studies to exploit bioactive microbial resources for increased production of Sanghuangporus via coculture, as well as to explore the relationship between macrofungi and their associated endophytes.

Identification and profiling of the community structure and potential function of bacteria from the fruiting bodies of Sanghuangporus vaninii.

Sanghuangporus sp., a medicinal and edible homologous macrofungus known as 'forest gold', which has good effects on antitumor, hypolipidemia and the treatment of gynecological diseases. However, the natural resources of fruiting body are on the verge of depletion due to its long growth cycle and over exploitation. The growth and metabolism of macrofungi are known to depend on the diverse bacterial community. Here, we characterized the diversity and potential function of bacteria inhabiting in the fruiting body of the most widely applied S. vaninii using a combination method of high-throughput sequencing with pure culturing for the first time, and tested the biological activities of bacterial isolates, of which Illumina NovaSeq provided a more comprehensive results on the bacterial community structure. Total 33 phyla, 82 classes, 195 orders, 355 families, 601 genera and 679 species were identified in the fruiting body, and our results revealed that the community was predominated by the common Proteobacteria, Gammaproteobacteria, Burkholderiales, Methylophilaceae (partly consistent with pure-culturing findings), and was dominated by the genera of distinctive Methylotenera and Methylomonas (yet-uncultured taxa). Simultaneously, the functional analysis showed that companion bacteria were involved in the pathways of carbohydrate transport and metabolism, metabolism of terpenoids and polyketides, cell wall/membrane/envelope biogenesis, etc. Hence, it was inferred that bacteria associated with fruiting body may have the potential to adjust the growth, development and active metabolite production of host S. vaninii combined with the tested results of indole-3-acetic acid and total antioxidant capacity. Altogether, this report first provided new findings which can be inspiring for further in-depth studies to exploit bioactive microbial resources for increased production of Sanghuangporus, as well as to explore the relationship between medicinal macrofungi and their associated endophytes.

Morel Production Associated with Soil Nitrogen-Fixing and Nitrifying Microorganisms.

True morels (Morchella, Pezizales) cultivated in soil are subject to complex influences from soil microbial communities. To explore the characteristics of soil microbial communities on morel cultivation, and evaluate whether these microbes are related to morel production, we collected 23 soil samples from four counties in Sichuan and Yunnan Provinces, China. Based on ITS and 16S rDNA amplicon sequencing, the alpha diversity analysis indicated that the biodiversity of morel cultivation soil showed a downward trend compared with the bare soil. The results also showed that there were no significant differences in soil microbial communities between OC (bare soil) and OO (after one-year suspension of sowing). This means that, after about one year of stopping sowing, the component and structure of soil that once cultivated morel would be restored. In co-occurrence networks, some noteworthy bacterial microbes involved in nitrogen fixation and nitrification have been identified in soils with high morel yields, such as Arthrobacter, Bradyhizobium, Devosia, Pseudarthrobacter, Pseudolabrys, and Nitrospira. In contrast, in soils with low or no morel yield, some pathogenic fungi accounted for a high proportion, including Gibberella, Microidium, Penicillium, Sarocladium, Streptomyces, and Trichoderma. This study provided valuable information for the isolation and culturing of some beneficial microbes for morel cultivation in further study and, potentially, to harness the power of the microbiome to improve morel production and health.

Clitopilus, Clitocella, and Clitopilopsis in China.

Within the family Entolomataceae, many reports of Entoloma and Rhodocybe have been published, but few reports on Clitopilus and allied genera are available. In this study, Clitopilus, Clitocella, and Clitopilopsis from China were studied by morphological and phylogenetic methods. Portions of nuc 28S rDNA (28S), RNA polymerase II second largest subunit (rpb2), translation elongation factor 1-alpha (tef1), and ATPase subunit 6 (atp6), were employed to elucidate the relationships of Clitopilus and allies. Results indicate that Clitopilus should be divided into three sections, namely, sect. Clitopilus, sect. Scyphoides, and sect. Crispi, the last of which is newly described. In total, 17 species of Clitopilus, Clitocella, and Clitopilopsis are found in China; 14 species belong to Clitopilus, of which 5 are new: C. yunnanensis, C. brunniceps, C. rugosiceps, C. sinoapalus, and C. umbilicatus. Two species of Clitocella, including one new species, Clitocella orientalis, are described. One new species of Clitopilopsis, namely, Clitopilopsis albida, is proposed.

Genome of lethal Lepiota venenata and insights into the evolution of toxin-biosynthetic genes.

BACKGROUND: Genomes of lethal Amanita and Galerina mushrooms have gradually become available in the past ten years; in contrast the other known amanitin-producing genus, Lepiota, is still vacant in this aspect. A fatal mushroom poisoning case in China has led to acquisition of fresh L. venenata fruiting bodies, based on which a draft genome was obtained through PacBio and Illumina sequencing platforms. Toxin-biosynthetic MSDIN family and Porlyl oligopeptidase B (POPB) genes were mined from the genome and used for phylogenetic and statistical studies to gain insights into the evolution of the biosynthetic pathway. RESULTS: The analysis of the genome data illustrated that only one MSDIN, named LvAMA1, exits in the genome, along with a POPB gene. No POPA homolog was identified by direct homology searching, however, one additional POP gene, named LvPOPC, was cloned and the gene structure determined. Similar to ApAMA1 in A. phalloides and GmAMA1 in G. marginata, LvAMA1 directly encodes α-amanitin. The two toxin genes were mapped to the draft genome, and the structures analyzed. Furthermore, phylogenetic and statistical analyses were conducted to study the evolution history of the POPB genes. Compared to our previous report, the phylogenetic trees unambiguously showed that a monophyletic POPB lineage clearly conflicted with the species phylogeny. In contrast, phylogeny of POPA genes resembled the species phylogeny. Topology and divergence tests showed that the POPB lineage was robust and these genes exhibited significantly shorter genetic distances than those of the house-keeping rbp2, a characteristic feature of genes with horizontal gene transfer (HGT) background. Consistently, same scenario applied to the only MSDIN, LvAMA1, in the genome. CONCLUSIONS: To the best of our knowledge, this is the first reported genome of Lepiota. The analyses of the toxin genes indicate that the cyclic peptides are synthesized through a ribosomal mechanism. The toxin genes, LvAMA1 and LvPOPB, are not in the vicinity of each other. Phylogenetic and evolutionary studies suggest that HGT is the underlining cause for the occurrence of POPB and MSDIN in Amanita, Galerina and Lepiota, which are allocated in three distantly-related families.

Species clarification of the culinary Bachu mushroom in western China.

The Bachu mushroom, previously identified as Helvella leucopus, is characterized by a saddle-shaped, to irregularly lobed pileus, with a gray, brown to blackish hymenium and a whitish to pale receptacle surface and white, terete stipe with enlarged basal grooves. It has high economic value, mostly as a dietary supplement in western China, and its medicinal functions have raised broad interest. In the present paper species of the Bachu mushroom in Xinjiang Autonomous Region, western China were investigated with morphology and DNA sequence data. Phylogenetic analyses inferred from ITS, 28S and TEF1 sequence data strongly supported lineages corresponding to morphological features. The Bachu mushroom, which differs from the European Helvella leucopus, comprises two distinct new species, namely Helvella bachu and Helvella subspadicea. In this paper we introduce the new species with descriptions and figures and compare them with similar taxa. The European Helvella spadicea is also re-examined, described and illustrated.

[Diversity of uncultured actinomycetes in saline-alkali soil from Jiuquan area of Hexi Corridor].

ABSTRACT: In order to more accurately understand community structure and diversity of actinomycetes in saline-alkali soil from Jiuquan area of Hexi Corridor, the community structure and diversity from three kinds of soil samples (primary, secondary saline alkali soil and farmland soil) were analyzed using uncultured methods. The results showed that the 16S rDNA clone library of actinomycetales from the primary saline-alkali soil belonged to 19 OTUs, Micrococcineae, Propionibacterineae, Corynebacterineae, Frankineae, Pseudonocardineae and unknown groups of Actinomycetales; the 16S r DNA clone library of actinomycetales from the secondary saline-alkali soil belonged to 14 OTUs, Micrococcineae, Propionibacterineae, Corynebacterineae, Frankineae, Pseudonocardineae and unknown groups of Actinomycetales; the 16S rDNA clone library of farmland soil belonged to 7 OTUs, Micrococcineae, Propionibacterineae, Corynebacterineae, Frankineae, Pseudonocardineae and unknown groups of Actinomycetales; Micrococcineae was the common population in the three soils, and also was the dominant population in primary saline alkali soil and farmland soil. The diversity index and rarefaction curves analysis showed that actinomycetes species richness was in order of primary saline-alkali soil > secondary saline-alkali soil > farmland soil. The dilution curves of primary saline-alkali soil and secondary saline-alkali soil were not leveled off, which indicated the actinomycetes diversity in saline-alkali soil was more enriched than the actual. The rich and diverse actinomycetes resources in saline-alkali soil from Jiuquan area of Hexi Corridor provide important data on the actinomycetes ecology distribution research, exploitation and utilization in saline-alkali soil.

[Allelopathy and chemical constituents of Ligularia virgaurea volatile].

Ligularia virgaurea is a noxious weed widely distributed on the psychro-grasslands of east Qinghai-Tibet Plateau of China, but the allelopathic effects of its volatile is less known. In this study, the allelopathy of L. virgaurea volatile to 5 kinds of grasses was examined, and its chemical constituents were analyzed by GC-MS. The results demonstrated that the volatile of growing L. virgaurea could inhibit the seed germination speed and germination rate of all tested grasses. The inhibitory effects on Elymus nutans and Bromus magnus were significant, while Poa annua and Festuca ovin were not very sensitive. 18 major constituents were identified from the essential oil of L. virgaurea, accounted for 68.24% of the total. The main chemical components were 2-methylheptane (9.84%), 3-methyl-heptane (8.25%), heptane (7.93%), 4-methyl-1-(methylethyl)-bicyclo [3, 1, 0] hex-2-ene (7.79%), 3-methyl-hexane (6.38%), 2-methyl-hexane (5.54%), and D-limonene (4.70%). Monoterpenoids accounted for 16.58% of the total, indicating that volatilization was one of the ways by which L. virgaurea released allelochemicals. The allelopathy of the volatile may play an important role in enhancing the competitive ability of the species, and be one of reasons of grassland degeneration.