This article is the 13th contribution in the Fungal Diversity Notes series, wherein 125 taxa from four phyla, ten classes, 31 orders, 69 families, 92 genera and three genera incertae sedis are treated, demonstrating worldwide and geographic distribution. Fungal taxa described and illustrated in the present study include three new genera, 69 new species, one new combination, one reference specimen and 51 new records on new hosts and new geographical distributions. Three new genera, Cylindrotorula ( Torulaceae ), Scolecoleotia ( Leotiales genus incertae sedis ) and Xenovaginatispora ( Lindomycetaceae ) are introduced based on distinct phylogenetic lineages and unique morphologies. Newly described species are Aspergillus lannaensis , Cercophora dulciaquae , Cladophialophora aquatica , Coprinellus punjabensis , Cortinarius alutarius , C. mammillatus , C. quercoflocculosus , Coryneum fagi , Cruentomycena uttarakhandina , Cryptocoryneum rosae , Cyathus uniperidiolus , Cylindrotorula indica , Diaporthe chamaeropicola , Didymella azollae , Diplodia alanphillipsii , Dothiora coronicola , Efibula rodriguezarmasiae , Erysiphe salicicola , Fusarium queenslandicum , Geastrum gorgonicum , G. hansagiense , Helicosporium sexualis , Helminthosporium chiangraiensis , Hongkongmyces kokensis , Hydrophilomyces hydraenae , Hygrocybe boertmannii , Hyphoderma australosetigerum , Hyphodontia yunnanensis , Khaleijomyces umikazeana , Laboulbenia divisa , Laboulbenia triarthronis , Laccaria populina , Lactarius pallidozonarius , Lepidosphaeria strobelii , Longipedicellata megafusiformis , Lophiotrema lincangensis , Marasmius benghalensis , M. jinfoshanensis , M. subtropicus , Mariannaea camelliae , Melanographium smilaxii , Microbotryum polycnemoides , Mimeomyces digitatus , Minutisphaera thailandensis , Mortierella solitaria , ...

Plants stably transformed to manipulate the expression of genes mediating ecological performance have profoundly altered research in plant ecology. Agrobacterium-mediated transformation remains the most effective method of creating plants harbouring a limited number of transgene integrations of low complexity. For ecological/physiological research, the following requirements must be met: (i) the regenerated plants should have the same ploidy level as the corresponding wild-type plant and (ii) contain a single transgene copy in a homozygous state; (iii) the T-DNA must be completely inserted without vector backbone sequence and all its elements functional; and (iv) the integration should not change the phenotype of the plant by interrupting chromosomal genes or by mutations occurring during the regeneration procedure. The screening process to obtain transformed plants that meet the above criteria is costly and time-consuming, and an optimized screening procedure is presented. We developed a flow chart that optimizes the screening process to efficiently select transformed plants for ecological research. It consists of segregational analyses, which select transgenic T₁ and T₂ generation plants with single T-DNA copies that are homozygous. Indispensable molecular genetic tests (flow cytometry, diagnostic PCRs and Southern blotting) are performed at the earliest and most effective times in the screening process. qPCR to quantify changes in transcript accumulation to confirm gene silencing or overexpression is the last step in the selection process. Because we routinely transform the wild tobacco, Nicotiana attenuata, with constructs that silence or ectopically overexpress ecologically relevant genes, the proposed protocol is supported by examples from this system.

Background: The involvement of small RNAs in cotton fiber development is under explored. The objective of this work was to directly clone, annotate, and analyze small RNAs of developing ovules to reveal the candidate small interfering RNA/microRNAs involved in cotton ovule and fiber development.

The whole-plant activation of defense responses to wounding and herbivory requires systemic signaling in which jasmonates (JAs) play a pivotal role. To examine the nature of the slower cell-nonautonomous as compared to the rapid cell-autonomous signal in mediating systemic defenses in Nicotiana attenuata, reciprocal stem graftingexperiments were used with plants silenced for the JA biosynthetic gene ALLENE OXIDE CYCLASE (irAOC) or plants transformed to create JA sinks by ectopically expressing Arabidopsis JA-O-methyltransferase (ovJMT). JA-impaired irAOC plants were defective in the cell-nonautonomous signaling pathway but not in JA transport. Conversely, ovJMT plants abrogated the production of a graft-transmissible JA signal. Both genotypes displayed unaltered cell-autonomous signaling. Defense responses (17-hydroxygeranyllinalool diterpene glycosides, nicotine, and proteinase inhibitors) and metabolite profiles were differently induced in irAOC and ovJMT scions in response to graft-transmissible signals from elicited wild type stocks. The performance of Manduca sexta larvae on the scions of different graft combinations was consistent with the patterns of systemic defense metabolite elicitations. Taken together, we conclude that JA and possibly MeJA, but not JA-Ile, either directly functions as a long-distance transmissible signal or indirectly interacts with long distance signal(s) to activate systemic defense responses.

The genus Agrilus comprises diverse exotic and agriculturally important wood-boring insects that have evolved efficient digestive systems. Agrilus mali Matsumara, an invasive insect, is causing extensive mortality to endangered wild apple trees in Tianshan. In this study, we present an in-depth characterization of the gut microbiota of A. mali based on high-throughput sequencing of the 16S rRNA gene and report the presence of lignocellulose-degrading bacteria. Thirty-nine operational taxonomic units (OTUs) were characterized from the larval gut. OTUs represented 6 phyla, 10 classes, 16 orders, 20 families, and 20 genera. The majority of bacterial OTUs belonged to the order Enterobacteriales which was the most abundant taxa in the larval gut. Cultivable bacteria revealed 9 OTUs that all belonged to Gammaproteobacteria. Subsequently, we examined the breakdown of plant cell-wall compounds by bacterial isolates. Among the isolates, the highest efficiency was observed in Pantoea sp., which was able to synthesize four out of the six enzymes (cellulase, cellobiase, β-xylanase, and β-gluconase) responsible for plant-cell wall degradation. One isolate identified as Pseudomonas orientalis exhibited lignin peroxidase activity. Our study provides the first characterization of the gut microbial diversity of A. mali larvae and shows that some cultivable bacteria play a significant role in the digestive tracts of larvae by providing nutritional needs.

Agrilus mali Matsumara (Coleoptera: Buprestidae) is a wood‐boring beetle distributed to eastern China that occasionally injures apple species. However, this wood‐boring beetle is new to the wild apple forests (Malus sieversii) of the Tianshan Mountains (western China) and has caused extensive tree mortality. The development of a biological control program for these wild apple forests is a high priority that requires exploration of the life cycle, DNA barcoding and taxonomic status of A. mali. In this study, to determine the diversity of invasive beetles, a fragment of the mitochondrial cytochrome oxidase gene was analyzed. Based on the results, beetles from Gongliu and Xinyuan counties of Xinjiang were identical but differed from those in the apple nursery of Gongliu by a single‐nucleotide substitution. We summarize the taxonomic status, relationships, and genetic distances of A. mali among other Agrilus species using the Tajima‐Nei model in maximum likelihood phylogeny. Analysis revealed that A. mali was closely related to Agrilus mendax and both belong to the Sinuatiagrulus subgenus. The life cycle of A. mali was investigated based on a monthly regular inspection in the wild apple forests of Tianshan. Similar to congeneric species, hosts are injured by larvae of A. mali feeding on phloem tissue, resulting in serpentine galleries constructed between bark and xylem that prevent nutrient transport and leading to tree mortality. Future studies will focus on plant physiological responses to the invasive beetles and include surveys of natural enemies for a potential classical biological control program.

BackgroundPlant microRNAs (miRNAs) play key roles in the transcriptional responses to environmental stresses. However, the role of miRNAs in responses to insect herbivory has not been thoroughly explored. To identify herbivory-responsive miRNAs, we identified conserved miRNAs in the ecological model plant Nicotiana attenuata whose interactions with herbivores have been well-characterized in both laboratory and field studies.ResultsWe identified 59 miRNAs from 36 families, and two endogenous trans-acting small interfering RNAs (tasiRNA) targeted by miRNAs. We characterized the response of the precursor and mature miRNAs to simulated attack from the specialist herbivore Manduca sexta by quantitative PCR analysis and used ir-aoc RNAi transformants, deficient in jasmonate biosynthesis, to identify jasmonate-dependent and -independent miRNA regulation. Expression analysis revealed that groups of miRNAs and tasiRNAs were specifically regulated by either mechanical wounding or wounding plus oral secretions from M. sexta larvae, and these small RNAs were accumulated in jasmonate-dependent or -independent manners. Moreover, cDNA microarray analysis indicated that the expression patterns of the corresponding target genes were correlated with the accumulation of miRNAs and tasiRNAs.ConclusionsWe show that a group of miRNAs and tasiRNAs orchestrates the expression of target genes involved in N. attenuata’s responses to herbivore attack.

Background: Wild apple, Malus sieversii, is an endangered species and a valuable genetic resource that requires a variety of conservation techniques. This study aimed to investigate the influence of different concentrations of hormones on wild apple regeneration from leaf and stem explants to establish an optimal regeneration system. Results:Leaves and stems derived from seedlings were cultured on several media supplemented with various concentrations of thidiazuron (TDZ) or 6-benzylaminopurine (BA) in different combinations with 1-naphthaleneacetic acid (NAA). The results showed that the most efficient shoot formation media (35% and 90%) were MS medium containing 4.0 mg L −1 TDZ and 1.0 mg L −1 NAA for leaf explants and MS medium containing 1.0 mg L −1 BA without NAA for stem explant. MS medium supplemented with 0.4 mg L −1 BA and 0.1 mg L −1 NAA (for shoot multiplication) and 1/2 MS + 0.1 mg L −1 NAA + 1.5% sucrose (for rooting) were effective media. Shoot regeneration from leaf explants was the most effective when the explants were placed abaxial side down onto the medium and were subjected to a pre-treatment of 3 weeks in darkness.Conclusions: An optimized regeneration system for M. sieversii that allowed regeneration within 2-3 months developed. The protocol developed herein can be used in large-scale clonal propagation for the conservation of wild apple, M. sieversii.