Additional file 3 of Fused expression of Sm1-Chit42 proteins for synergistic mycoparasitic response of Trichoderma afroharzianum on Botrytis cinerea

Additional file: Figure S3. Hydrophobicity modulation ability of TaSm1, MaChi42, and SCf expressing in T. afroharzianum. (A) Pictures and (B) box plot of a water droplet in the surface of T. afroharzianum wild-type (T30), OE:TaSm1, OE:MaChi42, and OE:SCf strains. Hydrophobicity of spores suspension of T. afroharzianum wild-type (T30), OE:TaSm1, OE:MaChi42, and OE:SCf strains in glass (C) and PET (D) slides

Biodiversity of <i>Trichoderma</i> Community in the Tidal Flats and Wetland of Southeastern China

<div><p>To investigate the biodiversity of <i>Trichoderma</i> (<i>Hypocreaceae</i>) and their relation to sediment physical and chemical properties, we collected a total of 491 sediment samples from coastal wetlands (tidal flat and wetland) in Southeast China. Further, we applied two types of molecular approaches such as culture dependent and independent methods for identification of <i>Trichoderma</i> spp. A total of 254 isolates were obtained and identified to 13 species such as <i>T</i>. <i>aureoviride</i>, <i>T</i>. <i>asperellum</i>, <i>T</i>. <i>harzianum</i>, <i>T</i>. <i>atroviride</i>, <i>T</i>. <i>koningiopsis</i>, <i>T</i>. <i>longibrachiatum</i>, <i>T</i>. <i>koningii</i>. <i>T</i>. <i>tawa</i>, <i>T</i>. <i>viridescens</i>, <i>T</i>. <i>virens</i>, <i>T</i>. <i>hamatum</i>, <i>T</i>. <i>viride</i>, and <i>T</i>. <i>velutinum</i> by the culture-dependent (CD) method of these, <i>T</i>. <i>tawa</i> was newly described in China. Subsequently, the culture indepented method of 454 pyrosequencing analysis revealed a total of six species such as <i>T</i>. <i>citrinoviride</i>, <i>T</i>. <i>virens</i>, <i>T</i>. <i>polysporum</i>, <i>T</i>. <i>harzianum</i>/<i>Hypocrea lixii</i> and two unknown species. Notably, <i>T</i>. <i>citrinoviride</i> and <i>T</i>. <i>polysporum</i> were not found by the CD method. Therefore, this work revealed that the combination of these two methods could show the higher biodiversity of <i>Trichoderma</i> spp., than either of this method alone. Among the sampling sites, Hangzhou, Zhejiang Province, exhibited rich biodiversity and low in Fengxian. Correlation and Redundancy discriminant analysis (RDA) revealed that sediment properties of temperature, redox potential (Eh) and pH significantly influenced the biodiversity of <i>Trichoderma</i> spp.</p></div

Additional file 4 of Fused expression of Sm1-Chit42 proteins for synergistic mycoparasitic response of Trichoderma afroharzianum on Botrytis cinerea

Additional file: Table S1 Primers used in this study

Additional file 2 of Fused expression of Sm1-Chit42 proteins for synergistic mycoparasitic response of Trichoderma afroharzianum on Botrytis cinerea

Additional file: Figure S2. Sm1 gene expressing in the process of T. afroharzianum engineered strains interact with B. cinerea

Identification of a Novel Fungus, <i>Leptosphaerulina chartarum</i> SJTU59 and Characterization of Its Xylanolytic Enzymes

<div><p>Xylanolytic enzymes are widely used in processing industries, e.g., pulp and paper, food, livestock feeds, and textile. Furthermore, certain xylanotic enzymes have demonstrated the capability to improve the resistance and immunity of plants. Screening of high-yield microbial xylanolytic enzyme producers is significant for improving large-scale cost-effective xylanolytic enzyme production. This study provided new evidence of high-level xylanolytic enzyme production by a novel fungus, designated <i>Leptosphaerulina chartarum</i> SJTU59. Under laboratory conditions, <i>L. chartarum</i> SJTU59 produced xylanolytic enzymes of up to 17.566 U/mL (i.e., 878.307 U/g substrate). The enzyme solution was relatively stable over a wide range of pH (pH 3.0 to pH 9.0) and temperature (40°C to 65°C) while showing high resistance to the majority of metal ions tested. Composition analysis of the hydrolytic products of xylan showed sufficient degradation by xylanolytic enzymes from <i>L. chartarum</i> SJTU59, mainly the monosaccharide xylose, and a small amount of xylobiose were enzymatically produced; whereas in the presence of sufficient xylan substrates, mainly xylooligosaccharides, an emerging prebiotic used in food industry, were produced. In addition, the xylanolytic enzyme preparation from <i>L. chartarum</i> SJTU59 could initiate tissue necrosis and oxidative burst in tobacco leaves, which may be related to enhanced plant defense to adversity and disease. <i>L. chartarum</i> SJTU59 possessed a complex xylanolytic enzyme system, from which two novel endo-β-1,4-xylanases of the glycoside hydrolase (GH) family 10, one novel endo-β-1,4-xylanase of the GH family 11, and one novel β-xylosidase of the GH family 43 were obtained via rapid amplification of complementary DNA ends. Given the high yield and stable properties of xylanolytic enzymes produced by <i>L. chartarum</i> SJTU59, future studies will be conducted to characterize the properties of individual xylanolytic enzymes from <i>L. chartarum</i> SJTU59. xylanolytic enzymes-encoding gene(s) of potential use for industrial and agricultural applications will be screened to construct genetically engineered strains.</p></div

Alignments of orthologous proteins

Firstly, 1328 orthologous proteins were screened out among Curvualria lunata and other 11 close fungi including Bipolaris maydis, Setosphaeria turcica, Pyrenophora tritici-repentis, Phaeosphaeria nodorum, Cercospora zea-madis, Aspergillus flavus, Magnaporthe oryzae, Fusarium graminearum, Saccharomyces cerevisiae, and Ustilago maydis with a cutoff E value of 1e-20. Secondly, these orthologous proteins were aligned using Clustal W 2.1. Lastly, the aligned proteins were concatenate

Univariate diversity indices analysis.

<p>Univariate diversity indices analysis.</p

The number of operational taxanomical units (OTUs) found in different samplescollected from Southeast China [Zhuhai (wt11), Hangzhou (wt12), Ningbo (wt13), Fengxian (wt14), Yantai (wt15), Shanghai (wt16), Shantou (wt17), Beihai (wt18), Fuzhou (wt19) and Lianyungang (wt20)].

<p>The number of operational taxanomical units (OTUs) found in different samplescollected from Southeast China [Zhuhai (wt11), Hangzhou (wt12), Ningbo (wt13), Fengxian (wt14), Yantai (wt15), Shanghai (wt16), Shantou (wt17), Beihai (wt18), Fuzhou (wt19) and Lianyungang (wt20)].</p

Distribution of midgut bacterial communities between clone libraries of SDT-1, FL-1, FL-2, and FL-3.

<p>(<b>A</b>) Predicted numbers of phylotypes based on the S_Chao1 of the 16S rRNA clone libraries. a, clone library of SDT-1; b, clone library of FL-1; c, clone library of FL-2; d, clone library of FL-3. X axis, Bacterial clone numbers in each library; Y axis, Predicted value of S_chao1. (<b>B</b>) The bacterial composition in the larval midgut of fourth instar at the family level. a, clone library of SDT-1; b, clone library of FL-1; c, clone library of FL-2; d, clone library of FL-3. (<b>C</b>) Comparison of genera <i>Achromobacter</i>, <i>Enterococcus</i>, and <i>Ochrobactrum</i> distribution in the larval midguts between clone libraries of SDT-1, FL-1, FL-2, and FL-3.</p

Additional file 1 of Fused expression of Sm1-Chit42 proteins for synergistic mycoparasitic response of Trichoderma afroharzianum on Botrytis cinerea

Additional file: Figure S1. Construction of chimeric protein engineered strains of T. afroharzianum. (A) Sm1 and Chit42 overlap fragments for chimeric protein and TaSm1 and MaChit42 overexpression vectors construction; (B) PCR verification of chimeric protein and TaSm1 and MaChit42 engineered strains by using hygromycin primer; (C) and (D) were PCR verification of chimeric protein and TaSm1 and MaChit42 engineered strains using by differential primer pairs (PC between trpC promoter and Chit42; CS between Chi42 and Sm1; ST between Sm1 and trpC terminator; PS between trpC promoter and Sm1; SC between Sm1 and Chit42; CT between Chit42 and trpC terminator); (E) Southern blot analysis of chimeric protein and TaSm1 and MaChit42 engineered strains; (F) qPCR results of Sm1 gene expressing in T. afroharzianum with different culture medium (PDA and PD)