Diversity of Endophytes in Panax quinquefolium Root / 中国实验方剂学杂志

ObjectiveTo analyze the community structure of endophytes in Panax quinquefolium root and explore the dominant endophytic bacteria and fungi， to provide scientific basis for the establishment of endophytic microbial bank in P. quinquefolium root. MethodInternal Transcribed Spacer （ITS） sequencing and 16S sequencing were performed on six P. quinquefolium root samples collected from Wendeng， Shandong province on PacBio Sequel Ⅱ. ResultA total of 8 phyla， 11 classes， 23 orders， 27 families and 53 genera of endophytic bacteria were identified in P. quinquefolium root， among which an unidentified Burkholderiaceae and an unidentified Rhizobiaceae were dominant. A total of 9 phyla， 23 classes， 35 orders， 43 families and 48 genera of endophytic fungi were identified in P. quinquefolium root， among which an unclassified Helotiales and Pseudogymnoascus were dominant. The community structure of endophytic bacteria revealed that the roots were selectively enriched with nitrogen-fixing bacteria such as unidentified Rhizobiaceae， Bradyrhizobium and Herbaspirillum， which suggested that nitrogen is important for the growth of P. quinquefolium root. The community structure of endophytic fungi indicated that P. quinquefolium in Shandong province might be infected by unclassified Helotiales. ConclusionThere is a rich diversity of endophytic bacteria and fungi in P. quinquefolium root， which provides scientific basis for studying the interaction of the plant with endophytic microorganisms and screening the endophytes to promote the growth of P. quinquefolium root.

Caracterização da microbiota de queijos artesanais provenientes da Serra da Canastra - MG e da cultura iniciadora natural utilizada em sua produção / Microbiota characterization of artisanal cheeses from Serra da Canastra - MG and their endogenous starter culture

O queijo Canastra possui grande importância na cultura e economia local, é parte do Patrimônio Imaterial do Brasil (IPHAN, 2014) e recebeu o selo de produto com designação de origem em 2012 (INPI, 2016). Sua produção utiliza leite, sal, coalho e uma cultura iniciadora natural, chamada popularmente de pingo. Esse estudo visou a caracterização da microbiota presente no queijo maturado da Serra da Canastra e no pingo utilizado em sua produção utilizando técnicas avançadas de sequenciamento em larga escala para identificação das bactérias e fungos ali presentes. Nossos dados da microbiota bacteriana foram comparados com dados da microbiota de outros queijos brasileiros e do mundo disponíveis na literatura. As principais bactérias encontradas em amostras de pingo pertencem aos gêneros Lactococcus (45.6%), Streptococcus (30.3%), Staphylococcus (5.1%), e em amostras de queijo aos gêneros Lactococcus (22.5%), Streptococcus (27.2%), Corynebacterium (18.8%), Staphylococcus (13.6%), Leuconostoc (6.3%) e Weissella (6%). Os principais gêneros de fungos encontrados nos queijos foram Debaryomycesa (78.6%), Trichosporona (7.8%). Nosso estudo foi capaz de separar a microbiota dos queijos produzidos na Serra da Canastra de outros queijos na Europa e América do Norte, sendo o pH um possível fator de segregação. Também foi observada uma diferença entre a microbiota do queijo Canastra com outros queijos Brasileiros. Além disso, visualizamos que a distância geográfica entre produtores e a sazonalidade possuem um efeito sobre a microbiota dos pingos e queijos. A partir da análise de todos os microrganismos encontrados na microbiota bacteriana, foram detectados táxons que discriminam produtores por suas aplicações de boas práticas de fabricação e por sua infraestrutura. Observamos proporções menores de um táxon de Kocuria Kristinae nos pingos e um de Streptococcus nos queijos e proporções maiores de um táxon de Staphylococcus nos queijos. Também pudemos observar uma diminuição nas proporções de táxons de Debaryomycesa e aumento na proporção de táxons de Trichosporona na composição fúngica dos queijos, possivelmente devido a transição sazonal do período seco para o chuvoso. Usando técnicas moleculares de sequenciamento em larga escala, demonstramos que há uma diferença na microbiota presente em diferentes áreas da Serra da Canastra, um possível efeito da sazonalidade na composição fúngica e bacteriana. E evidenciamos que táxons de Streptococcus, Staphylococcus e Kocuria estão correlacionados às boas práticas de produção e elucidamos a conexão existente entre a microbiota do pingo e a do queijo. Estes resultados podem influenciar o desenvolvimento de métodos de rastreamento de sub-regiões específicas da Canastra e auxiliar os produtores na produção de queijos de boa qualidade, mantendo as características específicas de sua região

The Canastra cheese has great importance for the local culture and economy, being part of the Intangible Heritage of Brazil (IPHAN, 2014). It has received the protected designation of origin certification in 2012 (INPI, 2016). It's made using milk, salt, rennet and a endogenous starter culture, popularly called as "pingo". This study aimed to characterize the microbiota present in the Serra da Canastra's cheese and the pingo used in its production. In order to conduct this research we used next generation sequencing to identify the bacteria and fungi present there. Our bacterial microbiota dataset was compared with microbiota datasets from other Brazilian and world cheeses available in the literature. The main bacteria found were Lactococcus (45.6%), Streptococcus (30.3%) and Staphylococcus (5.1%) in the endogenous starter samples and Lactococcus (22.5%), Streptococcus (27.2%), Corynebacterium (18.8 %), Staphylococcus (13.6%), Leuconostoc (6.3%) and Weissella (6%) in cheese samples. The main fungi found in the cheeses were Debaryomycesa (78.6%) and Trichosporona (7.8%). We were able to separate the microbiota from Serra da Canastra cheeses and other cheeses in Europe and North America, being the pH a possible segregation factor. Furthermore, a difference was also observed between the microbiota of Canastra and other Brazilian cheeses. In addition, we observed that the geographical distance between producers and the seasonality could be affecting the pingos and cheeses microbiota. We found bacterial taxa that could discriminate producers by their good manufacturing practices and their local infrastructure. Low levels of good manufacturing practices (GMPs) were assigned to bigger proportions of a Kocuria Kristinae taxon in the pingos and a Staphylococcus taxon in the cheeses. Also, higher levels of GMPs were assigned to smaller proportions of Streptococcus taxons in the cheeses. Furthermore We could observe a decrease of Debaryomycesa and an increase of Trichosporona proportions in the fungal composition of cheeses. This could be due to a climate transition: from the dry season to the rainy season. Using large-scale sampling coupled with molecular sequencing techniques, we observe a connection between pingo and cheeses microbiota. We show that the microbiota of different areas in Serra da Canastra is different, also, there is a possible effect of seasonality on fungal and bacterial composition. Furthermore, we could see that Streptococcus, Staphylococcus and Kocuria taxons are correlated with good practices. These results may influence the development of tracking methods for specific Canastra subregions and assist producers to manufacture good quality cheeses while maintaining the specific characteristics of their region

Molecular characterization of phosphate solubilizing fungi Aspergillus niger and its correlation to sustainable agriculture

Aim: The present study aimed to investigate the phosphate solubilization potential of agriculturally important fungi, i.e., Aspergillus sp. isolated from the rhizosphere of healthy plants in Abha city, Saudi Arabia.Methodology: Sixteen Aspergillus sp. isolated and tested for phosphate solubilization potential were identified by 5.8S-ITS region sequencing and characterized by 11 ISSR-PCR markers. Finally, the highest phosphate solubilization potential isolates were used in field experiments on cucumber and tomato plants. Results: All Aspergillus niger isolates showed 96–100% similarity to A. niger strains available at GenBank database, Isolate ASAB-5 was most efficient at solubilizing phosphate on Pikovskaya’s medium, with a solubilization index of 2.67, and 235.22 mg l-1 of solubilized phosphate. ISSR-PCR markers revealed is total 142 bands in all isolates, with about 32.3% showing monomorphism and 67.6% polymorphism. Based on genetic similarity and intraspecies variability, the Aspergillus isolates were grouped into two different clusters with about 67.9% genetic similarity. The results of field experiments showed no significant difference between seeds treated with culture filtrate or conidial suspension of ASAB-5; however, both differed remarkably from untreated seeds. Interpretation: The current study confirms the existence of several useful phosphate solubilizing fungi in plants, which may serve as potential biological fertilizers. They are safer than chemical fertilizers and increase the bioavailability of soil phosphates for plants

Molecular Identification of Mycorrhizae of Cymbidium kanran (Orchidaceae) on Jeju Island, Korea

A fungal internal transcribed spacer region was used to identify the mycorrhizae of Cymbidium kanran. The family Russulaceae was found to be the most frequently occurring group in both root and soil samples. In phylogenetic analyses, the majority of the Russulaceae clones were clustered with Russula brevipes and R. cyanoxantha. Therefore, C. kanran may form symbiotic relationships with the genus Russula.

Characterization of cytotoxic compound from mangrove derived fungi Irpex hydnoides VB4

<p><b>OBJECTIVE</b>To investigate the cytotoxic activity of endophytic fungi isolated from mangrove fungi.</p><p><b>METHODS</b>In the present study the DNA was isolated and the ITS region of 5.8s rRNA was amplified using specific primers ITS 1 and ITS4 and sequence was determined using automated sequencers. Blast search sequence similarity was found against the existing non redundant nucleotide sequence database thus, identified as Aspergilus flavus, Hyporcaea lixii, Aspergillus niger, Eutorium amstelodami, Irpex hydnoides and Neurospora crassa. Among the seven isolates, one fungi Irpex hydnoides was selected for further studies. The fungi were grown in sabouraud broth for five days and filtrate were separated and subjected to ethyl acetate for further studies.</p><p><b>RESULTS</b>Nearly half (49.25%) of the extracts showed activity (IC50 of 125µg/mL). These values were within the cutoff point of the National Cancer Institute criteria for cytotoxicity (IC50<20 µg/mL) in the screening of crude plant extracts. The GC MS analysis revealed that the active principals might be Tetradecane (6.26%) with the RT 8.606.</p><p><b>CONCLUSIONS</b>It is clear from the present study that mangrove fungi with bioactive metabolites can be expected to provide high quality biological material for high throughout biochemical, anti cancer screening programmes. The results help us conclude that the potential of using metabolic engineering and post genomic approaches to isolate more novel bioactive compounds and to make their possible commercial application is not far off.</p>

Identification and Characterization of the Causal Organism of Gummy Stem Blight in the Muskmelon (Cucumis melo L.)

Gummy stem blight is a major foliar disease of muskmelon (Cucumis melo L.). In this study, morphological characteristics and rDNA internal transcribed spacer (ITS) sequences were analyzed to identify the causal organism of this disease. Morphological examination of the Jeonbuk isolate revealed that the percentage of monoseptal conidia ranged from 0% to 10%, and the average length x width of the conidia was 70 (+/- 0.96) x 32.0 (+/- 0.15) microm on potato dextrose agar. The BLAST analysis showed nucleotide gaps of 1/494, 2/492, and 1/478 with identities of 485/492 (98%), 492/494 (99%), 491/494 (99%), and 476/478 (99%). The similarity in sequence identity between the rDNA ITS region of the Jeonbuk isolate and other Didymella bryoniae from BLAST searches of GenBank was 100% and was 95.0% within the group. Nucleotide sequences of the rDNA ITS region from pure culture ranged from 98.2% to 99.8%. Phylogenetic analysis with related species of D. bryoniae revealed that D. bryoniae is a monophyletic group distinguishable from other Didymella spp., including Ascochyta pinodes, Mycosphaerella pinodes, M. zeae-maydis, D. pinodes, D. applanata, D. exigua, D. rabiei, D. lentis, D. fabae, and D. vitalbina. Phylogenetic analysis, based on rDNA ITS sequence, clearly distinguished D. bryoniae and Didymella spp. from the 10 other species studied. This study identified the Jeonbuk isolate to be D. bryoniae.