Phylogenetic Tree Construction Research Articles

Unveiling the Arsenal of Apple Bitter Rot Fungi: Comparative Genomics Identifies Candidate Effectors, CAZymes, and Biosynthetic Gene Clusters in Colletotrichum Species.

The bitter rot of apple is caused by Colletotrichum spp. and is a serious pre-harvest disease that can manifest in postharvest losses on harvested fruit. In this study, we obtained genome sequences from four different species, C. chrysophilum, C. noveboracense, C. nupharicola, and C. fioriniae, that infect apple and cause diseases on other fruits, vegetables, and flowers. Our genomic data were obtained from isolates/species that have not yet been sequenced and represent geographic-specific regions. Genome sequencing allowed for the construction of phylogenetic trees, which corroborated the overall concordance observed in prior MLST studies. Bioinformatic pipelines were used to discover CAZyme, effector, and secondary metabolic (SM) gene clusters in all nine Colletotrichum isolates. We found redundancy and a high level of similarity across species regarding CAZyme classes and predicted cytoplastic and apoplastic effectors. SM gene clusters displayed the most diversity in type and the most common cluster was one that encodes genes involved in the production of alternapyrone. Our study provides a solid platform to identify targets for functional studies that underpin pathogenicity, virulence, and/or quiescence that can be targeted for the development of new control strategies. With these new genomics resources, exploration via omics-based technologies using these isolates will help ascertain the biological underpinnings of their widespread success and observed geographic dominance in specific areas throughout the country.

Four New Species of Jelly Fungi from Northeastern China.

Four new species of jelly fungi were described from northeastern China based on morphological and molecular evidence. These new species were classified into the four genera Sirobasidium (Sirobasidium jilinense), Calocera (Calocera velutina), Dacrymyces (Dacrymyces jauensis), and Dacryopinax (Dacryopinax manghanensis). Maximum likelihood and Bayesian analyses were performed using a combined nuc rDNA internal transcribed spacer region (ITS) and nuc 28S rDNA (nrLSU) dataset for the construction of phylogenetic trees. Morphological descriptions, line illustrations, and the ecological habits of these new species are provided.

A re-evaluation of Diaporthe: refining the boundaries of species and species complexes

AbstractDiaporthe is an important plant pathogenic genus, which also occurs as endophytes and saprobes. Many Diaporthe species that are morphologically similar proved to be genetically distinct. The current understanding of Diaporthe taxonomy by applying morphological characters, host associations and multi-gene phylogeny are problematic leading to overestimation/underestimation of species numbers of this significant fungal pathogenic genus. Currently, there are no definite boundaries for the accepted species. Hence, the present study aims to re-structure the genus Diaporthe, based on single gene phylogenies (ITS, tef, tub, cal and his), multi-gene phylogeny justified by applying GCPSR (Genealogical Concordance Phylogenetic Species Recognition) methodology as well as the coalescence-based models (PTP—Poisson Tree Processes and mPTP—multi-rate Poisson Tree Processes). Considering all available type isolates of Diaporthe, the genus is divided into seven sections while boundaries for 13 species and 15 species-complexes are proposed. To support this re-assessment of the genus, 82 Diaporthe isolates obtained from woody hosts in Guizhou Province in China were investigated and revealed the presence of two novel species and 17 previously known species. Synonymies are specified for 31 species based on molecular data and morphological studies. Dividing Diaporthe into several specific sections based on phylogenetic analyses can avoid the construction of lengthy phylogenetic trees of the entire genus in future taxonomic studies. In other words, when one conducts research related to the genus, only species from the appropriate section need to be selected for phylogenetic analysis.

Comparative Analysis and Phylogeny of the Complete Chloroplast Genomes of Nine Cynanchum (Apocynaceae) Species.

Cynanchum belongs to the Apocynaceae family and is a morphologically diverse genus that includes around 200 shrub or perennial herb species. Despite the utilization of CPGs, few molecular phylogenetic studies have endeavored to elucidate infrafamilial relationships within Cynanchum through extensive taxon sampling. In this research, we constructed a phylogeny and estimated divergence time based on the chloroplast genomes (CPGs) of nine Cynanchum species. We sequenced and annotated nine chloroplast (CP) genomes in this study. The comparative analysis of these genomes from these Cynanchum species revealed a typical quadripartite structure, with a total sequence length ranging from 158,283 to 161,241 base pairs (bp). The CP genome (CPG) was highly conserved and moderately differentiated. Through annotation, we identified a total of 129-132 genes. Analysis of the boundaries of inverted repeat (IR) regions showed consistent positioning: the rps19 gene was located in the IRb region, varying from 46 to 50 bp. IRb/SSC junctions were located between the trnN and ndhF genes. We did not detect major expansions or contractions in the IR region or rearrangements or insertions in the CPGs of the nine Cynanchum species. The results of SSR analysis revealed a variation in the number of SSRs, ranging from 112 to 150. In five types of SSRs, the largest number was mononucleotide repeats, and the smallest number was hexanucleotide repeats. The number of long repeats in the cp genomes of nine Cynanchum species was from 35 to 80. In nine species of Cynanchum, the GC3s values ranged from 26.80% to 27.00%, indicating a strong bias towards A/U-ending codons. Comparative analyses revealed four hotspot regions in the CPG, ndhA-ndhH, trnI-GAU-rrn16, psbI-trnS-GCU, and rps7-ndhB, which could potentially serve as molecular markers. In addition, phylogenetic tree construction based on the CPG indicated that the nine Cynanchum species formed a monophyletic group. Molecular dating suggested that Cynanchum diverged from its sister genus approximately 18.87 million years ago (Mya) and species diversification within the Cynanchum species primarily occurred during the recent Miocene epoch. The divergence time estimation presented in this study will facilitate future research on Cynanchum, aid in species differentiation, and facilitate diverse investigations into this economically and ecologically important genus.

Sonic hedgehog (shh) gene from Pseudopleuronectes yokohamae (Teleostei: Pleuronectidae): Molecular cloning, characterization, and expression profile during early embryonic, juvenile, and adult stages.

The hedgehog signaling pathway plays an important role in early development and growth of most vertebrates. Sonic hedgehog (shh) gene is a critical regulator of embryonic development in many species, including humans. However, it is not clear what roles shh can play in the development of fish. In this paper, shh gene was cloned from Pseudopleuronectes yokohamae. The full-length complementary DNA (cDNA) of P. yokohamae sonic hedgehog gene (Pyshh) comprises 3194 bp, with a 1317-bp open reading frame (ORF) that encodes a polypeptide of 438 amino acids with a typical HH-signal domain and Hint-N domain. The conserved sequences of the protein among species were predicted by using multiple sequence comparison. The phylogenetic tree construction showed that PySHH is clustered in a branch of Pleuronectidae. To explore the expression of Pyshh gene in various tissues of P. yokohamae, we used real-time fluorescence quantitative PCR technology to detect it. The results showed that Pyshh gene is widely distributed in various tissues of P. yokohamae juveniles, different tissues of adult males and females, and is particularly expressed in immune organs. The Pyshh gene expression was higher in the muscle and brain of juvenile fish, and higher in bone, gill, and skin of male fish than that of female fish, suggesting that Pyshh might be involved in the formation of immune organs of P. yokohamae. The expression of Pyshh gene significantly upregulated from the gastrula stage to the hatching stage. Western blotting of the expression levels of PySHH during different embryonic development stages revealed that PySHH levels increased gradually during development stages from oosperm stage to hatching stage. These results indicate that Pyshh is highly conserved among species and plays a critical role in the complex process of embryonic development. Its precise regulation is essential for the proper formation of many organs and tissues in the body, and disruptions in its function may have serious consequences for the formation of immune organs in fish.

The effective and green biodegradation of polyethylene microplastics by the screening of a strain with its degrading enzymes

The polyethylene microplastics (MPs) degrading fungi named the strain DL-1 were isolated and screened to reduce the pollution of polyethylene MPs. The strain was preliminarily identified as Aspergillus niger by the sequence homology comparison and phylogenetic tree construction. The biodegradation rate of polyethylene MPs by the strain DL-1 was 7.65 ± 0.92 % after 30 days. The grooves and wrinkles appeared on the surface of MPs after the degradation. During the effective degradation process, the strain DL-1 produced the enzymes (plastic degrading enzymes) to biodegrade the polyethylene MPs. It was showed that the plastic degrading enzymes increased under the induction of polyethylene MPs in the medium, indicating that the enzymes played the important role in biodegradation of polyethylene MPs. Moreover, the ability of strain to biodegrade polyethylene MPs and the mechanism of biodegradation were investigated with varied measurements and characteristics. It enriched the way of microbial degradation of polyethylene MPs and provided the environmentally-friendly biodegradation with the green treatment for MPs.

Characterising the molecular epidemiology of human parechovirus in young infants in the UK and Canada

ObjectivesWe evaluated the extent of virus heterogeneity in PeV infected infants in the UK, Canada and Australia. MethodsSamples were collected from PeV infected infants during 2013–16. Next generation sequencing was used to obtain sequencing data and construct phylogenetic trees based on analysis of the VP1 region. Comparison was made with sequencing data available from an outbreak in Australia. ResultsWe amplified and sequenced 58 samples. All obtained PeV sequences were genotype 3 apart from one UK sample which was PeV-A5. Phylogenetic analysis revealed that all strains clustered together on the same clade and showed no significant genetic variation. We saw no significant evidence of association between sequence and either clinical severity (defined by admission to paediatric intensive care), geographical origin (compared between Canada and U.K) or year of sample collection (samples sequenced during 2013 – 2018). ConclusionsIn this small cohort, sequencing data indicate that PeV circulating in the UK and Canada from 2013 to 18 are derived from a common ancestor. No association between disease severity and genetic sequence was seen in the UK or Canadian cohorts. Larger studies are required to support these findings.

Advancements in Phylogenetic Techniques: Revolutionizing Microbiology and Evolutionary Framework. A Review

Phylogenetic trees describe the evolutionary relationships among different types of organisms. The selection of methods used in the construction of these trees is not easy because the data used is often incomplete, meaning it comes from studies of both modern organisms and fossils. Also, the morphological data can be misleading ; when closely related organisms may not look the same and distantly related organisms may share certain features, all these reasons make it difficult to determine the true relationships between these organisms. Therefore, the genetic data is more dependable, so it's usually a preferred method used to construct phylogenetic trees. Despite that, the genetic data can be affected by several factors, such as gene transfer, loss, and the choice of genes that are used. It's important to note that phylogenetic trees do not account for the length of time over which evolution happened ; they only reflect the order of divergence among lineages. Despite all these challenges, phylogenetic trees are still a crucial tool in understanding the evolution of life and in identifying new relationships among different organisms. Recently, the use of phylogenetic analysis in microbiological research has increased significantly. This review provides an overview of the concept of phylogeny, the different methods used in the construction of phylogenetic trees, the applications of phylogenetic analysis in microbiology, and the steps involved in building a suitable phylogenetic tree.

Investigation on the Potential Functions of ZmEPF/EPFL Family Members in Response to Abiotic Stress in Maize.

Maize is an important crop used for food, feed, and fuel. Abiotic stress is an important factor affecting maize yield. The EPF/EPFL gene family encodes class-specific secretory proteins that play an important role in the response to abiotic stress in plants. In order to explore and utilize the EPF/EPFL family in maize, the family members were systematically identified, and their chromosomal localization, physicochemical properties, cis-acting element prediction in promoters, phylogenetic tree construction, and expression pattern analysis were carried out using bioinformatics techniques. A total of 18 ZmEPF/EPFL proteins were identified in maize, which are mostly alkaline and a small portion acidic. Subcellular localization results showed that ZmEPF6, ZmEPF12, and ZmEPFL2 are localized in the nucleus and cytoplasm. Analysis of cis-acting elements revealed that members of the ZmEPF/EPFL family contain regulatory elements such as light response, anoxic, low temperature, and hormone response regulatory elements. RT-qPCR results showed that these family members are indeed responding to cold stress and hormone treatments. These results of this study provide a theoretical basis for improving the abiotic stress resistance of maize in future research.

A Comparative Assessment Study on the Prolactin Gene as a Genetic Marker for Milk Production in High Milk-Producing Mammals through Phylogenetic Analysis

The prolactin (PRL) gene is a focus in milk-related studies for high-yielding ruminants like dairy cows, water buffaloes, dairy goats, and sheep. Despite its recognition as a genetic marker for milk traits, little is known about the specific genetic features enabling heightened milk production. The PRL nucleotide sequence of the ruminants and the human outgroup were obtained from the GenBank database. Using the MEGA 7 software, amino acid translation was conducted and multisequence analysis was done, allowing for the construction of phylogenetic trees. In both trees, ruminants were consistently clustered together forming pairings of the dairy cow: water buffalo and the dairy goat: sheep. In the nucleotide sequences of all the involved taxa, cytosine and guanine were the unanimous most and least comprised nucleotide. In the amino acid composition, the unanimous most and least amino acid percentages were leucine and tryptophan, respectively. Leucine, being the highest comprised, was substantiated by related studies stating that such amino acid is highly involved in milk production and yield. Glycine percentages were found to be higher in ruminants than in humans, providing a possible reasoning for such milk-yield disparity in the involved taxa.

A Comparative Assessment Study on the Prolactin Gene as a Genetic Marker for Milk Production in High Milk-Producing Mammals through Phylogenetic Analysis

The prolactin (PRL) gene is a focus in milk-related studies for high-yielding ruminants like dairy cows, water buffaloes, dairy goats, and sheep. Despite its recognition as a genetic marker for milk traits, little is known about the specific genetic features enabling heightened milk production. The PRL nucleotide sequence of the ruminants and the human outgroup were obtained from the GenBank database. Using the MEGA 7 software, amino acid translation was conducted and multisequence analysis was done, allowing for the construction of phylogenetic trees. In both trees, ruminants were consistently clustered together forming pairings of the dairy cow: water buffalo and the dairy goat: sheep. In the nucleotide sequences of all the involved taxa, cytosine and guanine were the unanimous most and least comprised nucleotide. In the amino acid composition, the unanimous most and least amino acid percentages were leucine and tryptophan, respectively. Leucine, being the highest comprised, was substantiated by related studies stating that such amino acid is highly involved in milk production and yield. Glycine percentages were found to be higher in ruminants than in humans, providing a possible reasoning for such milk-yield disparity in the involved taxa.

Complete S1 glycoprotein analysis of two distinct groups of GI-19 (QX) infectious bronchitis virus from Indonesia: Molecular phylogenetic and in-silico approaches

Infectious bronchitis (IB) is an important viral disease that impacts poultry productivity, according to many studies. The main important infectious bronchitis virus (IBV) protein, spike glycoprotein subunit 1 (S1), plays a role in pathogenicity and immunogenicity. Conventional molecular detection and analysis of the S1 gene still face many challenges due to its hypervariability. In Indonesia, there were only a few reports on partial S1 gene analysis of IBVs; thus far, there have been no phylogenetic or in-silico approaches to their S1 complete gene. This study aims to reveal molecular pathogenicity and immunogenicity features on the S1 complete gene of IBVs from Indonesia. We analyzed fourteen IBV isolates using conventional RT-PCR and Sanger sequencing and then aligned their verified S1 complete sequences with other Indonesian GI-19 IBVs from Genbank for molecular phylogenetic and in-silico analysis. Phylogenetic analysis consisted of amino acid polymorphism and identity, as well as phylogenetic tree construction. As part of the in-silico analysis, we used proteins that have homology and immunoinformatic predictions (B-cell epitope and N-glycosylation). Molecular phylogenetic analysis revealed the division of the Indonesian GI-19 IBVs into two distinct groups. These groups had different amino acid variations at receptor binding sites, hypervariable antigenic sites, and cleavage sites. We also calculated the amino acid identity at 95.4–98.3%. In-silico studies revealed significant similarities in B-cell epitopes, distinct N-glycosylation sites, and the suggestion of a homotrimer protein structure. We found that the S1 full gene of two different groups of GI-19 IBVs (QX strain) from Indonesia predicted changes in their pathogenicity and immunogenicity.

First Report of Diaporthe tectonendophytica Causing Leaf Spot on Dalbergia odorifera in Guangxi, China.

Dalbergia odorifera T. Chen (Family: Fabaceae) is a national level II protected plant in China, with extremely high economic value and medical properties (Zhao et al. 2020). In June 2023, an unknown leaf spot was found in a garden land of Pingxiang city, Guangxi, China, and approximately 80% of the plants covered an area of 500 m2 displayed similar symptoms. The spots were grey to white, 4~6 mm in diameter (n=30) with black pycnida on the spots surface (Fig S1, A-D). Multiple disease spots were observed on a single leaf. The pycnida on the lesion were picked and mashed, to make a conidia suspension using sterile water. The conidial solution was then spread onto a potato dextrose agar (PDA) plate containing streptomycin, with 10 mg of streptomycin per 100 mL, and incubated for 3 days at 28°C with a 12 hour photoperiod. Three isolates (GXPX01, GXPX02 and GXPX03) were obtained by re-culturing the colonies on fresh PDA plates. The colony on PDA were white with aerial mycelia (Fig S1, E-F). Black conidiomata developed at 28°C with a 12 hour photoperiod in 20 days (Fig S1, G-H). Alpha conidia were 4.2~6.4 μm × 1.8~2.6 μm (average =5.1 × 2.3 μm, n = 30), mostly bi-guttulate, hyaline, ellipsoid, apex bluntly rounded, base obtuse to subtruncate, smooth (Fig S1, I). Beta conidia were 15.1~33.5 μm × 1~1.8 μm (average = 24.5 × 1.5 μm, n = 30), filiform, hyaline, curved or hamate, aseptate, base subtruncate (Fig S1, J). Morphological characteristics of the three isolates matched those of Diaporthe spp.(Gomes et al. 2013). The rDNA internal transcribed spacer (ITS) region, the translation elongation factor 1-α (TEF1), the calmodulin (CAL), the histone H3 (HIS) and the β-tubulin (TUB2) genes of the three isolates were amplified using the primer pairs ITS4/ITS5, EF1-728F/EF1-986R, CAL-228F/CAL2Rd, CYLH3F/H3-1B, and T1 /CYLTUB1R, respectively (Crous et al. 2004, Sun et al. 2021). The sequences were all deposited in GenBank (accession numbers OR437511 to OR437513 for ITS, OR454965 to OR454967 for TEF1, OR454968 to OR454970 for CAL, OR454971 to OR454973 for TUB2, OR454974 to OR454976 for H3). Sequences had 98.36% to 100% homology with the corresponding sequences of known Diaporthe tectonendophytica strains MFLUCC 13-0471 in the NCBI database. Phylogenetic analysis was based on combined ITS, TEF1, TUB2 and CAL sequences data using MEGA 11 software to construct phylogenetic tree with Maximum Likelihood (Doilom et al. 2017). In the phylogenetic tree, the combined sequences attributed the three isolates to the D. tectonendophytica (Fig S2). The pathogenicity was tested on leaves of 1.5-year-old D. odorifera seedlings. Three leaves were wounded with a sterile needle and individually inoculated with a 5 mm mycelial disk of PDA culture from each isolate. Sterile PDA disks inoculated leaves as a control. The test was repeated three times. The inoculated plants were placed in a greenhouse at 25℃ and 90% humidity, with a photoperiod of 12 hours. Five days after inoculation, necrotic lesions appeared on inoculated leaves and symptoms from all three isolates were the same as those form natural infections ( Fig S1, K-N), whereas all the control remained symptomless (Fig S1, P). The pathogen was reisolated from the inoculated leaves and again identified as D. tectonendophytica, with the same methodology used for the initial identification. D. tectonendophytica was reported to cause plant diseases, such as stem gray blight of red-fleshed dragon fruit (Hylocereus polyrhizus) (Rahim et al. 2021), leaf spots disease on Elaeagnus conferta and Pometia pinnata (Sun et al. 2021). To our knowledge, this is the first report of D. ctonendophytica causing leaf spot disease on D. odorifera.

Functional genomic analysis of the isolated potential probiotic Lactobacillus delbrueckii subsp. indicus TY-11 and its comparison with other Lactobacillus delbrueckii strains.

This study aimed to conduct functional genomic analysis to uncover the probiotic potential of Lactobacillus delbrueckii subsp. indicus TY-11 isolated from native yogurt in Bangladesh. We also performed transmission electron microscopic (TEM) analysis, comparative genomic study as well as phylogenetic tree construction with 332 core genes from 262 genomes. In our current investigation, we revealed a number of common and unique excellences of the probiotic Lactobacillus delbrueckii subsp. indicus TY-11 that are likely to be important to illustrate its intestinal residence and probiotic roles. This is the first study to explore the molecular insights into intestinal residence and probiotic roles, including antimicrobial activities and antibiotic sensitivity, of a representative strain (TY-11) of Lactobacillus delbrueckii subsp. indicus.

Integrative multi-region molecular profiling of primary prostate cancer in men with synchronous lymph node metastasis

Localized prostate cancer is frequently composed of multiple spatially distinct tumors with significant inter- and intra-tumoral molecular heterogeneity. This genomic diversity gives rise to many competing clones that may drive the biological trajectory of the disease. Previous large-scale sequencing efforts have focused on the evolutionary process in metastatic prostate cancer, revealing a potential clonal progression to castration resistance. However, the clonal origin of synchronous lymph node (LN) metastases in primary disease is still unknown. Here, we perform multi-region, targeted next generation sequencing and construct phylogenetic trees in men with prostate cancer with synchronous LN metastasis to better define the pathologic and molecular features of primary disease most likely to spread to the LNs. Collectively, we demonstrate that a combination of histopathologic and molecular factors, including tumor grade, presence of extra-prostatic extension, cellular morphology, and oncogenic genomic alterations are associated with synchronous LN metastasis.

RsRbohD1 Plays a Significant Role in ROS Production during Radish Pithiness Development.

Pithiness is one of the physiological diseases of radishes, which is accompanied by the accumulation of reactive oxygen species (ROS) during the sponging of parenchyma tissue in the fleshy roots. A respiratory burst oxidase homolog (Rboh, also known as NADPH oxidase) is a key enzyme that catalyzes the production of ROS in plants. To understand the role of Rboh genes in radish pithiness, herein, 10 RsRboh gene families were identified in the genome of Raphanus sativus using Blastp and Hmmer searching methods and were subjected to basic functional analyses such as phylogenetic tree construction, chromosomal localization, conserved structural domain analysis, and promoter element prediction. The expression profiles of RsRbohs in five stages (Pithiness grade = 0, 1, 2, 3, 4, respectively) of radish pithiness were analyzed. The results showed that 10 RsRbohs expressed different levels during the development of radish pithiness. Except for RsRbohB and RsRbohE, the expression of other members increased and reached the peak at the P2 (Pithiness grade = 2) stage, among which RsRbohD1 showed the highest transcripts. Then, the expression of 40 genes related to RsRbohD1 and pithiness were analyzed. These results can provide a theoretical basis for improving pithiness tolerance in radishes.

Common Methods for Phylogenetic Tree Construction and Their Implementation in R.

A phylogenetic tree can reflect the evolutionary relationships between species or gene families, and they play a critical role in modern biological research. In this review, we summarize common methods for constructing phylogenetic trees, including distance methods, maximum parsimony, maximum likelihood, Bayesian inference, and tree-integration methods (supermatrix and supertree). Here we discuss the advantages, shortcomings, and applications of each method and offer relevant codes to construct phylogenetic trees from molecular data using packages and algorithms in R. This review aims to provide comprehensive guidance and reference for researchers seeking to construct phylogenetic trees while also promoting further development and innovation in this field. By offering a clear and concise overview of the different methods available, we hope to enable researchers to select the most appropriate approach for their specific research questions and datasets.

A α-L-rhamnosidase from Echinacea purpurea endophyte Simplicillium sinense EFF1 and its application in production of Calceorioside B

Calceorioside B, a multifunctional phenylethanol glycosides (PhGs) derivative, exhibits a variety of notable properties, such as antithrombotic, anti-tumorigenic, anti-neocoronavirus, anti-inflammatory, and neuroprotective effects. However, the large-scale production of calceorioside B is routinely restricted by its existence as an intermediary compound derived from plants, and still unachieved through excellent and activity chemical synthesis. Here, a total of 51 fungal endophytes were isolated from four PhGs-producing plants, and endophyte Simplicillium sinense EFF1 from Echinacea purpurea was identified with the ability to de-rhamnosing isoacteoside to generate calceorioside B. According to the RNA-transcription of EFF1 under the various substrates, a key gene CL1206.Contig2 that undertakes the hydrolysis function was screened out and charactered by heterologous expression. The sequence alignment, phylogenetic tree construction and substrate specificity analysis revealed that CL1206 was a novel α-L-rhamnosidase that belongs to the glycosyl hydrolase family 78 (GH78). The optimum catalytic conditions for CL1206 were at pH 6.5 and 55 °C. Finally, the enzyme-catalyzed approach to produce calceorioside B from 50 % crude isoacteoside extract was explored and optimized, with the maximum conversion rate reaching 69.42 % and the average producing rate reaching 0.37 g-1.L-1.h−1, which offered a great biocatalyst for potential industrial calceorioside B production. This is the first case for microorganism and rhamnosidase to show the hydrolysis ability to caffeic acid-modified PhGs.

A re-evaluation of Diaporthe: refining the boundaries of species and species complexes

Diaporthe is an important plant pathogenic genus, which also occurs as endophytes and saprobes. Many Diaporthe species that are morphologically similar proved to be genetically distinct. The current understanding of Diaporthe taxonomy by applying morphological characters, host associations and multi-gene phylogeny are problematic leading to overestimation/underestimation of species numbers of this significant fungal pathogenic genus. Currently, there are no definite boundaries for the accepted species. Hence, the present study aims to re-structure the genus Diaporthe, based on single gene phylogenies (ITS, tef, tub, cal and his), multi-gene phylogeny justified by applying GCPSR (Genealogical Concordance Phylogenetic Species Recognition) methodology as well as the coalescence-based models (PTP—Poisson Tree Processes and mPTP—multi-rate Poisson Tree Processes). Considering all available type isolates of Diaporthe, the genus is divided into seven sections while boundaries for 13 species and 15 species-complexes are proposed. To support this re-assessment of the genus, 82 Diaporthe isolates obtained from woody hosts in Guizhou Province in China were investigated and revealed the presence of two novel species and 17 previously known species. Synonymies are specified for 31 species based on molecular data and morphological studies. Dividing Diaporthe into several specific sections based on phylogenetic analyses can avoid the construction of lengthy phylogenetic trees of the entire genus in future taxonomic studies. In other words, when one conducts research related to the genus, only species from the appropriate section need to be selected for phylogenetic analysis.

The characterization of Apis cerana and Apis mellifera mrjp2 gene in Indonesia and the phylogeny relationship with mrjp family genes

The major royal jelly protein 2 (mrjp2) gene is one of the molecular markers that can discriminate between Apis cerana Fabricius and A. mellifera Linnaeus. Due to the lack of mrjp2 gene sequences registered in GenBank for Indonesian A. cerana and A. mellifera, DNA characterization and bioinformatics analysis were needed. This research aimed to characterize the exon-intron organization of mrjp2 genes for both Indonesian bee species and analyze the phylogenetic relationship with other mrjp family genes. DNA samples of Apis cerana and A. mellifera, collected from Bogor, were amplified using MF-MR primer at annealing temperatures of 47 oC and 50 oC, respectively. The length of A. cerana and A. mellifera DNA sequences were 579 and 597 bp, respectively. The DNA sequences of both species were comprised of partial exons 1, introns 1, exons 2, introns 2, and partial exon 3. The number of putative amino acids of A. cerana and A. mellifera mrjp2 genes were 111 and 123, respectively. We confirmed that the partial MRJP2 putative amino acids of both honey bees belong to the MRJP family and contained the peptide signal in the 14 first amino acid sites. Nucleotide variation of the mrjp2 gene in A. cerana was higher than A. mellifera. Phylogenetic tree construction showed that A. cerana and A. mellifera form a monophyletic clade with the A. mellifera mrjp7 gene and another mrjp family gene clustered as reported in a previous study.