Isolation and screening of fungi for enhanced agarwood formation in Aquilaria sinensis trees.

Agarwood is a resinous heartwood of Aquilaria sinensis that is formed in response to mechanical wounding. In the present study pre-treatment of Aquilaria sinensis was carried out, and then the dominant fungi were isolated and purified from the surface and electroshock holes of trees. The isolated Trichoderma sp. and Neurospora sp. were then screened for resistance against benzyl acetone and then inoculated into healthy Aquilaria sinensis trees. After six months, the agarwood was collected for analysis. The chemical composition of incense was analyzed using gas chromatography-mass spectroscopy, and 82 chemical constituents were identified. Agarwood products formed by using Trichoderma sp. and Neurospora sp. consisted of 50.22% and 48.71% ether extracts, respectively, which surpassed the 10% threshold specified by the Chinese Pharmacopoeia. Similarly, relative aromatic contents in the two agarwood products were 30.1% and 32.86%, while proportions of sesquiterpene constituents were 10.21% and 11.19%, respectively. These two agarwood-specific chemical constituents accounted for a large proportion of the total chemical composition, which showed that the generated agarwood was of good quality. The results of the study revealed that both Trichoderma sp. and Neurospora sp. were able to effectively induce agarwood production in Aquilaria sinensis trees in 6 months. This study expands the library of fungi that promote the production of agarwood from Aquilaria sinensis trees.

The Research of Antagonistic Endophytic Bacterium Bacillus velezensis CSUFT-BV4 for Growth Promotion and Induction of Resistance to Anthracnose in Camellia oleifera.

Camellia oleifera (C. oleifera) is one of the four main, woody, edible oil tree species in the world, while C. oleifera anthracnose is mainly caused by the fungus Colletotrichum fructicola (C. fructicola), which severely affects the yield of C. oleifera and the quality of tea oil. Bacillus velezensis (B. velezensis) CSUFT-BV4 is an antagonistic endophytic bacterium isolated from healthy C. oleifera leaves. This study aimed to investigate the biocontrol potential of strain CSUFT-BV4 against C. oleifera anthracnose and its possible functional mechanism, and to determine its growth-promoting characteristics in host plants. In vitro, CSUFT-BV4 was shown to have efficient biofilm formation ability, as well as significant functions in the synthesis of metabolic substances and the secretion of probiotic substances. In addition, the CSUFT-BV4 fermentation broth also presented efficient antagonistic activities against five major C. oleifera anthracnose pathogens, including C. fructicola, C. gloeosporioides, C. siamense, C. camelliae, and C. kahawae, and the inhibition rate was up to 73.2%. In vivo, it demonstrated that the growth of C. oleifera treated with CSUFT-BV4 fermentation broth was increased in terms of stem width, plant height, and maximum leaf area, while the activities of various defense enzymes, e.g., superoxide dismutase (SOD), phenylalanine aminotransferase (PAL), and polyphenol oxidase (PPO), were effectively increased. The remarkable antagonistic activities against C. oleifera anthracnose, the growth-promoting characteristics, and the induction of host defense responses indicate that endophytic bacterium CSUFT-BV4 can be effectively used in the biological control of C. oleifera anthracnose in the future, which will have a positive impact on the development of the C. oleifera industry.

Research progress and management strategies of fungal diseases in Camellia oleifera.

Camellia oleifera Abel, a woody oil plant, that is endemic to China. Tea oil, also referred to as "oriental olive oil," is a superior quality plant-based cooking oil. The production of tea oil accounts for 8% of the total edible vegetable oil production in the country. Since 2022, the annual output value of C. oleifera industry has exceeded 100 billion yuan, making it one of the major economic contributors to China's rural revitalization development strategy. In recent years, demand and production have grown in parallel. However, this has led to an increase in the incidence levels of pest and diseases. Pests and diseases significantly reduce the quality and yield of C. oleifera. C. oleifera diseases are mainly caused by pathogenic fungi. C. oleifera anthracnose, soft rot, leaf spot, coal stain, leaf gall disease, and root rot are the most important fungal diseases affecting the C. oleifera industry. However, the same disease may be caused by different pathogenic fungi. C. oleifera can be found in half of China and is found in several climatic zones. The geographical distribution of woody plant diseases is consistent with the distribution of the tree species and the ecology of the range, which also results in a highly complex distribution of fungal diseases of C. oleifera. The management of fungal diseases in C. oleifera is extremely challenging due to the variety of pathogenic fungal species, multiple routes of transmission, the lack of resistant plants, and the environmental safety of chemical measures. The optimal strategy for addressing fungal diseases in C. oleifera is to develop and apply an integrated disease management plan. This review provides a brief overview of the pathogenic species, pathogenesis, pathogenesis, geographical distribution, current management strategies, and potentially new methods of C. oleifera fungal diseases, to provide direction for the development of comprehensive management measures for C. oleifera fungal diseases in the future.

Plant immune receptors interact with hemibiotrophic pathogens to activate plant immunity.

Phytopathogens pose a devastating threat to the productivity and yield of crops by causing destructive plant diseases in natural and agricultural environments. Hemibiotrophic pathogens have a variable-length biotrophic phase before turning to necrosis and are among the most invasive plant pathogens. Plant resistance to hemibiotrophic pathogens relies mainly on the activation of innate immune responses. These responses are typically initiated after the plant plasma membrane and various plant immune receptors detect immunogenic signals associated with pathogen infection. Hemibiotrophic pathogens evade pathogen-triggered immunity by masking themselves in an arms race while also enhancing or manipulating other receptors to promote virulence. However, our understanding of plant immune defenses against hemibiotrophic pathogens is highly limited due to the intricate infection mechanisms. In this review, we summarize the strategies that different hemibiotrophic pathogens interact with host immune receptors to activate plant immunity. We also discuss the significant role of the plasma membrane in plant immune responses, as well as the current obstacles and potential future research directions in this field. This will enable a more comprehensive understanding of the pathogenicity of hemibiotrophic pathogens and how distinct plant immune receptors oppose them, delivering valuable data for the prevention and management of plant diseases.

Utilization of current pyrolysis technology to convert biomass and manure waste into biochar for soil remediation: A review.

Traditional disposal of animal manures and lignocellulosic biomass is restricted by its inefficiency and sluggishness. To advance the carbon management and greenhouse gas mitigation, this review scrutinizes the effect of pyrolysis in promoting the sustainable biomass and manure disposal as well as stimulating the biochar industry development. This review has examined the advancement of pyrolysis of animal manure (AM) and lignocellulosic biomass (LB) in terms of efficiency, cost-effectiveness, and operability. In particular, the applicability of pyrolysis biochar in enhancing the crops yields via soil remediation is highlighted. Through pyrolysis, the heavy metals of animal manures are fixated in the biochar, thereby both soil contamination via leaching and heavy metal uptake by crops are minimized. Pyrolysis biochar is potentially use in soil remediation for agronomic and environmental co-benefits. Fast pyrolysis assures high bio-oil yield and revenue with better return on investment whereas slow pyrolysis has low revenue despite its minimum investment cost because of relatively low selling price of biochar. For future commercialization, both continuous reactors and catalysis can be integrated to pyrolysis to ameliorate the efficiency and economic value of pyrolysis biochar.

Recent progress on harm, pathogen classification, control and pathogenic molecular mechanism of anthracnose of oil-tea.

Oil tea (Camellia oleifera), mainly used to produce high-quality edible oil, is an important cash crop in China. Anthracnose of oil tea is a considerable factor that limits the yield of tea oil. In order to effectively control the anthracnose of oil tea, researchers have worked hard for many years, and great progress has been made in the research of oil tea anthracnose. For instance, researchers isolated a variety of Colletotrichum spp. from oil tea and found that Colletotrichum fructicola was the most popular pathogen in oil tea. At the same time, a variety of control methods have been explored, such as cultivating resistant varieties, pesticides, and biological control, etc. Furthermore, the research on the molecular pathogenesis of Colletotrichum spp. has also made good progress, such as the elaboration of the transcription factors and effector functions of Colletotrichum spp. The authors summarized the research status of the harm, pathogen types, control, and pathogenic molecular mechanism of oil tea anthracnose in order to provide theoretical support and new technical means for the green prevention and control of oil tea anthracnose.

Selection of potential reference genes for RT-qPCR in the plant pathogenic fungus Colletotrichum fructicola.

Colletotrichum is widespread, and these pathogenic fungi can cause various plant diseases. Studies have shown that Colletotrichum fructicola cause oil-tea (Camellia oleifera) anthracnose and is widely distributed as a dominant fungus in all Ca. oleifera-producing regions. Real-time quantitative PCR(RT-qPCR) is considered the most reliable technique for simultaneously measuring relative gene expression levels in different tissues. Target genes are typically quantified using RT-qPCR to explore gene function, and reliable RT-qPCR results require data normalization using stable reference genes. No studies have reported a suitable reference gene in C. fructicola. This study has eight candidate reference genes (CfCk, CfRpp, CfUce, CfRrp, CfAdrh, CfDd, CfAct, and CfTub) which were selected from C. fructicola-Ca. oleifera transcriptome data and evaluated and sequenced using geNorm, NormFinder, and BestKeeper algorithms. The results showed that CfRrp had better stability in C. fructicola, both during the growth of pure pathogenic fungi and during the invasion of different oil-tea leaves. After normalization with CfRrp, the differentially expressed target genes were similar to the transcriptome. Our work provides suitable reference genes for future studies to quantify target gene expression levels in C. fructicola.

A perspective on the interaction between biochar and soil microbes: A way to regain soil eminence.

Soil ecosystem imparts a fundamental role in the growth and survival of the living creatures. The interaction between living and non-living constituents of the environment is important for the regulation of life in the ecosystem. Biochar is a carbon rich product present in the soil that is responsible for various applications in diversified fields. In this review, we focused on the collaboration between the soil, biochar and microbial community present in the soil and consequences of it in the ecosystem. Herein, it primarily discusses on the different approaches of the production and characterization of biochar. Furthermore, this review also discusses about the optimistic interaction of biochar with soil microbes and their role in plant growth. Eventually, it reveals the various physio-chemical properties of biochar, including its specific surface area, porous nature, ion exchange capacity, and pH, which aid in the modification of the soil environment. Furthermore, it elaborately discloses the impact of the biochar addition in the soil focusing mainly on its interaction with microbial communities such as bacteria and fungi. The physicochemical properties of biochar significantly interact with microbes and improve the beneficial microbes growth and increase soil nutrients, which resulting reasonable plant growth. The main focus remains on the role of biochar-soil microbiota in remediation of pollutants, soil amendment and inhibition of pathogenicity among plants by promoting resistance potential. It highlights the fact that adding biochar to soil modulates the soil microbial community by increasing soil fertility, paving the way for its use in farming, and pollutant removal.

Research on Forest Conversation Analysis Using Autoregressive Neural Network-Based Model.

Forest biodiversity is an important component of biological diversity that should not be disregarded. The question of how to evaluate it has sparked scholarly inquiry and discussion. The purpose of this paper is to describe the principles of general linear regression, the selection of model variables in OLS autoregressive modelling, model coefficient testing, analysis of variance of autoregressive models, and model evaluation indicators in order to clarify the suitability of GWR models for solving biomass-related data problems. The GWR 4.0 program was used to create a spatially weighted autoregressive model. Model testing and an accuracy analysis were performed on the model. Following a comparison and study with the general linear regression model, it was discovered that the geographically weighted autoregressive model is better suited to defining spatially correlated data than the general linear regression model.

The Multifunctions and Future Prospects of Endophytes and Their Metabolites in Plant Disease Management.

Endophytes represent a ubiquitous and magical world in plants. Almost all plant species studied by different researchers have been found to harbor one or more endophytes, which protect host plants from pathogen invasion and from adverse environmental conditions. They produce various metabolites that can directly inhibit the growth of pathogens and even promote the growth and development of the host plants. In this review, we focus on the biological control of plant diseases, aiming to elucidate the contribution and key roles of endophytes and their metabolites in this field with the latest research information. Metabolites synthesized by endophytes are part of plant disease management, and the application of endophyte metabolites to induce plant resistance is very promising. Furthermore, multi-omics should be more fully utilized in plant-microbe research, especially in mining novel bioactive metabolites. We believe that the utilization of endophytes and their metabolites for plant disease management is a meaningful and promising research direction that can lead to new breakthroughs in the development of more effective and ecosystem-friendly insecticides and fungicides in modern agriculture.

Transcriptome Analysis of Colletotrichum fructicola Infecting Camellia oleifera Indicates That Two Distinct Geographical Fungi Groups Have Different Destructive Proliferation Capacities Related to Purine Metabolism.

Anthracnose, caused by Colletotrichum spp., is a significant disease affecting oil tea (Camellia oleifera Abel.). Extensive molecular studies have demonstrated that Colletotrichum fructicola is the dominant pathogen of oil tea anthracnose in China. This study aims to investigate differences in molecular processes and regulatory genes at a late stage of infection of C. fructicola, to aid in understanding differences in pathogenic mechanisms of C. fructicola of different geographic populations. We compared the pathogenicity of C. fructicola from different populations (Wuzhishan, Hainan province, and Shaoyang, Hunan province) and gene expression of representative strains of the two populations before and after inoculation in oil tea using RNA sequencing. The results revealed that C. fructicola from Wuzhishan has a more vital ability to impact oil tea leaf tissue. Following infection with oil tea leaves, up-regulated genes in the strains from two geographic populations were associated with galactosidase activity, glutamine family amino acid metabolism, arginine, and proline metabolism. Additionally, up-regulated gene lists associated with infection by Wuzhishan strains were significantly enriched in purine metabolism pathways, while Shaoyang strains were not. These results indicate that more transcriptional and translational activity and the greater regulation of the purine metabolism pathway in the C. fructicola of the Wuzhishan strain might contribute to its stronger pathogenicity.

Functional analysis of CfSnf1 in the development and pathogenicity of anthracnose fungus Colletotrichum fructicola on tea-oil tree.

BACKGROUND: Tea-oil tree (Camellia oleifera) is a unique edible-oil tree in China, and anthracnose occurs in wherever it is cultivated, causing great economic losses each year. We have previously identified the Ascomycete fungus Colletotrichum fructicola as the major pathogen of anthracnose in Ca. oleifera. The purpose of this study was to characterize the biological function of Snf1 protein, a key component of the AMPK (AMP-activated protein kinase) pathway, for the molecular pathogenic-mechanisms of C. fructicola. RESULTS: We characterized CfSnf1 as the homolog of Saccharomyces cerevisiae Snf1. Targeted CfSNF1 gene deletion revealed that CfSnf1 is involved in the utilization of specific carbon sources, conidiation, and stress responses. We further found that the ΔCfSnf1 mutant was not pathogenic to Ca. oleifera, resulting from its defect in appressorium formation. In addition, we provided evidence showing crosstalk between the AMPK and the cAMP/PKA pathways for the first time in filamentous fungi. CONCLUSION: This study indicate that CfSnf1 is a critical factor in the development and pathogenicity of C. fructicola and, therefore, a potential fungicide target for anthracnose control.

New Mycodiplosis gall midge (Diptera: Cecidomyiidae) feeding on fungal rusts (Fungi: Pucciniomycetes) that are pathogenic on cultured plants.

Larvae of a gall midge were found feeding on the fungal rust Maravalia pterocarpi (Pucciniomycetes: Pucciniales: Chaconiaceae) infesting the economically important sua tree Dalbergia tonkinensis (Fabaceae) on Hainan Island, China. The adults, pupae and larvae were collected, their morphology was studied and a segment of the Cytochrome Oxidase unit I (COI) mitochondrial gene was sequenced. The gall midge proved to be a species new to science that belongs to the genus Mycodiplosis (Diptera: Cecdiomyiidae). Comparison of the sequence to published Cecdiomyiidae sequences revealed that, despite being undescribed and unnamed, it was previously found in east and south-east Asia to feed on several rust species: Puccinia coronata (Pucciniomycetes: Pucciniales: Pucciniaceae) that develops on Lolium multiflorum (Poaceae), Puccinia sp. on Zea mays (Poaceae), Puccinia arachidis on Arachis hypogaea (Fabaceae) and Puccinia allii on Allium fistulosum (Amaryllidaceae). The new species is described and named here Mycodiplosis puccinivora Jiao, Bu Kolesik. It occurs in China, Japan, Thailand, Bangladesh and possibly Malaysia and Australia. In Hainan it has four to five generations per year.

Population Genetic Analyses of the Fungal Pathogen Colletotrichum fructicola on Tea-Oil Trees in China.

The filamentous fungus Colletotrichum fructicola is found in all five continents and is capable of causing severe diseases in a number of economically important plants such as avocado, fig, cocoa, pear, and tea-oil trees. However, almost nothing is known about its patterns of genetic variation and epidemiology on any of its host plant species. Here we analyzed 167 isolates of C. fructicola obtained from the leaves of tea-oil tree Camellia oleifera at 15 plantations in seven Chinese provinces. Multilocus sequence typing was conducted for all isolates based on DNA sequences at fragments of four genes: the internal transcribed spacers of the nuclear ribosomal RNA gene cluster (539 bp), calmodulin (633 bp), glutamine synthetase (711 bp), and glyceraldehyde-3-phosphate dehydrogenase (190 bp), yielding 3.52%, 0.63%, 8.44%, and 7.89% of single nucleotide polymorphic sites and resulting in 15, 5, 12 and 11 alleles respectively at the four gene fragments in the total sample. The combined allelic information from all four loci identified 53 multilocus genotypes with the most frequent represented by 21 isolates distributed in eight tea-oil plantations in three provinces, consistent with long-distance clonal dispersal. However, despite evidence for clonal dispersal, statistically significant genetic differentiation among geographic populations was detected. In addition, while no evidence of recombination was found within any of the four gene fragments, signatures of recombination were found among the four gene fragments in most geographic populations, consistent with sexual mating of this species in nature. Our study provides the first insights into the population genetics and epidemiology of the important plant fungal pathogen C. fructicola.

[Soil microfauna diversity among Cunninghamia lanceolata plantations based on pyrosequencing].

In order to study the function of soil microfauna and its responses to environmental changes, we used metagenome analyses of the 18S rDNA gene region to identify differences in microfauna diversity and community structure among fifteen soil samples belonging to five different Cunninghamia lanceolate plantations. The plantations were located in Youxian County, Hunan Province in central China. The trees in these plantations were of different ages (3, 13, and 26 years) and belonged to different ecological successions (first, second, and third successions). The total dataset comprised 94922 high quality sequences with an average length of 436 bp. The dominant taxonomic groups across all samples were Chordata, Annelida, Arthropoda, Nematoda, Rotifera and Platyhelminthes with each accounting for 60.8%, 24.0%, 7.4%, 3.6%, 1.5% and 1.2% of the sequences, respectively. There were significant differences in ACE index and Shannon index among the five plantations. The lowest diversity of soil microfauna was in the 13-year old plantation of the first ecological succession. The correlation analysis showed that both ACE and available potassium concentration were negatively correlated to the Chaol index. However, there were no significant correlations between the Shannon, Simpson indices and the physical-chemical properties of soil. Overall, the Jaccard's similarity coefficient was less than 0.4 among samples at each site, and significant differences were found among plantations.

Optimization of medium components for production of chitin deacetylase by Bacillus amyloliquefaciens Z7, using response surface methodology.

Plackett-Burman design and Box-Behnken response surface methodology (RSM) was employed to optimize the medium components for the chitin deacetylase (CDA) activity from Bacillus amyloliquefaciens Z7. Plackett-Burman design was applied to determine the specific medium components affecting CDA activity and found that starch, chitin and MgSO4 were critical in augmenting CDA activity. These significant parameters were further optimized using Box-Behnken RSM and the optimum concentrations of starch, chitin and MgSO4 were found to be 24.4, 8.8 and 0.19 g/L, respectively. The optimum medium composition was chitin 8.8 g/L, starch 24.4 g/L, yeast extract 10g/L, MgSO4 0.19 g/L, K2HPO4 0.3 g/L and NaCl 5 g/L. Under these optimal conditions, the CDA activity of Bacillus amyloliquefaciens Z7 increased distinctly from 18.75 to 27.48 U/mL (46.6% increase in total yield).

[Prediction of chlorophyll content of leaves of oil camelliae after being infected with anthracnose based on Vis/NIR spectroscopy].

The prediction model of chlorophyll content of leaves in canopies of oil camelliae under disease was explored and built by analyzing the Vis/NIR spectroscopy characteristics of oil camelliae canopies after being injected with anthracnose. Through field survey of disease index (DI), chlorophyll content and spectral data of leaves in canopies surviving different severity of disease were acquired. The first order differential of spectral data combined with moving average filter was pretreated. The prediction model of BP neural network of chlorophyll content was built by extracting sensitive wave band from spectral resample data. The results showed that with the disease being aggravated, reflection peaks and valleys of spectra of oil camelliae canopies in visible-light region vanished gradually, steep red edges from red light to near infrared leveled little by little, and reflectivity of healthy oil camelliae was far larger than that of ill ones. The sensitive wave band of absorption and reflection of chlorophyll lay in the region of 84-512, 533-565, 586-606 and 672-724 nm. The correlation coefficient r and RMSE between predictive values calculated from BP neural network using sensitive wave band as input variables and observed values was 0.9921 and 0.0458 respectively. It was therefore feasible to utilize Vis/NIR spectroscopy technology to forecast the chlorophyll content of oil camelliae after being infected with anthracnose.