Apple mold heart disease, primarily caused by Trichothecium roseum, is the most severe disease affecting stored apples. Developing biocontrol resources as an alternative to chemical pesticides is crucial for the advancement of green agriculture. This study demonstrated that a pathogenic fungus isolated from sunflower leaves exhibited specific inhibitory effects against T. roseum. Through morphological observation, identification, and the construction of a phylogenetic tree analysis, the fungus was identified as Alternaria angustiovoidea. GW2A was found to inhibit the spread of diseases on apple twigs. Additionally, GW2A has significant preventive and therapeutic effects on apple mold heart disease. Furthermore, GW2A can induce apple trees to upregulate defense-related genes, thereby enhancing resistance. Transcriptome analysis revealed that GW2A inhibits T. roseum growth by suppressing the function of polysaccharide hydrolases, oxidoreductases, and intermediate steps in carbohydrate metabolism. In conclusion, our research has effectively isolated and characterized A. angustiovoidea, revealing its significant potential as a biocontrol agent against T. roseum and apple mold heart disease, particularly in areas where sunflowers and apple trees are not co-cultivated. Additionally, we demonstrated its ability to induce resistance in apple trees, offering a sustainable approach to disease management in apple cultivation.

As a crucial and integral adaptation for thriving in diverse habitats, whether for survival or disease prevention and control, biofilm plays a vital role for most biocontrol bacteria, such as B. subtilis , Bacillus amyloliquefaciens, and plant-growth-promoting rhizobacteria (PGPR). However, the process of biofilm formation is intricate, and its regulatory mechanism remains unclear. In this study, we discovered that the regulatory protein MutL significantly influenced biofilm formation and exhibited a diminished colonization effectiveness on rice leaves. The mutant, lacking protein MutL expression, was unable to form biofilm with normal morphology and yielded only a quarter of the biofilm weight observed in the wild type B.subtilis YT1. In a petri dish confrontation assay examining the inhibitory effects on Rhizoctonia solani, no significant differences were observed between the mutant strain and the wild type YT1. Furthermore, through GFP fluorescent labeling technology, we conducted additional colonization tests, which demonstrated that the mutant failed to colonize rice stems effectively in the presence of R. solani. We hypothesize that the negative impact on biofilm formation resulted in inadequate colonization of rice stems, this combination accounts for the poor biocontrol efficacy against rice sheath blight, but it does not affect the normal growth of the strain or other biological phenotypes.

Leaf spot, a major apple disease, manifests in diverse symptoms. In this study, the pathogen was isolated from diseased ‘Yanfu 3’ apple leaves in Yantai, Shandong Province, and identified as Neopestalotiopsis clavispora through morphological observation, molecular identification, and multi-gene (ITS, TEF1α, and TUB2) phylogenetic analysis. Three isolates (YTNK01, YTNK02, and YTNK03) were selected for pathogenicity tests to verify Koch’s postulates. To our knowledge, this is the first report of N. clavispora being responsible for apple leaf spots in China, and the disease has been named ‘apple Neopestalotiopsis leaf spot’. Additionally, N. clavispora was found to infect crabapple, sweet cherry, grape, peach, and pear under laboratory conditions, indicating that these fruit trees may be potential hosts for N. clavispora in the field. The in vitro toxicity of ten fungicides to the pathogen was assessed using the mycelial growth rate method. All ten fungicides were effective in inhibiting the growth of N. clavispora. Among them, those based on pylocyanonitrile, propiconazole, pyraclostrobin, tebuconazole, diphenoxazole, and osthole showed higher toxicity to N. clavispora, with EC50 values of 0.11, 0.41, 0.47, 1.32, 1.85, and 3.82 µg/mL, respectively. These fungicides could be used as alternatives to prevent this disease in production. Overall, these findings provide valuable insights into the characteristics of N. clavispora causing apple leaf spot and are crucial for developing effective management strategies.

The purpose of this study is to discover and excavate more key factors and signaling pathways that regulate the formation intensity of biofilms and to fully reveal the possible models affecting biofilm formation. By using gene homologous recombination and bioinformatics technology, a MutL protein-directed deletion mutant strain was successfully constructed. The growth status of the mutant strain was observed, and it was confirmed that, except for the change in cell morphology, there were no significant differences in growth and reproduction between the mutant strain and the wild-type strain. By using the induced biofilm formation technique, the significant decrease in biofilm formation in the MutL mutant strain was successfully verified. The plate confrontation test confirmed that the inhibitory ability of the mutant strain against rice blast fungus was not significantly different from that of the WT strain. The colonization ability of the mutant strain on rice stems was tested, and it was confirmed that the colonization ability of the mutant strain was significantly lower than that of the WT strain. In terms of the prevention and control effect of rice blast disease, the mutant strain showed a significant decrease. By using transcriptomic big data, the gene and pathway expression differences between the mutant strain and the WT strain during biofilm formation were analyzed. The analysis revealed no significant correlation with the previously reported spo0A and tapA-sipW-tasA pathways. The key factor capB of the polyglutamic acid signaling pathway, which affects the formation of the biological model, was found to have a significant decrease in expression. A mechanical hypothesis was proposed: MutL may participate in regulating the formation intensity of Bacillus biofilms by regulating the formation of glutamic acid to polyglutamic acid.

With the development of material science and increasing awareness of ecological environmental protection, liquid biodegradable mulch films (LBDMs) have garnered significant public interest. In this research, new LBDMs were developed using hydrophobically modified polymer materials, surfactants, and photosensitive catalysts. Characterization by scanning electron microscopy (SEM) revealed good material compatibility. LBDMs exhibited excellent wettability and degradability, effectively covering soil surfaces and enhancing soil moisture conservation, with a degradation rate of 76.09% after 80 days of burial. The field performance experiment was conducted over two consecutive years, 2021 and 2022, to assess differences in soil temperature and moisture, peanut agronomic traits, pod traits, and yield under four treatments: non-mulching (CK), LBDMs, clear polyethylene mulch films (CPEMs), and black polyethylene mulch films (BPEMs). LBDMs increased soil temperature by 0.56 °C and soil moisture by 19.25%, accelerated the seedling stage by 4-to-6 days, and improved the average emergence rate by 15.91%. Furthermore, LBDMs significantly promoted peanut growth, and it increased yield by 14.34% compared to CK. LBDMs performed comparably to the two types of PE films in maintaining soil conditions and different crop phenotype traits, including plant height, branch number, yield, and quality, and they even outperformed PE films in productivity per plant and 100-kernel weight. These findings suggest that LBDMs are a promising eco-friendly alternative to traditional PE films.

Apple Alternaria blotch disease, caused by Alternaria alternata (Fr.) Keissl, is one of the most famous leaf diseases. When the disease is prevalent, it causes leaf abscission and influences the formation of flower buds and photosynthesis. Therefore, a simple, rapid, high-specificity and sensitivity method for monitoring infected leaves at early developmental stages is urgently needed, so that the occurrence and expansion of A. alternata can be controlled in time. In our research, a rapid, specific and efficient loop-mediated isothermal amplification (LAMP) method was developed to detect A. alternata within 60 min. Six primers of LAMP detection can only specifically amplify the aapg-1 gene in A. alternata but not in four other important fungi in apples. The aapg-1 gene encodes endopolygalacturonase in A. alternata, and there are significant differences among different species. Thus, it was applied as the target for LAMP primers. Compared to conventional PCR detection, our LAMP method had the same sensitivity as that of detecting as little as 1 fg of pure genomic DNA of A. alternata. When leaves were inoculated with A. alternata conidia, LAMP detected 1 × 102 conidia/mL as the minimum concentration. However, the traditional tissue isolation and identification method only isolated A. alternata from leaves inoculated with 1 × 105 and 1 × 106 conidia/mL, indicating that the LAMP method was more sensitive than the traditional tissue isolation and identification method for A. alternata before symptoms. Further tests also indicated that LAMP detection was more accurate and sensitive than the traditional tissue isolation and identification method for A. alternata in leaves with the Alternaria blotch symptom collected from the field. Our results showed that the LAMP-targeting the aapg-1 gene has the advantages of high sensitivity, specificity and simplicity and can be used for rapid detection and early monitoring of A. alternata in the field. LAMP is instructive for us to effectively prevent and control apple Alternaria blotch disease.

Background Anastomosis group 1 IA (AG1-IA) of Rhizoctonia solani is the major agent of banded leaf and sheath blight (BLSB) disease that causes severe yield loss in many worldwide crops. MicroRNAs (miRNAs) are ~ 22 nt non-coding RNAs that negatively regulate gene expression levels by mRNA degradation or translation inhibition. A better understanding of miRNA function during AG1-IA infection can expedite to elucidate the molecular mechanisms of fungi-host interactions. Results In this study, we sequenced three small RNA libraries obtained from the mycelium of AG1-IA isolate, non-infected maize sheath and mixed maize sheath 3 days after inoculation. In total, 137 conserved and 34 novel microRNA-like small RNAs (milRNAs) were identified from the pathogen. Among these, one novel and 17 conserved milRNAs were identified as potential virulence-associated (VA) milRNAs. Subsequently, the prediction of target genes for these milRNAs was performed in both AG1-IA and maize, while functional annotation of these targets suggested a link to pathogenesis-related biological processes. Further, expression patterns of these virulence-associated milRNAs demonstrated that theyparticipate in the virulence of AG1-IA. Finally, regulation of one maize targeting gene, GRMZM2G412674 for Rhi-milRNA-9829-5p, was validated by dual-luciferase assay and identified to play a positive role in BLSB resistance in two maize mutants. These results suggest the global differentially expressed milRNAs of R. solani AG1-IA that participate in the regulation of target genes in both AG1-IA and maize to reinforce its pathogenicity. Conclusions Our data have provided a comprehensive overview of the VA-milRNAs of R. solani and identified that they are probably the virulence factors by directly interfered in host targeting genes. These results offer new insights on the molecular mechanisms of R.solani -maize interactions during the process of infection.

In a recent study, anthocyanins, which have a strong free radical-scavenging activity, were examined for their potential to effectively prevent cancer. However, clinical trials are limited by the purity of the anthocyanin. Multiple methods are used to extract and purify anthocyanins. Based on previous work on Solanum nigrum, which is a widely distributed plant, in this study, DM130 macroporous resin, Sephadex LH20, and a C18 column were used to separate cis-trans anthocyanin isomers. These anthocyanins constitute the majority of total S. nigrum anthocyanins. The results showed that this "DM130-LH20-C18 system" can be used to obtain a cinnamic acid-derived cistrans anthocyanin, petunidin-3-(p- coumaroyl)-rutinoside-5-glucoside, with a purity of 98.5%, for effective quantitation. In order to determine the antioxidant ability of the petunidin-3-(p- coumaroyl)-rutinoside-5-glucoside cis-trans isomers, three ordinary methods were adopted. The maximum antioxidant ability of the cis-trans anthocyanin was dozens of times higher than that of vitamin C.

Cherry tomato ( Lycopersicon esculentum M.) is considered a healthy fruit worldwide due to its wide range of nutrients. Flavonol, one of the major nutrients in cherry tomato, has antioxidant and cell-modulating properties. In this study, we showed a correlation between the expression of SlMYB12 and flavonol content (R 2 = 0.922). To characterize the function of SlMYB12 , SlMYB12 -overexpressing transgenic tomato plants were generated in three different cherry tomato varieties. Significant increases in flavonol content and flavonol biosynthetic gene expression were identified in SlMYB12 -overexpressing plants. Therefore, we suggest that SlMYB12 plays a positive role in the flavonol biosynthesis pathway in cherry tomatoes, which further indicates a potential role as a marker in analyzing flavonol content in different cherry tomato varieties.

Solanum nigrum fruits have been conventionally used in beverages due to their nutritional substances such as minerals, vitamins, amino acids, proteins, sugars, polyphenols, and anthocyanins. The characterization of components and regulatory mechanism of anthocyanins in S. nigrum fruits have rarely been reported. In this study, we determined that the peel and flesh of S. nigrum fruits shared similar HPLC profiles but different contents and total antioxidant activities for anthocyanins. After an efficient purification method, mainly including extraction with pH 1.0 distilled water and then desorption with pH 1.0 95% ethanol after a DM-130 resin adsorption step to obtain more pure anthocyanin extracts, the purity of anthocyanins extracted from S. nigrum fruits reached 56.1%. Moreover, eight anthocyanins from S. nigrum fruit were identified with HPLC-MS/MS for the first time. A typical R2R3-MYB transcription factor gene, SnMYB, was also cloned for the first time by rapid amplification of cDNA ends (RACE)-PCR from S. nigrum. Moreover, the contents of anthocyanins were shown to correlate well (r = 0.93) with the expression levels of SnMYB gene during the fruit's developmental stages. Most significantly, SnMYB gene successfully produced high anthocyanin content (1.03 mg/g) when SnMYB gene was transiently expressed in tobacco leaves. Taken together, S. nigrum fruits are a promising resource for anthocyanin extraction, and SnMYB gene is an activator that positively regulates anthocyanin biosynthesis in S. nigrum.

Potato (Solanum tuberosum L.) is a major crop worldwide that meets human economic and nutritional requirements. Potato has several advantages over other crops: easy to cultivate and store, cheap to consume, and rich in a variety of secondary metabolites. In this study, we generated three marker-free transgenic potato lines that expressed the Arabidopsis thaliana flavonol-specific transcriptional activator AtMYB12 driven by the tuber-specific promoter Patatin. Marker-free potato tubers displayed increased amounts of caffeoylquinic acids (CQAs) (3.35-fold increases on average) and flavonols (4.50-fold increase on average). Concentrations of these metabolites were associated with the enhanced expression of genes in the CQA and flavonol biosynthesis pathways. Accumulation of CQAs and flavonols resulted in 2-fold higher antioxidant capacity compared to wild-type potatoes. Tubers from these marker-free transgenic potatoes have therefore improved antioxidant properties.

Interest in so-called functional foods, i.e., foods that promote health and disease prevention, is increasing. Consumption of plant polyphenols has been implicated in the prevention of age-related diseases, cardiovascular disease and cancer. AtMYB12 and AtMYB111, which were originally identified as flavonol-specific transcriptional activator in Arabidopsis thaliana, induces the accumulation of high levels of polyphenols in tobacco and tomato plants. In this study, we determined that the transcription factor AtMYB11 as its close homologs, AtMYB12 and AtMYB111, regulate flavonoid and caffeoylquinic acid biosynthesis in tomato and tobacco plants. Constitutive expression of AtMYB11 enhanced the expression of genes in the phenylpropanoid biosynthesis pathway, resulting in the accumulation of flavonoids and chlorogenic acid in tobacco and tomato plants. Increases in dicaffeoylquinic acid and tricaffeoylquinic acid content were only detected in AtMYB11-expressing tomato fruits. These results suggest the potential of transgenic tomatoes with improved antioxidant properties as functional foods.