Fusarium wilt of banana, caused by Fusarium oxysporum f. sp. cubense (Foc), especially Tropical Race 4 (TR4), seriously threatens banana production worldwide. There is no single effective control measure, although certain Bacillus strains secrete antibiotics as promising disease-biocontrol agents. This study identified five Bacillus strains displaying strong antibiotic activity against TR4, using a systemic assessment for presence/absence of genetic markers at genome level, and expression profiles at transcriptome level. A conventional PCR with 13 specific primer pairs detected biocontrol-related genes. An accurate, quantitative real-time PCR protocol with novel designed specific primers was developed to characterise strain-specific gene expression, that optimises strain-culturing and RNA-isolation methodologies. Six genes responsible for synthesising non-ribosomal peptide synthetase biocontrol metabolites were detected in all five strains. Three genes were involved in synthesising three Polyketide synthetase metabolites in all five strains, but the macrolactin synthase gene mln was only detected in WBN06 and YN1282-2. All five Bacillus strains have the genes dhb and bioA, essential for synthesising bacillibactin and biotin. However, the gene sboA, involved in subtilisin synthesis, is absent in all five strains. These genes’ expression patterns were significantly different among these strains, suggesting different mechanisms involved in TR4 biocontrol. Results will help elucidate functional genes’ biocontrol mechanisms.
Banana is one of the most produced and consumed fruits in the world and its fruit peel accounts for about 40% of the total fresh quantity of ripe fruit, which is usually regarded as waste and poses serious environmental hazards. However, it is a promising source of natural bioactive compounds including phenolic compounds. Determination of the phenolic compounds in fruit peel from different cultivars and subgroups over a range of maturities provides convincing information for making full use of them. This study developed a sensitive and reliable analytical method—ultra-high performance liquid chromatography—coupled with electrospray ionization tandem mass spectrometry (UPLC-MS/MS) for measuring phenolic compounds in fruit peel from different ecotype cultivars and subgroups with different maturity. The results showed that quinic acid had the highest concentration ratio among the main phenolic compounds in the green/ripe peel of all banana cultivars; among all banana cultivars, the total phenolic compound contents of green banana peel were significantly higher than that of ripe banana peel; the total phenolic compound contents in the green/ripe fruit peel of non-dessert bananas were significantly higher than that of dessert bananas (green: non-dessert banana 1.48 ± 0.44 mg/g vs. dessert banana 0.97 ± 0.12 mg/g; ripe: non-dessert banana 0.26 ± 0.13 mg/g vs. dessert banana 0.19 ± 0.06 mg/g). These data provide a basis for the rational utilization of phenolic compound extractions from banana peel with huge biomass in the next step.
Magnaporthe oryzae, the causal agent of rice blast disease, produces devastating damage to global rice production. It is urgent to explore novel strategies to overcome the losses caused by this disease. 9-phenanthrol is often used as a transient receptor potential melastatin 4 (TRPM4) channel inhibitor for animals, but we found its fungal toxicity to M. oryzae. Thus, we explored the antimicrobial mechanism through transcriptome and metabolome analyses. Moreover, we found that overexpression of a gene encoding 4-hydroxyphenylpyruvate dioxygenase involved in the tyrosine degradative pathway enhanced the tolerance of 9-phenanthrol in M. oryzae. Thus, our results highlight the potential fungal toxicity mechanism of 9-phenanthrol at metabolic and transcriptomic levels and identify a gene involving 9-phenanthrol alleviation. Importantly, our results demonstrate the novel mechanism of 9-phenanthrol on fungal toxicity that will provide new insights of 9-phenanthrol for application on other organisms.
Manganese, as an essential trace element, participates in many physiological reactions by regulating Mn associated enzymes. Magnaporthe oryzae is a serious pathogen and causes destructive losses for rice production. We identified a cytochrome P450 gene, MoMCP1, involving the alleviation of manganese toxin and pathogenicity. To identify the underlying mechanisms, transcriptomics were performed. The results indicated that many pathogenicity related genes were regulated, especially hydrophobin related genes in ∆Momcp1. Furthermore, the Mn2+ toxicity decreased the expressions of genes involved in the oxidative phosphorylation and energy production, and increased the reactive oxygen species (ROS) levels, which might impair the functions of mitochondrion and vacuole, compromising the pathogenicity and development in ∆Momcp1. Additionally, our results provided further information about Mn associated the gene network for Mn metabolism in cells.
Magnaporthe oryzae caused huge losses in rice and wheat production worldwide. Comparing to long-term co-evolution history with rice, wheat-infecting isolates were new-emerging. To reveal the genetic differences between rice and wheat blast on global genomic scale, 109 whole-genome sequences of M. oryzae from rice, wheat, and other hosts were reanalyzed in this study. We found that the rice lineage had gone through stronger selective sweep and fewer conserved genes than those of Triticum and Lolium lineages, which indicated that rice blast fungi adapted to rice by gene loss and rapid evolution of specific loci. Furthermore, 228 genes associated with host adaptation of M. oryzae were found by presence/absence variation (PAV) analyses. The functional annotation of these genes found that the fine turning of genes gain/loss involved with transport and transcription factor, thiol metabolism, and nucleotide metabolism respectively are major mechanisms for rice adaption. This result implies that genetic base of specific host plant may lead to gene gain/loss variation of pathogens, so as to enhance their adaptability to host. Further characterization of these specific loci and their roles in adaption and evaluation of the fungi may eventually lead to understanding of interaction mechanism and develop new strategies of the disease management.
Fusarium wilt disease severely constrains the global banana industry. The highly destructive disease is caused by Fusarium oxysporum f. sp. cubense, especially its virulent tropical race 4 (Foc TR4). Selenium (Se), a non-essential mineral nutrient in higher plants, is known to enhance plant resistance against several fungal pathogens. The experiments we conducted showed that selenium (≥10 mg/L) dramatically inhibited the growth of Foc TR4 mycelia and promoted plant growth. The further study we performed recorded a substantial reduction in the disease index (DI) of banana plants suffering from Foc TR4 when treated with selenium. The selenium treatments (20~160 mg/L) demonstrated significant control levels, with recorded symptom reductions ranging from 42.4% to 65.7% in both greenhouse and field trials. The DI was significantly negatively correlated with the total selenium content (TSe) in roots. Furthermore, selenium treatments enhanced the antioxidant enzyme activities of peroxidase (POD), polyphenol oxidase (PPO), and glutathione peroxidase (GSH-Px) in banana. After two applications of selenium (100 and 200 mg/plant) in the field, the TSe in banana pulps increased 23.7 to 25.9-fold and achieved the Se enrichment standard for food. The results demonstrate that selenium applications can safely augment root TSe levels, both reducing Fusarium wilt disease incidence and producing Se-enriched banana fruits. For the first time, this study has revealed that selenium can significantly reduce the damage caused by soil-borne pathogens in banana by increasing the activities of antioxidant enzymes and inhibiting fungal growth.