The soil-borne fungi Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) causes banana Fusarium wilt, which seriously threats global banana production. Biocontrol has been considered viable alternative method to manage banana Fusarium wilt. In previous study, we found that novel S. luomodiensis SCA4-21 exhibited antifungal activity. Here, we revealed that the extracts of strain SCA4-21 significantly inhibited the growth of Foc TR4 hyphae and spore germination, severely disrupting the ultrastructure of Foc TR4 hyphal cells and spore morphology. These extracts also exhibited broad-spectrum antifungal activity against eight other phytopathogenic fungi. Furthermore, we demonstrated that strain SCA4-21 produce 32 volatile organic compounds, including five antifungal compounds. In a pot experiment, we discovered that the inoculation of strain SCA4-21 significantly inhibited the infection of Foc TR4 in banana seedling corms, achieving a biocontrol efficiency of 59.3%, and promoted the growth of banana seedlings. Additionally, this inoculation significantly enhanced the abundances of beneficial bacterial genera Streptomyces, Bacillus, Sphingomonas, and Massilia, as well as fungal genera Mortierella, Purpureocillium, Gibellulopsis, and Xenomyrothecium, while significantly reduced the abundances of pathogenic bacteria genus Pantoea and fungal genus Fusarium, in the banana rhizosphere soil. Moreover, the inoculation of strain SCA4-21 significantly increased the enrichment of pathways such as carbohydrate metabolism, amino acid metabolism, and metabolism of terpenoids and polyketides. Therefore, we postulated that strain SCA4-21 may synergistically combat banana Fusarium wilt by producing antifungal compounds and enriching beneficial bacteria and fungi. Our findings present a promising biocontrol agent for the management of banana Fusarium wilt.
Banana anthracnose caused by Colletotrichum musae is a serious postharvest disease. Use of fungicides has potential adverse effects on the environment safety and human health. Application of biological control agents (BCA) is a promising strategy for managing postharvest fruit diseases. In this study, Streptomyces sp. XT34 was isolated from the banana rhizosphere soil and exhibited a strong antifungal activity against C. musae. Combining the phenotypic and whole-genomic alignment, strain XT34 was identified as Streptomyces diastatochromogenes. Strain XT34 extracts reduced the disease incidence of anthracnose and maintained the fruit quality of banana. Treatment of extracts reduced the colonization of C. musae on fruit surface. The activity inhibition of cell wall-degrading enzymes maintained the cell-wall strength of fruit. The activity increase of defense enzymes contributed to the low disease symptom. It was supported by the differentially expressed genes (DEGs) related to supramolecular fiber organization and flavonoid biosynthesis. In addition, spore germination of C. musae was effectively inhibited by extracts. The morphology of mycelia and spore become wrinkled and ruptured. The treatment of extracts increased the optical density of A260 and soluble sugar and protein contents of C. musae. The integrity of cell membrane was also destroyed, reflecting on an increase in electrical conductivity and malondialdehyde. Several metabolites were found in the genome of strain XT34 using antiSMASH. Eleven volatile compounds were identified by gas chromatography-mass spectrometer (GC-MS). Notably, strain XT34 and extracts exhibited a broad-spectrum antifungal activity against eight phytopathogenic fungi. Hence, S. diastatochromogenes XT34 provides a potential BCA to control banana anthracnose.
Nitrogen is a crucial element for the growth and development of plants, directly affecting crop growth and yield. To investigate the physiological and molecular mechanism of nitrogen-deficiency stress, we conducted an investigation into the effects of different nitrogen levels on the growth, photosynthetic characteristics, and gene transcription levels of banana seedlings. Compared with the control group with normal nitrogen levels (NN), the height of plants receiving Reduced-N (NR), Low-N (LN), and N-Free (NF) treatments was decreased by 0.45 cm, 2.5 cm, and 3.25 cm, respectively. Their dry weight was reduced by 1.63 g, 2.99 g, and 2.88 g, respectively. Conversely, the dry weight of the underground plant part in the LN and NF treatment groups exhibited an increase of 0.13 g and 0.16 g, respectively. Regarding photosynthetic characteristics, the Specialty Products Agricultural Division (SPAD) values of the NR, LN, and NF treatments showed reductions of 15.5%, 30.4%, and 35.9%, respectively, compared with those of the control treatments. The values of maximum photosynthetic efficiency (Fv/Fm), actual photosynthetic efficiency (Y(Ⅱ)), and relative electron transfer (ETR) of the banana seedlings decreased to different degrees after NR, LN, and NF treatment, and their values were positively correlated with N levels. Gene transcription analysis showed that N transport-related proteins, including NRT1.7, NRT2.3a, NRT2.3b, and NRT2.5, were significantly up-regulated to increase the nitrogen absorption capacity of plant roots. On the other hand, various transcription factors including GRAS, MYB, and WRKY were notably up-regulated, facilitating root growth and the expanding root absorption area, thereby enhancing nitrogen uptake. Furthermore, genes associated with endogenous hormone metabolic pathways such as gibberellin (GA), strigolactone (SL), and brassinosteroids (BR) were activated in banana plants subjected to low nitrogen stress, enhancing the plant’s ability to adapt to nitrogen-deficient conditions. These findings offer valuable insights into understanding the transcriptional regulatory mechanisms governing banana responses to low nitrogen stress and breeding new varieties with improved nutrient utilization.
Pineapple is the third most crucial tropical fruit worldwide and available in five varieties. Genomes of different pineapple varieties have been released to date; however, none of them are complete, with all exhibiting substantial gaps and representing only two of the five pineapple varieties. This significantly hinders the advancement of pineapple breeding efforts. In this study, we sequenced the genomes of three varieties: a wild pineapple variety, a fiber pineapple variety, and a globally cultivated edible pineapple variety. We constructed the first gap-free reference genome (Ref) for pineapple. By consolidating multiple sources of evidence and manually revising each gene structure annotation, we identified 26,656 protein-coding genes. The BUSCO evaluation indicated a completeness of 99.2%, demonstrating the high quality of the gene structure annotations in this genome. Utilizing these resources, we identified 7,209 structural variations across the three varieties. Approximately 30.8% of pineapple genes were located within ±5 kb of structural variations, including 30 genes associated with anthocyanin synthesis. Further analysis and functional experiments demonstrated that the high expression of AcMYB528 aligns with the accumulation of anthocyanins in the leaves, both of which may be affected by a 1.9-kb insertion fragment. In addition, we developed the Ananas Genome Database, which offers data browsing, retrieval, analysis, and download functions. The construction of this database addresses the lack of pineapple genome resource databases. In summary, we acquired a seamless pineapple reference genome with high-quality gene structure annotations, providing a solid foundation for pineapple genomics and a valuable reference for pineapple breeding.
Mycotoxin contamination is a serious threat to the world agriculture and health. Fusarium oxysporum f. sp. cubense (Foc) is a soil-borne fungal pathogen that causes Fusarium wilt of crops. Foc produces highly toxic mycotoxins such as fusaric acid (FA), beauvericin (BEA) and fusarenone (FUS), which are cytotoxic and carcinogenic. It is urgent to explore biofungicides to inhibit the secretion and accumulation of mycotoxin. Here, a bioactive compound fluvirucin B6 was obtained from Streptomyces solisilvae with strong antifungal activity. Metabolomics revealed that fluvirucin B6 could reduce the production of FA, BEA and FUS of Foc. This mechanism attributes to the alteration of sugar, protein and fat content in Foc cells and the activity increase of chitinase and β-1,3-glucanase, thereby inhibiting mycelial growth and spore germination. Furthermore, fluvirucin B6 disrupted the morphology and function of Foc mitochondria, resulting in the decrease of enzyme activities in the tricarboxylic acid (TCA) cycle and the electron transport chain (ETC). Microbiome analysis revealed that the application of fluvirucin B6 decreased the incidence of Fusarium wilt and reduced Foc density in banana, and enriched beneficial microbes in rhizosphere. Therefore, fluvirucin B6 was a potential biofungicide to manage Foc infection and detoxify FA, BEA and FUS.
Colletotrichum gloeosporioides is a fungal disease of strawberry fruit. Biocontrol strategies holds tremendous promise in alleviating fruit decay. Here, 30 actinomycetes were isolated from rhizosphere soil of Calotropis gigantea. A strain labeled with CG-G2 exhibited the strongest antagonistic activity against C. gloeosporioides and was assigned as Streptomyces corchorusii. Compared to strain CG-G2 extracts, the volatile organic compounds (VOCs) had a high antifungal activity against anthracnose. These volatiles effectively inhibited mycelial growth and spore germination of C. gloeosporioides. The hyphal and conidial structure was severely destroyed. Metabolomics analysis revealed that VOCs inhibited C. gloeosporioides via inducing flavonoids metabolism contributing to antifungal activity. Three main antagonistic compounds in VOCs were identified as methyl 2-methyl butyrate, hexanenitrile and methyl 2-Ethyl hexanoate. Especially, methyl 2-methyl butyrate demonstrated a remarkable efficacy in inhibiting fruit decay and preserving fruit quality. Hence, S. corchorusii CG-G2 will be a potential biocontrol agent for controlling anthracnose on harvested fruits.
Tomato fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici (Fol) is a highly destructive disease, resulting in severe economic losses of global tomato production annually. An eco-friendly alternative to chemical fungicide using biological control agents (BCAs) is urgently needed. Here, Bacillus siamensis QN2MO-1 was isolated from Noli fruit and had a strong antagonistic activity against Fol in vitro and in vivo. Strain QN2MO-1 also exhibited a broad-spectrum antifungal activity against the selected 14 phytopathogenic fungi. The crude protein produced by strain QN2MO-1 could inhibit the spore germination of Fol and destroy the spore structure. It was closely related with the generation of chitinase and β-1,3-glucanase secreted by strain QN2MO-1. In a pot experiment, the application of B. siamensis QN2MO-1 effectively alleviated the yellowing and wilting symptoms of tomato plants. The disease index and incidence rate were decreased by 72.72% and 80.96%, respectively. The rhizospheric soil in tomato plants owed a high abundance of microbial community. Moreover, strain QN2MO-1 also enhanced the plant growth and improved the fruit quality of tomato. Therefore, B. siamensis QN2MO-1 will be explored as a potential biocontrol agent and biofertilizer.
Banana Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (Foc TR4) is the most destructive soil-borne fungal disease. Until now, there has been a lack of effective measures to control the disease. It is urgent to explore biocontrol agents to control Foc TR4 and the secretion of mycotoxin. In this study, fluvirucin B6 was screened from Streptomyces solisilvae using an activity-guided method. Fluvirucin B6 exhibited strong antifungal activity against Foc TR4 (0.084 mM of EC50 value) and significantly inhibited mycelial growth and spore germination. Further studies demonstrated that fluvirucin B6 could cause the functional loss of mitochondria, the disorder of metabolism of Foc TR4 cells, and the decrease of enzyme activities in the tricarboxylic acid cycle and electron transport chain, ultimately inhibiting mycotoxin metabolism. In a pot experiment, the application of fluvirucin B6 significantly decreased the incidence of banana Fusarium wilt and the amount of Foc TR4 and controlled fungal toxins in the soil. Additionally, fluvirucin B6 could positively regulate the changes in the structure of the banana rhizosphere microbial community, significantly enriching beneficial microbes associated with disease resistance. In summary, this study identifies fluvirucin B6, which plays versatile roles in managing fungal diseases and mycotoxins.
Banana wilt caused by Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) is a devastating fungal disease. Biocontrol strategies hold immense potential for inhibiting the spread of Foc TR4. Here, 30 actinobacteria were isolated from soils and screened for their antagonistic activity against Foc TR4. Strain SCA4-21T was selected due to its strongest antagonistic activity against Foc TR4. Strain SCA4-21T also exhibited strong antagonistic activity against the other eight phytopathogenic fungi. The strain was identified as the genus Streptomyces according to its physiological, biochemical, and phenotypic characteristics. The phylogenetic trees of 16S rRNA sequences demonstrated that strain SCA4-21T formed a subclade with S. iranensis HM 35T and/or S. rapamycinicus NRRL B-5491T with low bootstrap values. Considering that 16S rRNAs did not provide sufficient resolution for species-level identification, the whole genome of strain SCA4-21T was sequenced. Multilocus sequence analysis (MLSA) based on five housekeeping gene alleles (atpD, gyrB, recA, rpoB, and trpB) revealed that strain SCA4-21T clustered into S. hygroscopicus subsp. hygroscopicus NBRC 13472T with 100% of bootstrap value. The analysis of the genome-based phylogeny also approved the results. Average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) were 91.26 and 44.30%, respectively, with values below the respective species level threshold of 95 and 70%. Hence, strain SCA 4–21T represented a novel species within the genus Streptomyces, named Streptomyces luomodiensis sp. nov. The type strain is SCA4-21T (=GDMCC4.340T = JCM36555T). By the CAZymes analysis, 348 carbohydrate-active enzymes (CAZymes) were detected, including 15 chitinases and eight β-1,3-glucanases. The fermentation broth of strain SCA4-21T, exhibiting strong antagonistic activity against Foc TR4, demonstrated high activities of chitinase and β-1,3-glucanase, which might be involved in antifungal activity. Our results showed an innovative potential biocontrol agent for managing plant fungal diseases, specifically banana fusarium wilt.
The Sapindaceae family, encompassing a wide range of plant forms such as herbs, vines, shrubs, and trees, is widely distributed across tropical and subtropical regions. This family includes economically important crops like litchi, longan, rambutan, and ackee. With the wide application of genomic technologies in recent years, several Sapindaceae plant genomes have been decoded, leading to an accumulation of substantial omics data in this field. This surge in data highlights the pressing need for a unified genomic data center capable of storing, sharing, and analyzing these data. Here, we introduced SapBase, that is, the Sapindaceae Genome Database. SapBase houses seven published plant genomes alongside their corresponding gene structure and functional annotations, small RNA annotations, gene expression profiles, gene pathways, and synteny block information. It offers user-friendly features for gene information mining, co-expression analysis, and inter-species comparative genomic analysis. Furthermore, we showcased SapBase's extensive capacities through a detailed bioinformatic analysis of a MYB gene in litchi. Thus, SapBase could serve as an integrative genomic resource and analysis platform for the scientific exploration of Sapinaceae species and their comparative studies with other plants.
Background: Fusarium wilt of banana, a soil-borne disease caused by Fusarium oxysporum (f.sp.) cubense, particularly the Tropical Race 4 (Foc TR4), severely hinders the progress of the banana industry. Utilizing antagonistic microorganisms to suppress pathogen growth and enhance banana production is seen as a promising strategy for disease management. Results: Strain XZ11-1, exhibiting the highest siderophore production ability, was isolated from the Qixianling Tropical Rainforest in Hainan province, China. Through a combination of colony morphology, physiological and biochemical analysis, as well as a dual-gene analysis of ITS4 and TEF1, the strain was identified as Trichoderma viride. Following optimization of fermentation conditions, strain XZ11-1 produced siderophores with a relative content of 79.45%, identified as both hydroxamate siderophores and carboxylate-type siderophores. Additionally, strain XZ11-1 demonstrated potent antifungal activity against 10 phytopathogenic fungi. Co-cultivation experiments revealed that under iron-limiting conditions, siderophores were the primary factor inhibiting the growth of Foc TR4. Notably, the siderophores produced by strain XZ11-1 exhibited a stimulating effect on maize seed germination and effectively colonized the roots of banana plants. Pot experiments further demonstrated that Trichoderma viride XZ11-1 could suppress the growth of Foc TR4 by competing for iron in the environment through siderophore mediation. Moreover, it could enhance iron absorption by banana plants by secreting siderophores to bind with Fe3+ in the environment. Conclusions: Therefore, the high-yield siderophores produced by Trichoderma viride XZ11-1 represent a valuable microbial resource for controlling plant pathogenic fungi and promoting plant growth.
Since the official release of the stand-alone bioinformatics toolkit TBtools in 2020, its superior functionality in data analysis has been demonstrated by its widespread adoption by many thousands of users and references in more than 5000 academic articles. Now, TBtools is a commonly used tool in biological laboratories. Over the past 3 years, thanks to invaluable feedback and suggestions from numerous users, we have optimized and expanded the functionality of the toolkit, leading to the development of an upgraded version—TBtools-II. In this upgrade, we have incorporated over 100 new features, such as those for comparative genomics analysis, phylogenetic analysis, and data visualization. Meanwhile, to better meet the increasing needs of personalized data analysis, we have launched the plugin mode, which enables users to develop their own plugins and manage their selection, installation, and removal according to individual needs. To date, the plugin store has amassed over 50 plugins, with more than half of them being independently developed and contributed by TBtools users. These plugins offer a range of data analysis options including co-expression network analysis, single-cell data analysis, and bulked segregant analysis sequencing data analysis. Overall, TBtools is now transforming from a stand-alone software to a comprehensive bioinformatics platform of a vibrant and cooperative community in which users are also developers and contributors. By promoting the theme "one for all, all for one", we believe that TBtools-II will greatly benefit more biological researchers in this big-data era.
Plant height is an important and valuable agronomic trait associated with yield and resistance to abiotic and biotic stresses. Dwarfism has positive effects on plant development and field management, especially for tall monocotyledon banana (Musa spp.). However, several key genes and their regulation mechanism of controlling plant height during banana development are unclear. In the present study, the popular cultivar ‘Brazilian banana’ (‘BX’) and its dwarf mutant (‘RK’) were selected to identify plant height-related genes by comparing the phenotypic and transcriptomic data. Banana seedlings with 3–4 leaves were planted in the greenhouse and field. We found that the third and fourth weeks are the key period of plant height development of the selected cultivars. A total of 4563 and 10507 differentially expressed genes (DEGs) were identified in the third and fourth weeks, respectively. Twenty modules were produced by the weighted gene co-expression network analysis (WGCNA). Eight modules were positively correlated with the plant height, and twelve other modules were negatively correlated. Combining with the analysis of DEGs and WGCNA, 13 genes in the signaling pathway of gibberellic acid (GA) and 7 genes in the signaling pathway of indole acetic acid (IAA) were identified. Hub genes related to plant height development were obtained in light of the significantly different expression levels (|log2FC| ≥ 1) at the critical stages. Moreover, GA3 treatment significantly induced the transcription expressions of the selected candidate genes, suggesting that GA signaling could play a key role in plant height development of banana. It provides an important gene resource for the regulation mechanism of banana plant development and assisted breeding of ideal plant architecture.
Banana Fusarium wilt caused by Fusarium oxysporum f. sp. cubense tropical race4 (Foc TR4) is one of the most destructive soil-borne fungal diseases and currently threatens banana production around the world. Until now, there is lack of an effective method to control banana Fusarium wilt. Therefore, it is urgent to find an effective and eco-friendly strategy against the fungal disease. In this study, a strain of Trichoderma sp. N4-3 was isolated newly from the rhizosphere soil of banana plants. The isolate was identified as Trichoderma parareesei through analysis of TEF1 and RPB2 genes as well as morphological characterization. In vitro antagonistic assay demonstrated that strain N4-3 had a broad-spectrum antifungal activity against ten selected phytopathogenic fungi. Especially, it demonstrated a strong antifungal activity against Foc TR4. The results of the dual culture assay indicated that strain N4-3 could grow rapidly during the pre-growth period, occupy the growth space, and secrete a series of cell wall-degrading enzymes upon interaction with Foc TR4. These enzymes contributed to the mycelial and spore destruction of the pathogenic fungus by hyperparasitism. Additionally, the sequenced genome proved that strain N4-3 contained 21 genes encoding chitinase and 26 genes encoding β-1,3-glucanase. The electron microscopy results showed that theses cell wall-degrading enzymes disrupted the mycelial, spore, and cell ultrastructure of Foc TR4. A pot experiment revealed that addition of strain N4-3 significantly reduced the amount of Foc TR4 in the rhizosphere soil of bananas at 60 days post inoculation. The disease index was decreased by 45.00% and the fresh weight was increased by 63.74% in comparison to the control. Hence, Trichoderma parareesei N4-3 will be a promising biological control agents for the management of plant fungal diseases.
Lychee is an exotic tropical fruit with a distinct flavor. The genome of cultivar ‘Feizixiao’ was assembled into 15 pseudochromosomes, totaling ~470 Mb. High heterozygosity (2.27%) resulted in two complete haplotypic assemblies. A total of 13,517 allelic genes (42.4%) were differentially expressed in diverse tissues. Analyses of 72 resequenced lychee accessions revealed two independent domestication events. The extremely early maturing cultivars preferentially aligned to one haplotype were domesticated from a wild population in Yunnan, whereas the late-maturing cultivars that mapped mostly to the second haplotype were domesticated independently from a wild population in Hainan. Early maturing cultivars were probably developed in Guangdong via hybridization between extremely early maturing cultivar and late-maturing cultivar individuals. Variable deletions of a 3.7 kb region encompassed by a pair of CONSTANS -like genes probably regulate fruit maturation differences among lychee cultivars. These genomic resources provide insights into the natural history of lychee domestication and will accelerate the improvement of lychee and related crops.
Small RNAs (sRNAs) are essential regulatory molecules, and there are three major sRNA classes in plants: microRNAs (miRNAs), phased small interfering RNAs (phased siRNAs or phasiRNAs), and heterochromatic siRNAs (hc-siRNAs). Excluding miRNAs, the other two classes are not well annotated or available in public databases for most sequenced plant genomes. We performed a comprehensive sRNA annotation of 143 plant species that have fully sequenced genomes and next-generation sequencing sRNA data publicly available. The results are available via an online repository called sRNAanno ( www.plantsRNAs.org ). Compared with other public plant sRNA databases, we obtained was much more miRNA annotations, which are more complete and reliable because of the consistent and highly stringent criteria used in our miRNA annotations. sRNAanno also provides free access to genomic information for >22,721 PHAS loci and >22 million hc-siRNA loci annotated from these 143 plant species. Both miRNA and PHAS loci can be easily browsed to view their main features, and a collection of archetypal trans-acting siRNA 3 ( TAS3 ) genes were annotated separately for quick access. To facilitate the ease of sRNA annotation, sRNAanno provides free service for sRNA annotations to the community. In summary, the sRNAanno database is a great resource to facilitate genomic and genetic research on plant small RNAs.
The pineapple (Ananas comosus) is cold sensitive. Most cultivars are injured during winter periods, especially in sub-tropical regions. There is a lack of molecular information on the pineapple's response to cold stress. In this study, high-throughput transcriptome sequencing and gene expression analysis were performed on plantlets of a cold-tolerant genotype of the pineapple cultivar 'Shenwan' before and after cold treatment. A total of 1,186 candidate cold responsive genes were identified, and their credibility was confirmed by RT-qPCR. Gene set functional enrichment analysis indicated that genes related to cell wall properties, stomatal closure and ABA and ROS signal transduction play important roles in pineapple cold tolerance. In addition, a protein association network of CORs (cold responsive genes) was predicted, which could serve as an entry point to dissect the complex cold response network. Our study found a series of candidate genes and their association network, which will be helpful to cold stress response studies and pineapple breeding for cold tolerance.