Codon usage bias of secretory protein in Fusarium oxysporum f. sp. cubense tropical race 4.

Banana Fusarium oxysporum f. sp. cubense tropical race 4 (Foc-TR4) is a highly destructive pathogen that infects nearly all major banana cultivars and has a tendency to spread further. Secreted proteins play a crucial role in the process of Fusarium wilt infection in bananas. In this study, we analyzed the codon usage bias (CUB) of the Foc-TR4 classical secretory protein genome for the first time and observed a strong bias toward codons ending with C. We found that 572 out of the 14,543 amino acid sequences in the Foc-TR4 genome exhibited characteristics of classical secretory proteins. The CUB was largely influenced by selection optimization pressure, as indicated by the ENC value and neutral plot analysis. Among the identified codons, such as UCC and CCC, 11 were found to be optimal for Foc-TR4 gene expression. Codons with higher GC content and a C base in the third position showed greater selectivity. The CUB in the secretory proteins encoded by Foc-TR4 provides insights into their evolutionary patterns, contributing to the development and screening of novel and effective antifungal drugs.

Promotion of bio-oil production from the microwave pyrolysis of cow dung using pretreated red mud as a bifunctional additive: Parameter optimization, energy efficiency evaluation, and mechanism analysis.

To address the issues of high oxygen content and energy consumption in the microwave-assisted pyrolysis of biomass for biofuel production, this study used high-temperature pretreated red mud (RM) as an additive. The pretreated RM exhibited dual functionalities, namely microwave absorption and catalytic properties, during the microwave-assisted pyrolysis of cow dung (CD). This study also evaluated the optimization potential of energy recovery efficiency. The results showed that the addition of pretreated RM significantly increased the oil yield during the microwave-assisted pyrolysis of CD. The highest oil yield (59.63%) was obtained via the microwave-assisted pyrolysis of CD over catalysis with RM pretreated at 750 °C (RM750). Through the optimization of the RM750-to-CD mixing ratio, optimal oil quality and energy recovery efficiency were achieved. At a mixing ratio of 1:1, the pyrolysis oil featured the highest aromatic hydrocarbon content and lowest acid content. The high-temperature pretreatment of RM increased the Fe2O3 content, which enhanced the dielectric properties and magnetic loss ability of the reactants. This resulted in localized high temperatures and the formation of "hot spots," which can promote the deoxygenation and hydrogenation reactions of oil. Consequently, the lower heating rate of oil increased from 35.12 to 40.11 MJ kg-1. The released oxygen escaped in the form of CO. In addition, pyrolytic char was used as an in situ microwave absorbing material owing to its increased Fe2O3 content and graphitization degree, leading to an increase in energy recovery efficiency from 4.71% to 9.98%. This study provides valuable guidance for the efficient utilization of diversified solid wastes and demonstrates the potential application of microwave-assisted pyrolysis technology in the resource utilization of solid wastes.

Synergistic optimization of bio-oil quality and heavy metal solidification during microwave co-pyrolysis of cow dung and red mud.

To decrease the environmental risks caused by heavy metals (HMs) in red mud (RM) and improve the quality of pyrolysis oil from biomass, high-temperature pretreated RM and cow dung (CD) were microwave co-pyrolyzed. Then, the optimization potential of energy consumption was evaluated and the interaction mechanism between RM and CD was explored. The results showed that the increase in transition metal oxides and specific surface area improved the microwave-absorption and catalytic capacity of the pretreated RM. By optimizing the parameters, a pretreatment temperature of 650 °C resulted in a 21.65% reduction in acid content of bio-oil, higher HMs immobilization rates (>91%) and a 7.44% reduction in energy consumption. The synergistic optimization of bio-oil quality, HMs immobilization and energy consumption was achieved. After microwave co-pyrolysis with cow dung, the larger specific surface area (92.90 m2 g-1) and higher carbon crystallinity (ID/IG = 1.02) of pyrolysis residues enhanced the physical adsorption to HMs. The complexation of HMs with -OH could further enhance the solidification of HMs. This work will provide support to efficient resource utilization of solid waste, and demonstrate the great potential of microwave co-pyrolysis in HMs immobilization.

Genetic Mapping, Candidate Gene Identification and Marker Validation for Host Plant Resistance to the Race 4 of Fusarium oxysporum f. sp. cubense Using Musa acuminata ssp. malaccensis.

Fusarium wilt of banana is a devastating disease that has decimated banana production worldwide. Host resistance to Fusarium oxysporum f. sp. Cubense (Foc), the causal agent of this disease, is genetically dissected in this study using two Musa acuminata ssp. Malaccensis segregating populations, segregating for Foc Tropical (TR4) and Subtropical (STR4) race 4 resistance. Marker loci and trait association using 11 SNP-based PCR markers allowed the candidate region to be delimited to a 12.9 cM genetic interval corresponding to a 959 kb region on chromosome 3 of 'DH-Pahang' reference assembly v4. Within this region, there was a cluster of pattern recognition receptors, namely leucine-rich repeat ectodomain containing receptor-like protein kinases, cysteine-rich cell-wall-associated protein kinases, and leaf rust 10 disease-resistance locus receptor-like proteins, positioned in an interspersed arrangement. Their transcript levels were rapidly upregulated in the resistant progenies but not in the susceptible F2 progenies at the onset of infection. This suggests that one or several of these genes may control resistance at this locus. To confirm the segregation of single-gene resistance, we generated an inter-cross between the resistant parent 'Ma850' and a susceptible line 'Ma848', to show that the STR4 resistance co-segregated with marker '28820' at this locus. Finally, an informative SNP marker 29730 allowed the locus-specific resistance to be assessed in a collection of diploid and polyploid banana plants. Of the 60 lines screened, 22 lines were predicted to carry resistance at this locus, including lines known to be TR4-resistant, such as 'Pahang', 'SH-3362', 'SH-3217', 'Ma-ITC0250', and 'DH-Pahang/CIRAD 930'. Additional screening in the International Institute for Tropical Agriculture's collection suggests that the dominant allele is common among the elite 'Matooke' NARITA hybrids, as well as in other triploid or tetraploid hybrids derived from East African highland bananas. Fine mapping and candidate gene identification will allow characterization of molecular mechanisms underlying the TR4 resistance. The markers developed in this study can now aid the marker-assisted selection of TR4 resistance in breeding programs around the world.

Revealing the different resistance mechanisms of banana 'Guijiao 9' to Fusarium oxysporum f. sp. cubense tropical race 4 using comparative proteomic analysis.

Fusarium wilt of banana caused by Fusarium oxysporum f. sp. cubense is a worldwide devastating fungal disease in the banana industry. The disease caused by Fusarium oxysporum f. sp. cubense is becoming more and more serious. The pathogen of Fusarium oxysporum f. sp. cubense tropical race 4 (Foc4) is the most harmful one. 'Guijiao 9' is a banana cultivar with good resistance to Foc4, which is identified by resistance screening of natural variant lines. It is of great significance to explore the resistance genes and key proteins of 'Guijiao 9' for banana cultivar improvement and disease resistance breeding. In this study, iTRAQ (isobaric Tags for Relative and Absolute quantitation) was used to analyze the xylem proteomic data of banana roots from the resistant variety 'Guijiao 9' and susceptible variety 'Williams', and the differences in protein accumulation profiles between these two varieties at 24, 48, and 72 h after infection with Foc4 were compared. The identified proteins were analyzed by the protein WGCNA (Weighted Gene Correlation Network Analysis), and the differentially expressed proteins (DEPs) were verified by qRT-PCR experiments. Proteomic analysis showed that there were differences in the protein accumulation profiles of the resistant cultivar 'Guijiao 9' and the susceptible cultivar 'Williams' after infection with Foc4, and there were differences in resistance-related proteins, biosynthesis of secondary metabolites, peroxidase, and pathogenesis-related proteins. The stress response of bananas to pathogens was affected by multiple factors. Protein co-expression analysis showed that there was a high correlation between the MEcyan module and resistance, and 'Guijiao 9' had a different resistance mechanism compared with 'Williams'. SIGNIFICANCE: 'Guijiao 9' is a banana variety with good resistance to Foc4, which is identified by screening the resistance of natural variant lines in the farmland where banana plants are seriously infected by Foc4. It is of great significance to excavate the resistance genes and key proteins of 'Guijiao 9' for banana variety improvement and disease resistance breeding. The aim of this paper is to identify the proteins and related functional modules controlling the pathogenicity differences of Foc4 by comparative proteomic analysis of 'Guijiao 9', so as to understand the resistance mechanism of banana to Fusarium wilt, and offer basis for the final identification, isolation and utilization of Foc4 resistance-related genes in banana variety improvement. The research results will also provide a basis for further understanding the host-pathogen interaction and revealing the resistance mechanism of bananas.

Identification of a Major QTL-Controlling Resistance to the Subtropical Race 4 of Fusarium oxysporum f. sp. cubense in Musa acuminata ssp. malaccensis.

Vascular wilt caused by the ascomycete fungal pathogen Fusarium oxysporum f. sp. cubense (Foc) is a major constraint of banana production around the world. The virulent race, namely Tropical Race 4, can infect all Cavendish-type banana plants and is now widespread across the globe, causing devastating losses to global banana production. In this study, we characterized Foc Subtropical Race 4 (STR4) resistance in a wild banana relative which, through estimated genome size and ancestry analysis, was confirmed to be Musa acuminata ssp. malaccensis. Using a self-derived F2 population segregating for STR4 resistance, quantitative trait loci sequencing (QTL-seq) was performed on bulks consisting of resistant and susceptible individuals. Changes in SNP index between the bulks revealed a major QTL located on the distal end of the long arm of chromosome 3. Multiple resistance genes are present in this region. Identification of chromosome regions conferring resistance to Foc can facilitate marker assisted selection in breeding programs and paves the way towards identifying genes underpinning resistance.

Effects of microwave and plastic content on the sulfur migration during co-pyrolysis of biomass and plastic.

In order to reduce the risks of sulfur-containing contaminants present in biofuels, the effects of microwave and content of hydrogen donor on the cracking of C-S bonds and the migration of sulfur were studied by co-pyrolysis of biomass and plastic. The synergistic mechanism of microwave and hydrogen donor was explored from the perspective of deducing the evolution of sulfur-containing compounds based on microwave thermogravimetric analysis. By combining temperature-weight curves, it was found that microwaves and hydrogen radicals promoted the cracking of sulfur-containing compounds and increased the mass loss of biomass during pyrolysis. The mixing ratio of hydrogen donor (plastic) was the key parameter resulting in the removal of sulfur from oil. By adjusting the mixing ratio, the yield of co-pyrolyzed oil was three times higher than that of cow dung pyrolysis alone and the relative removal rate of sulfur reached 73.67%. The relative content of sulfur in the oil was reduced by 73.77% due to the escape of sulfur-containing gases (H2S, COS and C2H5SH) and the formation of sulfate crystals in the char. Microwave selectively heated sulfur-containing organics and hydrogen radicals stimulated the breaking of C-S bonds, which improved the cracking efficiency of the oil. This breaking will provide a theoretical and technological reference for the environmentally friendly treatment of biomass and biofuels.

In-depth exploration of mechanism and energy balance characteristics of an advanced continuous microwave pyrolysis coupled with carbon dioxide reforming technology to generate high-quality syngas.

This study firstly coupled advanced continuous microwave pyrolysis with CO2 reforming technology to recover syngas from cow manure and CO2. The contribution of CO2 to syngas, pyrolysis mechanism, and energy balance characteristics were analyzed thoroughly. The results showed that continuous microwave pyrolysis coupled with CO2 reforming technology has superiorities over other pyrolysis methods in bio-gas generation. The bio-gas yield, syngas content, and heating value of syngas reached the maximum value of 71.02 wt%, 85.70 vol%, and 10.87 MJ/Nm3, respectively. CO2 strengthened pyrolysis and reacted with pyrolysis products to produce high-quality syngas and reduce H2S. Due to the limited substances that can react with CO2 and excessive energy consumption with increasing CO2 concentration, the utilization efficiencies of CO2 and energy decreased from 36.31% and 27.27% to 31.16% and 24.24%, respectively. This work provides basic theory and technical support for advanced technology to recover high-quality syngas from biomass with low energy consumption.

Assessing Variations in Host Resistance to Fusarium oxysporum f sp. cubense Race 4 in Musa Species, With a Focus on the Subtropical Race 4.

Fusarium oxysporum f. sp. cubense (Foc) has severely curtailed banana production in the tropical regions of the world. The tropical race 4 (TR4) of Foc was detected in Australia in the 1990s and it is virulent to all Cavendish type banana cultivars, which represents the majority of banana production in Australia. Genetic resistance to Foc race 4 is urgently needed. To characterize sources of resistance, we have assessed the Foc resistance response of 34 Musa cultivars with plants grown under controlled settings. Amongst diploid banana cultivars carrying the AA genome, resistance is found in Musa acuminata sub-species including malaccensis 'Pahang' and burmannica 'Calcutta4.' In the polyploid group, the hybrids such as 'FHIA-18' and 'FHIA-25' are highly resistant against both Foc-TR4 and subtropical race 4 (Foc-STR4). Interestingly, 'FHIA-2' and 'CAM020' appear to be resistant to Foc-TR4 but susceptible to Foc-STR4, suggesting potential differences in the resistance mechanisms against the different race 4 strains. Using a GFP tagged Foc-STR4 strain challenged onto both resistant and susceptible M. a. malaccensis lines, a high inoculum dosage rapidly induced vascular wilt in the susceptible M. a. malaccensis lines at 2.5 weeks. This was associated with an accumulation of micro-conidia in the rhizome and the movement of the fungus through the xylem vessels. In contrast, the fungal movement was restrained in the rhizome of the resistant M. a. malaccensis lines and no sporulation was observed. Overall, this research suggests that the resistance response is dependent to an extent on inoculum dosage and that the plant host's response, in the rhizome, plays an important role in inhibiting the fungus from spreading to the rest of the plant. Identifying race 4 resistant accessions can help to understand mechanisms of resistance and provide banana breeders with the genetic resources to integrate resistance genes into commercial varieties.

Comparative transcriptome analysis reveals resistance-related genes and pathways in Musa acuminata banana 'Guijiao 9' in response to Fusarium wilt.

Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (Foc), is one of the most devastating diseases in bananas resulting in significant loss of Cavendish bananas production worldwide. Here we show the agronomic traits and the resistance of 'Guijiao 9' in the field trials from 2012 to 2017. And then we dissect and compare the transcriptome response from these two cultivars (cv. 'Guijiao 9' and cv. Williams) in an attempt to understand the molecular basis that contribute to the enhanced Foc tropical race 4 (Foc-TR4) resistance. 'Guijiao 9' is a Cavendish cultivar with strong resistance to Foc-TR4, which was reflected in a lower disease severity and incidence in glasshouse and field trails, when compared to the susceptible cultivar Williams. Gene expression profiles of 'Guijiao 9' and Williams were captured by performing RNA-Seq analysis on 16 biological samples collected over a six day period post inoculation with Foc-TR4. Transcriptional reprogramming in response to Foc-TR4 was detected in both genotypes but the response was more drastic in 'Guijiao 9' than in Williams. Specific genes involved in plant-pathogen interaction and defense signaling including MAPK, calcium, salicylic acid, jasmonic acid and ethylene pathways were analyzed and compared between 'Guijiao 9' and Williams. Genes associated with defense-related metabolites synthesis such as NB-LRR proteins, calmodulin-binding protein and phenylpropanoids biosynthesis genes were significantly up-regulated in 'Guijiao 9' resistant to Foc-TR4 infection. Taken together, this study highlights the important roles of plant hormone regulation and defense gene activation in mediating resistance in 'Guijiao 9'.

Proteomic analysis of differentially expressed proteins induced by salicylic acid in suspension-cultured ginseng cells.

In this study, optimized 2-DE sample preparation methodologies were established for suspension-cultured ginseng cells. Three commonly used protein extraction methods (Trichloroacetic acid-acetone, urea/thiourea and phenol extraction method) were evaluated for proteomic analysis of suspension cultures of ginseng. A comparative analysis of suspension-cultured ginseng cells proteome induced by salicylic acid (SA) was reported. The results demonstrated that phenol extraction method was the best method based on protein extraction efficiency and the good quality of 2-DE patterns for suspension-cultured ginseng cells. Fifteen differentially expressed proteins induced by salicylic acid in suspension-cultured ginseng cells were identified by MALDI-TOF-MS. These identified proteins were involved in defense and stress response, energy metabolism, signal transduction/transcription, protein synthesis and metabolism, and photosynthesis. Chaperonin 60, related to defense responses, was more abundant in suspension-cultured ginseng cells after application of SA. Vacuolar ATPase subunit B was newly induced in SA treatment.