Identification of ARF transcription factor gene family and its defense responses to bacterial infection and salicylic acid treatment in sugarcane

Auxin response factor (ARF) is a critical regulator in the auxin signaling pathway, involved in a variety of plant biological processes. Here, gene members of 24 SpapARFs and 39 SpnpARFs were identified in two genomes of Saccharum spontaneum clones AP85-441 and Np-X, respectively. Phylogenetic analysis showed that all ARF genes were clustered into four clades, which is identical to those ARF genes in maize (Zea mays) and sorghum (Sorghum bicolor). The gene structure and domain composition of this ARF family are conserved to a large degree across plant species. The SpapARF and SpnpARF genes were unevenly distributed on chromosomes 1–8 and 1–10 in the two genomes of AP85-441 and Np-X, respectively. Segmental duplication events may also contribute to this gene family expansion in S. spontaneum. The post-transcriptional regulation of ARF genes likely involves sugarcane against various stressors through a miRNA-medicated pathway. Expression levels of six representative ShARF genes were analyzed by qRT-PCR assays on two sugarcane cultivars [LCP85-384 (resistant to leaf scald) and ROC20 (susceptible to leaf scald)] triggered by Acidovorax avenae subsp. avenae (Aaa) and Xanthomonas albilineans (Xa) infections and salicylic acid (SA) treatment. ShARF04 functioned as a positive regulator under Xa and Aaa stress, whereas it was a negative regulator under SA treatment. ShARF07/17 genes played positive roles against both pathogenic bacteria and SA stresses. Additionally, ShARF22 was negatively modulated by Xa and Aaa stimuli in both cultivars, particularly LCP85-384. These findings imply that sugarcane ARFs exhibit functional redundancy and divergence against stressful conditions. This work lays the foundation for further research on ARF gene functions in sugarcane against diverse environmental stressors

Sustained organic amendments utilization enhances ratoon crop growth and soil quality by enriching beneficial metabolites and suppressing pathogenic bacteria

IntroductionOrganic soil amendments such as filter mud (FM) and biochar (BC) can potentially influence the abundance and composition of metabolites. However, our current understanding of the stimulatory effects of FM and BC’s long-term impact on stress-regulating metabolites, such as abscisic acid (ABA), jasmonic acid (JA), melatonin, and phenyllactic acid (PLA), and these substrates regulatory effects on disease-causing bacteria in sugarcane ratooning field, which is susceptible to nutrients depletion, diseases, etc., remain poorly understood. Additionally, little is known about how the long-term interaction of these substrates and compounds influences sugarcane ratooning soil enzyme activities, nutrient cycling, and crop growth performance.MethodsTo answer these questions, we adopted metabolomics tools combined with high-throughput sequencing to explore the stimulatory effects of the long-term addition of FM and BC on metabolites (e.g., PLA and abscisic aldehyde) and quantify these substrates’ regulatory effects on disease-causing bacteria, soil enzyme activities, nutrient cycling, and crop growth performance.ResultsThe result revealed that ratoon crop weight, stem diameter, sugar content, as well as soil physico-chemical properties, including soil nitrate (NH3+-N), organic matter (OM), total nitrogen (TN), total carbon (TC), and β-glucosidase, marked a significant increase under the BC and FM-amended soils. Whereas soil available potassium (AK), NO3–N, cellulase activity, and phosphatase peaked under the BC-amended soil, primarily due to the enduring effects of these substrates and metabolites. Furthermore, BC and FM-amended soils enriched specific stress-regulating metabolites, including JA, melatonin, abscisic aldehyde, etc. The sustained effects of both BC and FM-amended soils suppressed disease-causing bacteria, eventually promoting ratooning soil growth conditions. A number of key bioactive compounds had distinct associations with several beneficial bacteria and soil physico-chemical properties.DiscussionThis study proves that long-term BC and FM application is one of the eco-friendly strategies to promote ratoon crop growth and soil quality through the enrichment of stress-regulating metabolites and the suppression of disease-causing bacteria

Plant growth and stress-regulating metabolite response to biochar utilization boost crop traits and soil health

IntroductionThe utilization of biochar (BC) as a soil amendment in agriculture has gained significant traction among many farmers and researchers, primarily due to its eco-friendly role in boosting crop output. However, the performance of specific metabolites (e.g., zeatin, melatonin, sucrose, and phenyllactic acid) in the various tissues of sugarcane plant (leaf, stem, and root) and rhizosphere soil-deemed plant growth and stress regulators in a long-term BC-amended field remains poorly understood. Additionally, literature on the shift in soil attributes and crop growth triggered by the strong response of these bioactive compounds to longterm BC utilization remains undocumented.MethodsMetabolome integrated with highthroughput sequencing analyses were conducted to identify and quantify the performance of plant growth and stress-regulating metabolites in a long-term BC-amended field. Additionally, we investigated how the response of these compounds to BC-treated soil influences crop traits and soil biochemical properties.ResultsWe also identified and quantified the performance of pathogenic bacteria and unraveled the association between these compounds and potential plant growth-promoting bacteria. The BC-supplemented soil significantly boosted the crop traits, including brix, sucrose content, and chlorophyll, as well as soil nutrients, such as soil total nitrogen (TN), ammonium (NH4+-N), and nitrate (NO3--N). We also noticed that metabolite-deemed plant growth and stress regulators, including melatonin and phenyllactic acid, were enriched considerably in the stem and root tissues of the BC-amended soil. Zeatin in the leaf, stem, and root tissues exhibited the same trend, followed by sucrose in the leaf tissue of the BC-treated soil, implying that the strong response of these compounds to BC utilization contributed to the promotion of crop traits and soil quality. Pathogenic bacteria belonging to Proteobacteria and Acidobacteria were suppressed under the BC-supplemented soil, especially in the root tissue and rhizosphere soil, whereas plant growth-regulating bacteria, mainly Bradyrhizobium, responded strongly and positively to several metabolites.DiscussionOur finding provides valuable information for agronomists, farmers, and environmentalists to make informed decisions about crop production, land use, and soil management practices. Proper soil assessment and understanding of the interaction between the attributes of soil, BC, and metabolites are essential for promoting sustainable agriculture practices and land conservation

Comparative Transcriptome Profiling of Resistant and Susceptible Sugarcane Cultivars in Response to Infection by Xanthomonas albilineans

International audienceSugarcane (Saccharum spp. hybrids) is a major source of sugar and renewable bioenergy crop worldwide and suffers serious yield losses due to many pathogen infections. Leaf scald caused by Xanthomonas albilineans is a major bacterial disease of sugarcane in most sugarcane-planting countries. The molecular mechanisms of resistance to leaf scald in this plant are, however, still unclear. We performed a comparative transcriptome analysis between resistant (LCP 85-384) and susceptible (ROC20) sugarcane cultivars infected by X. albilineans using the RNA-seq platform. 24 cDNA libraries were generated with RNA isolated at four time points (0, 24, 48, and 72 h post inoculation) from the two cultivars with three biological replicates. A total of 105,783 differentially expressed genes (DEGs) were identified in both cultivars and the most upregulated and downregulated DEGs were annotated for the processes of the metabolic and single-organism categories, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the 7612 DEGs showed that plant-pathogen interaction, spliceosome, glutathione metabolism, protein processing in endoplasmic reticulum, and plant hormone signal transduction contributed to sugarcane's response to X. albilineans infection. Subsequently, relative expression levels of ten DEGs determined by quantitative reverse transcription-PCR (qRT-PCR), in addition to RNA-Seq data, indicated that different plant hormone (auxin and ethylene) signal transduction pathways play essential roles in sugarcane infected by X. albilineans. In conclusion, our results provide, for the first time, valuable information regarding the transcriptome changes in sugarcane in response to infection by X. albilineans, which contribute to the understanding of the molecular mechanisms underlying the interactions between sugarcane and this pathogen and provide important clues for further characterization of leaf scald resistance in sugarcane

Complete Genome Sequence Reveals Evolutionary and Comparative Genomic Features of Xanthomonas albilineans Causing Sugarcane Leaf Scald

International audienceLeaf scald (caused by Xanthomonas albilineans) is an important bacterial disease affecting sugarcane in most sugarcane growing countries, including China. High genetic diversity exists among strains of X. albilineans from diverse geographic regions. To highlight the genomic features associated with X. albilineans from China, we sequenced the complete genome of a representative strain (Xa-FJ1) of this pathogen using the PacBio and Illumina platforms. The complete genome of strain Xa-FJ1 consists of a circular chromosome of 3,724,581 bp and a plasmid of 31,536 bp. Average nucleotide identity analysis revealed that Xa-FJ1 was closest to five strains from the French West Indies and the USA, particularly to the strain GPE PC73 from Guadeloupe. Comparative genomic analysis between Xa-FJ1 and GPE PC73 revealed prophage integration, homologous recombination, transposable elements, and a clustered regulatory interspaced short palindromic repeats (CRISPR) system that were linked with 16 insertions/deletions (InDels). Ten and 82 specific genes were found in Xa-FJ1 and GPE PC73, respectively, and some of these genes were subjected to phage-related proteins, zona occludens toxin, and DNA methyltransferases. Our findings highlight intra-species genetic variability of the leaf scald pathogen and provide additional genomic resources to investigate its fitness and virulence

Molecular Detection and Quantification of Xanthomonas albilineans in Juice from Symptomless Sugarcane Stalks Using a Real-Time Quantitative PCR Assay

International audienceLeaf scald, a bacterial disease caused by Xanthomonas albilineans (Ashby) Dowson, is a major limiting factor for sugarcane production worldwide. Accurate identification and quantification of X. albilineans is a prerequisite for successful management of this disease. A sensitive and robust quantitative PCR (qPCR) assay was developed in this study for detection and quantification of X. albilineans using TaqMan probe and primers targeting a putative adenosine triphosphate–binding cassette (ABC) transporter gene (abc). The novel qPCR assay was highly specific to the 43 tested X. albilineans strains belonging to different pulsed-field gel electrophoresis groups. The detection thresholds were 100 copies/µl of plasmid DNA, 100 fg/µl of bacterial genomic DNA, and 100 CFU/ml of bacterial suspension prepared from pure culture. This qPCR assay was 100 times more sensitive than a conventional PCR assay. The pathogen was detected by qPCR in 75.1% (410/546) of symptomless stalk samples, whereas only 28.4% (155/546) of samples tested positive by conventional PCR. Based on qPCR data, population densities of X. albilineans in symptomless stalks of the same varieties differed between two sugarcane production areas in China, Beihai (Guangxi Province) and Zhanjiang (Guangdong Province), and no significant correlation between these populations was identified. Furthermore, no relationship was found between these populations of the pathogen in asymptomatic stalks and the resistance level of the sugarcane varieties to leaf scald. The newly developed qPCR assay proved to be highly sensitive and reliable for the detection and quantification of X. albilineans in sugarcane stalks

Identification of Differentially Expressed Proteins in Sugarcane in Response to Infection by Xanthomonas albilineans Using iTRAQ Quantitative Proteomics

Sugarcane can suffer severe yield losses when affected by leaf scald, a disease caused by Xanthomonas albilineans. This bacterial pathogen colonizes the vascular system of sugarcane, which can result in reduced plant growth and plant death. In order to better understand the molecular mechanisms involved in the resistance of sugarcane to leaf scald, a comparative proteomic study was performed with two sugarcane cultivars inoculated with X. albilineans: one resistant (LCP 85-384) and one susceptible (ROC20) to leaf scald. The iTRAQ (isobaric tags for relative and absolute quantification) approach at 0 and 48 h post-inoculation (hpi) was used to identify and annotate differentially expressed proteins (DEPs). A total of 4295 proteins were associated with 1099 gene ontology (GO) terms by GO analysis. Among those, 285 were DEPs during X. albilineans infection in cultivars LCP 85-384 and ROC20. One hundred seventy-two DEPs were identified in resistant cultivar LCP 85-384, and 113 of these proteins were upregulated and 59 were downregulated. One hundred ninety-two DEPs were found in susceptible cultivar ROC20 and half of these (92) were upregulated, whereas the other half corresponded to downregulated proteins. The significantly upregulated DEPs in LCP 85-384 were involved in metabolic pathways, the biosynthesis of secondary metabolites, and the phenylpropanoid biosynthesis pathway. Additionally, the expression of seven candidate genes related to photosynthesis and glycolytic pathways, plant innate immune system, glycosylation process, plant cytochrome P450, and non-specific lipid transfer protein was verified based on transcription levels in sugarcane during infection by X. albilineans. Our findings shed new light on the differential expression of proteins in sugarcane cultivars in response to infection by X. albilineans. The identification of these genes provides important information for sugarcane variety improvement programs using molecular breeding strategies

DataSheet_1_Plant growth and stress-regulating metabolite response to biochar utilization boost crop traits and soil health.docx

IntroductionThe utilization of biochar (BC) as a soil amendment in agriculture has gained significant traction among many farmers and researchers, primarily due to its eco-friendly role in boosting crop output. However, the performance of specific metabolites (e.g., zeatin, melatonin, sucrose, and phenyllactic acid) in the various tissues of sugarcane plant (leaf, stem, and root) and rhizosphere soil-deemed plant growth and stress regulators in a long-term BC-amended field remains poorly understood. Additionally, literature on the shift in soil attributes and crop growth triggered by the strong response of these bioactive compounds to longterm BC utilization remains undocumented.MethodsMetabolome integrated with highthroughput sequencing analyses were conducted to identify and quantify the performance of plant growth and stress-regulating metabolites in a long-term BC-amended field. Additionally, we investigated how the response of these compounds to BC-treated soil influences crop traits and soil biochemical properties.ResultsWe also identified and quantified the performance of pathogenic bacteria and unraveled the association between these compounds and potential plant growth-promoting bacteria. The BC-supplemented soil significantly boosted the crop traits, including brix, sucrose content, and chlorophyll, as well as soil nutrients, such as soil total nitrogen (TN), ammonium (NH4+-N), and nitrate (NO3--N). We also noticed that metabolite-deemed plant growth and stress regulators, including melatonin and phenyllactic acid, were enriched considerably in the stem and root tissues of the BC-amended soil. Zeatin in the leaf, stem, and root tissues exhibited the same trend, followed by sucrose in the leaf tissue of the BC-treated soil, implying that the strong response of these compounds to BC utilization contributed to the promotion of crop traits and soil quality. Pathogenic bacteria belonging to Proteobacteria and Acidobacteria were suppressed under the BC-supplemented soil, especially in the root tissue and rhizosphere soil, whereas plant growth-regulating bacteria, mainly Bradyrhizobium, responded strongly and positively to several metabolites.DiscussionOur finding provides valuable information for agronomists, farmers, and environmentalists to make informed decisions about crop production, land use, and soil management practices. Proper soil assessment and understanding of the interaction between the attributes of soil, BC, and metabolites are essential for promoting sustainable agriculture practices and land conservation.</p

Table_8_Plant growth and stress-regulating metabolite response to biochar utilization boost crop traits and soil health.xlsx

IntroductionThe utilization of biochar (BC) as a soil amendment in agriculture has gained significant traction among many farmers and researchers, primarily due to its eco-friendly role in boosting crop output. However, the performance of specific metabolites (e.g., zeatin, melatonin, sucrose, and phenyllactic acid) in the various tissues of sugarcane plant (leaf, stem, and root) and rhizosphere soil-deemed plant growth and stress regulators in a long-term BC-amended field remains poorly understood. Additionally, literature on the shift in soil attributes and crop growth triggered by the strong response of these bioactive compounds to longterm BC utilization remains undocumented.MethodsMetabolome integrated with highthroughput sequencing analyses were conducted to identify and quantify the performance of plant growth and stress-regulating metabolites in a long-term BC-amended field. Additionally, we investigated how the response of these compounds to BC-treated soil influences crop traits and soil biochemical properties.ResultsWe also identified and quantified the performance of pathogenic bacteria and unraveled the association between these compounds and potential plant growth-promoting bacteria. The BC-supplemented soil significantly boosted the crop traits, including brix, sucrose content, and chlorophyll, as well as soil nutrients, such as soil total nitrogen (TN), ammonium (NH4+-N), and nitrate (NO3--N). We also noticed that metabolite-deemed plant growth and stress regulators, including melatonin and phenyllactic acid, were enriched considerably in the stem and root tissues of the BC-amended soil. Zeatin in the leaf, stem, and root tissues exhibited the same trend, followed by sucrose in the leaf tissue of the BC-treated soil, implying that the strong response of these compounds to BC utilization contributed to the promotion of crop traits and soil quality. Pathogenic bacteria belonging to Proteobacteria and Acidobacteria were suppressed under the BC-supplemented soil, especially in the root tissue and rhizosphere soil, whereas plant growth-regulating bacteria, mainly Bradyrhizobium, responded strongly and positively to several metabolites.DiscussionOur finding provides valuable information for agronomists, farmers, and environmentalists to make informed decisions about crop production, land use, and soil management practices. Proper soil assessment and understanding of the interaction between the attributes of soil, BC, and metabolites are essential for promoting sustainable agriculture practices and land conservation.</p

Table_4_Plant growth and stress-regulating metabolite response to biochar utilization boost crop traits and soil health.xlsx

IntroductionThe utilization of biochar (BC) as a soil amendment in agriculture has gained significant traction among many farmers and researchers, primarily due to its eco-friendly role in boosting crop output. However, the performance of specific metabolites (e.g., zeatin, melatonin, sucrose, and phenyllactic acid) in the various tissues of sugarcane plant (leaf, stem, and root) and rhizosphere soil-deemed plant growth and stress regulators in a long-term BC-amended field remains poorly understood. Additionally, literature on the shift in soil attributes and crop growth triggered by the strong response of these bioactive compounds to longterm BC utilization remains undocumented.MethodsMetabolome integrated with highthroughput sequencing analyses were conducted to identify and quantify the performance of plant growth and stress-regulating metabolites in a long-term BC-amended field. Additionally, we investigated how the response of these compounds to BC-treated soil influences crop traits and soil biochemical properties.ResultsWe also identified and quantified the performance of pathogenic bacteria and unraveled the association between these compounds and potential plant growth-promoting bacteria. The BC-supplemented soil significantly boosted the crop traits, including brix, sucrose content, and chlorophyll, as well as soil nutrients, such as soil total nitrogen (TN), ammonium (NH4+-N), and nitrate (NO3--N). We also noticed that metabolite-deemed plant growth and stress regulators, including melatonin and phenyllactic acid, were enriched considerably in the stem and root tissues of the BC-amended soil. Zeatin in the leaf, stem, and root tissues exhibited the same trend, followed by sucrose in the leaf tissue of the BC-treated soil, implying that the strong response of these compounds to BC utilization contributed to the promotion of crop traits and soil quality. Pathogenic bacteria belonging to Proteobacteria and Acidobacteria were suppressed under the BC-supplemented soil, especially in the root tissue and rhizosphere soil, whereas plant growth-regulating bacteria, mainly Bradyrhizobium, responded strongly and positively to several metabolites.DiscussionOur finding provides valuable information for agronomists, farmers, and environmentalists to make informed decisions about crop production, land use, and soil management practices. Proper soil assessment and understanding of the interaction between the attributes of soil, BC, and metabolites are essential for promoting sustainable agriculture practices and land conservation.</p