Identification and fine mapping of qSW2 for leaf slow wilting in soybean

Drought is one of the abiotic stresses limiting the production of soybean(Glycine max).Elucidation of the genetic and molecular basis of the slow-wilting(SW)trait of this crop offers the prospect of its genetic improvement.A panel of 188 accessions and a set of recombinant inbred lines produced from a cross between cultivars Liaodou 14 and Liaodou 21 were used to identify quantitative-trait loci(QTL)associated with SW.Plants were genotyped by Specific-locus amplified fragment sequencing and seedling leaf wilting was assessed under three water-stress treatments.A genome-wide association study identified 26 SW-associated single-nucleotide polymorphisms(SNPs),including three located in a 248-kb linkage-disequilibrium(LD)block on chromosome 2.Linkage mapping revealed a major-effect QTL,qSW2,associated with all three treatments and adjacent to the LD block.Fine mapping in a BC_(2)F_(3) population derived from a backcross between Liaodou 21 and R26 confined qSW2 to a 60-kb interval.Gene expression and sequence variation analysis identified the gene Glyma.02 g218100,encoding an auxin transcription factor,as a candidate gene for qSW2.Our results will contribute significantly to improving drought-resistant soybean cultivars by providing genetic information and resources.

Expression analysis of the R2R3-MYB gene family in upland cotton and functional study of GhMYB3D5 in regulating Verticillium wilt resistance

Improving plant resistance to Verticillium wilt(VW),which causes massive losses in Gossypium hirsutum,is a global challenge.Crop plants need to efficiently allocate their limited energy resources to maintain a balance between growth and defense.However,few transcriptional regulators specifically respond to Verticillium dahliae and the underlying mechanism has not been identified in cotton.In this study,we found that the that expression of most R2R3-MYB members in cotton is significantly changed by V.dahliae infection relative to the other MYB types.One novel R2R3-MYB transcription factor(TF)that specifically responds to V.dahliae,GhMYB3D5,was identified.GhMYB3D5 was not expressed in 15 cotton tissues under normal conditions,but it was dramatically induced by V.dahliae stress.We functionally characterized its positive role and underlying mechanism in VW resistance.Upon V.dahliae infection,the up-regulated GhMYB3D5 bound to the GhADH1 promoter and activated GhADH1expression.In addition,GhMYB3D5 physically interacted with GhADH1 and further enhanced the transcriptional activation of GhADH1.Consequently,the transcriptional regulatory module GhMYB3D5-GhADH1 then promoted lignin accumulation by improving the transcriptional levels of genes related to lignin biosynthesis(GhPAL,GhC4H,Gh4CL,and GhPOD/GhLAC)in cotton,thereby enhancing cotton VW resistance.Our results demonstrated that the GhMYB3D5 promotes defense-induced lignin accumulation,which can be regarded as an effective way to orchestrate plant immunity and growth.

Fusarium wilt of banana:Current update and sustainable disease control using classical and essential oils approaches

Fusarium species were reported to produce biofilms.Biofilms are superficial societies of microbes bounded and endangered by being situated or taking place outside a cell or cells.The most destructive fungal diseases caused by phytopathogens are as a result of biofilms formation.Fusarium wilt of banana(Panama disease)is caused by a soil-borne pathogen called Fusarium oxysporum f.sp.cubense.Fusarium oxysporum occurs in a form of a species complex(FOSC)which encompasses a crowd of strains.Horizontal genetic factor transfer may donate to the observed assortment in pathogenic strains,while sexual reproduction is unknown in the FOSC.Fusarium wilt is a notorious disease on several crops worldwide.Yield loss caused by this pathogen is huge,and significant to destroy crop yields annually,thereby affecting the producer countries in various continents of the world.The disease is also resistant to various synthetic chemical fungicides.However,excessive use of synthetic fungicides during disease control could be lethal to humans,animals,and plants.This calls for alternative eco-friendly management of this disease by targeting the biofilms formation and finally suppressing this devastating phytopathogen.In this review,we,therefore,described the damage caused by Fusarium wilt disease,the concept of filamentous fungal biofilms,classical control strategies,sustainable disease control strategies using essential oils,and prevention and control of vegetables Fusarium wilt diseases.

Association mapping of lignin response to Verticillium wilt through an eight-way MAGIC population in Upland cotton

Lignin metabolism plays a pivotal role in plant defense against pathogens and is always positively correlated as a response to pathogen infection. Thus, understanding resistance genes against plant pathogens depends on a genetic analysis of the lignin response. This study used eight Upland cotton lines to construct a multi-parent advanced generation intercross(MAGIC) population(n=280), which exhibited peculiar characteristics from the convergence of various alleles coding for advantageous traits. In order to measure the lignin response to Verticillium wilt(LRVW), the artificial disease nursery(ADN) and rotation nursery(RN) were prepared for MAGIC population planting in four environments. The stem lignin contents were collected, and the LRVW was measured with the lignin value of ADN/RN in each environment, which showed significant variations. We employed 9 323 high-quality single-nucleotide polymorphism(SNP) markers obtained from the Cotton-SNP63K array for genotyping the MAGIC population. The SNPs were distributed through the whole genome with 4.78 SNP/Mb density, ranging from 1.14(ChrA06) to 10.08(ChrD08). In addition, a genome-wide association study was performed using a Mixed Linear Model(MLM) for LRVW. Three stable quantitative trait loci(QTLs), qLRVW-A04, qLRVW-A10, and qLRVW-D05, were identified in more than two environments. Two key candidate genes, Ghi_D05G01046 and Ghi_D05G01221, were selected within the QTLs through the combination of variations in the coding sequence, induced expression patterns, and function annotations. Both genes presented nonsynonymous mutations in coding regions and were strongly induced by Verticillium dahliae. Ghi_D05G01046 encodes a leucine-rich extensin(LRx) protein involved in Arabidopsis cell wall biosynthesis and organization. Ghi_D05G01221 encodes a transcriptional co-repressor novel interactor of novel interactor of jasmonic acid ZIM-domain(JAZ–NINJA), which functions in the jasmonic acid(JA) signaling pathway. In summary, the study creates valuable genetic resources for breeding and QTL mapping and opens up a new perspective to uncover the genetic basis of VW resistance in Upland cotton.

Integrative analysis of the transcriptome and metabolome reveals the response mechanism to tomato spotted wilt virus

Tomato spotted wilt virus(TSWV)is an important virus that has rapidly spread throughout the world.TSWV seriously hinders the production of tomato(Solanum lycopersicum)and other plants.In order to discover more new genes and metabolites related to TSWV resistance in tomato plants,the genes and metabolites related to the resistance of tomato plants inoculated with TSWV were identified and studied herein.The tomato TSWV-resistance line YNAU335(335)and TSWV-susceptible lines NO5 and 96172I(961)were used as the transcriptome and metabolome research materials.Transcriptomic and metabolomic techniques were used to analyze the gene and metabolite response mechanisms to TSWV inoculation.A total of 3566,2951,and 2674 differentially expressed genes(DEGs)were identified in lines 335,NO5,and961,respectively.Meanwhile,208,228,and 273 differentially accumulated metabolites(DAMs)were identified in lines 335,NO5,and 961,respectively.In line 335,the number of DEGs was the highest,but the number of DAMs was lowest.Furthermore,903 DEGs and 94 DAMs were common to the response to TSWV in the three inbred lines.The 903 DEGs and 94 DAMs were mainly enriched in the plant hormone signal transduction and flavonoid synthesis pathways.In addition,many nucleotide-binding site-leucine-rich repeat genes and transcription factors were found that might be involved in the TSWV response.These results provide new insights into TSWV resistance mechanisms.

Assessing the severity of cotton Verticillium wilt disease from in situ canopy images and spectra using convolutional neural networks

Verticillium wilt(VW)is a common soilborne disease of cotton.It occurs mainly in the seedling and bollopening stages and severely impairs the yield and quality of the fiber.Rapid and accurate identification and evaluation of VW severity(VWS)forms the basis of field cotton VW control,which has great significance to cotton production.Cotton VWS values are conventionally measured using in-field observations and laboratory test diagnoses,which require abundant time and professional expertise.Remote and proximal sensing using imagery and spectrometry have great potential for this purpose.In this study,we performed in situ investigations at three experimental sites in 2019 and 2021 and collected VWS values,in situ images,and spectra of 361 cotton canopies.To estimate cotton VWS values at the canopy scale,we developed two deep learning approaches that use in situ images and spectra,respectively.For the imagery-based method,given the high complexity of the in situ environment,we first transformed the task of healthy and diseased leaf recognition to the task of cotton field scene classification and then built a cotton field scenes(CFS)dataset with over 1000 images for each scene-unit type.We performed pretrained convolutional neural networks(CNNs)training and validation using the CFS dataset and then used the networks after training to classify scene units for each canopy.The results showed that the Dark Net-19 model achieved satisfactory performance in CFS classification and VWS values estimation(R^(2)=0.91,root-mean-square error(RMSE)=6.35%).For the spectroscopy-based method,we first designed a one-dimensional regression network(1D CNN)with four convolutional layers.After dimensionality reduction by sensitive-band selection and principal component analysis,we fitted the 1D CNN with varying numbers of principal components(PCs).The 1D CNN model with the top 20 PCs performed best(R^(2)=0.93,RMSE=5.77%).These deep learning-driven approaches offer the potential of assessing crop disease severity from spatial and spectral perspectives.

Comparative Assessment of Genetic Diversity of Peanut (Arachis hypogaea L.)Genotypes with Various Levels of Resistance to Bacterial Wilt Through SSR and AFLP Analyses

Bacterial wilt （BW） caused by Ralstonia solanacearum is an important constraint to peanut （Arachis hypogaea L.） production in several Asian and African countries, and planting BW-resistant cultivars is the most feasible method for controlling the disease. Although several BW-resistant peanut germplasm accessions have been identified, the genetic diversity among these has not been properly investigated, which has impeded efficient utilization. In this study, the genetic relationships of 31 peanut genotypes with various levels of resistance to BW were assessed based on SSR and AFLP analyses. Twenty-nine of 78 SSR primers and 32 of 126 AFLP primer combinations employed in this study were polymorphic amongst the peanut genotypes tested. The SSR primers amplified 91 polymorphic loci in total with an average of 3.14 alleles per primer, and the AFLP primers amplified 72 polymorphic loci in total with an average of 2.25 alleles per primer. Four SSR primers （14H06, 7G02, 3A8, 16C6） and one AFLP primer （P1M62） were found to be most efficient in detecting diversity. The genetic distance between pairs of genotypes ranged from 0.12 to 0.94 with an average of 0.53 in the SSR data and from 0.06 to 0.57 with an average of 0.25 in the AFLP data. The SSR-based estimates of the genetic distance were generally larger than that based on the AFLP data. The genotypes belonging to subsp, fastigiata possessed wider diversity than that of subsp, hypogaea. The clustering of genotypes based on the SSR and AFLP data were similar but the SSR clustering was more consistent with morphological classification ofA. hypogaea. Optimum diverse genotypes of both subsp, hypogaea and subsp.fastigiata can be recommended based on this analysis for developing mapping populations and breeding for high yielding and resistant cultivars.

Screening of Soil Antagonistic Bacteria for Watermelon Fusarium Wilt

[Objective] The aim of this study was to screen bacteria strains with stable antagonistic effect against watermelon fusarium wilt from soil and investigate the biological control of watermelon fusarium wilt by applying the antagonistic bacteria strains into soil.[Method] Actinomycete strains,fluorescent bacteria strains and bacillus strains were isolated from soil samples by the dilution-plate method,then its resistance was screened respectively by the improved confront culture method after colonies were purified.Finally,bacteria strains with better antagonistic effect were identified.[Result] 29 bacteria strains with stable antagonistic effect against watermelon fusarium wilt were screened from 39 soil samples,which contained 15 fluorescent bacteria strains,5 bacillus strains and 9 actinomycete strains.Furthermore,three antagonistic bacteria strains of FM2,FM3 and FM4 with the strongest antagonism were identified primarily.[Conclusion] According to cultural characteristics,morphological observation,biochemical and physiological tests,FM2 belongs to bacillus subtilis,while FM3 and FM4 belong to micrococcus.

Resistance Evaluation of Eggplant Resources to Verticillium wilt in Sichuan Province

[Objective] Aimed to lay a scientific foundation for eggplant resistant breeding, the resistant field identification of eggplant resources to Verticillium wilt was carried out. [Method] Test materials were 35 eggplant resources, which were collected from various places in Sichuan province since 2001. These eggplant resources were made resistance evaluation to Verticillium wilt by watering injured roots with Verticillium dahliae spore suspension. [Result] The eggplant resources had good resistance to Verticillium wilt. Among 35 evaluated resources, 10 resources like 242-9OP, 41-1,35-R5, etc. were resistant, while 23 resources like 68 ×47, Zizaoqie, 32-1, etc. were tolerant. Some resistant resources like 41-1,35-R5, 47-R1, etc. had been transferred into new resistant varieties like EF9910, etc. by breeding institutes. [Conclusion] The resistant level and future resistance breeding direction of eggplant resources in Sichuan were clarified by the resistance evaluation of eggplant resources so as to provide a technical guidance for eggplant resistance breeding.

Preliminary Study on Garlic Root Exudates Influences to the Growth of Pomegranate Wilt Pathogen(Ceratocystis fimbriata)and Bacillus subtilis

[Objectives] This paper aims to explore the possibility to intercrop garlic with pomegranate tree to control pomegranate wilt.[Methods] Root exudates of garlic is cultivated in 1/5 concentration of MS solution and distilled water is examined in lab to test their effect to growth of mycelia of pomegranate wilt pathogen（Ceratocystis fimbriata）and multiplication of Bacillus subtilis.[Results] The result shows that garlic root exudates whatever cultivated in MS solution or distilled water could not inhibit or promote mycelia growth of C.fimbriata.However,garlic root exudates cultivated in both methods effectively promote multiplication of B.subtilis.[Conclusions] It is suggested that intercropping garlic with pomegranate tree by combining application B.subtilis could be a promising way to prevent pomegranate wilt spread in practice.

Isolation and Identification of Antifungal Bacterial Strain KL-1 against Plant Wilt Disease

[Objective] The antifungal bacteria of plant wilt disease was screened and identified to provide foundation for the study on bio-control preparation of plant wilt disease.[Method] Confrontation culture method was adopted to screen the bio-control bacteria with good antifungal effect against plant wilt disease,Biolog bacteria automatic identification system and 16S rDNA sequence analysis method were selected to identify its taxonomic status,the biological safety of the strain towards cotton and mice was also determined.[Result] 12 bacterial strains were isolated from rhizosphere of cotton.Among those strains,5 isolates showed antifungal activity against F.decemcellulare Brick,F.oxysporum f.sp.Diathi,F.oxysporum f.sp.vasinfectum.The antifungal effect of KL-1 strain against three target strains of pathogen reached 69.09%,80.78% and 78.89% respectively.Identification results of Biolog bacteria automatic identification system and 16S rDNA sequence analysis method showed that KL-1strain was Bacillus amyloliquefaciens;primary determination results of biological safety also showed that the strain KL-1 was safe and non-toxic towards cotton and mice.[Conclusion] KL-1strain of B.amyloliquefaciens had antifungal effect against several pathogens of plant wilt diseases,which was safe and non-toxic towards cotton and mice,being the bio-control strain with research and development potential.

Effects of Seedling-cake Integrated with Arbuscular Mycorrhizal Fungi on Resistance of Bacterial Wilt in Pepper (Capsicum annuum L.)

[Objective] The aim was to investigated the effects of seedling-cake with AMF on resistance of pepper （Capsicum annuum L.） to bacterial wilt. [Methods] A pot experiment was conducted to evaluate the dynamics of mycorrhizal colonization, disease index, morbidity rate, control effect and several agronomic traits of mycorrhizal seedlings after inoculation with RS in two kinds of seeding-cakes with AMF. [Results] Two kinds of seedling-cakes formed steadily mycorrhizal colonization after inoculation, which enhanced disease-resistance and decreased morbidity rate and disease index in different degrees. What’s more, the performance of self-made seedling-cakes was obviously better than that of commercial seedling-cakes. [Conclusions] Self-made seedling-cakes with AMF have superior performance on bacterial wilt resistance of pepper, which should be made use of in other crops extensively.

Molecular Marker Assisted Selection for Fusarium Wilt Resistance Breeding in Watermelon(Citrullus lanatus)

Molecular identification on diploid and tetraploid watermelon breeding lines which were resistant to Fusarium wilt was carried out with the published dCAPS marker ＂4451_fon＂ which was closely linked with resistance gene of Fusarium wilt race 1. The results showed that all the diploid and tetraploid lines expressed as re- sistant genotype, which were defined as Fusarium wilt-resistant materials. The re- sults were consistent with that of artificial inoculation identification. Molecular identifi- cation results also indicated that the resistant lines were homozygote, and the Fusarium wilt-resistant gene would not separate or lose during the future self- crossed purification. Therefore, resistance selection would not be necessary in their progeny populations. The study results thought that dCAPS marker ＂4451_fon＂ could be applied on molecular marker assisted selection for Fusarium wilt resistance breeding in watermelon to increase breeding selection efficiency and accelerate breeding progress.

Bioinformatics Analysis of Antifungal Peptide 25a2 of Cotton Verticillium Wilt

Protein 25a2 is the antifungal peptide of cotton Verticillium wilt which was isolated from Bacillus amyloliquefaciens. The amino acid sequence of antifungal peptide 25a2 was analyzed using bioinformatics tools, and the characters of signal peptides, transmembrane topological structura, physicochemical signatures, protein domain, secondary and tertiary structure of protein were predicted. The results showed that 25a2 was a secreted protein, the sequence of which included a signal peptide in N end and a transmembrane domain in C end. The predicted secondary structure showed that the antibacterial peptide was mainly free random coils, belonging to mixed protein, three-dimensional model of 25a2 was a compact ball. These results showed that the most possible action mechanism of antifungal peptide 25a2 might be ＂carpet＂ model.

Inhibitory Effects of Extracts from Marigold(Tagetes patula) against Tomato Fusarium Wilt

[ Objective ] The study aimed to explore a new way for the control of Tomato Fusarium Wilt. [ Method ] Different solvents were used to prepare the ex-tracts of marigold, and the inhibitory effects of different extraction solvents and different extraction parts of marigold against Tomato Fusar/um Wilt were compared. [ Result ] Among different solvent extracts of marigold, chloroform extracts had the strongest inhibitory effects against the growth of the pathogen; among the chloro- form extracts from different parts of marigold, root extract had the most obvious inhibitory effect against the disease, followed by flower and leaf extracts, and the in- hibitory effect of stem extract was the weakest. [ Conclusion ] The active components of marigold have inhibitory effect against Tomato Fusarium Wilt, and the plant has good development prospects and application value.

Purification of Antifungal Protein from Biocontrol Strain XG-1 against Fusarium Wilt of Watermelon

[Objective] This study aimed to isolate and purify the antifungal protein against Fusarium oxysporum f. sp niveum （FON） from Bacillus subtilis XG-I. [Method] The crude protein was obtained by using fractional precipitation with am- monium sulfate, then chromatography using both DEAE-Sepharose FF anion ex- change and Sephacryl S-100 gel filtration columns was adopted for further purifica- tion of the protein, followed by MALDI-TOF-MS analysis of the structure of the at- tained protein which was antagonistic to FON. [Result] The result from MALDI-TOF- MS analysis suggested that the antifungal protein extracted from strain XG-1 was highly homologous to the flagellin （gi114278900） deriving from B. subtilis, with a pro- tein score of 248 and a coverage rate of 63%. It was thus speculated that this an- tagonistic protein was a kind of flagellin with a molecular mass of 30.6 kD. [Conclusion] The results from this study provide theoretical basis for clarifying the mechanisms of the action of strain XG-1 as well as important references for the preven- tion and control of watermelon Fusarium wilt.

Induction of Tolerance to Fusarium Wilt and Defense-Related Mechanisms in the Plantlets of Susceptible Berangan Banana Pre-Inoculated with Pseudomonas sp.(UPMP3) and Burkholderia sp.(UPMB3)

This study is aimed at assessing the ability of two endophytic bacteria originally isolated from healthy oil palm roots, Pseudomonas sp. （UPMP3） and Burkholderia sp. （UPMB3） to induce resistance in susceptible Berangan banana against Fusarium oxysporum race 4 （FocR4）. Increased accumulation of resistance-related enzymes such as peroxidase （PO）, phenylalanine ammonia lyase （PAL）, lignithioglycolic acid （LTGA）, and pathogenesis-related （PR） proteins （ehitinase and β-1,3-glucanase） has been observed in plantlets treated with endophytic bacteria UPMP3 and UPMB3 singly or as mixture under glasshouse conditions. Pre-inoculation of banana plantlets with UPMP3 showed a significant reduction in Fusarium wilt incidence 72 d after challenged inoculation with FocR4. UPMB3 was less effective in suppressing Fusarium wilt compared to UPMP3, whereas, the mixture of both endophytes showed an intermediate effect. Based on these results, it is concluded that UPMP3 could be a promising biological control agent that can trigger resistance against Fusarium wilt in susceptible Berangan banana.

The effects of Trichoderma on preventing cucumber fusarium wilt and regulating cucumber physiology

In our previous studies, we identified 3 Trichoderma strains with anti-Fusarium oxysporum activity, including T. asperellum 525, T. harzianum 610, and T. pseudokoningii 886. Here, we evaluated the effects of these 3 Trichoderma strains on preventing cucumber fusarium wilt through pot culture and greenhouse culture experiments. All 3 Trichoderma strains demonstrated higher control effects toward cucumber fusarium wilt than previous studies, with efficacies over 78%. Additionally, inoculation with the 3 Trichoderma strains significantly promoted the quality and yield of cucumbers. Among the 3 strains, Trichoderma 866 was the most effective, with disease control efficacy of 78.64% and a cucumber yield increase of 33%. Furthermore, seedlings inoculated with Trichoderma exhibited significantly increased measures of plant height, stem diameter, leaf area, aboveground fresh weight, underground fresh weight, chlorophyll content, and nitric nitrogen content, as well as the activities of several stress-resistance enzymes, including superoxide dismutase(SOD), peroxidase(POD), catalase(CAT), polyphenol oxidase(PPO), and ascorbate oxidase(AAO). In addition, the plants inoculated with Trichoderma showed decreased cell membrane permeability and malondialdehyde(MDA) content in the leaves. Together, our results suggest that T. asperellum 525, T. harzianum 610, and T. pseudokoningii 886 inoculations inhibit F. oxysporum infection, stimulate the metabolism in cucumbers, and enhance the activities of stress-resistance enzymes, which consequently promote the growth of cucumber plants, prevent cucumber fusarium wilt, and improve the yield and quality of cucumbers. T. harzianum is a commonly used biocontrol fungus, while few studies have focused on T. asperellum or T. koningense. In this study, strains of T. asperellum and T. pseudokoningii showed excellent plant disease prevention and growth promoting effects on cucumber, indicating that they also have great potential as biocontrol fungi.

Pine wilt disease: a short review of worldwide research

This article summarizes the results of the research papers presented at the International Symposium on pine wilt disease （IUFRO Working Party Meeting 4.04.03） held in July 2009, at Nanjing, China. The general topics covered were on pine wilt disease （PWD）, its causal organism, the pinewood nematode （PWN） Bursaphelenchus xylophilus, plus other PWN-associated microorganisms that play a significant role in PWD such as bacteria （e.g. Pseudomonasfluorescens）. Most of the papers that are reviewed are based on work on PWD-PWN in East Asia and Russia. Specific topics covered include： 1） the fundamental conceptions of PWD development, 2） pathogenicity, 3） host-parasite relationships including the histopathology of diseased conifers and the role of toxins from bacteria-nematode ecto-symbionts, 4） PWN life cycle and transmission, 5） B. xylophilus dissemination models, 6） associations （with other nematodes）, 7） diagnostics, 8） quarantine and control of the PWN and 9） biocontrol of the PWN.

Advances in Cotton Breeding for Resistance to Fusarium and Verticillium Wilt in the Last Fifty Years in China

This review summarizes the main advances in cotton breeding for resistance to fusarium and verticillium wilt in the last fifty years in China. The topics discussed include main achievements, experiences, problems and countermeasures for solving.