Investigation of the epidemiology,pathogenicity and immunogenicity of Bordetella bronchiseptica isolated from cats and dogs in China from 2021 to 2023

Bordetella bronchiseptica(Bb)is recognized as a leading cause of respiratory diseases in dogs and cats.However,epidemiological data on Bb in dogs and cats in China are still limited,and there is no commercially available vaccine.Live vaccines containing Bb that are widely used abroad are generally efective but can establish latency and potentially reactivate to cause illness in some immunodefcient vaccinated recipients,raising safety concerns.In this study,34 canine-derived and two feline-derived Bb strains were isolated from 1809 canine and 113 feline nasopharyngeal swab samples collected from eight provinces in China from 2021 to 2023.The PCR results showed that the percentage of positive Bb was 22.94%(441/1922),and more than 90%of the Bb isolates had four virulence factor-encoding genes(VFGs),namely,fhaB,prn,betA and dnt.All the isolated strains displayed a multidrug-resistant phenotype.The virulence of 10 Bb strains isolated from dogs with respiratory symptoms was tested in mice,and we found that eight isolates were highly virulent.Furthermore,the eight Bb isolates with high virulence were inactivated and intramuscularly injected into mice,and three Bb strains(WH1218,WH1203 and WH1224)with the best protective efcacy were selected.Dogs immunized with these three strains exhibited strong protection against challenge with the Bb feld strain WH1218.Ultimately,the WH1218 strain with the greatest protection in dogs was selected as the vaccine candidate.Dogs and cats that received a vaccine containing 109 CFU of the inactivated WH1218 strain showed complete protection against challenge with the Bb feld strain WH1218.This study revealed that Bb is an important pathogen that causes respiratory diseases in domestic dogs and cats in China,and all the isolates exhibited multidrug resistance.The present work contributes to the current understanding of the prevalence,antimicrobial resistance,and virulence genes of Bb in domestic dogs and cats.Additionally,our results suggest that the WH1218 strain is a promising candidate safe and efcacious inactivated Bb vaccine.

SsdchA is a novel secretory cellobiohydrolase driving pathogenicity in Sclerotinia sclerotiorum

The necrotrophic fungus, Sclerotinia sclerotiorum, employs an array of cell wall-degrading enzymes(CWDEs), including cellulase, to dismantle host cell walls. However, the molecular mechanisms through which S. sclerotiorum degrades cellulose remain elusive. Here, we unveil a novel secretory cellobiohydrolase, SsdchA, characterized by a signal peptide and a Glyco_hydro_7(GH7) domain. SsdchA exhibits a robust expression of during early infection stages. Interestingly, colony morphology and growth rates remain unaffected across the wild-type, SsdchA deletion strains and SsdchA overexpression strains on potato dextrose agar(PDA) medium. Nevertheless, the pathogenicity and cellobiohydrolase activity decreased in the SsdchA deletion strains, but enhanced in the SsdchA overexpression strains. Moreover,the heterologous expression of SsdchA in Arabidopsis thaliana leads to reduced cellulose content and heightened susceptibility to S. sclerotiorum. Collectively, our data underscore the pivotal role of the novel cellobiohydrolase SsdchA in the pathogenicity of S. sclerotiorum.

Identification and characterization of FpRco1 in regulating vegetative growth and pathogenicity based on T-DNA insertion in Fusarium pseudograminearum

Fusarium pseudograminearum is a devastating pathogen that causes Fusarium crown rot(FCR)in wheat and poses a significant threat to wheat production in terms of grain yield and quality.However,the mechanism by which F.pseudograminearum infects wheat remains unclear.In this study,we aimed to elucidate these mechanisms by constructing a T-DNA insertion mutant library for the highly virulent strain WZ-8A of F.pseudograminearum.By screening this mutant library,we identified nine independent mutants that displayed impaired pathogenesis in barley leaves.Among these mutants,one possessed a disruption in the gene FpRCO1 that is an ortholog of Saccharomyces cerevisiae RCO1,encoding essential component of the Rpd3S histone deacetylase complex in F.pseudograminearum.To further investigate the role of FpRCO1 in F.pseudograminearum,we employed a split-marker approach to knock out FpRCO1 in F.pseudograminearum WZ-8A.FpRCO1 deletion mutants exhibit reduced vegetative growth,conidium production,and virulence in wheat coleoptiles and barley leaves,whereas the complementary strain restores these phenotypes.Moreover,under stress conditions,the FpRCO1 deletion mutants exhibited increased sensitivity to NaCl,sorbitol,and SDS,but possessed reduced sensitivity to H_(2)O_(2)compared to these characteristics in the wild-type strain.RNA-seq analysis revealed that deletion of FpRCO1 affected gene expression(particularly the downregulation of TRI gene expression),thus resulting in significantly reduced deoxynivalenol(DON)production.In summary,our findings highlight the pivotal role of FpRCO1 in regulating vegetative growth and development,asexual reproduction,DON production,and pathogenicity of F.pseudograminearum.This study provides valuable insights into the molecular mechanisms underlying F.pseudograminearum infection in wheat and may pave the way for the development of novel strategies to combat this devastating disease.

The DNA damage repair complex MoMMS21-MoSMC5 is required for infection-related development and pathogenicity of Magnaporthe oryzae

The conserved DNA damage repair complex,MMS21-SMC5/6(Methyl methane sulfonate 21-Structural maintenance of chromosomes 5/6),has been extensively studied in yeast,animals,and plants.However,its role in phytopathogenic fungi,particularly in the highly destructive rice blast fungus Magnaporthe oryzae,remains unknown.In this study,we functionally characterized the homologues of this complex,MoMMS21 and MoSMC5,in M.oryzae.We first demonstrated the importance of DNA damage repair in M.oryzae by showing that the DNA damage inducer phleomycin inhibited vegetative growth,infection-related development and pathogenicity in this fungus.Additionally,we discovered that MoMMS21 and MoSMC5 interacted in the nuclei,suggesting that they also function as a complex in M.oryzae.Gene deletion experiments revealed that both MoMMS21 and MoSMC5 are required for infection-related development and pathogenicity in M.oryzae,while only MoMMS21 deletion affected growth and sensitivity to phleomycin,indicating its specific involvement in DNA damage repair.Overall,our results provide insights into the roles of MoMMS21 and MoSMC5 in M.oryzae,highlighting their functions beyond DNA damage repair.

Pathogenicity Analysis on Magnaporthe oryzae from Hybrid Combination Wuyou 308

Eighteen blast isolates were obtained from hybrid combination Wuyou308 using the Magnaporthe oryzae pathogen isolation method.Race identification of these isolates was conducted based on seven Chinese blast differentials and 11 blast monogenic lines.The results indicated that the isolates were identified as the races of ZB13,ZB15 and ZC13,accounting for 66.67%,27.78%,5.56%,respectively,and the resistance genes including Pi-ta2 and Pi-sh,Pi-i were highly susceptible to these isolates,while the resistance genes like Pi-kh,Pi-1,Pi2,Pi-9 and Pi-50 showed good resistance to tested pathogens.All isolates were compatible to the original rice hybrid Wuyou308.Three isolates including GDHY-308-1401 were used for testing their pathogenicity to 45 local varieties.The results demonstrated that 13 varieties appeared highly susceptible to the tested isolates,accounting for 28.89%;two varieties appeared moderately susceptible to the tested isolates,accounting for 4.44%;30 varieties showed moderately/highly resistance,accounting for 66.67%.Among them,some of new hybrid combinations such as Wufengyou 9802,Wuyou 613,Wuyou 1179 showed good resistance to the inoculated strains,and they were recommended to be candidates in the rice region where Wuyou308 showed susceptibility.

Characteristics of Mycobacterium tuberculosis serine protease Rv1043c in enzymology and pathogenicity in mice

The serine proteases of Mycobacteria tuberculosis(Mtb)are important contributors to the process of bacterial invasion and its pathogenesis.In the present study,we systematically characterized the role of the Rv1043c protein in Mycobacterium infection by purifying the Rv1043c protein in Escherichia coli and constructing a Mycobacterium smegmatis(Msg)strain overexpressing Rv1043c(Msg_Rv1043c).We found that Rv1043c had serine protease activity and localized to the surface of Mtb.We determined that the optimal pH and temperature for the Rv1043c serine protease were 9.0 and 45°C,respectively.Moreover,the serine protease activity of Rv1043c was enhanced by divalent metal ions of Ca^(2+)and Mg^(2+).Site-directed mutagenesis studies demonstrated that the serine 279 residue in Rv1043c plays a catalytic role.Additionally,mouse model studies confirmed that Rv1043c significantly enhanced the survival of Msg in vivo,induced pulmonary injury and lung cell apoptosis,and promoted the release of pro-inflammatory cytokines interleukin-1βand interleukin-6 in mice.This study presents novel insights into the relationship between mycobacterial serine protease and the pathogenesis of the disease.

Analysis of Subcellular Localization and Pathogenicity of Plum Bark Necrosis Stem-Pitting Associated Virus Protein P6

Infection of plum bark necrosis stem pitting associated virus(PBNSPaV)has been reported in many Prunus species in several countries,causing significant economic losses.The very small proteins encoded by plant viruses are often overlooked due to their short sequences and uncertain significance.However,numerous studies have indicated that they might play important roles in the pathogenesis of virus infection.The role of small hydrophobic protein P6,encoded by the open reading frame 2 of PBNSPaV,has not been well explored.In this study,we amplified the P6 fragment from a PBNSPaV isolate by RT-PCR using specific primers and found that it is 174 bp long and encodes a protein of approximately 6.3 kD with a transmembrane domain.Subcellular localization analysis of P6 proteins in tobacco leaves showed that P6 localizes to the cytomembrane and nuclear membrane.To further clarify the pathogenicity of P6 proteins,we constructed a PVX-P6 expression vector by inserting the p6 fragment into a potato virus X(PVX)-based vector and transformed it into Agrobacterium tumefaciens GV3101.Infiltration of Nicotiana benthamiana(N.benthamiana)with the PVX vector-transformed A.tumefaciens led to slight mosaic symptoms at 14 days of post-inoculation.Meanwhile,infiltration with the PVX-P6 vector-transformed A.tumefaciens resulted in no significant symptoms.These results demonstrated that heterologous expression of P6 in N.benthamiana could not enhance the pathogenicity of PVX.Our study indicates that P6 may not be a potential pathogenic factor associate with the causing of symptoms,and the mode of action of PBNSPaV-P6 protein remains to be further studied.

Production profile and comparison analysis of main toxin components of Fusarium oxysporum f.sp.sesami isolates with different pathogenicity levels

Fusarium wilt is a common fungal disease in sesame caused by Fusarium oxysporum f.sp.sesami(FOS).To determine the toxin production profiles of the FOS isolates with different pathogenicity levels under various culture conditions,we assessed the content variation of fusaric acid(FA)and 9,10-dehydrofusaric acid(9,10-DFA)produced by the four representative FOS isolates.Results indicated that the concentration of FA reached to a maximum of 2848.66μg/mL in Czapek medium,while 9,10-DFA was mainly produced in Richard and Lowcarbon Richard medium.The concentration of 9,10-DFA on Richard culture medium varied from 0μg/mL to 716.89μg/mL.Of the five culture media used in this study,Czapek culture medium was the most conductive to produce FA.FA production was significantly affected by culture medium,culture time,and their interactions.Results suggest that there is no correlation between toxin production and pathogenicity level of FOS isolates.These findings provide key information for the mechanism analysis of FOS-sesame interaction and pathogen control.

Molecular Characterization,Morphology and Pathogenicity of Curvularia pseudobrachyspora,A New Causal Agent of Leaf Spot on Banana

[Objectives]The paper was to study molecular characterization,morphology and pathogenicity of Curvularia pseudobrachyspora,a new causal agent of leaf spot on banana.[Methods]Banana(Musa acuminate)leaves with streaks or long ellipse-shaped lesions were sampled in an orchard of Danzhou City,Hainan Province,China in 2021.Fungus isolates were isolated from the diseased tissues and further identified as C.pseudobrachyspora based on morphological characteristics of colony,conidiophore and spore,phylogenetic analyses of the ITS region,GAPDH and TEF-1αgenes.[Results]In the pathogenicity test,the fungus re-isolated from inoculated leaves with necrotic lesions was identified morphologically and molecularly,fulfilling Koch's postulates.[Conclusions]C.pseudobrachyspora is a new pathogen causing leaf spot of banana in China and the world.

Evaluation under Semi-Controlled Conditions of the Pathogenicity of Three Isolates of Phaeoisariopsis personata (Berk. & M.A Curt.)

Peanut (Arachis hypogaea L.) late leaf spot is an important disease caused by Phaeoisariopsis personata (Berk. Et M. A Curt.). This fungus is responsible for the most damaging leaf spots in peanut production. The present experiment was undertaken to evaluate the pathogenic variability of Phaeoisariopsis personata in Burkina Faso. To this end, detached leaves and healthy plants of three peanut varieties were inoculated. Isolates I3TF, I2TG and I1TK of the pathogen (105 conidia/ml), collected respectively in the western, central and eastern agroecological zones of country, were used. The inoculated leaves were kept in Petri dishes on moist blotting paper and stored in the laboratory during the experimental period. The inoculated plants were grown under glass in pots containing a mixture of sterilized sand and clay. The development of disease was monitored and severity was scored every 15 days using rating scale. The results obtained in the laboratory and in the greenhouse revealed that there is pathogenic variability in the isolates tested. Indeed, for each variety, the highest severity score was recorded in plants inoculated with isolate I3TF and the lowest severity score with isolate I1TG. In the laboratory the severity scores ranged from 6.76 to 8.80 in TS32-1, 6.18 to 8.29 in SH70P and 5.98 to 7.92 in PC79-79. In the greenhouse, the average severity scores ranged from 5.61 to 8.33 in TS32-1, from 5.19 to 8.00 in SH70P, from 4.90 to 7.50 in PC79-79. Thus, the variety TS32-1 was the most susceptible to all three isolates of the pathogen.

A Preliminary Study on Genetic Variation of g E Gene of an Epidemic Pseudorabies Virus Strain and Its Pathogenicity to Piglets

[Objective] This study aimed to investigate the genetic variation of g E gene of an epidemic pseudorabies virus（PRV） strain and its pathogenicity to piglets. [Method] By serial passage in Vero cells, a PRV strain was isolated from the brain tissues of stillborn fetuses delivered by sows with suspected PRV infection and preliminarily identified by PCR. g E gene of the isolated PRV strain was amplified and sequenced for phylogenetic analysis. In addition, the pathogenicity of the isolated PRV strain to 6-week-old piglets was evaluated. [Result] A PRV strain was successfully isolated and named PRV N5 B strain, which could proliferate in Vero cells and TCID50 of the 15 thgeneration virus liquid reached 10^7.125/0.1 ml. Specific bands could be amplified by PCR. g E gene in the isolated PRV strain was 1 740 bp in length. A phylogenetic tree was constructed based on full-length g E sequences, which showed that PRV N5 B strain and PRV strains isolated since 2012 were clustered into the same independent category and shared 99.7%-100% homology of nucleotide sequences. Compared with related sequences published previously, there were insertions of three consecutive bases at two loci. Animal experiments showed that intranasal inoculation of 6-week-old piglets with 2 ml of PRV N5 B strain（10^6/0.1 ml） led to a mortality rate of 100%. [Conclusion] In this study,genetic variability of g E gene in PRV N5 B isolate and its pathogenicity to piglets were analyzed, which provided a theoretical basis for the development of new vaccines to prevent and control porcine pseudorabies.

Study on Pathogenicity Difference of Plasmodiophora brassicae Under Different Temperature and pH Conditions

[Objective] This study was conducted to investigate the pathogenicity of Plasmodiophora brassicae on cabbage grown under different temperature and soil pH conditions. [Method] The pathogenicity of P. brassicae were tested at seven different temperatures and at six different soil pH values with the resting spore concentration of lx108 （spores/g） in the soil. The plant survival rate and incidence rate of clubroot were investigated after 90 d. [Result] The incidence rate of clubroot on cabbage among the different temperature sets varied in a descending order as follows： 30 ℃〉25 ℃〉20 ℃〉35 ℃〉15 ℃〉10 ℃〉5 ℃ at soil pH value of 6, indicating that the pathogenicity of P. brassicae was weak at 5 and 10 ~（3. The incidence rate increased with soil temperature increasing from 15 to 30 ℃, but decreased at 35 ℃. The incidence rates of clubroot were 80.36%, 100%, 65%, 10.77%, 3.23% and 0% at soil pH 4, 5, 6, 7, 8 and 9 at 25 ℃, respectively. The growth of cabbage was inhibited and the survival rate was reduced at pH 4.The incidence rates of clubroot were low at pH value of 7 and 8, and was 0% at pH 9. The Chinese cabbage grew better at pH value of 5 and 6, but had high incidence rates of clubroot. [Conclusion] The results revealed that the incidence rate of clubroot on cabbage was closely related to the temperature and soil pH.

Study on Population Genetic Structure and Pathogenicity of Ustilaginoidea virens from Anhui Province

In order to determine population genetic structure and pathogenicity of Ustilaginoidea virens in the major rice-growing areas of Anhui Province, total 92 U. virens strains were collected from 28 rice-planting counties （cities） of Anhui Province. Their genetic diversity was analyzed by using REP-PCR （repetitive extragenic palindromic sequence PCR）, and pathogenicity was determined with artificial inoculation method. The results showed that U. virens in rice-growing regions of Anhui Province had a rich genetic diversity. At the similarity level of 0.76, the 92 U. virens strains could be classified into 7 groups. Significant differences were found in pathogenicity among the 24 U. virens strains belonging to different groups, which showed no association with territorial source of U. virens strain or cluster method adopted by this study. Strain pathogenicity and rice varieties showed significant specificity.

Biological Characteristics and Pathogenicity of Verticillium lecanii Isolated from Naturally Died Boettcherisca peregrine

[Objective] In order to find the pathogenic microorganisms suitable for biological control of filth flies, the pathogenic microorganism was isolated from the dead fly, Boettcherisca peregrine. [Method] The conidia and mycelia were observed by optical microscope. The pathogenic microorganism was identified on the basis of its culture characters and the optical morphologies of the conidia and mycelia, and its biological characteristics and pathogenicity were preliminarily studied. [Result] The pathogenic microorganism isolated from the dead fly was a new strain of Verticillium lecanii. The new strain of V. lecanii was numbered as KMZW-1. The colonies of V. lecanii KMZW-1 grew fastest on potato dextrose agar medium （PDA） at 29 ℃ and pH 6.0. The LC50 of its spore suspension to the adults of B. peregrine, Lucilia sericata, Musca domestic, Piophila casei and Drosophila melanogaster were 9.50×10^5, 4.58×10^7, 4.06×10^7, 4.10×10^3 and 1.05×10^7 conidia/ml, respectively. The LT50 were 6.86, 8.17, 8.16, 8.12 and 3.22 d, respectively. [Conclusion] V. lecanii KMZW-1 is an active pathogenic microorganism to control the adults of five fly species.

Research on the Pathogen of Rice Sheath Blight(Rhizoctonia solani Kühn)and Its Pathogenicity in Sichuan Regions

[ Objective] The paper was to study the cultural characteristic of the pathogen of Rice Sheath Blight （ Rhizoctonia solani Kuhn） and its pathogenicity in Sichuan regions. [ Method] The samples of rice sheath blight collected from six main rice planting areas in Sichuan regions were separated. The separated pathogen of rice sheath blight was cultured on PDA medium, and its cultural characteristic was recorded. Meanwhile, the pathogenicity of the obtained 23 strains was determined. [ Result] The growth rates among different strains had significant difference. According to the growth rate, only one strain belonged to medium strain（colony diameter： 40 mm≤（Ф≤60 mm）, the rest were all slow-type strains （colony diam- eter Ф〈40 mm）, and there was no strain with fast growth rate （colony diameter Ф 〉 60 mm）. Pathogenicity test showed that the pathogenicity among strains was significantly different, only one strain had strong pathogenicity, and the others all had moderate or weak pathogenicity. [ Conclu- sion] The study confirmed the basic biological characteristics of the pathogen of rice sheath blight in Sichuan region, which would provide theoretical basis for effective control of rice sheath blight in the region.

Relationship between Biological Characteristics of Beauveria bassiana (Bals.) Vuill and Pathogenicity to Bombyx mori L.

[Objective] This study was to investigate the relationship between biological characteristics of Beauveria bassiana （Bals.） Vuill and pathogenicity to Bombyx rnori L, with the aim to provide scientific basis for the control of white muscardine in Bombyx mori L. [Method] The strains isolated and purified from the 6 Beauveria bassiana biocontrol agents from all over the country and the 3 white muscardine silkworms collected from Guangxi provincial silkworm rearing areas were identified by the morphological observation and molecular biology technology. The pathogenicity of B. bassaina to silkworms was determined, and the biological characteristics such as growth diameter, sporulation and the extracellular protease activity of the different B. bassiana strains were compared. [Result] The isolated 9 strains were all B. bassaina （Bals.） Vuillemin, and all strains had high pathogenicity to silkworm, but with different pathogenicities. The growth diameter, sporulation and extracellular protease activity of different B. bassiana strains were also different, and showed correlation with the patheogenicity to silkworms. [Conclusion] B. bassiana spores production amount and exocellular protease activity had significant positive correlation with their pathogenicity to silkworm.

hrpZ_(Psg12) Gene of Pseudomonas syringae pv.glycinea can Enhance Pathogenicity of the Pathogen on Soybean and Cause the Hypersensitive Response of Tobacco

[ Objective ] The paper was to confirrm the effect of hrpZpsg12 gene on the pathogenicity of Pseudomonas syringae pv. glycinea. [ Method ] hrpZpsg12 gene was cloned from P. syringae using PCR method. The knockout plasmid pKNOCK-Cm with suicide characteristics and cosmid pUFR034 with complementation func- tion were used to construct the mutation vector pKNOCK477-7 and complementary vector pUFR1026-68 of hrpZpsg12 gene, the mutant 477-1 and the functional com- plementation unit 1026-5 of the gene was also screened out. Three strains including wild-type Psg12, mutant 477-1 and complementary unit 1026-5 were simultane- ously inoculated into soybean leaves and tobacco leaves, then pathogenicity determination and hypersensitive reaction analysis were carried out. [ Result] All the inoculated leaves of soybean and tobacco produced reaction lesion. However, the sizes of reaction lesion were different. The lesion in the leaves inoculated with Psgl2 was relatively large, while the lesion in the leaves inoculated with 477-1 was relatively small; the lesion of complementary unit 1026-5 was similar to wild- type Psgl2. Analysis of reproduction quantity of bacteria in lesions showed that the reproduction quantity of wild-type Psg12 was the highest, while that of mutant 477-1 was the lowest. The reproduction quantity of complementary unit 1026-5 was similar to that of wild-type Psg12. [ Conclusion] hrpZpsg12 gene could enhance the pathogenicity of P. syrimgae on Soybean and produce hypersensitive response in tobacco.

Isolation and Identification of Pandora neoaphidis and Its Pathogenicity against Turnip Aphid(Lipaphis erysimi)

[Objective] The study was to isolate and identify the pathogen of Pandora neoaphidis and to determine its pathogenicity against turnip aphid （Lipaphis erysimi）.[Method] Based on the morphological characteristics,the species of pathogenic fungus was identified.Spore shower method was used to carry out bioassay on the pathogen against turnip aphid.[Result] Primary conidia were ovoid,bitunicate and uninucleate,（24.7±1.4）μm×（10.7±0.9）μm,L/D=2.3±0.2.Secondary conidia had the similar shape with the primary ones,（18.6±2.1）μm×（13.3±1.3）μm,L/D=1.4±0.2.Hyphal body was like mycelium with the diameter of （10.6±0.8）μm.Conidiophores had palmate branch with the diameter of （10.0±0.9）μm.Pseudocystidia was not branched,which had rough base with the diameter of （19.2±1.7）μm,and gradually became more angular towards the apex with the diameter of （8.0±0.9）μm at tips.Rhizoid was like monohyphal shape with the diameter of （21.0±3.0）μm at base,the terminal apex had regular discoid holdfast.No resting spores were observed.The lethal dose of the pathogen against turnip aphid was 18.2/mm2.[Conclusion] The entomopathogenic fungus against turnip aphid was identified to be Pandora neoaphidis,and the pathogen was confirmed to have strong pathogenicity against turnip aphid.

Biological Concept of Bacterial Pathogenicity (Theoretical Review)

Biological nature of the bacterial pathogenicity phenomenon is based on the interaction of prokaryotic and eukaryotic organisms. The phenomenon is the poly-functional biological potency of germs that are realized by factors (determinants) of pathogenicity. Some fundamental biological functions are responsible for bacterial pathogenicity in a multi-cellular host organism: the adhesive function, the function of invasion and penetration into the cell, the function of evasion of host defense, and the damage function. The action of adhesion, invasion and evasionis directed to towards establishing an ecological niche in multi-cellular host while the aim of the damaging function is destruction of the environment.

MAP kinase gene STK1 is required for hyphal, conidial, and appressorial development, toxin biosynthesis, pathogenicity, and hypertonic stress response in the plant pathogenic fungus Setosphaeria turcica

The mitogen-activated protein kinase （MAPK）, a key signal transduction component in the MAPK cascade pathway, regulates a variety of physiological activities in eukaryotes. However, little is known of the role MAPK plays in phytopathogenic fungi. In this research, we cloned the MAPK gene STK1 from the northern corn leaf blight pathogen Setosphaeria turcica and found that the gene shared high homology with the high osmolality glycerol （HOG） MAPK gene HOG1 of Saccharomy- ces cerevisiae. In addition, gene knockout technology was employed to investigate the function of STKI. Gene knockout mutants （KOs） were found to have altered hyphae morphology and no conidiogenesis, though they did show similar radial growth rate compared to the wild-type strain （WT）. Furthermore, microscope observations indicated that STK1 KOs did not form normal appressoria at 48 h post-inoculation on a hydrophobic surface. STK1 KOs had reduced virulence, a significantly altered Helminthosporium turcicum （HT）-toxin composition, and diminished pathogenicity on the leaves of susceptible inbred corn OH43. Mycelium morphology appeared to be significantly swollen and the radial growth rates of STK1 KOs declined in comparison with WT under high osmotic stress. These results suggested that STK1 affects the hyphae development, conidiogenesis, and pathogenicity of S. turcica by regulating appressorium development and HT-toxin biosynthesis. Moreover, the gene appears to be involved in the hypertonic stress response in S. turcica.