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  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Preliminary functional analysis of T106 gene specifically induced by Meloidogyne enterolobii in root-knot in tomato plants
    YAN Xirui, GAO Zewen, DONG Ying, WU Wentao, ZENG Yuanling, DUAN Shanquan, WANG Yang
    Acta Phytopathologica Sinica. 2024, 54(5): 950-960. https://doi.org/10.13926/j.cnki.apps.001629
    Abstract (22) PDF (12)   Knowledge map   Save
    Meloidogyne enterolobii, which is highly pathogenic to a wide range of host plants and spreads rapidly, can cause devastating damage to many crops. To deeply analyze the pathogenic mechanism of this nematode, here we take T106, a gene specifically induced in tomato roots in response to M. enterolobii infection based on previous transcriptome data, as our target. We silenced T106 in tomato plants via TRV virus-induced gene silencing technology, and then inoculated tomato seedlings with M. enterolobii to observe the difference in nematode and giant cell development in root system between T106-silenced and T106-unsilenced plants. The results showed that the silencing vector we constructed could effectively silence T106 gene in tomato plants, with a silencing efficiency of 85%; compared with T106-unsilenced control plants, there was no significant decrease in the percentage of root knots in T106-silenced plants, but the development of M. enterolobii in root knots was inhibited, and the number of eggs produced by M. enterolobii was reduced by 79.3%; meanwhile, the area occupied by giant cells was also decreased. In summary, T106 might be a susceptible gene targeted by M. enterolobii. Exploration of such susceptible genes in plants is vital for finding new ways to control root-knot nematodes including M. enterolobii.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Functions of HMG-box family genes MfHMG5 and MfHMG6 on growth, sporulation and pathogenicity in Monilinia fructicola
    YANG Jingya, ZENG Zhezheng, XIAO Yuanling, CAI Minzheng, WU Peishan, WEI Wenkai, YIN Weixiao, LUO Chaoxi
    Acta Phytopathologica Sinica. 2024, 54(5): 961-973. https://doi.org/10.13926/j.cnki.apps.000919
    Abstract (15) PDF (10)   Knowledge map   Save
    Brown rot, caused by Monilinia spp., is a serious threat to both stone fruit and pome fruit, greatly affecting the long-distance transportation and exportation of fruits. Based on genomic and transcriptomic analysis of infection of Monilinia fructicola on peach fruit, it was detected that the expression patterns of MfHMG5 and MfHMG6 genes in early stages of infection were similar, and both down-regulated significantly at 1 h after inoculation and then gradually increased. In order to investigate the biological functions of these two genes, the knockout and overexpression transformants of MfHMG5, and knockout and complemented transformants of MfHMG6 were obtained and the corresponding phenotypes were investigated. It was found that the knockout and overexpression of MfHMG5 gene decreased the growth rate and sporulation ability, but did not affect the pathogenicity of M. fructicola. Knock out of MfHMG6 gene reduced the growth rate, virulence and sporulation ability of M. fructicola, and led to the increased expression of MfHMG5 gene. These results indicated that HMG-box family genes MfHMG5 and MfHMG6 were involved in regulating the growth and pathogenesis of M. fructicola.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Identification and functional analysis of dynamin-like GTPase protein FpSey1 in Fusarium pseudograminearum
    WANG Limin, KANG Jiangang, LI Haiyang, CHEN Linlin, XING Xiaoping, DING Shengli, LI Honglian
    Acta Phytopathologica Sinica. 2024, 54(5): 974-984. https://doi.org/10.13926/j.cnki.apps.001635
    Abstract (27) PDF (22)   Knowledge map   Save
    The occurrence of Fusarium crown rot (FCR) caused by Fusarium pseudograminearum has been becoming increasingly serious in China, which has posed a severe threat to wheat yield and quality. The SEY1 belongs to the RHD3 (Root Hair Defective 3) family and encodes a dynamin-like GTPase protein participating in endoplasmic reticulum (ER) fusion. The ER is involved in the synthesis of deoxynivalenol (DON) in different pathogenic fungi, while its function in F. pseudograminearum has not been reported. In this study, subcellular localization of GFP-tagged Sey1 (FpSey1) protein in F. pseudograminearum was observed, and the results showed that FpSey1 was localized in the ER. The FpSEY1 deletion mutant (ΔFpSey1) was generated through PEG-mediated protoplast transformation and verified by Southern blot analysis, and complemented strains were obtained as well. Compared with the wild-type strain, the ΔFpSey1 mutant exhibited significant reduction in vegetative growth, conidiation, relative expression of DON biosynthesis related genes (TRI1, TRI5, TRI10) , and the virulence on wheat coleoptiles and barley leaves. In addition, the ΔFpSey1 mutant is more sensitive to salt stress, hydrogen peroxide (H2O2), but more tolerant to dithiothreitol (DTT) than the wild-type and complemented strains. These results indicate that FpSey1 localized in the ER plays important roles in the growth and infection of F. pseudograminearum.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Preliminary study on the phoq-mediated pathogenicity of Xanthomonas albilineans causing sugarcane leaf scald disease
    CHEN Wenhan, LI Meilin, WU Deng, DU Jinxia, XIONG Liya, HU Qin, ZHANG Muqing
    Acta Phytopathologica Sinica. 2024, 54(5): 985-994. https://doi.org/10.13926/j.cnki.apps.001633
    Abstract (20) PDF (14)   Knowledge map   Save
    Leaf scald, caused by Xanthomonas albilineans (Xa), is a bacterial disease that seriously affects sugarcane production. Understanding the pathogenicity of this bacterial pathogen is crucial to preventing and controlling sugarcane leaf scald disease. Previous studies have shown that Phoq, a transmembrane histidine protein kinase in the two-component system of Xanthomonas, is a very important transduction factor, sensing extracellular signals, activating intracellular kinase activity, and subsequently regulating downstream gene expression. In this study, we collected Phoq protein sequences from eight pathogenic bacteria in the genus Xanthomonas, including Xa. The results of phylogenetic analysis and motif composition prediction showed that these Phoq proteins have conserved structure and similar physicochemical properties. To further investigate the biological role of phoq in Xa, we produced phoq knockout mutant in Xa-JG43 strain using the homologous recombination method. Compared with the wild-type strain Xa-JG43, the swimming ability and pathogenicity of the Xa-phoq knockout mutant were seriously weakened, but the swarming and stress response ability were not affected; In Xa-phoq complementary strain, the pathogenicity and swimming ability were restored to the level of the wild-type strain. This study provides a theoretical basis for further determination of the pathogenic mechanism of Xa.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Rapid and specific identification of Fusarium pseudograminearum by qPCR based on RPB sequence
    HE Yanqiu, JIANG Qi, CHI Yuankai, WANG Tao, QI Rende, ZHAO Wei
    Acta Phytopathologica Sinica. 2024, 54(5): 995-1007. https://doi.org/10.13926/j.cnki.apps.001630
    Abstract (24) PDF (11)   Knowledge map   Save
    Fusarium crown rot, mainly caused by Fusarium pseudograminearum, is a destructive disease in wheat production. To establish a rapid and reliable detection method for F. peasudeograminearum, the specific PCR primer pair (Fpg-F1/R2) was designed based on the RPB sequence, and real-time fluorescence quantitative PCR (qPCR) was used to validate the efficiency of the primer. The results showed that the primer pair had high specificity and sensitivity of 100 pg of DNA. Furthermore, the qPCR system for early and rapid detection of F. peasudeograminearum had an amplification efficiency of 87.5% and correlation coefficient of 0.99, and the pathologic threshold of F. pseudograminearum in soil was determined by using this detection system. It was found that F. pseudograminearum could cause Fusarium crown rot when the DNA concentration of F. pseudograminearum in field soil exceeded 213 pg·g-1. Hence, the qPCR-based method we developed for F. pseudograminearum detection has the advantages of high specificity and sensitivity, and can be used for rapid and early detection of F. pseudograminearum even in field soils.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Functional study of Botrytis cinerea secreted protein BcSGP1
    SONG Xuantong, NIE Xiaofei, WANG Ziyao, BI Kai, ZHU Wenjun
    Acta Phytopathologica Sinica. 2024, 54(4): 726-737. https://doi.org/10.13926/j.cnki.apps.001040
    Abstract (68) PDF (57)   Knowledge map   Save
    Gray mold disease caused by Botrytis cinerea leads to severe crop yield reduction, and the secreted proteins play significant roles in the fungal infection. However, the functional mechanisms of these secreted proteins in B. cinerea remain largely unknown. In this study, a secreted protein, BcSGP1, from the secretome of B. cinerea during infection stages was identified. The expression level of BcSGP1 was upregulated during infection stages. Deletion of BcSGP1 caused reduction in pathogenicity, but not in growth rate, conidial production, or stress resistance. Transient expression of BcSGP1 in Nicotiana benthamiana leaves using agroinfiltration induced necrosis, and this necrosis-inducing activity depended on the plant receptor-like kinase BAK1, but not the SOBIR1. Furthermore, BcSGP1 could induce resistance against B. cinerea in N. benthamiana leaves. These results suggest that BcSGP1 is a pathogenesis-related secreted protein and involved in inducing plant resistance during the interaction between B. cinerea and plants. This study enhances our understanding of the pathogenic mechanisms of B. cinerea, providing a theoretical basis and genetic resources for effective control of gray mold disease.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Exploring key pathogenic genes in Fusarium graminearum based on genetic diver-sity and selection elimination analysis
    ZHEN Jinpeng, LIU Jianhu, CAO Hongzhe, XING Jihong, DONG Jingao, SHI Chengmin, ZHANG Kang
    Acta Phytopathologica Sinica. 2024, 54(4): 738-745. https://doi.org/10.13926/j.cnki.apps.001624
    Abstract (81) PDF (44)   Knowledge map   Save
    In recent years, wheat scab, corn stalk rot and ear rot caused by Fusarium graminearum have led to substantial losses in crop yields. To investigate the genetic diversity and identify pathogenicity-related genes of F. graminearum, we performed population genetic diversity analysis and selective elimination analysis on 93 F. graminearum strains with released genome-wide resequencing data, using single nucleotide polymorphism (SNP) technology. The resequencing data of these F. graminearum strains were meticulously processed by using the Genome Analysis Toolkit 4 (GATK4), yielding a collection of 3,817,652 SNP markers. Based on these markers, a phylogenetic tree was constructed, and principal component analysis (PCA) and population structure analysis were conducted, effectively partitioning the 93 F. graminearum into 3 distinct groups. The selection elimination analysis of group 1 and group 2 revealed that group 1 exhibited a more pronounced response to selection pressure. A total of 70 regions were identified as candidate sites within the top 5% intersection region of population polymorphism (θπ) and population differentiation index (Fst). Furthermore, 76 protein-coding genes were identified in F. graminearum by leveraging the genomic location information. The results of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that these 76 candidate genes are mainly involved in metabolic pathways. Among them, eight genes (FGSG_05447, FGSG_05610, FGSG_10272, FGSG_10313, FGSG_01353, FGSG_05545, FGSG_10858 and FGSG_12745) are closely related to the pathogenicity of F. graminearum through further gene expression analysis. The result lays a basis for clarifying the pathogenic mechanism of F. graminearum and breeding F. graminearum-resistant wheat and maize varieties.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Jasmonic acid biosynthetic genes positively regulate age-related resistance of tomato plants to Phytophthora infestans
    JIANG Yuxia, DAI Lili, XUE Xiaojing, YU Yue, ZHANG Jianli, DING Xinhua, ZHONG Weiran, CHU Zhaohui, LIU Haifeng
    Acta Phytopathologica Sinica. 2024, 54(4): 746-755. https://doi.org/10.13926/j.cnki.apps.001628
    Abstract (87) PDF (65)   Knowledge map   Save
    Late blight caused by Phytophthora infestans seriously affects the yield and quality of tomato. Previous research found that tomato plants contain age-related resistance (ARR) to P. infestans, but the underlying mechanisms are still unclear. Here, we used tomato variety ‘Micro Tom’ as the tested material and found that younger (4-week-old) plants are more resistant while older (8-week-old) plants are more susceptible to late blight. Through RNA sequencing and real-time quantitative PCR (qPCR) analysis, we observed that the transcription levels of genes involved in jasmonic acid (JA) synthesis, such as AOS1, AOS2 and AOC, are higher in 4-week-old plants than those in 8-week-old plants. We further examined the levels of several phytohormones and found that the concentration of JA in 4-week-old plants is significantly higher than that in 8-week-old plants. Transient expression of AOS1, AOS2 or AOC in tobacco leaves made them more resistant to late blight, suggesting that these JA biosynthetic genes can enhance tomato resistance to late blight. Tomato plants sprayed with MeJA were more resistant whereas tomato plants sprayed with JA synthesis inhibitor DIECA were more susceptible to late blight, suggesting that JA positively regulates tomato resistance to late blight. Thus, we provide evidence supporting a model in which genes involved in JA synthesis play important roles in the age-related resistance to late blight in tomato. Our results lay an important basis for using ARR to control tomato late blight.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Identification and characterization of PilZ domain-containing proteins in pathogenicity and motility of Pseudomonas syringae pv.actinidiae
    MA Jiabing, YANG Mingming, ZHAO Yingjia, TAN Yunxiao, CHANG Bangshuai, WANG Nana, HUANG Lili
    Acta Phytopathologica Sinica. 2024, 54(4): 756-768. https://doi.org/10.13926/j.cnki.apps.000882
    Abstract (61) PDF (28)   Knowledge map   Save
    PilZ domain-containing proteins are the largest known receptors of second messenger c-di-GMP in bacteria, but the functions and underlying mechanisms have not been reported in Pseudomonas syringae pv. actinidiae (Psa). To reveal the contribution and regulation mechanism of PilZ domain-containing proteins to the pathogenicity of Psa and to provide new ideas for controlling kiwifruit bacterial canker. Firstly, genome analysis and sequence alignment of Psa M228 were performed to identify the PilZ domain-containing proteins of Psa and analyze the conserved c-di-GMP binding motif. Then, homologous recombination was used to construct deletion mutants, and the pathogenicity, motility and growth between mutants and wild type were determined by leaf discs vacuum infiltration, soft agar plate assays and growth curve measurment, respectively. The transcripts of pathogenicity- and motility-relative genes in WT and ΔPsa_2195 mutant were measured by qRT-PCR. The results show that there are eight PilZ domain-containing proteins in Psa M228, among them PSA_2195 and PSA_1975 have neither a conserved RxxxR motif nor a (D/N)xSxxG binding motif that binds to c-di-GMP. The pathogenicity of ΔPsa_1116, ΔPsa_2195, ΔPsa_2203, ΔPsa_762, ΔPsa_4490 and ΔPsa_4763 were significantly reduced. Deletion of PSA_1116 and PSA_2195, PSA_3989, PSA_762 affected swimming motility and swarming motility, respectively. The growth curve of all mutants are no significant difference with wild type M228. Among all PilZ domain proteins, PSA_2195 regulates the transcription of flagella genes flgA, filE and T3SS genes. Taking together, our research revealed the function of eight PilZ domain-containing proteins in regulating pathogenicity and motility of Psa and the simple molecular mechanisms of PSA_2195.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    An atypical chemotactic receptor gene of Xanthomonas oryzae pv. oryzae is associated with bacterial swimming and early infection
    LEI Shimin, OUYANG Yanfei, WU Qiaozhi, XU Lidan, ZENG Chen, WU Huihong, XIE Yunqiao, HE Yongqiang
    Acta Phytopathologica Sinica. 2024, 54(3): 522-534. https://doi.org/10.13926/j.cnki.apps.001617
    Abstract (85) PDF (37)   Knowledge map   Save
    After annotating the genome of Xanthomonas oryzae pv. oryzae strain PXO99A-GX, we found an atypical chemoreceptor gene PXO_01024, which encodes a protein with two transmembrane domains (TMD) and a methyl-accepting domain (MA) but without the ligand-binding domain (LBD). To understand the biological function of PXO_01024, we constructed the PXO_01024 deletion mutant DM01024 and its complemented strain CDM01024 by homologous double exchange method. Deletion of PXO_01024 resulted in the reduced formation of biofilm and almost loss of bacterial swimming, while in CDM01024 the swimming motility and biofilm formation ability were restored to wild-type levels. The virulence of DM01024 was not significantly different from that of the wild-type strain when inoculation of wounded host plants was performed. However, when unwounded plants were inoculated with the spraying inoculation method, the disease index caused by DM01024 was significantly reduced compared with that caused by the wild-type strain and complemented strain CDM01024, indicating that PXO_01024 played a role in early infection of Xoo. The chemotaxis of these strains was subsequently detected by capillary method, and the results showed that DM01024 showed significantly reduced chemotaxis to methionine, alanine, leucine, glycine, asparagine, phenylalanine, isoleucine, glucose, maltose, xylose, fructose, succinic acid, and tartaric acid compared with the wild-type strain. This study demonstrated that the atypical chemotactic receptor gene PXO_01024 is associated with chemotaxis, swimming motility, and early infection of Xoo.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Functional analysis of a single-domain response regulator in Xanthomonas citri subsp. citri
    LI Ruifang, LIU Qianqian, YAO Ziting , XIAN Xiaoyong, LU Guangtao, ZHU Gui-ning
    Acta Phytopathologica Sinica. 2024, 54(3): 535-543. https://doi.org/10.13926/j.cnki.apps.001619
    Abstract (77) PDF (42)   Knowledge map   Save
    Citrus bacterial canker caused by Xanthomonas citri subsp. citri (Xcci) is an important bacterial disea-se of citrus plants. Our previous work revealed that the homologous of XAC3126 (Accession no. AAM37971.1) of Xcci strain 306, which was predicted to be a single-domain response regulator, might be involved in the bacterial virulence in Xcci Guangxi wild-type strain N8. To investigate the biological functions of this gene (named embR), a deletion mutant was constructed with strain N8 as the starting strain. Simultaneously, a complemented strain was constructed using a recombinant plasmid harboring this innate gene. Phenotypic analysis revealed that the embR deletion mutant ΔembR showed obviously reduced virulence on the host plant Citrus reticulata Blanco 'Orah' compared with the wild-type strain, while the complemented strain exhibited similar virulence with that of the wild-type strain. Additionally, the ΔembR mutant displayed an obvious reduction in extracellular polysaccharide (EPS) production, cell motility and cell aggregation. Reverse transcription quantitative real-time PCR (RT-qPCR) revealed that the transcript level of a set of genes involved in EPS production and cell motility in the ΔembR mutant was decreased compared with that in the wild-type strain. These combined data indicate that the embR gene is required for multiple cellular processes including virulence in Xcci and modulates the expression of a series of virulence factor-related genes.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Codon optimization of CI protein coding sequence of potato virus Y and establishment of eukaryotic expression system and preparation of monoclonal antibody
    YIN Guoying, JIA Mengao, CHANG Yongchun, ZHANG Pan, LI Zhen, CAI Changchun
    Acta Phytopathologica Sinica. 2024, 54(3): 544-554. https://doi.org/10.13926/j.cnki.apps.001335
    Abstract (79) PDF (28)   Knowledge map   Save
    The nucleic acid sequence encoding the CI protein of potato virus Y (PVY) had multiple regions similar to prokaryotic promoter elements. It was possible to translate toxic proteins in prokaryotic cells, so it was difficult to construct vectors. According to the codon bias of prokaryotes, the CI sequence was modified without changing the amino acid sequence. The expression vector containing CI open reading frame (ORF) was successfully constructed, and CI protein was successfully expressed through the eukaryotic cell-free protein expression system. Six cell lines were prepared by immunizing Balb/c mice with purified protein using hybridoma technology. Indirect ELISA and western blot showed that the prepared CI monoclonal antibody 4B7_2D6(IgG1) had high sensitivity and specificity. The successful expression of CI protein provides a prerequisite for the purification of CI protein and the subsequent study of the structure and function of CI protein, which is of great significance for further exploring the interaction mechanism between CI and plant proteins.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Preparation and application of antiserum to soybean stay-green associated virus coat protein
    ZHANG Yuyang, LI Qinglun, YU Lianwei, XIE Lina, JIANG Xingling, WANG He, LI Honglian, SHI Yan, YANG Xue, YUAN Hongxia
    Acta Phytopathologica Sinica. 2024, 54(3): 555-560. https://doi.org/10.13926/j.cnki.apps.001332
    Abstract (65) PDF (29)   Knowledge map   Save
    Soybean stay-green associated virus (SoSGV) is a newly discovered plant virus that can cause soybean stay-green. Soybean stay-green is widespread in the soybean producing area of China and seriously threatens the development of soybean industry. Therefore, it is of great importance to establish a rapid and effective SoSGV detection method for the monitoring and control of soybean stay-green. In this study, SoSGV CP primers were designed according to the nucleotide sequence of the SoSGV coat protein (CP) gene, CP gene of 786 bp length was amplified from infected soybean, and cloned into prokaryotic expression vector pET-28a to obtain the recombinant plasmid pET-28a-CP. The recombinant vector pET-28a-CP was transformed into Escherichia coli strain Rosseta. After induction with IPTG, a 30 kDa protein was obtained, which was consistent with the molecular weight of SoSGV CP. SoSGV CP antiserum was prepared by immunizing mice with recombinant SoSGV CP protein. The In-ELISA test results showed that the serum titer of the SoSGV CP antiserum was ≥3.2×104. Western blot analysis showed that the SoSGV CP antiserum could specifically recognize the SoSGV CP protein, which was 30 kDa, in the SoSGV-infected tobacco and soybean plants. These results indicate that the SoSGV CP antiserum provides favorable conditions for rapid detection of soybean stay-green in soybeans-infected by SoSGV.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Cloning and functional analysis of the Nicotiana benthamiana NbMybl gene related to late blight resistance
    ZHAO Di, CHEN Shengnan, ZHANG Die, WANG Hongyang, CHEN Aie
    Acta Phytopathologica Sinica. 2024, 54(2): 318-331. https://doi.org/10.13926/j.cnki.apps.001607
    Abstract (100) PDF (74)   Knowledge map   Save
    Myb transcription factors play important roles in the regulation of various biological processes in plants. However, the molecular mechanism underlying their roles in regulating late blight resistance remains elusive. Here, we report the cloning of NbMybl, a Myb-like gene from Nicotiana benthamiana, which has an open reading frame of 753 bp and encodes a protein of 250 aa. NbMybl contains a Myb-like DNA-binding domain. Real-time quantitative PCR (qPCR) revealed that NbMybl was induced by infection with Phytophthora infestans. Subcellular localization analysis showed that NbMybl is located in both the nucleus and the cytoplasm. Silencing of NbMybl by virus-induced gene silencing (VIGS) significantly increased the susceptibility of plants to P. infestans. Transcriptome profiling by RNA sequencing identified 8468 differentially expressed genes (DEGs) with fold change ≥ 2 and FDR < 0.01 between NbMybl silenced and non-silenced control lines in response to P. infestans infection, and the result of RNA-seq was further validated by qPCR with 10 randomly selected DEGs. KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis revealed that a total of 373 DEGs were involved in plant-pathogen interaction, 308 DEGs and 216 DEGs were involved in MAPK signaling pathway and plant hormone signal transduction, respectively. We speculated that these DEGs might be closely related to the reduced resistance of NbMybl-silenced N. benthamiana lines to P. infestans. Our study provides valuable insights into the molecular mechanisms of NbMybl in regulating resistance to P. infestans.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Biological function of the caspase FgCas4 in Fusarium graminearum
    WANG Xiaoyan, CONG Weiwei, CHEN Li, ZHANG Chengqi
    Acta Phytopathologica Sinica. 2024, 54(2): 332-342. https://doi.org/10.13926/j.cnki.apps.000876
    Abstract (74) PDF (53)   Knowledge map   Save
    Caspase is known to be involved in modulating apoptosis in mammals,controlling the occurrence and development of various diseases. In present study, the null mutant ΔFgCas4 and complemented strain ΔFgCas4-C of the caspase gene FgCas4 were obtained by gene disruption and complementation respectively in Fusarium graminearum. We observed that the FgCas4 deletion mutant (ΔFgCas4) did not affect the growth rate, colony morphology, conidiation, virulence and DON production. However, the deletion mutant ΔFgCas4 exhibited more hyphal branching and percentage of conidia with 3 septa increased by 8.7% compared to wild type PH-1 and complemented strain ΔFgCas4-C. External environmental stress assays showed that the gene disrupt mutant ΔFgCas4 became more sensitive to tested fungicides and metal ions. In addition, the lack of FgCas4 led to dramatically increased lipid droplet biosynthesis as well as increased resistance to osmotic stress agents. Subcellular localization showed that the caspase FgCas4 localized in vacuoles. On the other, the loss of FgCas4 resulted in earlier process of autophagy. Taken together, our study provides evidences that the caspase FgCas4 of Fusarium graminearum plays important roles in asexual reproduction, various environmental stress responses and auto-phagy regulation.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Transcriptome analysis of coronatine-induced resistance to Ralstonia solanacearum in tomato plants
    LI Na, ZOU Lifang, XIN Yidong, SUN Sheng, GENG Xueqing
    Acta Phytopathologica Sinica. 2024, 54(2): 343-354. https://doi.org/10.13926/j.cnki.apps.001611
    Abstract (97) PDF (35)   Knowledge map   Save
    The bacterial wilt caused by Ralstonia solanacearum has brought a serious threat to tomato production. Coronatine (COR), structurally similar to JA-Ile (JA-isoleucine), is a compound produced by Pseudomonas syringae. In this study, we observed that pre-treatment of tomato seedlings with COR alleviated the symptoms caused by R. solanacearum. To investigate how COR affects tomato resistance to R. solanacearum, transcriptome sequencing of tomato seedlings inoculated with the pathogen 24 h after COR treatment was carried out. Analysis of RNA-seq data showed that COR treatment induced a total of 2122 differentially expressed genes (DEGs), including 998 up-regulated genes and 1124 down-regulated genes. DEGs annotation and pathway enrichment were conducted using GO database and KEGG database, and the results showed that COR affected the expression of genes related to plant-pathogen interaction pathway and plant hormone signaling pathways. Meanwhile, COR induced the up-regulation of genes involved in the jasmonic acid synthesis pathway and inhibited the expression of photosynthesis-related genes. Our results provide a theoretical basis for revealing the role of COR in plant-microbe interaction.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Histopathological assessment on the lotus rhizome tissue infected by Fusarium spp. causing lotus rhizome rot
    TENG Hui, YANG Jishuo, LI Siyu, ZHOU Dongmei, WANG Nan, SUN ShuYan, LIU Huaqi, WEI Lihui, WANG Yunpeng, DENG Sheng
    Acta Phytopathologica Sinica. 2024, 54(1): 82-91. https://doi.org/10.13926/j.cnki.apps.001023
    Abstract (171) PDF (139)   Knowledge map   Save
    Lotus is the most important aquatic vegetable in China and has the largest cultivation area. With the expansion of cultivation and the simplification of agricultural practices, various lotus diseases have been becoming more serious, among which lotus rhizome rot disease is the most harmful and the most difficult to control. At present, there are few research reports on the pathogenesis of Fusarium spp. causing lotus rhizome rot disease around the world. From the microscopic and ultra-microscopic levels, this study revealed the changes to the rhizome and its vascular tissues during Fusarium infection. The observation of paraffin-embedded tissue sections showed that with the development of the disease, necrotic cells stained by toluidine blue appeared around the vascular bundles, and the closer to the bundles, the more numerous necrotic cells and the more densely distributed. In order to further reveal the influence of pathogen infection on vascular bundles, brightfield optical and scanning electron microscopy were used to observe the lotus fibers in the diseased lotus rhizomes. The results showed that the smooth and compact lotus fibers became rough and dispersed with the disease tending to be more severe, and a large number of galls-like protrusions and other possible metabolic attachments were attached to the surface. These changes to the vascular bundles are likely to reduce the water and nutrient transportation, and finally the aboveground parts of the plant exhibited the symptoms of water loss and wilting. The results of this study revealed the cellular biological mechanism of the disease symptom development, and provided a basis of histology for the breeding of resistant varieties in the future.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Cytopathological changes of different tobacco varieties infected with tomato mottle mosaic virus
    LI Yueyue, ZHANG Shuiying, ZHOU Wenpeng, ZHANG Tianyi, LAN Pingxiu, TAN Guanlin, LI Fan
    Acta Phytopathologica Sinica. 2024, 54(1): 92-101. https://doi.org/10.13926/j.cnki.apps.001324
    Abstract (127) PDF (117)   Knowledge map   Save
    Tomato mottle mosaic virus (ToMMV), a member of the genus Tobamovirus, showed stronger pathogenicity than tobacco mosaic virus (TMV) and tomato mosaic virus (ToMV) in most solanaceae crops. To figure out the cytopathological changes of tobacco plants that were infected by ToMMV, we constructed a full-length infectious cDNA clone of ToMMV, and carried out transmission electron microscopy observation. The results showed that the rod-shaped virions of ToMMV were 220-310 nm in length and 16-18 nm in diameter. ToMMV virions were also observed in the seeds of ToMMV-infected Nicotiana tabacum var. Xanthi nc. ToMMV virions were dispersed or angularly-layered in cytoplasm or vacuoles. Multivesicular bodies, small vesicles, myeloid bodies, mitochondrial cristae were observed to be reduced or disappeared in both N. tabacum var. Xanthi nc and N. rustica infected with ToMMV. In addition, broken chloroplast membranes and large amount of small peripheral vesicles protruding into the disintegrated chloroplasts were observed in the infected N. tabacum var. Xanthi nc showing mosaic and distortion symptoms, while large amount of small vesicles were observed in the cells of infected N. tabacum var. Xanthi nc showing leaf narrowing. However, chloroplasts changes were less frequent in N. rustica with systemic necrosis symptoms. In this study, the construction of ToMMV infectious clone and the observation of cytopathological changes in tobacco plants infected with ToMMV, provided important bases for the further study of the pathogenesis of ToMMV and its interaction with the host.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Genome-wide prediction and pathogenic analysis of non-classical secreted proteins of Lasiodiplodia theobromae
    XING Qikai, WANG Xinfang, PENG Junbo, ZHANG Wei, YAN Jiye, LI Yonghua
    Acta Phytopathologica Sinica. 2024, 54(1): 102-115. https://doi.org/10.13926/j.cnki.apps.001625
    Abstract (177) PDF (169)   Knowledge map   Save
    Lasiodiplodia theobromae is an important plant pathogenic fungus, infecting more than 500 species of woody plants and causing serious damages worldwide. The secreted proteins of pathogenic fungi play important roles in pathogen invasion, expansion, and colonization in plants. Although there are more reports on the involvement of classical secreted proteins in fungal pathogenicity, the roles of non-classical secreted proteins in the pathogenesis of plant pathogenic fungi have rarely been investigated. In this study, 238 candidate non-classical secreted protein-coding genes were obtained by bioinformatic prediction based on the whole genome sequence of L. theobromae. Gene functional prediction and GO enrichment analysis showed that these genes are enriched in the carbon-sulfur lyase activity pathway. The annotation results based on PHI-base showed that 15 non-classical protein-coding genes might be related to the pathogenicity of L. theobromae. Predication of subcellular location using ApoplastP showed that 4 of them were located in plant apoplast and 11 were located in plant proplastids. Reverse transcription quantitative PCR (RT-qPCR) analysis results further verified that the transcriptional expression levels of the above candidate non-classical secreted protein-coding genes were significantly changed after treatment with grapevine branch tissues, indicating that these non-classical secreted proteins may play important roles in grapevine-L. theobromae interaction. The results lay a foundation for further understanding of the pathogenic mechanism of non-classical secreted proteins of L. theobromae.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Screening and identification of effector candidates in the late stage of interaction between Alternaria solani and potato
    JIANG Jia, TIAN Mengjun, NIE Tingbin, YANG Zhihui, ZHU Jiehua
    Acta Phytopathologica Sinica. 2024, 54(1): 116-126. https://doi.org/10.13926/j.cnki.apps.001631
    Abstract (151) PDF (167)   Knowledge map   Save
    The potato early blight caused by Alternaria solani is an important disease in potato production, which seriously affects the yield and quality of potatoes. To reveal the interaction mechanism between A. solani and its host plant potato, we screened the effector candidates produced by A. solani during its interaction with potato plant using RNA transcriptome sequencing technology and biological software (signalp-4.1, tmhmm-2.0c, phobius101 and ProtComp v3) combined with characteristics analysis of effector proteins. As a result, 137 differentially expressed candidate proteins were obtained, and four (Gene03396, Gene06670, Gene02858 and Gene06508)of them were selected for further analysis. Based on the result of RNA-seq analysis, Gene03396, Gene06670 and Gene02858 had an up-regulated expression pattern at three time points (3, 4 and 5 dpi) compared with that at 0 dpi, whereas Gene06508 had a downregulated expression pattern. The four proteins do not have a common motif according to the MEME analysis result. Transient overexpression of the target effector candidate genes in tobacco showed that Gene03396 and Gene02858 directly induced tobacco cell necrosis, and Gene06670 inhibited programmed cell death (PCD) induced by BCL2-associated X, indicating that effector candidates function by stimulating plant immunity or inhibiting host defense response. The subcellular localization results revealed that Gene03396 was located on the cell membrane, while Gene06670 and Gene02858 may be located in the intercellular space. These results provide a basis for further identifying the effector proteins of A. solani and revealing the underlying pathogenic mechanism of the pathogen.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Microscopic and ultrastructural characteristics of the root of Potentilla anserine infected by Fusarium avenaceum
    LI Junqiao, LI Chenqin
    Acta Phytopathologica Sinica. 2023, 53(5): 841-851. https://doi.org/10.13926/j.cnki.apps.000839
    Abstract (116) PDF (146)   Knowledge map   Save
    This study aimed to comparatively investigate the morphology of the diseased fern tubers caused by pathogenic fungus Fusarium avenaceum with the healthy root tuber of Potentilla anserina L. as a control. Paraffin section, ultrathin section, and micrograph methods were used to study the anatomical structure and surface ultrastructure of the diseased root. The explored invasion and colonization rules of this fungus at the cell and subcellular level would provide a theoretical ground to understand the cellular mechanism of Potentilla anserine root rot. The results showed that the fungal infection could significantly affect the morphology of Potentilla anserinaroots. The fungus invaded through the natural pores and mechanical wounds of the root tube and destroyed the epidermis and internal structure from the outside to the inside. The root tuber appeared disease spotson the root surface and putrefactied later, and the internal cells were deformed. In addition, the infection of pathogenic fungus was selective and the fungal mycelia with varied morphologies were mainly concentrated in the phloem parenchyma cells of the tuberous roots of Potentilla anserine. The infection gradually deepened from the outside to the inside, and the infected cells appeared plasmolysis and decreased contents, which affected the appearance and quality of Potentilla anserine root tuber.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    The role of Ralstonia solanacearum type III effector RipAF1 in the induction of host PTI response
    WU Wei, ZOU Huasong
    Acta Phytopathologica Sinica. 2023, 53(5): 852-862. https://doi.org/10.13926/j.cnki.apps.000844
    Abstract (120) PDF (242)   Knowledge map   Save
    Ralstonia solanacearum injects a repertoire of effectors into host cells to modulate plant immune response via a type III secretion system. Our work reports the role of type III effector RipAF1 from GMI1000 in the induction of host immune response. Transient overexpression of RipAF1 in Nicotiana benthamiana resulted in the enhanced expression levels of PTI marker genes NbAcre31, NbPti5 and NbGras2. Simultaneously, a large amount of callose was accumulated. In a yeast two-hybrid experiment, a RipAF1-interacting C-terminal of BAK1 was screened from a cDNA library of N. benthamiana. Subsequent split-luciferase and bimolecular fluorescence complementation experiments showed that RipAF1 was interacted with NbBAK1 in vivo. A deletion mutant ΔripAF1 was constructed in GMI1000, which showed no difference from wild type in growth in both nutrient rich and minimal media. However, the mutant showed increased virulence on tomato plants, as well as the replication in planta. These results demonstrated that RipAF1 is an effector able to induce plant PTI and exerts a negative role for virulence.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Prokaryotic expression and antiserum preparation of CP protein of strawberry mild yellow edge virus
    LU Jing, HAN Jincheng, SUN Yixin, JIANG Lei, JIANG Tong
    Acta Phytopathologica Sinica. 2023, 53(5): 863-869. https://doi.org/10.13926/j.cnki.apps.001308
    Abstract (96) PDF (283)   Knowledge map   Save
    The coat protein (CP) gene of the Chinese isolate of strawberry mild yellow edge virus (SMYEV) was amplified by PCR. The gene was cloned into the prokaryotic expression vector pET-32a. The recombinant plasmid pET-cp was transformed into Escherichia coli strain BL21, and then incubated at 37 °C with 1 mmol·L-1 IPTG to express the target protein. The result showed that fusion protein CP was present abundantly in the cell lysate supernatant. The fusion protein CP was purified by Ni2+-NTA affinity resin, and western blot exhibited a single specific band with an approximate molecular weight of 46 kDa. The purified fusion protein CP was used to immunize rabbits and obtain polyclonal antibody. Indirect ELISA detection showed that the titer of antiserum to fusion protein reached 1: 512 000. Dot-ELISA and colloidal gold strip assays were established with the polyclonal antibody. These methods can be used not only for early diagnosis of SMYEV-infected strawberry samples, but also for the analyses of transient local and systemic expression of SMYEV CP protein.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Determination of critical amino acids in potato virus Y coat protein involved in the formation of vein necrosis symptom in Nicotiana tabacum cv. Xanthi plants
    JI Liyun, ZHAO Pingxiang, YAN Zhiyong, GENG Chao, TIAN Yanping, LI Xiangdong, WANG Fenglong
    Acta Phytopathologica Sinica. 2023, 53(5): 870-880. https://doi.org/10.13926/j.cnki.apps.001304
    Abstract (109) PDF (133)   Knowledge map   Save
    Potato virus Y (PVY)can infect many crops including potato and tobacco, and cause huge economic losses. PVY-encoded coat protein (CP) interacts with helper component proteinase (HC-Pro) to regulate the aphid transmission of PVY. However, its role in the formation of PVY vein necrosis symptoms is unclear. Here, we analyzed the symptoms of 84 PVY CP mutants on Nicotiana tabacum cv. Xanthi plants and found two mutants, PVY-71 and PVY-82, which have lost their ability to induce vein necrosis in N. tabacum cv. Xanthi plants. Site-directed mutagenesis was carried out to identify the critical amino acids in CP in the formation of vein necrosis on N. tabacum cv. Xanthi plants. Two mutants, namely pCamPVY-CPD6N and pCamPVY-CPG9R, in which the codons for the sixth amino acid, Aspartic acid (D6), and the ninth amino acid, Glycine (G9),of CP were mutated to those for asparagine (N) and arginine (R), respectively, were obtained. Both mutants can infect N. tabacum cv. Xanthi plants systemically and have similar virus accumulation levels to wild-type PVYN605. However, they could not induce vein necrosis in N. tabacum cv. Xanthi plants. Luciferase complementation assay showed that the interaction strength between mutant CP and HC-Pro was also reduced. It is the first report that PVY CP is one of the vein necrosis determinants on N. tabacum cv. Xanthi plants, and the conserved amino acids D6 and G9 played an essential role during this process. These findings will help us to understand the role of CP in the PVY vein necrosis symptom formation in N. tabacum cv. Xanthi plants and the role of CP in PVY infection.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Virions distribution and sub-cellular pathological characteristics at different stages of Hymenocallis littoralis infected with Hippeastrum chlorotic ringspot orthotospovirus
    ZHANG Han, WANG Maosen, WANG Tiantian, ZHU Ruomeng, TONG Jinjin, LI Yu
    Acta Phytopathologica Sinica. 2023, 53(4): 563-569. https://doi.org/10.13926/j.cnki.apps.000652
    Abstract (201) PDF (606)   Knowledge map   Save
    In this study, leaf tissues of Hymenocallis littoralis (Jacq.) Salisb. suspected to be infected with Hippeastrum chlorotic ringspot orthotospovirus (HCRV) were collected at different stages of the disease. The infection of HCRV was confirmed by RT-PCR amplification, cloning and sequencing analysis of HCRV-N gene-specific primers. Further, negative staining, ultra-thin section sample preparation and transmission electron microscopy (TEM) analyses were performed. The results showed that HCRV virions at different stages (symptomless or with symptoms of chlorosis, yellowing and necrosis) showed the characteristics of dispersion or aggregation. The dispersed virions with a double-membrane structure, and virions in the late stage of diseased cells showed a beaded shape and gathered in tubular vesicles. During the incubation period of virus infection, the nucleus, chloroplast and other subcellular structures remained intact. At the early and middle stages of the diseased cells, the nucleus structure was intact, the chloroplast grana lamella dissolved, and the number of mitochondria increased exhibiting an intact structure. At the late stage of the infection, most of the cells were necrotic, and there were residual endosomes in the vacuoles, and the vacuoles of a few cells contained beaded virions clustered in tubular vesicles, suggesting that the characteristics of HCRV were obviously different from viruses of the same genus and could be used as the basis for diagnosis and identification.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Preliminary study of the TaRLK3D.2 function in the interaction between wheat and Puccinia striiformis f. sp. tritici
    WANG Congnawei, CUI Zhijun, LI Xiang, WEI Jiajing, ZHANG Jiazhen, YI Ke, YUAN Fengping, KANG Zhensheng, HAN Dejun, ZENG Qingdong
    Acta Phytopathologica Sinica. 2023, 53(4): 570-579. https://doi.org/10.13926/j.cnki.apps.000827
    Abstract (302) PDF (727)   Knowledge map   Save
    Wheat stripe rust is an airborne fungal disease that occurs in all the wheat-growing areas worldwide and seriously threatens the safety of wheat production. Deploying the resistance cultivar is the most effective method to control this disease. Leucine-rich repeat receptor-like protein kinase (LRR-RLK), as the pattern recognition receptor of the largest subfamily of RLK, plays an essential role in preventing pathogen invasion. The present study systematically identified the wheat LRR-RLK family members and screened 43 significantly differentially expressed LRR RLKs from the transcriptome data in the competable manner of wheat vs Pst. Finally, TaRLK3D.2, which was significantly up-regulated both in compatible and incompatible interaction, was selected for functional analysis in the interaction processing through the real-time qRT-PCR, the subcelluar localization of its encoding protein, and barley stripe mosic virus-mediated silencing technology. Our results showed that the TaRLK3D.2 was located in the plant cell membrane. Based on the transiently silencing induced by barley stripe mosaic virus, TaRLK3.2 knocked down plant inhibited the growth of Pst resulting the significantly reduced disease severity, indicating that TaRLK3D.2 encoding a typical LRR-RLK and might act as a susceptibility gene in the process of wheat stripe rust fungous infection. Our preliminary study explored the function of TaRLK3D.2 gene that could be as a potential target used in resistance breeding of the durable stripe rust resistant material created through stable gene editing or mutation.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Identification and functional analysis of cupin domain-containing protein Vmcupin1 in Valsa mali
    ZHANG Qiong, LIU Zhaoyang, GAO Chengyu, DU Xuan, FENG Hao, HUANG Lili
    Acta Phytopathologica Sinica. 2023, 53(4): 580-588. https://doi.org/10.13926/j.cnki.apps.000653
    Abstract (261) PDF (512)   Knowledge map   Save

    Apple tree valsa canker, caused by Valsa mali, is a serious branch disease threatening the healthy development of the apple industry in China. Revealing its pathogenic mechanism has important theoretical significance for formulating new strategies for disease control. Cupin domain-containing proteins belong to a large protein family, and are involved in many biological processes in plants, such as development and stress-related responses. There are few studies on this kind of protein in plant pathogenic fungi, and its biological function in mycelial growth and infection is still unclear. Based on the transcriptome analysis between V. mali and apple twig bark tissues, a candidate gene was significantly up-regulated during pathogen infection. The gene encoded a cupin domain-containing protein, which was highly homologous with cupin domain-containing proteins from other organisms, and it was named Vmcupin1. Gene expression analysis by qRT-PCR revealed that Vmcupin1 was significantly up-regulated during pathogen infection. Furthermore, Vmcupin1 deletion mutants and complement transformants were created, respectively. It was found that the growth rate of mutants was slightly reduced while the pathogenicity and ability to adapt to H2O2 and NaCl stress were significantly reduced, indicating that Vmcupin1 plays an important role in the vegetative growth, pathogenicity and stress response of V. mali. These results lay an important foundation for the comprehensive analysis of pathogenic mechanism of V. mali and the enrichment of functional cognition of fungal cupin domain-containing proteins.

  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Genome-wide identification and expression analysis of TGA family genes in birch
    FENG Siyu, BI Delong, SUN Jingjing, NING Xiaomeng, LI Ranhong
    Acta Phytopathologica Sinica. 2023, 53(4): 589-600. https://doi.org/10.13926/j.cnki.apps.001008
    Abstract (197) PDF (497)   Knowledge map   Save
    TGA transcription factor, a member of the basic region-leucine zippers (bZIP) transcription factor family, is one of the earliest identified transcription factors in plants. It can regulate the transcription level of target genes through binding to the TGACG sequence of the gene promoters, and plays an important role in plant defense responses and floral organ development. In this study, six TGA genes were identified from the genome database of Betula platyphylla, named BpTGA1-BpTGA6. Bioinformatics analysis showed that the birch TGA family proteins were mainly rich in acidic amino acids, subcellular localization in the nucleus, and no signal peptide structure; the secondary structures of birch TGA protein were mainly α-helix, and all of the birch TGA family genes were hydrophilic proteins which have no transmembrane structure. There were 10~40 enhanced promoter elements (CAAT-box) and core promoter elements (TATA-box) in the promoter region of birch TGA family genes. Also, these regions have abiotic stress and biotic stress response-related elements, indicating that members of this family may be involved in environmental responses. The results of phylogenetic analysis indicated that the birch TGA family genes were closely related to those of Arabidopsis thaliana. The results of real-time fluorescence quantitative PCR (RT-qPCR) suggested that the expression level of birch TGA family genes in the roots, stems and leaves were different; The expression level of BpTGA3 was higher in roots, and BpTGA1, BpTGA2, BpTGA4, and BpTGA6 were higher in stems, whereas BpTGA5 was highly expressed in leaves. Among the members of TGA family genes, BpTGA2, BpTGA4, and BpTGA5 responded to the infection of Alternaria alternata and Rhizoctonia solani. Among them, BpTGA5 responded most strongly to both pathogens, and the expression reached the highest level at 24 h and 48 h after infection, which were 17.45 times and 328.76 times of the control, respectively. This study lays a foundation for further exploring the role of birch TGA transcription factor in birch.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Functional analysis of the polyketide synthase gene BmPKS18 in Bipolaris maydis
    LIU Qingqiang, SUN Bingjian, XING Xiaoping, DING Shengli, LI Haiyang, YUAN Hongxia
    Acta Phytopathologica Sinica. 2023, 53(4): 601-612. https://doi.org/10.13926/j.cnki.apps.000833
    Abstract (158) PDF (490)   Knowledge map   Save
    Southern corn leaf blight (SCLB) caused by Bipolaris maydis is a serious disease on corn, which affects serious loss of the yield and the quality of corn. Polyketide synthase plays an important role in the growth, development and pathogenesis of plant pathogens by regulating the synthesis of various toxins and melanins. It is important to elucidate the roles of polyketide synthase genes in B. maydis, which can provide insights for SCLB disease control. In this study, BmPKS18 encoding polyketide synthase was knocked out, and then the complementary strain was obtained. Compared with the wild-type and complementary strain, the ΔBmPKS18 mutant showed albino phenotype, significantly reduction in mycelium growth, spore production and germination rate, increasing conidia length with no pigmentation, defect in sexual reproduction and significantly reduction of pathogenicity to corn. These results demonstrated that BmPKS18 plays an important role in regulating vegetative growth, sexual and asexual reproduction, and pathogenicity of B. maydis. These results provide important basis for further elucidation the pathogenic mechanism of B. maydis.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Molecular cloning of WRKY15 transcription factor gene in Fagopyrum tataricum and analysis of its expression induced by leaf spot pathogens and hormones
    ZENG Lipei, PENG Xixu, WU Qingtao, TIAN Jianhong, TANG Xinke, WANG Haihua
    Acta Phytopathologica Sinica. 2023, 53(4): 613-623. https://doi.org/10.13926/j.cnki.apps.000824
    Abstract (144) PDF (518)   Knowledge map   Save
    Tartary buckwheat (Fagopyrum tataricum) leaf spot is caused by Nigrospora osmanthi and Alternaria alternata. WRKY transcription factors play an important regulatory role in plant immunity. Expression analysis of F. tataricum WRKY15 (FtWRKY15) induced by leaf spot fungi and hormones will provide a foundation for insights into functions and underlying mechanisms of FtWRKY15 in plant responses to pathogens. In this study, the entire coding sequence (CDS) of FtWRKY15 was cloned using reverse transcription PCR. The FtWRKY15 CDS is 537 bp in length and encodes a polypeptide of 178 amino acids. FtWRKY15 had a WRKY conserved domain, belonging toWRKY group IIc subgroup with zinc finger type CCHH. Homologous sequence alignment and phylogenetic analysis revealed that FtWRKY15 shared the highest identity at amino acid level with FtWRKY76 (64.0%), followed by Chenopodium quinoa WRKY50 (52.3%). Transient expression assay in protoplasts showed that FtWRKY15 was localized in nucleus, consistent with the prediction of subcellular localization. Yeast one-hybrid assay revealed that FtWRKY15 had transcription-activating activity. The acidic region (30th~60th amino acid residue) in FtWRKY15 N-terminus was rich in putative serine, threonine, and tyrosine phosphorylation sites, possibly responsible for the transcription-activating activity. Real-time fluorescence quantitative PCR analysis showed that the transcript abundance of FtWRKY15 was relatively higher in leaves and flowers, next was in stems and roots, whereas almost no expression in fruits. The expression of FtWRKY15 was significantly induced byleaf spot pathogens, N. osmanthi and A. alternata, and defense-related phytohormones, such as salicy-lic acid (SA), jasmonic acid (JA) and ethylene (ET). Taken together,FtWRKY15 possesses basic structural and biochemical characteristics as a putative transcription factor, andmay be involved in defense response to Tartary buckwheat leaf spot.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Sequence analysis of eukaryotic translation initiation factor eIF4E from two varieties of sugarcane with differential resistance to sugarcane streak mosaic virus
    SHAN Hongli, CHEN Du, WANG Xiaoyan, ZHANG Rongyue, LI Jie, WANG Changmi, LI Yinhu, HUANG Yingkun
    Acta Phytopathologica Sinica. 2023, 53(4): 624-632. https://doi.org/10.13926/j.cnki.apps.000826
    Abstract (163) PDF (621)   Knowledge map   Save
    Sugarcane streak mosaic virus (SCSMV) is the main pathogen causing sugarcane mosaic disease in recent years. Breeding and using resistant varieties is the most economical and effective means to control sugarcane mosaic disease. Different varieties have different resistance to sugarcane mosaic disease. It is important to identify the source of resistance difference for breeding resistant varieties. In this study, the sequences of pathogenic viruses and eukaryotic translation initiation factor 4E (eIF4E) were analyzed in the highly resistant variety CP94-1100 and the highly susceptible variety ROC22. The results showed that SCSMV isolated from CP94-1100 and ROC22 were similar in the nucleotide sequences and amino acid sequences for cp gene and vpg gene, and were the same variant. The eIF4E gene was obtained from CP94-1100 and ROC22, which contained a 663 bp ORF encoding a polypeptide with 220 amino acid residues. The amino acid sequence homology of the eIF4Es from these two varieties was 98.2%, indicating a possible correlation between the eIF4E polymorphism and difference in resistance to SCSMV. Bioinformatics analysis showed that the IF4E family proteins encoded by the eIF4E genes in CP94-1100 and ROC22 were stable hydrophilic proteins. In addition, this IF4E family protein encoded by the eIF4E gene in CP94-1100 also contains a CDC33 or PTZ00040 domain. Genetic analysis showed that these eIF4E genes in CP94-1100 and ROC22 were highly similar to other eIF4E family genes from grami-neae, and the eIF4E gene of CP94-1100 was most closely related to that of Saccharum spontaneum, suggesting that they had similar functions.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Cytological study on the spots of rice leaf smut infected by Entyloma oryzae
    ZHANG Xiaomin, XIE Li, WANG Weilan, LAI Chaohui, LIANG Wusheng, LIN Fucheng, HU Dongwei
    Acta Phytopathologica Sinica. 2023, 53(3): 395-400. https://doi.org/10.13926/j.cnki.apps.001009
    Abstract (207) PDF (344)   Knowledge map   Save
    Rice leaf smut is a fungal disease caused by a basidiomycete, Entyloma oryzae, which infects rice leaves and leaf sheaths forming black strip spot. However, the cytological mechanism of the interaction between pathogen and host is still poorly understood. Here the preliminary cytological analysis was carried out on diseased leaves collected from the field. The results showed that the morphology of host epidermal cells on the spots remained basically intact after pathogen infection. The pathogen produced a large number of chlamydospores in the position of host mesophyll cells, and gradually replaced all the mesophyll cells. The hyphae of the pathogen expanded outside the host cell and did not penetrate into the host cell walls, nor did not produce typical fungal haustorium. The organelles in the host cells close to the hyphae of the pathogen were degraded, and the lipids produced by degradation condensed into large lipid spheres. The cell wall of the host vascular bundle tissue remained intact, and no pathogenic hypha was found to invade the vascular bundle. The pathogenic mycelia and spores were confined between two adjacent vascular bundles of the host. In the late stage of the infection, a large number of bacteria entered due to the structural integrity destruction of host leaf epidermis, which accelerated the senescence and death of host leaves. The results of this study showed that the pathogen infection mode is extracellular, similar to obligate parasitic fungi. On the other hand, pathogen infection leads to the degradation and death of nearby host cells as saprophytic fungi.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    The apoplastic effector Ps140300 inhibits innate immunity activated by XEG1
    WANG Lan, LIU Hanmei, ZHANG Mingmei, ZHU Jinyi, WANG Yan, ZHENG Xiaobo, WANG Yuanchao
    Acta Phytopathologica Sinica. 2023, 53(3): 401-411. https://doi.org/10.13926/j.cnki.apps.000799
    Abstract (221) PDF (213)   Knowledge map   Save
    The apoplastic effectors play important roles in promoting microbial colonization by interfering with plant immune responses. XEG1, an apoplastic effector secreted by Phytophthora sojae, belongs to the glycoside hydrolase 12 family. XEG1 is recognized by the pattern recognition receptor RXEG1, triggering immune response and cell death in plants. In this study, we found that the XEG1 homologous protein Ps140300 could suppress XEG1 induced-cell death. Ps140300 was a virulence factor of P. sojae and up-regulated at the early infection stage. Unlike XEG1, Ps140300 did not trigger cell death in plants. Ps140300 could inhibit XEG1-induced cell death and reactive oxygen species burst. Further studies found that Ps140300 did not affect RXEG1 interaction with XEG1, or the signaling transduction sectors BAK1 and SOBIR1. Ps140300 was conserved in oomycetes and fungi and homologues can also suppress XEG1-triggered cell death. These results suggest that the apoplastic effectors can promote microbial infection by inhibiting host basal immunity.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Functional analysis of autophagy related gene ChAtg26 in Colletotrichum higginsianum
    DUAN Lingtao, ZENG Miaolin, WANG Li, ZHU Yiming, HE Jiuqing, ZHOU Erxun
    Acta Phytopathologica Sinica. 2023, 53(3): 412-423. https://doi.org/10.13926/j.cnki.apps.001002
    Abstract (207) PDF (191)   Knowledge map   Save
    CSCD(1)
    Colletotrichum higginsianum is an important plant pathogenic fungus that can cause severe cruciferous vegetable anthracnose, which has a great impact on vegetable production. In order to explore the function of the autophagy-related gene ChAtg26 in the pathogenesis of C. higginsianum, the cDNA of Arabidopsis thaliana Col-0 infected by C. higginsianum was used as a template in this study, and the expression pattern of the gene ChAtg26 during the infection process was determined by qRT-PCR. To characterize the function of ChAtg26, the gene knockout and mutant complementation were performed through homologous recombination technology, and the effects of the gene deletion on the growth, development and pathogenicity of C. higginsianum was evaluated. The result of expression pattern showed that the gene ChAtg26 reached the maximum expression level after 40 h of infection. When the deletion of this gene, no significantly effects were observed on the mycelial growth rate, conidium germination, appressorium formation, and sensitivity to oxidative stress, but the mutant showed the reduction of melanin accumulation in the colony, the sensitivity to cell wall stress, and the significantly decreased sporulation and pathogenicity. Taken together, our results suggest that ChAtg26 in C. higginsianum was involved in the melanin synthesis, cell wall stress response, conidia production and pathogenesis.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Inhibitory effect of fifteen small molecular compounds on the type Ⅲ secretion system of Xanthomonas oryzae pv. oryzicola
    LIU Jie, ZHU Kaixuan, WANG Guohua, ZHANG Linlin, YAN Xi, LIANG Xiangdong, HANG Xiaohong, JIANG Bole, YANG Mei
    Acta Phytopathologica Sinica. 2023, 53(3): 424-435. https://doi.org/10.13926/j.cnki.apps.000643
    Abstract (219) PDF (176)   Knowledge map   Save
    Type III secretion system (T3SS) is the virulence determinant of Xanthomonas oryzae pv. oryzicola (Xoc). A series of small molecule inhibitors of type III secretion system of Xanthomonas campestris pv. campestris (Xcc) have been identified in our laboratory before. Whether these inhibitors have the effect on T3SS of Xoc is worth investigating. In this study, the fusion reporter systems were constructed by fusing the luciferase gene (Lux) and the GUS gene with the promoter regions of T3SS-related genes respectively in Xoc. The small molecules inhibiting T3SS of Xoc were identified with the reporter system and quantitative PCR (qPCR). The results showed that Carmofur, 5-Fluorocytosine, A-3, and Dealkaline lignin also inhibit the expression of Xoc T3SS. Among them, A-3 and Dealkaline inhibit the growth of bacteria in XOM3. However, none of the four compounds could significantly reduce the disease symptoms caused by Xoc. In order to determine the quantitative hypersensitive response (HR) triggered by Xoc, and to check the biocontrol effects of the above inhibitors, the reporter strain GX01/avrBs1 was constructed by fusion PCR and homologous double exchange principle. The results indicated that GX01/avrBs1 could rapidly trigger HR on pepper ECW-10R, and Dealkaline lignin could significantly attenuate HR induction of Xoc on pepper ECW-10R, further indicating that Dealkaline lignin inhi-bits Xoc T3SS.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Functional study of effector protein AopK of Acidovorax citrulli
    LIU Jing, ZHAO Liping, GUAN Wei, YANG Yuwen, BAI Qingrong, ZHAO Tingchang
    Acta Phytopathologica Sinica. 2023, 53(3): 436-446. https://doi.org/10.13926/j.cnki.apps.000644
    Abstract (194) PDF (185)   Knowledge map   Save
    CSCD(1)
    The type Ⅲ secreted effectors (T3Es) play important roles in the pathogenesis of Acidovorax citrulli, the causal agent of bacterial fruit blotch in watermelon and melon plants. XopK is a T3E with E3 ubiquitin ligase activity in Xanthomonas spp. The homolog of XopK, designated as AopK, in A. citrulli has not been investigated. To explore the function of AopK, the aopK deletion mutant and complementary strain were constructed in A. citrulli wild-type strain AAC00-1. The pathogenicity assay showed that the deletion of the gene aopK significantly reduces the virulence of A. citrulli on watermelon seedlings. Subcellular localization analysis of AopK was performed using Nicotiana benthamiana. Results revealed that AopK was delivered to the cytoplasmic membrane of N. benthamiana. The transcriptional expression level, secretory and BAX co-expression analyses showed that the expression of aopK was positively regulated by HrpX and HrpG. AopK was secreted into extracellular matrix through the Type Ⅲ secretion system. AopK can inhibit the production of reactive oxygen species, callose deposition and cell necrosis in N. benthamiana, and affects the virulence via inhibiting plant defense responses. In summary, our data indicated that AopK is a T3E in A. citrulli. Our data laid a foundation for further exploring the mechanism of interaction between A. citrulli and host plants.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Utilization of GFP tagged Verticillium dahliae strain to evaluate dynamic infection on resistant/susceptible potato varieties
    KANG Liru, GAO Jing, JIA Ruifang, LIU Yan, FAN Junchen, ZHANG Zhiwei, ZHAO Jun
    Acta Phytopathologica Sinica. 2023, 53(2): 164-172. https://doi.org/10.13926/j.cnki.apps.001004
    Abstract (307) PDF (362)   Knowledge map   Save
    In order to study the systemic infection of Verticillium dahliae in potato, we firstly established a Petri dish filter paper system by which the evaluation of dynamic roots infection in resistant/susceptible potato varieties was conducted with the GFP fluorescence labelled Verticillium dahliae strain. The results showed that after soaking for 2 hours in a conidia suspension of 107conidia·mL-1, conidia could attach to the root hairs of resistant/susceptible potato roots, but a large amount of conidia was attached to the roots of susceptible varieties in which 65 conidia were on susceptible cultivar, while 23 conidia on resistant cultivar at average view of microscope. After 12 hours of inoculation, there was no significant difference in the germination rate of conidia on the roots surface of both resistant and susceptible varieties. Three days after inoculation, the mycelium could penetrate the root epidermis and enter the intercellular space of tissues. However, compared with the resistant varieties, the mycelium in the root system of the susceptible varieties grew faster and reached the inner vascular bundle of the root system at 7 dpi, and the green fluorescence signal was observed in the vascular bundle of resistant varieties only at 9 dpi. At 10 dpi, new conidia stalk and conidia formation could be seen in the roots of susceptible varieties, while no conidia formation in the resistant varieties. The amount of pathogen colonization in the aerial stems of susceptible potato varieties at 30 dpi was five times more than that of the resistant varieties. At the same time, the number of pathogens in the petioles of susceptible varieties was also significantly higher than that of resistant varieties.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Functional characterization of a strain-specific alcohol dehydrogenase in Verticillium dahliae JR2
    JIN Xianjiang, ZHANG Jiayi, LIU Tingli, WANG Yonglin
    Acta Phytopathologica Sinica. 2023, 53(2): 173-182. https://doi.org/10.13926/j.cnki.apps.000642
    Abstract (244) PDF (285)   Knowledge map   Save
    Verticillium dahliae is a worldwide phytopathogenic fungus which can infect more than 660 plant species, causing serious economic losses annually. Genomic analysis of V. dahlia revealed that different strains contained many strain-specific genes, however, the biological functions of most genes were not clear. In our study, by comparing the genomes of V. dahliae VdLs17 and JR2, a JR2 strain-specific gene Chr6g02370 was identified, which encodes alcohol dehydrogenase and was induced during infection. The enzymatic activity of ethanol dehydrogenase in the knockout mutant decreased significantly, but the utilization of carbon sources such as ethanol was not affected. In addition, the virulence of knockout mutants to tobacco and tomato was significantly reduced. The phenotypic analysis also showed that knockout of this gene reduced melanin synthesis ability, but did not affect mycelial growth and sporulation. In conclusion, our results expand the understanding of the biological functions of strain-specific genes in V. dahlia and provide genetic material for further functional genomics.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Functional characterization of wheat CAMTA gene family in the interaction between wheat and stripe rust fungus
    ZHANG Yanqin, ZENG Peng, GUO Shuangyuan, GAN Pengfei, WANG Xiaojie, KANG Zhensheng, XU Quanle, ZHANG Xinmei
    Acta Phytopathologica Sinica. 2023, 53(2): 183-194. https://doi.org/10.13926/j.cnki.apps.000789
    Abstract (305) PDF (288)   Knowledge map   Save
    Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), seriously harms wheat production in China. Calmodulin-binding transcription activator (CAMTA) is the most vital one regulated by Ca2+/CaM in plants playing a significant role to biological stress. In this study, four wheat CAMTA gene family members down-regulated after Pst treatment were screened out from transcriptome database of the wheat-Pst interaction, and their functions in the interaction between wheat and stripe rust were preliminarily analyzed following real-time quantitative PCR (qRT-PCR) and virus-mediated gene silencing (VIGS) technologies. The results showed that in the compatible interaction, TaCAMTA1-A, TaCAMTA1-B, and TaCAMTA3-D were significantly down-regulated at different time points, while TaCAMTA2-B was obviously up-regulated at 12 h. VIGS-based knockdown of TaCAMTA2-B in wheat leaves was significantly reduced the number of necrotic spots and spores on the leaves, indicating that the disease resistance was enhanced. Therefore, the wheat TaCAMTA2-B gene was cloned, and tobacco subcellular localization assay found that TaCAMTA2-B was localized in the nucleus. In addition, self-activation experiment in the yeast indicated that TaCAMTA2-B has transcriptional activation activity. The exogenous hormone SA, ABA, JA treatments showed that TaCAMTA2-B was up-regulated when ABA treatment, and expression of TaCAMTA2-B was detected in wheat roots, stems and leaves, especially in the roots with the highest level. The above results prove that TaCAMTA2-B may be a susceptibility-related gene involved in the interaction between wheat and stripe rust. Further exploring the possible susceptibility mechanism of wheat TaCAMTAs would provide a theoretical basis for the creation of broad-spectrum and durable wheat varieties with resistance to stripe rust.
  • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
    Function analysis of haloacid dehalogenase type Ⅱ gene FoHAD-typeⅡ in Fusarium oxysporum f. sp. momordicae
    ZHANG Yuanyuan, WEN Caiyi, LI Fengxin, WEI Chenxing, DU Hongyan, ZHONG Rongrong, ZHAO Ying
    Acta Phytopathologica Sinica. 2023, 53(2): 195-206. https://doi.org/10.13926/j.cnki.apps.000797
    Abstract (234) PDF (203)   Knowledge map   Save
    Fusarium wilt of bitter gourd is a soil-borne disease caused by Fusarium oxysporum f. sp. momordicae Sun & Huang (FOM), which seriously affects the production of bitter gourd. Clarifying the pathogenic mechanism of FOM has important guiding significance for the prevention and control of Fusarium wilt of bitter gourd, but the pathogenic mechanism of FOM is still unclear. In the previous analysis of the differentially expressed genes in the transcriptomes of the strong pathogenic strain SD-1 and the weak pathogenic strain SD-V (carrying mycoviruses) of FOM, it was found that the expression level of FoHAD-type II was significantly down-regulated in SD-V strain. Therefore, we speculated that this gene may be involved in the pathogenic process of FOM. In this study, based on the principle of homologous recombination, the fusion fragment of FoHAD-typeⅡ was obtained by Split-Marker PCR, the gene knockout mutant was obtained by PEG-mediated protoplast transformation, and the complementary mutant was obtained by Agrobacterium transformation method. The biological characteristics and pathogenicity of the mutants were tested and analyzed. The results showed that the growth rate and spore production of the FoHAD-type Ⅱ gene knockout mutant on the PDA medium were not significantly different from the wild-type strain, and there were no significant differences in the morphology of hyphal tips and spores. However, the vegetative growth of the knockout mutant was defective, manifested as different colony morphology, reduced aerial hyphae, significantly reduced virulence, and increased sensitivity to osmotic stress. The pathogenicity of the complemented mutant was consistent with that of the wild-type strain. Therefore, these findings suggested that FoHAD-type Ⅱ is not involved in the growth and development of FOM, but plays an important role in the pathogenic process of FOM, which may be related to the decline of pathogenicity of FOM caused by mycovirus. The results of this study provide a basis for the subsequent exploration of the pathogenic mechanism of FOM and a reference for the study of the hypovirulence mechanism of mycoviruses.
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ISSN 0412-0914
CN 11-2184/Q
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