20 December 2024, Volume 54 Issue 6
    

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    REVIEW
  • Research progress in leafy gall pathogen Rhodococcus fascians
    YANG Panpan, HAO Zehui, WANG Jiawen, XU Leifeng, MING Jun
    Acta Phytopathologica Sinica. 2024, 54(6): 1069-1082. https://doi.org/10.13926/j.cnki.apps.001036
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Rhodococcus fascians is a gram-positive actinomycete that can cause plant diseases with a very wide host range. The pathogenic strains of Rhodococcus fascians make abnormal plant tissues form, such as leaf gall, cluster, flat stem, which affects plant growth and causes serious economic losses. As the identification and classification of the genus Rhodococcus were complex and is often confused with other plant diseases with similar symptoms, resulting in few studies being carried out. This review outlines the classification status, biological characteristics, pathogenic mechanisms, disease symptoms, isolation and identification methods, transmission routes of infection and prevention and control measures of R. fascians. It is expected to provide a reference for related research and precise prevention and control of leafy gall disease caused by R. fascians.
  • Research progress of potato common scab
    HUANG Xun, LIU Xia, DENG Linmei, XU Gaihuan, YANG Yanli
    Acta Phytopathologica Sinica. 2024, 54(6): 1083-1090. https://doi.org/10.13926/j.cnki.apps.001039
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    Potato common scab is caused by Streptomyces spp. Potato common scab disease is widely distributed in potato producing areas of the world. For many years, common scab has hampered the potato industry, especially the developing of seed potatoes industry. In this paper, the occurrence of common scab, pathogen distribution, disease detection, pathogenic mechanism, cultivar resistance research and green prevention and control technology were reviewed, and the urgent problems in potato common scab research were put forward in order to provide new ideas for in-depth research and green prevention and control of potato common scab.
    • CELL BIOLOGY, PHYSIOLOGY, BIOCHEMISTRY, AND MOLECULAR BIOLOGY
  • Transcriptome analysis in potato cultivar ‘Cooperation-88’ upon infection of early blight pathogen Alternaria solani
    LI Qing, HOU Xiaoxue, ZHANG Xiangdong, WANG Rongyan, TANG Wei, LI Canhui
    Acta Phytopathologica Sinica. 2024, 54(6): 1091-1102. https://doi.org/10.13926/j.cnki.apps.001029
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    Potato Early Blight (PEB) is an important disease of the foliage of potatoes during the growing season and is widespread in all major potato producing areas worldwide. At present, there are no specific agents and potato varieties resistant to the disease completely. In this study, we identified the main cultivar ‘Cooperation-88’ (C88) in Yunnan Province was resistant to the potato early blight caused by Alternaria solani inoculated on leaves in comparison with the susceptible variety ‘Désirée’ by AUDPC. High-throughput RNA-seq in ‘Cooperation-88’ after infection at the early (A. so_e, 0-72 h), middle (A. so_m, 73-120 h) and late (A. so_l, >120 h) stages was performed on Illumina HiSeq PE150 platform. Transcriptome analysis revealed a total of 13 083 genes expressed differentially at A. so_e, of which 7 438 were up-regulated and 5 645 were down-regulated. At A. so_m, a total of 12 121 genes was differentially expressed, of which 3 299 were up-regulated and 8 822 were down-regulated. At A. so_l, a total of 10 530 genes was differentially expressed, of which 1 686 were up-regulated and 8 844 were down-regulated. A set of 2 720 identical genes was found in all the three periods, while 4 997 genes specific to A. so_e, 3 975 genes specific to A. so_m and 3 230 genes specific to A. so_l. Based on gene ontology (GO) term and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and qRT-PCR validation along with the results of electron microscopic observations, it was speculated that the increased synthesis of pectin lyase and cellulose synthetase was involved in cell wall remodeling of ‘C88’ in the early stage of A. solani infection. The expression levels of glutathione S-transferase and cytochrome P450 were significantly up-regulated in ‘C88’ at mid-infestation, which were involved in the detoxification pathway. Late synthesis of large amounts of antioxidants to stimulate defence mechanisms. In the three periods, high expression levels of transcription factors, such as bHLH, ZIP, MYB, ERF, etc, associated with disease resistance and extensive involvement in the ubiquitination pathway. Overall, our findings could provide a theoretical basis for research on the early blight resistance in potato and accelerate the resistance breeding.
  • Construction and application of a yeast two-hybrid library of rubber tree leaves infected with Oidium heveae
    WANG Jiali, LIANG Xiaoyu, KE Yuhang, XIAN Xuemei, WANG Lifeng, WANG Meng, ZHANG Yu
    Acta Phytopathologica Sinica. 2024, 54(6): 1103-1113. https://doi.org/10.13926/j.cnki.apps.001637
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    Powdery mildew of rubber tree caused by Oidium heveae is one of the major foliar diseases that affect rubber production, resulting in extensive leaf shedding and significant reduction in latex production. Breeding and planting O. heveae-resistant rubber varieties is the most economic and effective measure for the prevention and control of the disease. Identification of resistant proteins in rubber trees and investigation of the corresponding regulatory networks are therefore of great significance in unraveling the mechanism underlying the resistance response of rubber tree to powdery mildew. In this study, total RNA was extracted from mature rubber leaves (leaves sampled at 48 hpi : leaves sampled at 0 dpi=2:1) inoculated with Oidium heveae HO-1 using the Trizol method. A cDNA library of rubber tree leaves infected with powdery mildew was subsequently constructed using the Gateway technology. The primary library was recombined with pGADT7-DEST and pPR3-N-DEST secondary library vectors through LR recombination, resulting in the construction of hybrid secondary libraries for membrane system and nuclear system yeast, respectively. Both libraries have a capacity of more than 1.0×10-7 CFU, with a recombination rate of 100%. The length of inserted fragments ranges from 0.1 to 2.0 kb, with an average length of 1.5 kb. Meanwhile, bait vector containing the full-length coding region or the leucine-rich repeat (LRR) domain of the disease-resistant protein HbCNL2 was generated and used as a bait to screen for interacting proteins in the yeast library, and as a result, 7 potentially interacting proteins were obtained. Further yeast two-hybrid point-to-point validation confirmed the interaction between HbKLCR1 or HbGAIP and the LRR domain of HbCNL2. GO (Gene Ontology) annotation analysis demonstrated that HbKLCR1 has protein binding activity and HbGAIP is involved in the gibberellin signaling pathway, suggesting their potential roles in disease resistance responses of rubber trees. These results indicate that the cDNA library constructed in this study has high quality and good integrity, which provides important technical support for the identification of O. heveae-resistant rubber proteins and unraveling the protein interaction networks.
  • Exploring LRR-RLK genes involved in resistance of foxtail millet to Sclerospora graminicola based on the weighted gene co-expression network analysis
    HAN Yanqing, WU Xiaoxiong, JIANG Siming, WEI Anqi, TIAN Nana, WANG He
    Acta Phytopathologica Sinica. 2024, 54(6): 1114-1128. https://doi.org/10.13926/j.cnki.apps.001643
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Leucine-rich repeat receptor-like protein kinases (LRR-RLKs), a typical type of receptor-like kinase in plant, play important roles in response to pathogen infection. To clarify basic characteristics of the LRR-RLK family members in foxtail millet and their roles in resistance to infection by Sclerospora graminicola, members of this gene family in foxtail millet were identified, and their evolutionary pattern, sequence characteristics, gene structure, promoter sequence and expression pattern were analyzed by bioinformatics method. Meanwhile, the transcriptome data of resistant and susceptible foxtail millet varieties infected with S. graminicola were obtained at 3 different growth stages, and the co-expression modules of resistance gene and the core genes were identified via the weighted gene co-expression network analysis (WGCNA). The results showed that the LRR-RLK genes were distributed on overall 9 chromosomes of foxtail millet. A phylogenetic analysis was conducted on LRR-RLK genes from foxtail millet and Arabidopsis, and the result indicated that they were mainly divided into 4 categories. Structural analysis displayed that their kinase domains were relatively conservative. The promoter regions of these LRR-RLK genes contained multiple cis-acting elements related to defense and stress responses as well as meristem expression, indicating their involvement in multiple biological processes. A co-expression network of resistance-related genes was developed by using WGCNA. Of 44 gene co-expression modules that were identified, 3 (Turquoise, Blue and Yellow) were specific modules associated with resistance to S. graminicola, from which 12 core genes were identified. Functional annotation showed that these genes were involved in plant disease resistance. Further RT-qPCR analysis of the 6 core genes (Seita.9G413000, Seita.9G296000, Seita.9G557200, Seita.9G493600, Seita.3G241700 and Seita.9G163200) confirmed that they were induced in response to S. graminicola infection, indicating that these core genes may play important roles in resistance to the pathogen infection. The results provide a valuable reference for further revealing the molecular mechanism underlying the resistance of foxtail millet to S. graminicola.
  • Functional analysis of DHN-melanin biosynthesis related enzyme ChScd in Colletotrichum higginsianum
    DUAN Lingtao, WANG Li, CHEN Weilun, LIU Xiaoxue, ZHU Yiming, HE Zhenrui, ZHOU Erxun
    Acta Phytopathologica Sinica. 2024, 54(6): 1129-1141. https://doi.org/10.13926/j.cnki.apps.000922
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    Colletotrichum higginsianum is the major pathogen of anthracnose in cruciferous plants, which can severely damage the production of Chinese flowering cabbage (Brassica parachinensis) in south China. Scytalone dehydratase (Scd) is a key enzyme in the biosynthesis of DHN-melanin that can affect pathogenicity by mediated DHN-melanin biosynthesis in many plant pathogenic fungi. In this study, we identified a conserved scytalone dehydratase ChScd in C. higginsianum. The expression of gene ChSCD was analysed by using RT-qPCR technology, and it was found that the expression level of this gene was significantly up-regulated during the melanization of the hyphae and appressoria of C. higginsianum. Meanwhile, in order to analyse the biological function of the gene ChSCD, the Agrobacterium tumefaciens-mediated transformation technology was used to construct knockout and complementation mutant strains of the gene ChSCD. The results showed that the knockout of gene ChSCD blocked the biosynthesis of DHN-melanin in C. higginsianum, resulting in the loss of the melanization ability in both hyphae and appressoria, and leading to the significantly reduced in tolerance to cell wall interfering substances and oxidative stresses, the appressorium formation rate, turgor pressure, and pathogenicity of the ChscdΔ mutants. In summary, ChScd plays a crucial role in the biosynthesis of DHN melanin in C. higginsianum, which in turn affects the stress resistance, the formation rate of appressorium, turgor pressure, and the pathogenicity of the pathogen.
  • Identification of citron peroxidase gene family and its response to infection and expression of citrus bacterial canker
    LUO Jianming, MIAO Chouyu, HAN Jian, HAO Chenxing, YE Rongchun, SHENG Ling, MA Xianfeng, JIN Yan
    Acta Phytopathologica Sinica. 2024, 54(6): 1142-1157. https://doi.org/10.13926/j.cnki.apps.001037
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    Most citrus germplasm resources are susceptible to Xanthomonas citri subsp. citri (Xcc), which has seriously hindered the development of the citrus industry in China. In this study, 88 POD genes were identified in the citron genome and divided into 2 groups by phylogenetic tree analysis. Through RNA-seq analysis of the leaves from Citron C-05 and ‘Bingtang’ sweet orange, which are resistant and susceptible to Xcc respectively, 16 POD genes with Xcc-induced expression were obtained. The expression of CmPOD05, which has a high expression level and a large difference in resistant and susceptible germplasm, was further verified by qRT-PCR, and its possible localization to the cytoplasmic membrane was determined by subcellular localization in tobacco. The protein differences of POD05 homologous genes among different citrus germplasm were also analyzed, as well as several disease-resistance-related elements such as defense and stress response regulation through the analysis of cis-acting elements of their promoter sequences. Transient overexpression of CmPOD05 in ‘Bingtang’ sweet orange leaves significantly reduced the growth of Xcc per unit leaf area on the third day after Xcc inoculation compared to the control. Transient overexpression of CmPOD05 in Citron C-05 and tobacco leaves increased the H2O2 and superoxide anion content of leaves. The results indicated that CmPOD05 may be involved in the resistance process of Citron C-05 to Xcc.
    • GENETICS OF DISEASERESISTANCE AND PATHOGENICITY
  • Identification and molecular detection of resistance to wheat stripe rust in 121 Australian wheat varieties
    HUANG Jie, YANG Jinye, Davinder Singh, ZHANG Peng, Robert F. Park, WANG Baotong, LI Qiang
    Acta Phytopathologica Sinica. 2024, 54(6): 1158-1166. https://doi.org/10.13926/j.cnki.apps.000924
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    Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most important diseases of wheat worldwide. Breeding and cultivating disease-resistant varieties is the most economical and effective strategy to control stripe rust. Eight Australian races of Pst were used to evaluate the seedling resistance of 121 wheat varieties under controlled greenhouse conditions. The presence of genes Yr5, Yr9, Yr10, Yr15, Yr17 and Yr26 was assessed by genotyping with linked molecular markers for each gene. A total of 21 (17.35%) of 121 wheat varieties were resistant to all tested Pst races, seven (5.79%) varieties were susceptible to all Pst races. Based on molecular detection and disease resistance identification results,3 (2.48%) wheat varieties may contain Yr5 gene, 15 (12.40%) may contain Yr9, 9 (7.44%) may contain Yr10, 6 (4.96%) may contain Yr17, 19 (15.70%) may contain Yr26 and no varieties were detected to carry Yr15. Two or more stripe rust resistance genes in combination were detected in 13 wheat varieties. In addition, 19 varieties may contain other known or unknown new stripe rust resistance genes, which need further studies. This study provided scientific basis for rational utilization of 121 wheat varieties and breeding of wheat disease resistance.
  • Identification of the causing agents of soybean anthracnose and evaluation of soybean germplasm for resistance to the main anthracnose pathogen
    LI Yiyang, WU Mian, WANG Xing, GU Heping, CHEN Xin, CUI Xiaoyan
    Acta Phytopathologica Sinica. 2024, 54(6): 1167-1178. https://doi.org/10.13926/j.cnki.apps.001636
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    Anthracnose is a common worldwide fungal disease in soybean production, seriously affecting the commercial value of soybean pods as well as seed yield and vigour. In this study, four fungal strains (XZ-1-XZ-4) were isolated from the stalk of a diseased soybean plant within soybean-producing regions of Xuzhou City, Jiangsu Province, using tissue isolation and single spore purification methods. According to morphological chara-cteristics and ITS-TUB2-based phylogenetic analysis results, XZ-1 and XZ-2 were identified as Colletotrichum truncatum, XZ-3 and XZ-4 were identified as Fusarium proliferatum and Fusarium equiseti, respectively. Pathogenicity test results showed that all the four fungi were pathogenic, indicating a complex infection of soybean plants by Colletotrichum and Fusarium phytopathogens in the field. Furthermore, the resistance of 56 soybean varieties/lines to C. truncatum strain XZ-1 was evaluated by using two different inoculation methods (i.e., inoculation of fresh soybean pod with mycelial suspension, inoculation of etiolated hypocotyl with mycelial plug). It was showed that disease indexes obtained by the two inoculation methods were slightly different, but the resistance grades were almost the same, with a match rate of 92.9%. Of these 56 soybean varieties/lines tested, 3 were identified as resistant and 19 moderately resistant; No highly resistant varieties were found, and only 39.3% of the varieties were resistant to soybean anthracnose. This study combines the results of the above two inoculation methods to rapidly and accurately identify the resistance level of soybean varieties (lines) to anthracnose, laying a foundation for screening and utilizing resistant soybean germplasms, and for identifying anthracnose-resistant genes in soybean.
    • PLANT DISEASE AND CONTROL
  • Identification and biological characteristics of the pathogen causing quinoa anthracnose and indoor toxicity determination of several fungicides to the pathogen
    XUE Jing, HOU Xueping, JIANG Xiaodong, YIN Hui, ZHAO Xiaojun, LI Xinfeng
    Acta Phytopathologica Sinica. 2024, 54(6): 1179-1187. https://doi.org/10.13926/j.cnki.apps.001638
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    To identify the pathogen causing anthracnose disease on quinoa plants and investigate its biological characteristics, we collected diseased quinoa plants with typical anthracnose symptoms and conducted pathogen isolation and purification experiments. A representative strain LMTJ was obtained and determined as the pathogen of quinoa anthracnose by completing Koch′s postulates. Combined with morphological characteristics and the result of multi-gene phylogenetic analyses (ACT, CHS-1, GAPDH, ITS and TUB2), the pathogen was identified as Colletotrichum spinaciae. The suitable culture condition for mycelial growth of C. spinaciae LMTJ strain is that with starch as carbon source, peptone as nitrogen source, temperature at 20~25 ℃ and pH value of 6.0~7.0, while for sporulation is with sucrose as carbon source, sodium nitrate as nitrogen source, temperature at 20~25 ℃ and pH value of 6.0. To screen effective fungicides for the prevention and control of quinoa anthracnose, the toxicity of 5 fungicides to C. spinaciae LMTJ strain was tested. The results showed that all the tested fungicides could inhibit mycelial growth of LMTJ, of which 92.8% iprodione exhibited the strongest inhibitory effect, with EC50 of 2.7654 mg·L-1. The results provide scientific basis for the diagnosis and control of quinoa anthracnose.
  • Identification of the pathogen causing gummy stem blight on Trichosanthes kirilo-wii and screening of Chinese herbal medicine extracts for effective inhibition of the disease
    CHEN Dongfang, CAO Shun, LI Weiwen, HE Yanqiu, WANG Chuanwen, CHENG Youyu, YIN Dengke, ZHAO Wei, XIE Dongmei
    Acta Phytopathologica Sinica. 2024, 54(6): 1188-1197. https://doi.org/10.13926/j.cnki.apps.001639
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    To investigate the occurrence of gummy stem blight (GSB) on Trichosanthes kirilowii and the causing pathogen of the disease, and to screen effective plant-derived antimicrobial agents, we collected the plants with symptoms of GSB at T. kirilowii-planting regions in Qianshan City, Anhui Province, and conducted pathogen isolation and purification experiments. Based on morphological characteristics and the result of molecular identification, the isolates were identified as Stagonosporopsis caricae. Meanwhile, the inhibitory effects of 20 Chinese herbal medicine extracts against S. caricae were determined, and over 90% inhibition rate was obtained when ethanol extract from Cnidii Fructus (CF) or Macleaya Fructus (MF) was used at a concentration of 15 mg·mL-1, while over 80% inhibition rate for Sophorae Flavescentis Radix (SFR) or Polygoni Cuspidati Rhizoma et Radix (PCRR) extract. Further tests showed that both CF and SFR extracts showed significant inhibitory effect on mycelial growth of S. caricae, with EC50 values of 0.1648-0.5289 and 0.7347-0.8332 mg·mL-1, respectively. This study provides a theoretical basis for the environment-friendly control of GSB on T. kirilowii.
  • Isolation and identification of a spontaneous streptomycin-resistant strain, Pectobacterium aroidearum NJAU2, based on genome-wide analysis
    YANG Liping, LOU Wangying, CHE Shu, TANG Lidong, DENG Wei, GU Anyu, GUAN Junjiao, KUI Limei, AN Hua, WANG Rui, LI Xiaolin, FAN Jiaqin
    Acta Phytopathologica Sinica. 2024, 54(6): 1198-1214. https://doi.org/10.13926/j.cnki.apps.000930
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Bacteria from the genus of Pectobacterium are the important plant pathogens causing significant economic losses worldwide, and the emergence of antibiotic-resistant strains further exacerbates the challenges in Pectobacterium-caused disease prevention and control.This study used Pectobacteriums isolated from different host plants as materials and used plate analysis to obtain streptomycin-resistant strains formed in the wild. Clarify its taxonomic status through genome sequencing and comparative genome research, and streptomycin resistance genes were predicted. The results shown that among the eight strains isolated from different hosts, NJAU2 obtained from Yunnan yellow-flowered calla lily disease strains is highly resistant to streptomycin. NJAU2 genome was 5 062 459 bp in length, with G+C content of 51.83% and N50 length of 306 378 bp, containing 4 568 protein-coding genes. NJAU2 clusters on the same branch as the type strain P. aroidearum SCRI 109T and the reported strain P. aroidearum PC1 based on the Multi-Locus Sequence Analysis results of 8 housekeeping genes (fliA, fusA, glyA, gyrB, purA, rpoA, rpoS, secY). This result is consistent with the phylogenetic relationship constructed based on the 16S rRNA gene sequence. Furthermore, the Average Nucleotide Identity (ANI) and DNA-DNA Hybridization (isDDH) analysis with various Pectobacterium strains also support this classification. Therefore, the naturally occurring streptomycin-resistant strain NJAU2 is classified as a new species within the Pectobacterium genus: Pectobacterium aroidearum, and named Pectobacterium aroidearum NJAU2. P. aroidearum NJAU2 carrying sets of genes encoding proteins for streptomycin-resistanc in its genome. Comparison with the Comprehensive Antibiotic Resistance Database (CARD) identified 328 genes belonging seven types of drug resistance mechanisms. Moreover, in NJAU2 genome, 15 genes were annotated encoding aminoglycoside antibiotic resistance, including rpsL and gidB that are related to streptomycin resistance. The discovery of P. aroidearum NJAU2 provides new material for research in the direction of streptomycin resistance in Pectobacterium spp..
    • EXPERIMENTAL METHOD
  • Research and development of real-time fluorescence quantitative PCR molecular detection technology for Marssonina coronaria
    ZHANG Lixia, PENG Lan, WANG Yinghao, WU Xiaozheng, XU Liangsheng, HUANG Lili
    Acta Phytopathologica Sinica. 2024, 54(6): 1215-1225. https://doi.org/10.13926/j.cnki.apps.000905
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    Marssonina leaf blotch of apple is a significant apple leaf disease that seriously impacts both the yield and quality of apples. To enhance early diagnosis and prediction of Marssonina leaf blotch, we aimed to establish a real-time fluorescence quantitative PCR (TaqMan qPCR) detection system for Marssonina coronaria. Based on the conserved rDNA-ITS gene of M. coronaria, specific primers (HB-Taq-F/R) and a specific probe (HB-Taq probe) for M. coronaria were designed, and the TaqMan qPCR reaction system was subsequently established and optimized. The established system was utilized to quantitatively detect the pathogen’s DNA in the leaves of artificially inoculated apple trees with M. coronaria during the incubation period. Furthermore, the relationship between inoculation time and the latent infection amount of M. coronaria in the leaves was analyzed. Through the analysis of test results from samples collected at different times, it was demonstrated that this detection method could identify M. coronaria a noteworthy 10 days prior to disease onset. This approach holds immense promise for early, asymptomatic disease detection, providing a valuable tool for the timely and effective monitoring of Marssonina leaf blotch in apple orchards. Consequently, it significantly contributes to the enhancement of apple crop management and production.
  • Polyclonal antibody preparation and establishment of antibody-based method for the detection of coat proteins of Japanese yam mosaic virus
    LU Xiaojing, MENG Jiaorong, XIAO Dong, HE Longfei, CHEN Baoshan, ZOU Chengwu
    Acta Phytopathologica Sinica. 2024, 54(6): 1226-1235. https://doi.org/10.13926/j.cnki.apps.001344
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    Japanese yam mosaic virus(JYMV) is one of the most important yam viruses.It has a great impact on the yield and quality of yam. In this study, by analyzing the amino acid sequence of the coat protein (CP) of JYMV isolates from Henan, Shandong and Jiangsu provinces, three JYMV isolates JYMV-WX3, JYMV-SG1 and JYMV-FX1 were screened and their CP gene were constructed into the pET-30a prokaryotic expression vector. The fusion protein was purified and used for immunization of New Zealand white rabbits to prepare polyclonal antiserum. The polyclonal antibody against JYMV was obtained by antigen affinity purification, and it was found that the antibody titer was 1:512 000 by using indirect enzyme-linked immunosorbent assay (ELISA). Based on this antibody, ELISA, dot-blot enzyme-linked immunosorbent assay(Dot-ELISA) and immuno-capture reverse transcription-polymerase chain reaction (IC-RT-PCR) for the virus were established. The sensitivity of IC-RT-PCR for detecting JYMV in yam leaf juice is two orders of magnitude higher than that of ELISA. IC-RT-PCRcould detect yam leaf crude extracts diluted at 1:100 000(w/v, g·mL-1), while Dot-ELISA and ELISA could detect yam leaf crude extracts diluted at 1:100 and 1:1 000 (w/v, g·mL-1), respectively. The successful development of a series of immune detection methods can meet the quarantine and monitoring needs of JYMV for yam germplasm exchange, healthy seedling production and field disease epidemic, and contribute to the study of the pathogenic mechanism of this virus.
  • Establishment of RT-LAMP rapid detection method for broad bean wilt virus 2
    QIN YanhongWANG Fengli, ZHANG Zhongyi, LIU Hongyan, LIU Yuxia, GAO Suxia, WEN Yi, LI Shaojian, LIU Yongkang, ZHANG Desheng, LU Shuhao, ZHAO Zhengwei, WANG fei, LU Chuantao
    Acta Phytopathologica Sinica. 2024, 54(6): 1236-1243. https://doi.org/10.13926/j.cnki.apps.001346
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    Broad bean wilt virus 2 (BBWV 2), a member of the genus Fabavirus, is one of the important viruses infecting Rehmannia glutinosa and Dioscorea oppositifolia. This virus seriously affects the yield and quality of these medicinal crops. In this study, the detection method of reverse transcription loop-mediated isothermal amplification (RT-LAMP) for BBWV 2 was established. The primers were designed according to the conserved region of small coat protein (SCP) gene sequence of BBWV 2 RNA2 segments. The amplification temperature and time of the RT-LAMP were optimized and the optimal reaction condition was 60-65 ℃ within 60 min. The sensitivity, specificity and accuracy were demonstrated. The detection limit of RT-LAMP was 10 times higher than that of the conventional PCR, and it can detect virus at 6.47×103 copies·μL-1. This method can specifically detect BBWV 2 and the accuracy of RT-LAMP was higher than conventional PCR used for field samples of Rehmannia Atractylodes macrocephala and Angelica dahurica. The delection rate was consistent with that of conventional PCR for Dioscorea samples. The RT-LAMP established in this study is a specific, sensitive, rapid and convenient method for BBWV 2 detection and is suitable for use in grass-roots scientific research for the accurate detection and diagnosis of BBWV 2.
    • RESEARCH NOTES
  • First report of southern blight of Morinda officinalis caused by Sclerotium rolfsii in China
    SU Hailan, NIU Yuqing, LIN Fengfang, ZHU Yujing, XIAO Rongfeng
    Acta Phytopathologica Sinica. 2024, 54(6): 1244-1247. https://doi.org/10.13926/j.cnki.apps.001645
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    Morinda officinalis is a Chinese traditional herbal medicine in the family Rubiaceae. From April to June 2023, the southern blight disease occurred seriously on M. officinalis in Zhangzhou City, Fujian Province, China. The white mycelia and sclerotia of the pathogen were initially observed at the stem-soil interface of diseased seedlings, and gradually spread to neighboring plants, which resulted in the death of patches of plants. Fungal strains with same morphological characteristics were obtained from the diseased samples by using the tissue isolation method. Based on morphological characteristics, ribosomal large subunit (LSU) sequences and pathogenicity test, the representative fungal strains were identified as Sclerotium rolfsii Sacc. To our knowledge, this is the first report of S. rolfsii causing southern blight on M. officinalis, which provides important information on the pathogen for disease management.
  • Identification of the pathogen causing anthracnose on Caiyunhong pear in Yunnan Province
    HE Yingyun, WANG Lina, YANG Guangzhu, ZHANG Wen, SHU Qun, SU Jun
    Acta Phytopathologica Sinica. 2024, 54(6): 1248-1251. https://doi.org/10.13926/j.cnki.apps.001661
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    Anthracnose is one of the most important fungal diseases that pose significant threat to the production of Yunnan red pear cultivar ‘Caiyunhong’. The disease mainly causes fruit rot, resulting in a disease incidence of more than 35% in normal conditions. The causal agent was identified as Colletotrichum fructicola based on the morphological characteristics, phylogenetic relationship inferred from ITS, ACT, TUB and CHS sequences, and the result of pathogenicity test. This is the first report of C. Fructicola causing anthracnose on ‘Caiyunhong’ fruit in China, which provides a theoretical basis for effective prevention and control of the disease.
  • Isolation, identification of pathogen causing leaf spot on Polygonatum kingianum
    XU Rong, WANG Siting, WANG Zhandi, YANG Shiwei, LIU Kexin, XU Yongyan, JIAO Rong
    Acta Phytopathologica Sinica. 2024, 54(6): 1252-1256. https://doi.org/10.13926/j.cnki.apps.000906
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    A leaf spot of Polygonatum kingianum was found in Jianxing Town, Xinping County, Yunnan Province. Leaf infections first appeared as small, pale brown spots with a yellow haloes. The spots extended or coalesced into larger lesions, causing the entire leaf to be necrotic. The pathogen causing this disease was isolated, and its pathogenicity was tested according to the Koch' s postulates. Based on morphological characteristics and phylogenetic analysis of combined the rDNA 18S and ITS regions as well as beta-tubulin (TUB) gene sequences, the isolate DYB-2 was identified as Neopestalotiopsis acrostichi. To our knowledge, this is the first report of N. acrostichi causing the leaf spot on Polygonatum kingianum in the world.
  • Identification and biological characterization of root-rot disease in Ficus virens
    FU Jinxin, LI Zengping, ZHANG Yu
    Acta Phytopathologica Sinica. 2024, 54(6): 1257-1262. https://doi.org/10.13926/j.cnki.apps.000893
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    To clarify the species and biological characteristics of the pathogenic fungi that cause root rot of Ficus virens in Hainan Province, pathogenic fungi were isolated from diseased tissue and basidiocarps collected from the root of the diseased tree of Ficus virens in March 2022 by tissue separation method. The pathogenic bacteria causing the root rot disease were identified through pathogenicity determination, combing with morphology identification and molecular identification. The effects of pH, temperature, medium and light on mycelium growth under different treatments were measured by cross crossing method. The results showed that the isolated pathogenic ganoderma strain was the pathogen causing the root rot of Ficus virens. After morphological characterization and jointly phylogenetic tree construction based on rDNA-ITS, rDNA-LSU and rDNA-SSU, the pathogen strain was identified as Ganoderma tropicum. The biological characteristics showed that the optimal medium was PDA medium, the optimal pH was 7.0, the optimal light condition was total darkness, and the optimal growth temperature was 32 ℃.It will provide scientific theoretical basis for the diagnosis and control of this disease.
  • Identification and biological characteristics of the pathogen causing shoot canker of Prunus serrulata in Henan Province
    GAO Wenkai, QIN Shijiao, XIE Yuxu, CAO Jiayuan, HOU Kangle, XU yili, MA Qingzhou, XU Chao, GUO Yashuang, ZHANG Meng
    Acta Phytopathologica Sinica. 2024, 54(6): 1263-1267. https://doi.org/10.13926/j.cnki.apps.000918
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    In recent years, the planting area of Prunus serrulata in Henan Province has been increasing, and the disease problem is becoming more and more serious. A canker disease that infects the branches of P. serrulata has been observed in many areas; it causes brown canker tissue on the shoots, twigs and kills the branches. To clarify the pathogen causing shoot canker of P. serrulata in Henan Province, isolates were obtained from diseased shoots and identified based on the morphological and molecular biological methods. In this study, a total of 21 strains with the same morphological characteristics were obtained and Koch’s postulates was verified. According to its morphological and molecular biological characteristics, the pathogen causing shoot canker of P. serrulata in Henan Province was finally identified as Diaporthe eres. This is the first report of D. eres responsible for P. serrulata shoot canker. The optimum growth temperature of D. eres was 25 ℃, and the optimum pH was 6, the optimum carbon source was sucrose, the optimum nitrogen source was yeast powder.
  • Isolation and identification of the pathogen causing basal stem rot of Achyranthes bidentata
    WANG Fei, YANG Jin, LI Shaojian, GAO Suxia, LI Xuemeng, QIN Yanhong, ZHAO Ying, LU Chuantao
    Acta Phytopathologica Sinica. 2024, 54(6): 1268-1272. https://doi.org/10.13926/j.cnki.apps.000920
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    From 2020 to 2022, a new disease, basal stem rot of Achyranthes bidentata was prevalent in the field in Wuzhi county, Jiaozuo city, Henan province. The incidence of severe disease plot was over 90%, that seriously affected the yield and quality of A. bidentata. Tissue isolation and culture method was used to isolate and purify the pathogen from disease samples of A. bidentata with the typical symptoms of stem rot. Based on the morphological identification, sequence analysis of ITS and TEF-1α, and pathogenicity assays, the pathogen were identified as Rhizoctonia solani, and belonged to anastomosis subgroup (AG)-4-HGI. This is the first report of R. solani AG-4-HGI causing basal stem rot on A. bidentata in China, the results of this study could provide useful guidance for the effective prevention and control of A. bidentatae basal stem rot.
  • Re-identification of a pathogen of pea root rot disease
    HE Suqin, WEN Zhaohui, BAI Bin, LIU Yonggang
    Acta Phytopathologica Sinica. 2024, 54(6): 1273-1279. https://doi.org/10.13926/j.cnki.apps.000901
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    Mycoleptodiscus terrestris is a pathogen on many legumes, but there are very few reports about it occurring on peas. In May 1989, the pathogenic fungus producing black microsclerotia was isolated from diseased roots of pea in Dingxi city, Gansu province. Due to limited knowledge and the lack of important literature at that time, the fungus was named as Burgoa pisi in 1998. In May 2018, several fungal strains with the same morphological characteristics as B. pisi were isolated from diseased pea roots and hypocotyls in Huining county, Gansu province. The optimum mycelial growth temperature of tested strains was 25-30 ℃. When cultured on PCA medium at 30 ℃ for 14 d, the strains produced abundant of microsclerotia, the microsclerotia were nearly spherical to irregular, (104.5-567.2) μm × (95.5-343.3) μm, and there were 0 to dozens of setae on the sclerotial surface, the setae were multicellular and columnar, the basal cells were colorless or pale, and the terminal cell was black thick-walled (sheath-like), straight, slightly curved or short forked. However, when on PDA and PCA media at 25-30℃ for 21 days, no conidia produce, and they also failed to induce conidia by soybean leaves inoculation in vivo. BLASTn analysis showed that the sequences of strain TCD-11 (GenBank accession No. ITS: OL981504; LSU: OL981961) and TCD-28 (GenBank accession No. ITS: OL981505) had a 99.45% similarity in the ITS region and 100% identity in the LSU region with M. terrestris ex-type strain CBS 231.53 (GenBank accession No. ITS: MK487754.1; LSU: MK487730.1). In the combined phylogenetic tree of rDNA-ITS and LSU sequences, tested strains TCD-11 and TCD-28 were clustered with M. terrestris strains in same group. Pathogenicity test showed that the tested strains had aggressive pathogenicity to the root and epicotyl of pea (Pisum sativum) (included edible pod pea and edible seed pea), the infected roots and hypocotyls showed extensive brown to dark brown spots; they had weak pathogenicity to broad bean (Vicia faba) of Fabaceae, only small restricted black necrotic spots were formed at the inoculation site. The pathogen was reisolated from all infected pea and broad bean plants and had the same morphology as isolates TCD-11 and TCD-28, fulfilling Koch’s postulates. According to morphological and molecular biological characters, tested strains are identified as M. terrestris, and B. pisi in essence is a synonym of M. terrestris.
  • Pathogen identification of Paris polyphylla var. yunnanensis (Franch) Hand.-Mazz gray spot disease in Yunnan Province
    LIAO Yongqin, WANG Nan, YI Xiaopeng, SHI Zhufeng, SHEN Yunxin, MO Yanfang, YANG Tongyu, HE Yonghong, YANG Peiwen
    Acta Phytopathologica Sinica. 2024, 54(6): 1280-1285. https://doi.org/10.13926/j.cnki.apps.000928
    Abstract ( ) Download PDF ( )   Knowledge map   Save
    Paris polyphylla var. yunnanensis (Franch) Hand.-Mazz is an important cash crop in Yunnan ethnic minority areas. In 2022, we discovered a new type of leaf spot disease in the Lijiang planting area, which can damage the entire growth period of the crop. The typical symptoms in the field were spots(diameter 2.4-6.7 mm) on the surface of the disease leaves, the leave become brown and necrosis in later stage thus resulting in serious economic losses. We isolated pathogenic bacteria by monosporophyte isolation,and identified by morphological observation and polygenic phylogenetic analysis, and its pathogenicity was verified using Koch’s rule.The final identification results showed that the pathogen was a new species of Cercospora and was named Cercospora polyphylla x.he & P. W. Yang, sp.11 according to the binomial nomenclature.This is the first report of Paris polyphylla var. yunnanensis (Franch) Hand.-Mazz leaf disease caused by C. polyphylla in China, and this study will provide a theoretical basis for the epidemic and comprehensive prevention and control of the disease.
  • Identification of the pathogen causing Chinese cabbage verticillium wilt in Qinghai Province
    LIU Shuangru, KONG Xiaoping, ZHANG Xiaomei, YU Zilin, DUAN Weijun
    Acta Phytopathologica Sinica. 2024, 54(6): 1286-1290. https://doi.org/10.13926/j.cnki.apps.000902
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    Verticillium wilt of Chinese cabbage is an emerging disease in Qinghai Province. The symptoms of verticillium wilt is discoloration of the vascular bundle of roots, blackening and discoloration of stems or leaf veins which may extend to the whole leaves and death in severe case. This study isolated and cultured 24 strains from 10 Chinese cabbage samples in Qinghai Province and detected the strains with morphological observation, DNA sequence analysis, and pathogenicity test to determine the pathogen. Morphological observation showed that the strain QH-87 grew very slowly on potato dextrose agar medium with a large number of black microsclerotia( (5.8-27.8) μm × (35.3-356.8) μm), and the length to width ratio was 2.3-25.6. Conidia were transparent and cylindrical and its size was (2.1-5.4) μm × (4.5-11.9) μm. Chlamydospores and dormant hyphae were not observed. The fungus was verified as Verticillium longisporum lineage A1/D1 according to the results of ITS, ACT and EF sequencing. The pathogenicity test showed that strain QH-87 could infect Chinese cabbage and caused discoloration and wilt. In a word, the pathogen was identified as V. longesporum lineage A1/D1. It is the first report of verticillium wilt of Chinese cabbage in Qinghai province, and V. longisporum lineage A1/D1 is first reported on Chinese cabbage in China.
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China Agricultural University
Editor in Chief: FAN Jun
Started in 1955
ISSN 0412-0914
CN 11-2184/Q
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