Acta Phytopathologica Sinica 2023 Vol.53
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Research progress of mulberry sclerotial disease
LÜ Zhiyuan, TIAN Lichao, HE Ningjia
Acta Phytopathologica Sinica    2023, 53 (1): 1-12.   DOI: 10.13926/j.cnki.apps.000628
Abstract （456）      PDF（pc） （8932KB）（818）       Save
Mulberry sclerotial disease, a general term for diseases with similar symptoms caused by three ascomycetous fungi, is a devastating fungal disease for fruit mulberry production, which seriously restricts the deve-lopment of the mulberry industry. The three pathogens are necrotrophic fungi, with various infection means and complicated infection mechanisms. The different degrees of difficulty in artificial cultivation of these three pathogens limit the research process of pathogens to a certain extent. For instance, Ciboria shiraiana and Ciboria carunculoides are difficult to cultivate whereas Scleromitrula shiraiana cannot complete the life cycle on artificial medium. This article summarized the research progress on disease cycle, pathogens, disease epidemic, pathogens-host interaction, and prospects for future research. The studies would provide a reference for further research on mulberry sclerotial disease.
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Pathogenicity and virulence of two different PVY isolates associated with three different hosts
LIU Qing, JIANG Xin, LIU Chunju, GUO Yongliang, YAN Zhiyong, GENG Chao, WANG Jie, XU Pengjun, LI Xiangdong, TIAN Yanping
Acta Phytopathologica Sinica    2023, 53 (1): 13-21.   DOI: 10.13926/j.cnki.apps.000625
Abstract （452）      PDF（pc） （7321KB）（590）       Save
Potato is one of the most important food and economic crops. Potato virus Y is an economically important virus affecting potato production. Recent studies have shown that the composition of PVY strain groups has changed. Recombinant PVY isolates including PVYNTN-NWSYRI and PVYNTN-NWSYRII have become the predominant strains. However, their pathogenicity and virulence were unclear compared with previous PVYN strain isolates. In this study, we compared the infectivity of isolates PVYN605 and PVYNTN-NWSYRI-GZ (originally isolated from tobacco plant) on Nicotiana benthamiana, N. tabacum and Capsicum annuum (Tedaniujiaowang) plants and their virulence on the plants of N. benthamiana and N. tabacum. Results showed that both isolates PVYN605 and PVYNTN-NWSYRI-GZ could infect N. benthamiana and N. tabacum systemically and induced vein necrosis in N. tabacum. Isolate PVYNTN-NWSYRI-GZ, but not PVYN605, could infect C. annuum (Tedaniujiao-wang) systemically. The cell-to-cell movement of PVYNTN-NWSYRI-GZ is slower than that of PVYN605. The reduction in the height of N. benthamiana plants caused by PVYNTN-NWSYRI-GZ was more severe than that by PVYN605. At 11 days post agroinfiltration, the accumulation level of PVYNTN-NWSYRI-GZ in N. benthamiana plants was lower than that of PVYN605. The results obtained in this study will help us understand the evolution of the PVY strains and design a new control strategy for PVY.
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Effects of TeMV and PWV infection on metabolic physiology and tissue microstructure of passion fruit
HUANG Chengmei, HU Chunjin, SHI Guoying, LUO Haibin, CAO Huiqing, WU Xingjian, JIANG Shengli, YE Liping, WEI Yuanwen
Acta Phytopathologica Sinica    2023, 53 (1): 22-30.   DOI: 10.13926/j.cnki.apps.000645
Abstract （281）      PDF（pc） （8812KB）（340）       Save
Passion fruit variety ‘Tainong 1’ was used as experimental material. The morphological structure of leaves and fruit exocarp of healthy and TeMV and PWV-infected plants was observed by scanning electron microscope. The physiological and biochemical changes of leaves were determined, and the effects of passion fruit virus infection on the morphological structure of leaves and young fruit exocarp, physiology and biochemistry were studied. The results showed that after TeMV and PWV infection, the spongy tissue cells of passion fruit leaves arranged in clusters with small spaces, and the palisade tissue cells atrophied in an irregular strip shape. The stone cell layer in the outer epidermis of young fruit was severely damaged and arranged loosely, while the sponge layer cells were relatively loose in structure with serious lignification. Additionally, the surface texture of plant leaves and young fruits exocarp was rough and wrinkled; the stomatal guard cells shrank, and the tissue around stomata shrank and appeared rough. Moreover, the glandular hairs on the epicarp surface of the susceptible fruit were raised and disordered. After virus infection, the sucrose content of No.1-3 leaves of susceptible plants was 23.11% higher than that of No. 4-6 leaves, while it remained relatively balanced in the healthy leaves. The starch accumulation, PG enzyme, cellulase and PPO enzyme activities were reduced; whereas, the activities of α-amylase, GS enzyme and POD enzyme in No.1-3 leaves of susceptible plants were increased. After being infected by TeMV and PWV, the distribution of nutrients in No.1-6 leaves of the plant was unbalanced. The synthesis and decomposition of related substances and antioxidant defense enzyme system were damaged, and the morphological structure of leaves and fruits were damaged, resulting in the disruption of the normal absorption function of the plant.
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Cloning and characterization of Mdtl2 gene encoding a thaumatin like protein in Malus domestica cv. Fuji
GONG Wan, WANG Weidong, LÜ Luqiong, WANG Shuaile, HUANG Lili
Acta Phytopathologica Sinica    2023, 53 (1): 31-40.   DOI: 10.13926/j.cnki.apps.000790
Abstract （293）      PDF（pc） （8159KB）（448）       Save
Thaumatin like proteins (TLPs) are a class of pathogenesis-related proteins with antifungal activities and resistance to abiotic stress. In this study, we cloned a TLP, named Mdtl2, which contains an open reading frame of 912 bp, a protein of 303 amino-acid residues. Bioinformatics analysis showed that Mdtl2 contains several conserved cysteines, TLPs family conserved signature sequence, and a typical three-dimensional structure of TLPs. Based on these messages, we speculate that Mdtl2 belongs to GH64-TLP-SF superfamily of TLPs family. Overexpression of Mdtl2 distinctly inhibited the infection of Phytophthora infestans in Nicotiana benthamiana leaves and Valsa mali in apple leaves, respectively. Besides, overexpression of Mdtl2 induced accumulation of callose in apple leaves. Taken together, Mdtl2, as a member of glycoside hydrolase superfamily, enhanced plant resistance to pathogens by inducing the production of callose.
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Functional verification of M35 metalloprotease FocM35_2 in Fusarium oxysporum f. sp. cubense tropical race 4
WU Bangting, LIU Siwen, ZHANG Xiaoxia, LI Chunyu, LIU Hong
Acta Phytopathologica Sinica    2023, 53 (1): 41-51.   DOI: 10.13926/j.cnki.apps.000617
Abstract （287）      PDF（pc） （9230KB）（278）       Save
Fusarium wilt caused by Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4) is the most devastating disease on banana plants. In our previous study, FocM35_2 of Foc TR4 was identified from an earlier transcriptomics study on the infection process of banana roots. The FocM35_2 gene knockout mutant demonstrated a slightly vegetative growth defect, decreased sporulation rate and attenuated virulence. In addition, the mutant was more sensitive to exogenous oxygen stress, osmotic pressure and cell wall stress. Furthermore, transient expression of FocM35_2 in Nicotiana benthamiana showed that FocM35_2 could be expressed in the apoplastic region and induced HR in leaves. The absence of FocM35_2 also enhanced the enzymatic activity of chitinase in banana. The results showed that FocM35_2 is an important virulence factor in Foc TR4.
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Cited: CSCD(1)
Cloning of VvSUC12 gene promoter and identification of upstream regulatory factors
WU Jiahong, XING Qikai, ZHOU Yueyan, BAI Qingrong
Acta Phytopathologica Sinica    2023, 53 (1): 52-60.   DOI: 10.13926/j.cnki.apps.000623
Abstract （299）      PDF（pc） （6101KB）（365）       Save
Botryosphaeria dieback is an important grapevine trunk disease that has seriously limited the development of grape industry in recent years. Studies have shown that grape sucrose transporters are involved in the interaction between host plants and pathogens. In order to analyze the regulatory pathway of sucrose transporter VvSUC12 in grape immune response, the promoter region of 1 500 bp upstream of translation initiation site of VvSUC12 gene was cloned in this study. Bioinformatics analysis showed that this region contained four Dof transcription factor binding sequences (A/TAAAG) and a variety of hormone regulation and defense-related cis-elements. Real-time quantitative PCR analysis showed that the expression of VvSUC12 and VvDof19 genes was significantly induced by inoculation with Lasiodiplodia theobromae. The transcription factor VvDof19 interacting with VvSUC12 promoter was screened by yeast one-hybrid assay. Yeast two-hybrid assay confirmed that Vv-Dof19 had self-activation activity, the transient transformation of tobacco showed that the relative GUS activity of Dof19-GFP was about 9 times that of the control GFP. It confirmed that transcription factor VvDof19 could activate the expression of VvSUC12 gene. The above results lay the foundation for further study on the function of sucrose transporters in the grape immune response.
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Identification and functional analysis of TaLecRLK1 in wheat resistance to stripe rust
WANG Jianfeng, GAN Pengfei, XU Jinghua, TANG Chunlei, KANG Zhensheng, WANG Xiaojie
Acta Phytopathologica Sinica    2023, 53 (1): 61-71.   DOI: 10.13926/j.cnki.apps.000840
Abstract （526）      PDF（pc） （8656KB）（455）       Save
Lectin receptor-like kinase (LecRLK) is an important class of receptor-like protein kinases in plants that plays vital roles in plant response to various biotic and abiotic stresses. In this study, by analyzing the transcriptome data of wheat plants infected by Puccinia striiformis f. sp. tritici (Pst), we obtained a wheat lectin-like receptor kinase gene TaLecRLK1 that was significantly up-regulated during the wheat-Pst incompatible interaction. TaLecRLK1 contains a N-terminal signal peptide, an extracellular B-lectin domain, a PAN_AP domain, a transmembrane domain and an intracellular Pkinase-Tyr domain. Quantitative real-time PCR verified that TaLecRLK1 expression was induced in the early stage of wheat resistance to avirulent Pst infection. TaLecRLK1-GFP was located on the cell cytoplasmic membrane of Nicotiana benthamiana and wheat protoplasts. Virus induced gene silencing (VIGS) of TaLecRLK1 by barley stripe mosaic virus (BSMV) weakened wheat resistance to stripe rust, leading to sporadic production of uredinospores on TaLecRLK1-silencing wheat leaves challenged with Pst CYR23 that is avirulent on the control plants. Histological observation revealed longer infection hyphae in TaLecRLK1-silencing plants, along with decreased reactive oxygen species (ROS) accumulation per infection site compared with that in the control plants. In addition, the expression of defense-related genes TaPR1, TaPR2 and TaPR5 were decreased in TaLecRLK1-silencing plants, while the expression of TaCAT and TaSOD responsible for ROS scavenge were rapidly induced. These results indicate that TaLecRLK1 functions as positive regulator of wheat resistance to Pst.
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Construction of gene knockout system in sugarcane pokkah boeng pathogen Fusa-rium proliferatum YN41 based on transformation of protoplast
WANG Shuang, PAN Kaiyuan, YAO Ziting, YAO Wei, ZHANG Muqing
Acta Phytopathologica Sinica    2023, 53 (1): 72-83.   DOI: 10.13926/j.cnki.apps.000616
Abstract （322）      PDF（pc） （11345KB）（305）       Save
Gene-knockout has been becoming an important tool to verify gene functions in vivo. In order to further study the function of pathogenic genes of sugarcane Pokkah boeng pathogen Fusarium proliferatum, a gene knockout system for YN41 strain was created. In this paper, we constructed a linear hph selective marker cassette flanking upstream and downstream homologous fragments, by fusion PCR; A mixture solution of 0.01 g·mL-1 driselase and 0.05 g·mL-1 yase was optimized to produce the most high yield of protoplast was 3 h at 28 ℃; Protoplast transformation was mediated by polyethylene glycol (PEG); The transformants were identified by PCR and enzyme digestion. The gene deletion mutant was further verified by fluorescence quantitative PCR and Southern Blot. The biological phenotype, (such as colony morphology, growth rate, sporulation, biomass and pathogenicity), indicated the successful construction of gene knockout system. 2×107 protoplasts per milliliter were obtained by enzymatic hydrolysis at 28 ℃ for 3 h; The PK gene encoding pyruvate kinase was knocked out and homozygous was obtained. Real-time PCR detects no expression at the transcription level. Southern blot showed that there was no band in the gene deletion mutant using PK gene probe, but the wild type hybridized to the target band The PK gene knockout mutant exhibited an increased spre production, enhanced pathogenicity, and the varied growth rate which was slower firster and compared with wild type. The gene knockout system for F. proliferatum YN41 was successfully developed which provided a technical platform for the functional research on the pathogenic genes.
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Spray-induced gene silencing of CYP51 and Sdh genes effected the growth and pathogenicity of Rhizoctonia cerealis
CHENG Yongjie, ZHANG Haotian, SUN Haiyan, CAO Shulin, MA Dongfang, CHEN Huaigu, LI Wei
Acta Phytopathologica Sinica    2023, 53 (1): 84-96.   DOI: 10.13926/j.cnki.apps.000791
Abstract （294）      PDF（pc） （9969KB）（330）       Save
Due to the lack of stable genetic transformation system, researches on the function of pathogenicity-related genes in Rhizoctonia fungi have been severely restricted. CYP51 and Sdh are the target genes of the commonly used fungicides to prevent and treat Rhizoctonia diseases. Wheat sharp eyespot caused by R. cerealis is a stem-base wheat disease that is widely distributed in China. In this study, the dsRNA of three homologous genes of RcCYP51 and B, C, D subunit genes of RcSdh were synthesized in vitro. The influence of these six target genes to growth and pathogenicity of R. cerealis was studied based on the spray-induced gene silencing (SIGS) method. The results indicated that the growth and pathogenicity of R. cerealis were suppressed by the dsRNA of all target genes. On the infected barley leaves, exogenous dsRNA can significantly inhibit the expression of the corresponding gene. Among them, the RcCYP51-3-dsRNA at the concentration of 100 ng·μL-1 had a significant inhibitory effect on barley leaves, and can simultaneously inhibit the expression of the three homologous genes of RcCYP51. The RcSdhD-dsRNA can also inhibit the gene expression of the three subunits of RcSdh, and the optimum concentration was 50 ng·μL-1. This study explored a method to inhibit the gene expression in R. cerealis, and provided a useful exploration for using the SIGS method to study the gene function of Rhizoctonia fungi.
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Screening of biocontrol strains against sunflower white mold and analysis of its control effect
ZHANG Yurong, HAN Shengcai, ZHANG Jian, ZHANG wenbing, LI Hao, YANG Jianfeng, ZHAO Jun
Acta Phytopathologica Sinica    2023, 53 (1): 97-109.   DOI: 10.13926/j.cnki.apps.000785
Abstract （296）      PDF（pc） （7954KB）（544）       Save
To exploit effective microbial resource for sunflower white mold control, 33 endogenous strains with different colony phenotypes were isolated from 12 sunflower cultivars. Their function of sunflower white mold control caused by Sclerotinia sclerotiorum were analyzed via a paper cup system combined with in vitro leaves inoculation method. The only strain named as KB3 (Bacillus subtilis) protected sunflower seedlings from the destruction by S. sclerotiorum absolutely during the experiment period. Its sunflower white mold control effect was 79.47% under greenhouse condition. Its culture filtrate inhibited the growth of S. sclerotiorum on antagonism plate significantly (68.46%) and suppressed the black sclerotia formation severely, which was probably attributed to its various category of secondary metabolites production, indicated by whole genome sequencing and unidentified volatiles respectively. The microbial screening method used in this study could be an important refe-rence for biocontrol agent exploration and strain KB3 was a potent resource for the suppression of sunflower white mold.
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Optimization of PEG-mediated genetic transformation system of Colletotrichum acutatum HHDL02 from infected Capsicum annuum L.
SHENG Jiawei, MAN Yilong, OUYANG Chao, LIU Sizhen, GUO Sheng, SUN Qianlong, CHEN Yue, ZHANG Xin, TAN Xinqiu, WANG Yunsheng
Acta Phytopathologica Sinica    2023, 53 (1): 110-118.   DOI: 10.13926/j.cnki.apps.000629
Abstract （295）      PDF（pc） （7533KB）（291）       Save
Pepper anthracnose, caused by Collectotrichum acutatum infection, is the most destructive fungal disease in pepper production, which seriously threatens yield and quality of pepper. The genetic transformation with higher efficiency for the pepper pathogen C. acutatum is still not available. Our research provided optimal conditions to obtain protoplasts with higher efficiency at 2% of the enzyme in 1 mol·L-1 NH4Cl solution , 2 h of germination for the germ tube of conidium, and 2-3 h of incubation with application of the protoplasts , the GFP gene was successfully introduced into the wild type C. acutatum strain HHDL02 through modified PEG-mediated transformation. We further evaluated some of the transformants for colony morphology, growth rate, conidium morphology, conidial germination, appressorium formation, conidial production quantity and pathogenicity, which exhibited no obvious differences as compared to the wild type strains. In addition, visualizing the fluorescence signals of their progenies of the transformants suggested that the GFP gene integrated in C. acutatum strain HHDL02 was genetically stable. These results indicated that the protoplasts produced efficiently with the optimal approach from germ tube and the subsequent transformation mediated by PEG was suitable for the genetic operation of C. acutatum, and also helpful for the study on the mechanism of pathogenesis on pepper plants.
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SYBR Green I quantitative real-time PCR for melon yellow spot virus
SUN Xiaohui, PAN Ruijing, LIU Yongguang, WANG Dan, SHI Zhaopeng, QIAO Ning, SUN Zuowen, ZHU Xiaoping
Acta Phytopathologica Sinica    2023, 53 (1): 119-125.   DOI: 10.13926/j.cnki.apps.000630
Abstract （351）      PDF（pc） （5196KB）（396）       Save
The aim of this study was to establish a SYBR Green I quantitative real-time PCR method (qPCR) for detection of melon yellow spot virus (MYSV). The specific primer pair for qPCR was designed based on the conserved sequence of MYSV nucleocapsid protein gene, and the annealing temperature, primer concentration, specificity and sensitivity of the primer were optimized. The results showed that the optimal annealing temperature of the optimized qPCR method was 61.3 ℃, the optimal primer concentration was 0.65 μmol·L-1, the specificity and repeatability were good and this method was 100 times more sensitive than PCR. The cycle threshold of the qPCR standard curve was linear with the template concentration, and the correlation coefficient was 0.999 7. The results of field samples showed that the established qPCR method can be used for the quantitative detection of MYSV.
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Cited: CSCD(2)
Identification and pathogenicity of Fusarium species causing soybean root rot in Heilongjiang Province
WANG Shuang, LI Xinmin, LIU Chunlai, YANG Fan, JIANG Xifeng, LI Min, CAO Dawei, XU Chong, GUO Yanan
Acta Phytopathologica Sinica    2023, 53 (1): 126-128.   DOI: 10.13926/j.cnki.apps.000635
Abstract （322）      PDF（pc） （2698KB）（614）       Save
Soybean root rot disease caused by the genus Fusarium was observed from 30 soybean fields during 2017 and 2018 in 12 counties or cities of Heilongjiang Province. A total of 250 Fusarium isolates were recovered from the root tissues of soybean seedling collected from those fields. Identification of the Fusarium species was performed by examining their morphological features and sequences analysis of internal transcribed spacers (ITS) of rDNA. Six species of Fusarium were identified, with F. oxysporum being the predominant species (72.8%), followed by F. solani (15.6%), F. tricinctum (7.2%), F. equiseti (2.8%), F. brachygibbosum (1.2%) and F. graminearum (0.4%). All Fusarium isolates were highly virulent on soybean (cv. Heinong 69), causing root rot disease symptoms, which were reisolated from diseased plants and identified, fulfilling the Koch’s postulates. The pathogenicity tests for representative strains of the six Fusarium species showed that at 14 days after inoculation by sorghum seeds (4 g) fully colonized by Fusarium, the disease indexes of F. oxysporum (m3-3xu), F. solani (p15-5zhu), F. tricinctum (pz2-2), F. equiseti (dy-mz2-1), F. brachygibbosum (p13-1), and F. graminearum (mx7-2) were 69.05, 42.86,78.57, 45.24, 58.96, and 35.94, respectively. Both conidia cultures and secretions of the six isolates could cause root rot on soybean at 14 days post inoculation. The rot was serious with increased concentration of conidial cultures and secretions. F. solani was the most virulent to soybean with a disease index of 56.2, followed by F. oxysporum (44.8), F. graminearum (39.1), F. equiseti (37.2), F. tricinctum (32.9) and F. brachygibbosum (16.2) at a concentration of 1×108 conidia·mL-1. The disease indexes of F. solani and F. equiseti were 41.0 and 27.7, respectively at concentration of 1×105 conidia·mL-1, which showed no significant difference related to the disease index at concentration of 1×108 conidia·mL-1. As for the virulence of fermentation filtrates, F. solani and F. oxysporum were the most virulent to soybean with the disease indexes of 48.6 and 48.6, respectively, followed by F. equiseti (44.8), F. tricinctum (41), F. graminearum (40.1) and F. brachygibbosum (27.2).
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Cited: CSCD(1)
Isolation and Identification of a new leaf spot pathogen causing leaf spot Nandina domestica
WANG Xingchen, SU Pan, XU Qiaolan, ZHANG Pinghua, WANG Zhiying, YAO Linbo, GUO Wei
Acta Phytopathologica Sinica    2023, 53 (1): 129-132.   DOI: 10.13926/j.cnki.apps.000818
Abstract （366）      PDF（pc） （3223KB）（506）       Save
A new leaf spot disease causing reddish brown or black necrosis on Nandina domestica was identified in Jinhua city, Zhejiang province. Using tissue isolation, Koch’s postulation, morphological observation, and maximum likelihood estimate, we determined that the causative pathogen of N. domestica leaf spot was Diaporthe eres. To our knowledge, it is the first report to show that phytopathogenic fungus D. eres causes leaf spot disease of N. domestica in China. This study may provide important basis for rapid diagnosis and prevention of this new disease on N. domestica.
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Cited: CSCD(2)
Identification of pathogen causing Common bean (Phaseolus vulgaris) wilt in Inner Mongolia
CHAI Ali, YANG Hongmin, LI Xin, SHI Yanxia, XIE Xuewen, LI Lei, FAN Tengfei, LI Baoju
Acta Phytopathologica Sinica    2023, 53 (1): 133-136.   DOI: 10.13926/j.cnki.apps.000633
Abstract （335）      PDF（pc） （3078KB）（406）       Save
Common bean (Phaseolus vulgaris), also known as the Kidney bean, is widely grown in China for its fresh pod or edible dry seeds, which consumed as vegetable and cereal. In July 2021, severe outbreaks of common bean wilt were observed in several commercial fields in Inner Mongolia. A fungus was isolated from dark brown lesions on diseased stems and roots. The pathogenicity of the isolated fungus was confirmed from inoculation experiments. On the basis of morphological features and EF-1α and β-tubulin sequences, the pathogen was identified as Fusarium oxysporum. This is the first report of F. oxysporum causing Fusarium wilt on common bean in Inner Mongolia. As a destructive plant pathogen, F. oxysporum has a wide host range and is distributed all over China, control measures should be taken in advance.
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Identification of Colletotrichum siamense causing anthracnose on medicinal Paeonia suffruticosa in Anhui
YANG Hongmei, SUN Zhenxin, WANG Tielin, HUANG Luqi, ZHANG Ruqin, YAN Honghai
Acta Phytopathologica Sinica    2023, 53 (1): 137-140.   DOI: 10.13926/j.cnki.apps.000794
Abstract （302）      PDF（pc） （2833KB）（455）       Save
From 2019 to 2021, the sample leaves with typical anthracnose disease symptoms were collected from tree peony (Paeonia suffruticosa Andr.) in Anhui of China. Thirty-one fungal isolates obtained from the diseased samples and the purified cultures exhibited the same morphological characteristics on potato sucrose agar. Pathogenicity tests for the representative AF614 revealed that that this fungus was the causal agent of the disease. Molecular analysis of the sequences for the internal transcribed spacer (ITS) regions of their rDNA and the partial DNA fragments of actin (ACT), β-tublin (TUB2), and chitin synthase (CHS) genes showed the orthologues with these of Colletotrichum siamense species. Combined with morphological and molecular characteristics, we named the fungal AF614 as Colletotrichum siamense. To the best of our knowledge, this is the first report of C. siamense causing anthracnose of tree peony in China.
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Identification of pathogen causing sugarcane brown stripe in Yunnan Province
WANG Xiaoyan, LI Yinhu, PU Jinan, ZHANG Rongyue, LONG Chaohua, PU Jiarong, LI Wenfeng, SHAN Hongli, HUANG Yingkun
Acta Phytopathologica Sinica    2023, 53 (1): 141-145.   DOI: 10.13926/j.cnki.apps.000637
Abstract （327）      PDF（pc） （2623KB）（584）       Save
In recent years, sugarcane brown stripe disease has spread across a large area in Yunnan province. To identify the pathogen causing sugarcane brown stripe, 58 diseased samples were collected from different areas and varieties. The pathogen causing sugarcane brown stripe was identified by morphological characteristics and sequences analysis of rDNA ITS region and GAPDH. The results showed that the pathogen was Bipolaris setariae, a new record species of sugarcane brown stripe in Yunnan, which provide scientific basis for epidemic prediction, disease-resistant breeding, and precise control of sugarcane brown stripe.
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Cited: CSCD(2)
Sequence diversity and evolutionary analysis of S6 segment of Rice black-streaked dwarf virus
WANG Xinyu, XU Zhennan, ZHOU Yu, WENG Jianfeng, GUO Changhong, LI Xinhai
Acta Phytopathologica Sinica    2023, 53 (1): 146-150.   DOI: 10.13926/j.cnki.apps.000634
Abstract （302）      PDF（pc） （3432KB）（392）       Save
In order to deeply understand the variation and evolution of RBSDV S6 segment, we analyzed the genetic variation of RBSDV S6 sequence in 26 maize and rice samples from 13 locations. The results showed that there were a large number of base mutations in the coding region, indicating the presence of variation and recombination between S6 sequences. At the same time, by analyzing the amino acids and codon usage preferences encoded by S6, we found that S6 population has other codon usage preferences in addition to GC3s, and the codon usage of this population is under the pressure of natural selection. The genetic recombination analysis showed that there were 2 recombination events (S6-M-13-ⅤM-2 and S6-M-13-ⅤM-3) in 26 sequences. Further, based on phylogenetic analysis, S6 sequences can be divided into four groups. The above results show that S6 segment plays an important role in the genetic variation and evolution of RBSDV population, which provides a theoretical basis for further research on the pathogenic mechanism of RBSDV S6 in maize rough dwarf disease and rice black-streaked dwarf disease.
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Viral pathogens detection of Passiflora edulis mosaic disease and sequence analysis of polyprotein gene of East Asian passiflora virus in some areas of South China
YE Ting, ZHENG Guohua, MENG Hongyan, ZHANG Yuehua, LAI Ruiyun, GUO Ying
Acta Phytopathologica Sinica    2023, 53 (1): 151-154.   DOI: 10.13926/j.cnki.apps.000632
Abstract （225）      PDF（pc） （2460KB）（339）       Save
Passiflora edulis mosaic disease is a serious constraint to the production of Passiflora edulis. in China. During 2016-2019, 170 samples of P. edulis with typical mosaic symptoms were collected from 7 planting areas in Fujian, Guangxi and Guangdong. Cucumber mosaic virus (CMV), Potyvirus and Tobamovirus were detected using RT-PCR with degenerate primers and/or specific primers. The results showed that 90 out of 170 samples were detected positive. The detection rate of CMV was 51.18% while the detection rate of Potyvirus was 10.00 %, however, Tobamovirus was not found in this research. CMV and Potyvirus showed severe mixed infection which account for 15.56% of total infection rate. Further identification of viruses in the genus Potyvirus was performed and the amplified fragment shared 98.00%-100.00% identity with East Asian passiflora virus aviliable in the GenBank. Additionally, 5 isolates with polyprotein gene sequences were obtained and phylogenetic analysis of these 5 isolates indicated a high genetic relationship to the AO stain reported from Japan and China (Taiwan). These results will be helpful for the detection and effective control of Passiflora edulis mosaic disease.
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Research advances on citrus leaf blotch virus
LIU Huan, LÜ Zhaorui, WU Wei, WU Yunfeng
Acta Phytopathologica Sinica    2023, 53 (2): 155-163.   DOI: 10.13926/j.cnki.apps.000801
Abstract （437）      PDF（pc） （5910KB）（571）       Save
Citrus leaf blotch virus (CLBV) was first discovered in California in the 1970s, and it has since been reported to infect various host plants in many countries. In 2016, CLBV was detected in kiwifruits for the first time in China, and then it was reported in sweet cherries, citruses, peonies, apples, mulberries and Nantian bamboos, and it is now widespread in many regions. Herein, the research progresses of CLBV, including those of the occurrence, distribution, transmission, damage, molecular variation, detection technology, genome characteristics and gene function of CLBV were reviewed, which will provide references for the study of antiviral breeding, synthetical prevention and pathogenic mechanisms of CLBV.
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Cited: CSCD(2)
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.   DOI: 10.13926/j.cnki.apps.001004
Abstract （307）      PDF（pc） （8408KB）（362）       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.
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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.   DOI: 10.13926/j.cnki.apps.000642
Abstract （244）      PDF（pc） （7485KB）（285）       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.
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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.   DOI: 10.13926/j.cnki.apps.000789
Abstract （305）      PDF（pc） （9296KB）（288）       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.
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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.   DOI: 10.13926/j.cnki.apps.000797
Abstract （234）      PDF（pc） （9190KB）（203）       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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Functional characterization of endocytic protein FgSyp1 in Fusarium graminearum
HAN Xueqin, DU Kaiyue, LI Qingyi, LI Xuenan, LÜ Xiang, SHANG Mingqing, ZHANG Li, YU Jinfeng, LIANG Yuancun
Acta Phytopathologica Sinica    2023, 53 (2): 207-216.   DOI: 10.13926/j.cnki.apps.001003
Abstract （241）      PDF（pc） （7724KB）（329）       Save
Endocytosis plays critical roles in cellular processes of nutrient uptake and signal transduction. Syp1 and Ede1 are early proteins in clathrin-mediated endocytosis. Fusarium head blight mainly caused by Fusarium graminearum seriously affects the yield and quality of wheat. F. graminearum contains one FgSYP1 that is highly homologous to Saccharomyces cerevisiae SYP1. Yeast two-hybrid showed that FgSyp1 interacted with F. graminearum FgEde1. The ΔSyp1 mutant and the ΔΔSyp1/Ede1 double mutant were obtained by homologous recombination, and the function of FgSyp1 was determined by experiments including hyphal growth, conidiation and pathogenicity. The results showed that FgSyp1 played an important role in hyphal growth, conidiation and pathogenicity of F. graminearum, and that the ΔΔSyp1/Ede1 mutant exhibited obvious defects in asexual and sexual reproduction compared with the ΔSyp1 mutant.
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Cited: CSCD(1)
Identification and functional analysis of sucrose transporter FgSut1 and FgSut2 in Fusarium graminearum
HUANG Panpan, ZHANG Yongkang, ZHOU Linrui, YANG Xinyi, ZHANG Xue
Acta Phytopathologica Sinica    2023, 53 (2): 217-228.   DOI: 10.13926/j.cnki.apps.000627
Abstract （235）      PDF（pc） （9140KB）（248）       Save
Fusarium head blight (FHB) is a devastating disease of wheat, causing yield losses and mycotoxin contamination in harvested grain. Fusarium graminearum is one of the predominant species causing FHB in wheat. Sugar transporters have not previously been identified in F. graminearum. Here, we identified two sucrose transporter genes FgSUT1 and FgSUT2 homologous to Schizosaccharomyces pombe Sut1. We obtained the Fgsut1, Fgsut2, and Fgsut1Fgsut2 double mutants. The phenotype analysis showed that the growth rate, conidiation, sexual reproduction and DON production were not affected in the Fgsut1, Fgsut2 and Fgsut1Fgsut2 double mutants. The growth rate of Fgsut1 was significantly reduced in plates with glucose as carbon source, while the Fgsut1Fgsut2 grew slower than Fgsut1. In addition, the growth rate of Fgsut1Fgsut2 also significantly reduced when fructose or maltose was used as the carbon source. These results suggested that FgSut1 and FgSut2 were functionally redundant in carbon source utilization, but they had different roles in glucose utilization. The virulence of Fgsut2 and Fgsut1Fgsut2 was decreased on wheat spikelets and coleoptiles, indicating FgSut2 may be important for virulence, and FgSut1 and FgSut2 had redundant function in fungal pathogenicity. This study established a relationship between F. graminearum carbon source utilization, toxin synthesis and pathogenicity, and enriched the research on the carbon source utilization mechanism of F. graminearum.
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The flavin-dependent brominase XanJ is involved in the biosynthesis of Xanthomonas yellow pigment xanthomonadin
ZHENG Zhelin, CAO Xueqiang, HU Wenda, SONG Kai, HAO Xiangrui, ZHANG Hongyan, HE Yawen
Acta Phytopathologica Sinica    2023, 53 (2): 229-244.   DOI: 10.13926/j.cnki.apps.000639
Abstract （234）      PDF（pc） （12531KB）（167）       Save
The phytopathogenic Xanthomonas can infect approximately 400 plant species, causing serious economic losses in agriculture. One of the most important characteristics of Xanthomonads is to produce xanthomonadin, a kind of protective yellow pigment. Xanthomonadins are phospholipids with di-brominated aryl-polyene, however, mechanisms underlying xanthomonadin bromination remain unclear. In this study, the function of xanJ, one of the key genes required for xanthomonadin biosynthesis in Xanthomoas campestris pv. campestris strain XC1 and Xanthomoas oryzae pv. oryzae strain PXO99A, was investigated using bioinformatics, genetic, biochemical and pathological methods. XanJ protein contains a flavin adenine dinucleotide (FAD)-binding domain and a tryptophan halogenase domain, and harbors GXGXXG and WXWXIP motifs, suggesting that this protein is a FAD-dependent phenolic halogenase. Deletion of xanJ abolished the biosynthesis of dibrominated aryl-polyene. However, the xanJ deletion mutant strain still produces low levels of non-brominated xanthomonadin. No accumulation of 3-hydroxybenzoic acid and 4-hydroxybenzoic acid, the precursors of xanthomonadin biosynthesis, was observed in the xanJ deletion mutant strains. Overexpression of the phenolic halogenases encoding genes ab10, bmp5, aerJ or mcnD, which encoding phenolic halogenases, in the xanJ deletion mutant strains failed to restore the biosynthesis of di-brominated xanthomonadin. Additionally, deletion of xanJ had no effects on exopolysaccharide biosynthesis, extracellular endoglucanase, protease activities and virulence on cabbage. These findings indicate that XanJ is a FAD-dependent brominase most likely involved in bromination of ß-carbon of aryl-polyene and the elongation of polyene chain in xanthomonadin biosynthesis.
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Identification of WRKY group Ⅲ family and the response to Plasmodiophora Brassicae and hormone in turnip
LEI Ting, LIU Ying, ZHOU Xiaoqing, HUI Maixia, ZHAO Limin
Acta Phytopathologica Sinica    2023, 53 (2): 245-257.   DOI: 10.13926/j.cnki.apps.000800
Abstract （203）      PDF（pc） （13099KB）（145）       Save
The WRKY transcription factors are a class of plant-specific transcription factor family. Studies have shown that many genes in the WRKY group III family are involved in defense response of pathogenic bacteria and other biotic stress. To study potential function of turnip WRKY group Ⅲ family genes in clubroot disease resistance, 21 WRKY genes were identified from the turnip genome using bioinformatics analysis and were distributed on the nine chromosomes. These WRKY proteins contain 258-916 amino acid residues with motif quantity ranging from 0-8, and most of them contain a motif structure with the sequence 4-5-7/(8-2)-1-3. Ana-lysis of cis-acting elements in promoter regions of WRKY group Ⅲ family gene found that there were a large number of light, hormone and stress responsive elements. Among hormone-responsive elements, Me-JA had the largest number. qPCR analysis showed that WRKY group Ⅲ family genes could respond to the induction of Plasmodiophora brassicae. Among them, BrWRKY46-2 and BrWRKY70-2 showed opposite patterns of up- and down-regulated expression in resistant and susceptible materials, respectively. The expression levels of BrWRKY46-2 and three BrWRKY70s were significantly down-regulated after exogenous SA induction, and were significantly down-regulated after MeJA induction. Subcellular localization showed that two genes, BrWRKY46-2 and BrWRKY70-2, mainly located in the nucleus. The WRKY group Ⅲ family genes in turnip were widely involved in the response process under P. brassicae stress. BrWRKY46-2 and BrWRKY70-2 genes were closely related to the resistance and susceptibility of turnip varieties and were induced by SA and JA. The above results imply that WRKY transcription factors have regulatory effects on clubroot resistance, and can be used as candidate genes for further needed on disease resistance and functional analysis.
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Cited: CSCD(1)
Characteristics of the MYB transcription factors family and their responses to RGSV and RRSV infection in rice
XU Rongrong, CHANG Qing, WEI Ying, LI Xia, WU Jianguo, ZHAO Shanshan
Acta Phytopathologica Sinica    2023, 53 (2): 258-266.   DOI: 10.13926/j.cnki.apps.000636
Abstract （308）      PDF（pc） （8756KB）（223）       Save
As one of the largest transcription factor families in plants, MYB transcription factors family play important roles in regulating plant growth and development and resisting to stress. However, the role of OsMYBs in rice antiviral pathway is still unclear. In order to explore the roles of OsMYBs family in rice antiviral pathway, we first analyzed their structure and characteristics, as well as the expression profiles of single-stranded RNA virus rice grassy stunt virus (RGSV) and double-stranded RNA virus rice ragged stunt virus (RRSV)-infected rice published by Mohammed et al in 2015, and found that 51 of the OsMYBs family genes responded to RGSV and 50 of them responded to RRSV. Further analysis revealed that 26 OsMYBs responded to both RGSV and RRSV, and 10 of them with relatively high expression and multiple differences were selected for real-time fluorescence quantitative PCR (qRT-PCR) verification. Our results showed that the expression of OsMYB30, LOC_Os02g42850, LOC_Os02g47744, LOC_Os05g37050, LOC_Os06g43090 and LOC_Os08g33150 was down-regulated and the expression of LOC_Os05g38460 was up-regulated upon RGSV and RRSV infection, respectively. Moreover, the expression of LOC_Os05g51160 and LOC_Os09g26170 was up-regulated after RGSV infection while down-regulated after RRSV infection; Additionally, the expression of LOC_Os08g05520 was down-regulated after RGSV infection while up-regulated after RRSV infection. Seven out of these ten OsMYBs were identical in expression variation trend after infection of these two viruses, suggesting that these OsMYBs may have a broad-spectrum response to rice virus infection, which lays a foundation for future research on the function of OsMYBs during rice-virus interactions.
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Cited: CSCD(1)
The biological function of Bxy-egl-30 in regulating early-stage development of Bursaphelenchus xylophilus
CHEN Xi, LIU Wenyi, ZHOU Lifeng, GUO Kai, YU Hongshi, HU Jiafu
Acta Phytopathologica Sinica    2023, 53 (2): 267-276.   DOI: 10.13926/j.cnki.apps.000624
Abstract （217）      PDF（pc） （10579KB）（157）       Save
Rapid development and efficient mating are the basis of population expansion and disease epidemic of B. xylophilus. Bioinformatics analysis, in situ hybridization and RNA interference were used to investigate the potential role of Bxy-egl-30 in the growth and development of B. xylophilus. The cloned coding region of the Bxy-egl-30 gene has a length of 1 128 bp, encoding 375 amino acids, and the phylogenetic tree analysis showed that Bxy-egl-30 belonged to the EGL-30 protein subunit family. The results of in situ hybridization showed that the expression of Bxy-egl-30 was specific in different developmental stages of B. xylophilus, and it was widely expressed in the gonads, gut and nearby body wall cells, during embryonic and larval stages, the gene is expressed in the vulva muscle of females and the gonad of males. The results of RT-qPCR showed that the expression level of Bxy-egl-30 reached the highest level at the second larval stage, followed by the embryonic stage, and in the third instar stage, the fourth instar stage, and the adult stage, the expression level of Bxy-egl-30 decreased in turn. Knockdown of Bxy-egl-30 decreased the hatching rate of pinewood nematode embryos by 10.88%, and the activity of the second and third instar larvae decreased obviously, which the head swing frequency decreased about 4 times per minute. The results of the inoculation experiment showed that the pathogenicity of nematodes was weakened by Bxy-egl-30 RNAi and the death time of trees was slowed down. The death time of pine trees in the interference group was 9 days later than that in the control group. This study investigated the spatiotemporal dynamic expression patterns and biological functions of the Bxy-egl-30 gene at different deve-lopmental stages of B. xylophilus, and revealed the important role of the gene in the movement and development of B. xylophilus.
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Brassica napus miR6030 negatively regulates resistance to Sclerotinia sclerotiorum via targeting two genes encoding CC-NBS-LRR proteins
YANG Yuhan, CAO Jiayi, CAI Xinzhong
Acta Phytopathologica Sinica    2023, 53 (2): 277-286.   DOI: 10.13926/j.cnki.apps.000500
Abstract （226）      PDF（pc） （7672KB）（179）       Save
The role of miRNAs in regulating resistance to Sclerotinia sclerotiorum in oilseed rape (Brassica napus) is largely unknown. In this study, functions and mechanisms of miR6030 in resistance to S. sclerotiorum were analyzed. The results showed that miR6030 was expressed differentially in various tissues, higher in roots and stems while lowest in flowers. Silencing and transient over-expression of miR6030 in oilseed rape plants demonstrated that miR6030 negatively regulates resistance to S. sclerotiorum. The degradome analysis and GUS staining assay revealed that miR6030 targets BnaCnng64090D and BnaA09g10520D, which encode CC-NBS-LRR (CNL) proteins and positively regulate resistance to S. sclerotiorum. All the results indicated that miR6030 negatively regulates resistance to S. sclerotiorum via cleaving transcripts of the two CNL protein-encoding genes BnaCnng64090D and BnaA09g10520D. This study provides some insights into functions of miR6030 and molecular mechanisms underlying the resistance to S. sclerotiorum and new strategy for molecular breeding for the resistance of Brassica napus to S. sclerotiorum.
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Analysis of tomato yellow leaf curl virus resistance genes in different tomato varieties and field resistance evaluation
ZHANG Aihong, YANG Fei, LI Xiwang, ZHAO Yuanye, CHEN Wei, DI Dianping
Acta Phytopathologica Sinica    2023, 53 (2): 287-295.   DOI: 10.13926/j.cnki.apps.000640
Abstract （221）      PDF（pc） （4999KB）（236）       Save
In order to analyze the resistant genotypes and field application of different cultivars, tomato yellow leaf curl virus (TYLCV) resistance genes Ty-1,Ty-2, Ty-3 and Ty-3a were tested from 43 tomato cultivars through molecular marker technology. At the same time, field natural identification methods were used to verify the resistance level of different genotypes. Results based on the molecular detection showed that 31 tested cultivars contained Ty genes, which accounted for 72.09% of the total varieties; During detection of the resistant genes, Ty-1 has the highest detection rate of 65.12%, which showed that Ty-1 was the most widely used gene for tomato virus disease resistance breeding. 10 resistance genotypes were detected in all tested cultivars which contain 1-3 resistance genes respectively. The resistance performance from high to low was: Ty-1/ty-1+Ty-2/ty-2+Ty-3a/ty-3、Ty-1/Ty-1+Ty-2/ty-2+Ty-3a/ty-3a、Ty-1/Ty-1、Ty-1/ty-1+Ty-3/ty-3、 Ty-1/ty-1+Ty-3a/ty-3a、Ty-1/Ty-1+Ty-3a/ty-3a、Ty-3a/ty-3a、Ty-1/ty-1+Ty-2/ty-2、Ty-1/ty-1 and Ty-2/ty-2, respectively. Resistance of the materials showed medium resistance or susceptibility when it carried one resistant gene, whereas materials carrying two or more resistant genes had more stable resistant levels. The analysis of variance showed that the mate-rials which carried 3 resistance genes had significantly higher resistance than that of Ty-1 or Ty-2 heterozygous disease resistance, but there were no significant differences between the loci with 2 resistance genes and single gene locus. The results of identification showed that there were 6 varieties with high resistance, 9 varieties showed resistance, 9 varieties showed medium resistance and 13 varieties showed susceptibility to TYLCV. The incidence of TYLCV in the identification nurseries was 58.03%, and tomato chlorosis virus (ToCV) was 4.67%. The resistance was significantly reduced after the co-infection with TYLCV and ToCV. The phenomenon implied that complex infection of TYLCV and ToCV may have potential adverse effects on the resistance genes. The above research results provide a theoretical basis for the application of disease resistance genes.
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Resistance risk assessment of mefentrifluconazole against Alternaria alternata from ginseng in China
ZHANG Jiayi, FENG Zhiwei, HOU Wanpeng, LU Baohui, REN Guangyong, SUN Guogang, GAO Jie
Acta Phytopathologica Sinica    2023, 53 (2): 296-306.   DOI: 10.13926/j.cnki.apps.000796
Abstract （233）      PDF（pc） （7541KB）（170）       Save
In order to understand the resistance risk of Alternaria alternata to mefentrifluconazole, four mutant strains with different resistance levels were obtained by chemical domestication in this study. The resistance risk of A. alternata was evaluated by detecting genetic stability, biological characteristics and cross resistance between mefentrifluconazole and other fungicides. The results showed that the high resistance mutant had high fitness and stable inheritance. Once it appears, it is at risk of becoming a dominant strain in the field. There was no positive cross resistance between the resistance of mefentrifluconazole resistant mutants to mefentrifluconazole and the resistance of other five fungicides, azoxystrobin, kersonxim-methyl, difenoconazole, pyraclostrobin, and hexaconazole. A. alternata can develop mefentrifluconazole-resistant mutants under the fungicide pressure, and there is a risk of fungicide resis-tance. The results of this study provide a scientific basis for the reasonable application of mefentrifluconazole for the control of ginseng Alternaria leaf blight.
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Cited: CSCD(3)
Sensitivity of Fusarium pseudograminis to epoxiconazole in Henan Province
HOU Ying, XIN Hewen, ZHANG Xin, FAN Xinnuo, LIU Shengming, XU Jianqiang
Acta Phytopathologica Sinica    2023, 53 (2): 307-316.   DOI: 10.13926/j.cnki.apps.000792
Abstract （335）      PDF（pc） （7117KB）（248）       Save
In order to detect the sensitivity of Fusarium pseudograminearum to epoxiconazole in Henan Pro-vince, 100 isolates were collected from 16 counties in 2019 and their sensitivity to epoxiconazole (EC50) were conducted by mycelial growth rate method. The results indicated that the EC50 of the tested isolates ranged from 0.002 4 to 0.505 4 μg·mL-1, and the average value was (0.109 3±0.095 7) μg·mL-1. The sensitivity frequency distribution showed that EC50 values of the 89 tested isolates were lied in a main peak; the average value of these isolates (0.082 1±0.046 8) μg·mL-1 could be used as the sensitivity baseline of F. pseudograminearum to epoxiconazole. The results of variance analysis showed that there was little difference in the sensitivity of isolates from different counties to epoxiconazole with average EC50 ranged from 0.050 8 to 0.296 8 μg·mL-1. However, the sensitivities were varied for the isolates from the same county, and the highest EC50 value was 210.58 times higher than that of the lowest isolates in Yucheng from Shangqiu. The results of cluster analysis showed that there was no obvious correlation between the sensitivity and the geographical origin of the isolates. There was no significant correlation between the toxicity of epoxiconazole and pyraclostrobin, fluoxonil, difenoconazole, carbendazim, phenamacril, tebuconazole. Seed dressing with epoxiconazole hindered wheat growth and inhibited root length, plant height and fresh weight. Control efficacy in greenhouse test illustrated that spraying 12.5% epoxiconazole suspension with active ingredient at 150 μL·mL-1had the highest control efficacy of 60.18%. In vitro test, 12.5% epoxiconazole suspension not only had the highest control efficacy (79.78%) when the active ingredient at 150 μL·mL-1, but had no effect on the growth of wheat when applied. The results of this study provided a theoretical basis for the application of epoxiconazole to control wheat stem rot and a basis for monitoring the sensitivity of the pathogen to epoxiconazole.
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Cited: CSCD(2)
Pathogen identification of the Corynespora leaf spot on Salvia miltiorrhiza Bge. in Henan
WANG Fei, LI Xuemeng, GAO Suxia, WEN Yi, QIN Yanhong, LIU Yuxia, YANG Jin, QI Wenping, LU Chuantao
Acta Phytopathologica Sinica    2023, 53 (2): 317-320.   DOI: 10.13926/j.cnki.apps.000811
Abstract （236）      PDF（pc） （2829KB）（275）       Save
From 2019 to 2021, a new leaf spot disease was detected on Salvia miltiorrhiza Bge. in Henan province. The leaf spot disease could be found in all 12 investigated fields, and the rate of diseased plants was 20% to 70%. The disease spot was round or oval, gray white in the center, and appeared perforation at later stages which eventually led to leaf wilt by meeting together with multiple spots. The pathogen was isolated and purified by conventional tissue isolation, and was verified according to Koch′s rule. Based on morphological observation and rDNA-ITS, EF-1α and β-tubulin gene sequence analysis, isolate DS1 was identified as Corynespora cassiicola, the causal agent of the new leaf spot disease. This is the first report of leaf spot disease on S. miltiorrhiza caused by C. cassiicola in China, which will provide theoretical basis and technical support for the effective control of the disease in the planting and production of S. miltiorrhiza in the future.
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Cited: CSCD(1)
Pestalotiopsis hispanica,a new pathogenic fungus responsible for Cryptomeria japonica red blight
FAN Shaobin, LIU Xiangguo, SU Jiyu, HU Yanping, JIA Yue, LIANG Xinyu, WANG Zonghua, HU Hongli
Acta Phytopathologica Sinica    2023, 53 (2): 321-325.   DOI: 10.13926/j.cnki.apps.000815
Abstract （391）      PDF（pc） （4534KB）（246）       Save
Recently, the needles of Cryptomeria japonica in parks at Fuzhou have suffered from red blight seriously, and this situation is even getting worse. Four diseased needle samples of C. japonica were collected from different parks around. Four Pestalotiopsis strains were obtained after isolation and purification. Based on morphological characters in couple with phylogenetic analyses of sequences of ITS, β-tubulin and tef1, all the strains were identified as Pestalotiopsis hispanica. The results of Koch′s postulates for the represent isolate LS-1 on the needles of healthy C. japonica confirmed P. hispanica as the pathogen of the disease. This is the first report of P. hispanica on C. japonica, even in China.
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Identification of the pathogen causing leaf sheath rot on Curcuma wenyujin
LI Jing, QIU Fang, XIE Chang-ping, LI Xi, ZHANG Chao
Acta Phytopathologica Sinica    2023, 53 (2): 326-329.   DOI: 10.13926/j.cnki.apps.000798
Abstract （268）      PDF（pc） （3488KB）（217）       Save
As a famous medicinal plant, the growth and quality of Curcuma wenyujin can be affected by leaf sheath rot disease, which severely occurs in Dongshan plantations, Haikou city, Hainan province of China in 2017. Based on morphological traits and phylogenetic analyses of multi-loci (ITS, Cox I and Cox II), the pathogen isolated from the diseased plants was identified as Pythium myriotylum. The results provide a theoretical basis for the field control of leaf sheath rot disease on C. wenyujin.
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Identification of a Ganoderma stem rot pathogen infecting Dimocarpus longana
LIU Zhiyu, LI Zengping, ZHANG Yu, WANG Xiaoyu
Acta Phytopathologica Sinica    2023, 53 (2): 330-334.   DOI: 10.13926/j.cnki.apps.000814
Abstract （407）      PDF（pc） （4517KB）（271）       Save
A Ganoderma stem rot disease was found to infect Dimocarpus longana in 2021 in Binqiao town of Guangxi province. In order to clarify the pathogen of this disease, ten isolates with the same morphology were isolated from basidiocarps by using aconventional tissue separation method. LYJF001, a representative strain with excellent growth, was selected for pathogenicity test,morphological identification, sequence analysis of rDNA-ITS and SSU, and phylogenetic tree construction. The results showed that the pathogen causing the stem rot of D. longana was Ganoderma australe.
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Identification of bacterial leaf spot pathogens of tomato caused by Erwinia persicina
CHEN Ruxing, XIE Xuewen, SHI Yanxia, CHAI Ali, FAN Tengfei, LI Lei, LI Baoju
Acta Phytopathologica Sinica    2023, 53 (2): 335-337.   DOI: 10.13926/j.cnki.apps.000638
Abstract （296）      PDF（pc） （2745KB）（387）       Save
Bacterial leaf spot of tomato is a worldwide disease, which seriously affects the quality and yield of tomato because of its diverse routes of infestation, rapid spread and difficulty in control. To identify the pathogen of the disease, samples of tomato with leaf spots were collected and prepared for pathogen analysis. Five representative bacterial strains FQ21051401-FQ21051405 strains were isolated and purified from diseased tomato plants. Pathogenicity tests and re-isolation and re-identification of the bacteria were performed to confirm the isolate and fulfill the Koch′ postulates. Based on morphological and biochemical characteristics, phylogenetic analysis, and Koch′s postulates, the bacterial isolates were identified as Erwinia persicina. The aim of this study was to isolate and identify the causal agent of bacterial leaf spot disease of tomato, which will provide a better understanding of pathogenesis of the E. persicina and provided a scientific basis for its control.
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Identification of pathogenic species of garlic virus diseases in Shandong Province
XIAO Li, CHENG Chao, SHI Zhaopeng, SUN Xiaohui, SUN Zuowen, WANG Shusen, ZHU Xiaoping
Acta Phytopathologica Sinica    2023, 53 (2): 338-342.   DOI: 10.13926/j.cnki.apps.000641
Abstract （295）      PDF（pc） （2290KB）（321）       Save
In order to clarify the occurrence and distribution of garlic virus in Shandong Province, 382 garlic samples were collected in 2019. PCR was used for virus detection with specific primers. Eleven viruses were detected: leek yellow stripe virus (LYSV), onion yellow dwarf virus (OYDV), garlic common latent virus (GCLV), shallot latent virus (SLV), garlic virus A (GarV-A), garlic virus B (GarV-B), garlic virus D (GarV-D), garlic virus E (GarV-E), garlic virus X (GarV-X), milk vetch dwarf virus (MDV)and tobacco mosaic virus (TMV). The co-infection rate of the viruses was 44.24%, and there was co-infection of six kinds of viruses at most. Phylogenetic trees were constructed with CP of LYSV and OYDV. LYSV isolates were closely related to the Chinese, Turkish and Mexican isolates. OYDV isolates were closely related to the Chinese, Indian and Polish isolates.
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