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Characterization of Bean Common Mosaic Virus Isolates Infecting Three Leguminous Bean Crops from South and Southeast Asia. 感染南亚和东南亚三种豆类作物的豆类普通花叶病毒分离物的特征。
IF 5.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-18 DOI: 10.1094/PDIS-05-24-1113-SR
Shimul Das, Sridhar Jarugula, Basavaraj Bagewadi, Amer Fayad, Alexander V Karasev, Rayapati A Naidu

Bean common mosaic virus (BCMV) is causing economically important diseases in leguminous crops worldwide. In this study, BCMV isolates from country bean (CB; Lablab purpureus), yard-long bean (YLB; Vigna unguiculata), and rajma bean (RB; Phaseolus vulgaris) collected from Bangladesh, Nepal, and Cambodia were characterized. Samples that tested positive for BCMV in serological assays were subjected to high-throughput sequencing to generate near-full-length genome sequences. In pairwise comparisons of the polyprotein open reading frame, 13 BCMV isolates from Bangladesh, Cambodia, and Nepal showed sequence identity of 92.1 to 98.8% at the nucleotide and 94.2 to 99% at the amino acid level among themselves and with corresponding sequences of BCMV reported previously. In phylogenetic analyses using the global BCMV sequences, they segregated into five distinct lineages, with RB isolates from Nepal clustering with US1/NL1-clade of common bean isolates from different countries, YLB isolates aligning with blackeye cowpea strain sequences reported from China, and CB isolates from Nepal and Bangladesh clustering with soybean isolates from China. One YLB isolate from Nepal was identified as a putative recombinant. None of the BCMV sequences aligned with isolates representing the RU1 or PStV clades. In grow-out tests, seed samples from local markets showed a 14.3 to 38.1% transmission efficiency rate of BCMV with CB seed lots and 9.5 to 33.3% with YLB seed lots.

豆类普通花叶病毒(BCMV)在全球豆类作物中引起重要的经济病害。本研究对从孟加拉国、尼泊尔和柬埔寨采集的菜豆(CB,Lablab purpureus)、长豆(YLB,Vigna unguiculata)和蚕豆(RB,Phaseolus vulgaris)中分离出的 BCMV 进行了鉴定。对血清学检测中BCMV呈阳性的样本进行了高通量测序,以生成接近全长的基因组序列。在多聚蛋白开放阅读框的配对比较中,来自孟加拉国、柬埔寨和尼泊尔的 13 个 BCMV 分离物的核苷酸序列同一性为 92.1%-98.8%,氨基酸序列同一性为 94.2%-99%,与之前报道的 BCMV 相应序列的同一性为 92.1%-98.8%。在使用全球 BCMV 序列进行的系统进化分析中,它们分离成五个不同的系,尼泊尔的 RB 分离物与来自不同国家的普通豆分离物 US1/NL1 支系聚类,YLB 分离物与中国报道的黑眼豇豆株序列一致,尼泊尔和孟加拉国的 CB 分离物与中国的大豆分离物聚类。尼泊尔的一个 YLB 分离物被鉴定为假定重组株。没有一个 BCMV 序列与代表 RU1 或 PStV 支系的分离物一致。在生长试验中,来自当地市场的种子样本显示,CB种子批次的BCMV传播效率为14.3%至38.1%,YLB种子批次的传播效率为9.5%至33.3%。
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引用次数: 0
A Multiplex RT-qPCR Assay for Simultaneous Detection of Cucurbit Viruses from Individual Whitefly and Plant Samples. 从粉虱个体和植物样本中同时检测葫芦病毒的多重 RT-qPCR 分析法。
IF 5.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-18 DOI: 10.1094/PDIS-09-23-1964-RE
A Abdul Kader Jailani, Mathews L Paret

Whiteflies (Bemisia tabaci) are a significant pest of cucurbits and vector many viruses, leading to substantial economic losses. Modern diagnostic tools offer the potential for early detection of viruses in the whiteflies before crop production. One such tool is the multiplex reverse transcriptase quantitative PCR (RT-qPCR) probe-based technique, which can detect multiple targets in a single reaction and simultaneously quantify the levels of each target, with a detection limit of 100 copies per target. In this study, a multiplex RT-qPCR-based detection system capable of identifying one DNA virus and three RNA viruses in whiteflies-cucurbit leaf crumple virus (CuLCrV), cucurbit chlorotic yellows virus (CCYV), cucurbit yellow stunting disorder virus (CYSDV), and squash vein yellowing virus (SqVYV)-was developed. To ensure the reliability of the assay, an internal gene control as the fifth target to monitor false-negative results was incorporated. This newly developed molecular diagnostic tool possesses several advantages. It can detect up to five desired targets from a single whitefly RNA sample, even at concentrations as low as 1 ng/μl. To evaluate its sensitivity, we conducted experiments using serially diluted cloned plasmids and in vitro transcribed RNA transcripts of the target viruses. We also assessed the specificity of the assay by including aphid-transmitted viruses and other viruses known to infect cucurbits. The diagnostic method successfully detected all five targets simultaneously and allowed for the quantification of up to 100 copies using a mixture of healthy RNA and in vitro transcribed RNA. Our aim with this study was to develop a highly specific and sensitive one-step multiplex RT-qPCR system for the simultaneous detection of viruses transmitted by whiteflies in cucurbits. This system offers significant advantages for early detection, enabling prompt control measures to mitigate the further spread of viral infections and reduce yield losses. Additionally, we demonstrated the ability to simultaneously detect mixed viruses (CCYV, CYSDV, CuLCrV, and SqVYV) in individual whiteflies and quantify the number of viral copies carried by each whitefly. The multiplex RT-qPCR assay outperforms currently available techniques for detecting many samples at a given time and can be effectively utilized for early monitoring of plant viruses in individual whiteflies and symptomless plants.

粉虱(Bemisia tabaci)是葫芦科植物的重要害虫,它传播多种病毒,造成重大经济损失。现代诊断工具为在作物生产前及早检测粉虱体内的病毒提供了可能。其中一种工具是基于探针的多重反转录酶定量 PCR(RT-qPCR)技术,该技术可在一次反应中检测多个靶标,并同时量化每个靶标的水平,每个靶标的检测限为 100 个拷贝。本研究开发了一种基于 RT-qPCR 的多重检测系统,该系统能够鉴定粉虱中的一种 DNA 病毒和三种 RNA 病毒:葫芦皱叶病毒 (CuLCrV)、葫芦萎黄病病毒 (CCYV)、葫芦黄矮病病毒 (CYSDV) 和南瓜叶脉黄化病毒 (SqVYV)。为确保检测的可靠性,还加入了一个内部基因对照作为监测假阴性结果的第五个靶标。这种新开发的分子诊断工具具有多项优势。即使浓度低至 1 纳克/微升,它也能从一份粉虱 RNA 样品中检测到多达五个所需的靶标。为了评估其灵敏度,我们使用经过系列稀释的克隆质粒和体外转录的目标病毒 RNA 转录本进行了实验。我们还将蚜虫传播的病毒和其他已知可感染葫芦科植物的病毒纳入检测范围,从而评估了该检测方法的特异性。该诊断方法成功地同时检测了所有五个目标病毒,并利用健康病毒 RNA 和体外转录病毒 RNA 的混合物对多达 100 个拷贝的病毒进行了定量。RNA 和体外转录 RNA 的混合物,定量检测多达 100 个拷贝。我们这项研究的目的是开发一种高特异性、高灵敏度的一步法多重 RT-qPCR 系统,用于同时检测由葫芦粉虱传播的病毒。该系统在早期检测方面具有显著优势,能够及时采取控制措施,减少病毒感染的进一步传播,降低产量损失。此外,我们还展示了同时检测单个粉虱体内混合病毒(CCYV、CYSDV、CuLCrV 和 SqVYV)并量化每只粉虱携带病毒拷贝数的能力。这种多重 RT-qPCR 检测方法在给定时间内检测多个样本方面优于现有技术,可有效用于早期监测单个粉虱和无症状植物中的植物病毒。
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引用次数: 0
Pathotype Identification and Host Resistance Evaluation of Clubroot in Zhejiang Province, China. 中国浙江省马尾松病的病原型鉴定与寄主抗性评价
IF 5.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-18 DOI: 10.1094/PDIS-12-23-2748-SR
Yi Zhang, Wenfeng Gong, Yingying Gao, Kun Zhao, Fangzhan Wang, Yapei Liu, Mei Zhang, Xiaolin Yu

Clubroot, caused by Plasmodiophora brassicae, is a globally destructive soilborne disease affecting cruciferous plants. In this study, the predominant pathotypes of P. brassicae in six cities within Zhejiang Province were identified using the Williams and European clubroot differential (ECD) systems. A phylogenetic analysis of P. brassicae isolates infecting cruciferous crops worldwide was conducted using MEGA, and their ITS2 secondary structures were predicted through the ITS2 database. Accessions of Brassica rapa, B. oleracea, B. juncea, and Eruca sativa Mill. were employed to assess clubroot resistance. The results revealed that the prevalent pathotypes in Zhejiang Province were pathotype 1, ECD20/31/12 and ECD24/16/30; pathotype 3, ECD20/15/4; pathotype 8, ECD16/0/0 and ECD24/0/0; and pathotype 2, ECD16/15/15. Isolates from distinct genera of Brassicaceae formed separate branches in the evolutionary tree. Moreover, isolates of Brassica crops from Zhejiang Province exhibited homology with those from other global regions, a finding corroborated by their ITS2 secondary structure. Approximately 80 and 95% of B. rapa and B. juncea crops displayed susceptible phenotypes for pathotype 8, ECD16/0/0, whereas approximately 60% of B. oleracea crops exhibited resistance. Furthermore, three Brassica crop accessions showed significant variation in resistance to the pathogen, both among morphological and geographical origin groups. This study contributes to understanding the distribution of diverse P. brassicae pathotypes in different regions of Zhejiang Province and facilitates the identification of Brassica crops with potential disease resistance suitable for cultivation in the province.

由黄铜疫霉菌(Plasmodiophora brassicae)引起的马铃薯病是一种影响十字花科植物的全球性破坏性土传病害。在此,我们利用威廉姆斯和欧洲铜锈病差异(ECD)系统鉴定了浙江省六个城市中铜锈病菌的主要病型。利用 MEGA 对全球感染十字花科作物的黄铜病菌分离株进行了系统进化分析,并通过 ITS2 数据库预测了它们的 ITS2 二级结构。采用 B. rapa、B. oleracea、B. juncea 和 Eruca sativa Mill.结果表明,浙江省流行的病原型为病原型 1(ECD20/31/12 和 ECD24/16/30)、病原型 3(ECD20/15/4)、病原型 8(ECD16/0/0 和 ECD24/0/0)和病原型 2(ECD16/15/15)。来自不同十字花科属的分离物在进化树中形成了不同的分支。此外,浙江省芸苔属作物的分离物与全球其他地区的分离物具有同源性,其 ITS2 二级结构也证实了这一发现。约 80% 和 95% 的 B. rapa 和 B. juncea 作物对病原型 8(ECD16/0/0)表现出易感表型,而约 60% 的 B. oleracea 作物表现出抗性。此外,三种芸苔属作物品种对病原体的抗性在形态组和地理起源组之间都有显著差异。这项研究有助于了解不同黄花芸苔属病原菌在浙江省不同地区的分布情况,并有助于鉴定适合该省种植的具有潜在抗病性的芸苔属作物。
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引用次数: 0
Genetic Analysis of Stripe Rust Resistance in the Chinese Wheat Cultivar Luomai 163. 中国小麦栽培品种洛麦 163 抗条锈病遗传分析。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-15 DOI: 10.1094/PDIS-06-24-1195-RE
Zimeng Li, Chan Yuan, Shunda Li, Yu Zhang, Bin Bai, Fangping Yang, Pengpeng Liu, Wei Sang, Yong Ren, Ravi Singh, Pingan Liao, Caixia Lan

Stripe or yellow rust (YR) caused by Puccinia striiformis tritici (Pst) is an important foliar disease affecting wheat production globally. Resistant varieties are the most economically and environmentally effective way to manage this disease. The common winter wheat (Triticum aestivum L.) cultivar Luomai 163 exhibited resistance to the Pst races CYR32 and CYR33 at the seedling stage and showed a high level of adult plant resistance in the field. To understand the genetic basis of YR resistance in this cultivar, 142 F5 recombinant inbred lines (RILs) derived from cross Apav#1 × LM163 and both parents were genotyped with the 16K SNP array and bulked segregant analysis sequencing. The analysis detected a major gene, YrLM163, at the seedling stage associated with the 1BL.1RS translocation. Additionally, three genes for resistance at the adult plant stage were detected on chromosome arms 1BL (Lr46/Yr29/Pm39/Sr58), 6BS, and 6BL in Luomai 163, whereas Apav#1 contributed resistance at a quantitative trait locus (QTL) on 2BL. These QTL explained YR disease severity variations ranging from 6.9 to 54.8%. The kompetitive allele-specific PCR (KASP) markers KASP-2BL, KASP-6BS, and KASP-6BL for the three novel loci QYr.hzau-2BL, QYr.hzau-6BS, and QYr.hzau-6BL were developed and validated. QYr.hzau-1BL, QYr.hzau-2BL, and QYr.hzau-6BS showed varying degrees of resistance to YR when present individually or in combination based on genotype and phenotype analysis of a panel of 570 wheat accessions. Six RILs combining resistance alleles of all QTL, showing higher resistance to YR in the field than Luomai 163 with disease severities of 10.7 to 16.0%, are important germplasm resources for breeding programs to develop YR-resistant wheat varieties with good agronomic traits.

由三尖杉条锈病菌(Pst)引起的条锈病或黄锈病(YR)是影响全球小麦生产的一种重要叶面病害。抗病品种是管理这种病害最经济、最环保的方法。普通冬小麦(Triticum aestivum L.)栽培品种鲁麦 163 在幼苗期就表现出对 Pst CYR32 和 CYR33 的抗性,在田间成株抗性水平也很高。为了解该栽培品种抗 YR 的遗传基础,利用 16K SNP 阵列和大量分离分析测序(BSA-Seq)对 Apav#1 × LM163 杂交育成的 142 个 F5 重组近交系(RIL)进行了基因分型。分析结果发现,幼苗期的一个主要基因 YrLM163 与 1BL.1RS 易位相关。此外,在罗麦 163 的染色体臂 1BL(Lr46/Yr29/Pm39/SR58)、6BS 和 6BL 上检测到三个成株期抗性基因,而 Apav#1 则在 2BL 上的 QTL 上产生抗性。这些 QTL 解释了 6.9% 至 54.8% 的 YR 病害严重程度变化。针对三个新基因座 QYr.hzau-2BL、QYr.hzau-6BS 和 QYr.hzau-6BL 开发并验证了 KASP 标记 KASP-2BL、KASP-6BS 和 KASP-6BL。根据对570个小麦品种的基因型和表型分析,QYr.hzau-1BL、QYr.hzau-2BL和QYr.hzau-6BS单独存在或组合存在时对YR表现出不同程度的抗性。6个RIL结合了所有QTL的抗性等位基因,在田间表现出比洛麦163更高的抗YR能力,病害严重程度为10.7-16.0%,是育种计划中培育具有良好农艺性状的抗YR小麦品种的重要种质资源。
{"title":"Genetic Analysis of Stripe Rust Resistance in the Chinese Wheat Cultivar Luomai 163.","authors":"Zimeng Li, Chan Yuan, Shunda Li, Yu Zhang, Bin Bai, Fangping Yang, Pengpeng Liu, Wei Sang, Yong Ren, Ravi Singh, Pingan Liao, Caixia Lan","doi":"10.1094/PDIS-06-24-1195-RE","DOIUrl":"10.1094/PDIS-06-24-1195-RE","url":null,"abstract":"<p><p>Stripe or yellow rust (YR) caused by <i>Puccinia striiformis tritici</i> (<i>Pst</i>) is an important foliar disease affecting wheat production globally. Resistant varieties are the most economically and environmentally effective way to manage this disease. The common winter wheat (<i>Triticum aestivum</i> L.) cultivar Luomai 163 exhibited resistance to the <i>Pst</i> races CYR32 and CYR33 at the seedling stage and showed a high level of adult plant resistance in the field. To understand the genetic basis of YR resistance in this cultivar, 142 F<sub>5</sub> recombinant inbred lines (RILs) derived from cross Apav#1 × LM163 and both parents were genotyped with the 16K SNP array and bulked segregant analysis sequencing. The analysis detected a major gene, <i>YrLM163</i>, at the seedling stage associated with the 1BL.1RS translocation. Additionally, three genes for resistance at the adult plant stage were detected on chromosome arms 1BL (<i>Lr46/Yr29/Pm39/Sr58</i>), 6BS, and 6BL in Luomai 163, whereas Apav#1 contributed resistance at a quantitative trait locus (QTL) on 2BL. These QTL explained YR disease severity variations ranging from 6.9 to 54.8%. The kompetitive allele-specific PCR (KASP) markers <i>KASP-2BL</i>, <i>KASP-6BS</i>, and <i>KASP-6BL</i> for the three novel loci <i>QYr.hzau-2BL</i>, <i>QYr.hzau-6BS</i>, and <i>QYr.hzau-6BL</i> were developed and validated. <i>QYr.hzau-1BL</i>, <i>QYr.hzau-2BL</i>, and <i>QYr.hzau-6BS</i> showed varying degrees of resistance to YR when present individually or in combination based on genotype and phenotype analysis of a panel of 570 wheat accessions. Six RILs combining resistance alleles of all QTL, showing higher resistance to YR in the field than Luomai 163 with disease severities of 10.7 to 16.0%, are important germplasm resources for breeding programs to develop YR-resistant wheat varieties with good agronomic traits.</p>","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":"PDIS06241195RE"},"PeriodicalIF":4.4,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141760236","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Identification, Pathogenicity, and Fungicide Sensitivity of Colletotrichum Species Associated with Anthracnose on Italian Ryegrass in Southwestern China. 中国西南地区意大利黑麦草炭疽病相关 Colletotrichum 菌种的鉴定、致病性和杀菌剂敏感性。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-14 DOI: 10.1094/PDIS-01-24-0206-RE
Lingling Xu, Zhiting Xu, Jiaqi Liu, Huawei Cui, Jiahui Long, Longhai Xue, Chunjie Li

Italian ryegrass (Lolium multiflorum L.) is widely cultivated as an important forage grass worldwide because of its high nutritional value and good palatability. Anthracnose caused by Colletotrichum species was a common and new emerging disease of Italian ryegrass. In this study, 88 Colletotrichum isolates were collected from diseased leaves of Italian ryegrass planting regions in Sichuan, Chongqing, and Guizhou provinces of southwestern China between 2019 and 2022. By the pure culture technique, 15 representative single-spore isolates were obtained for further study. Multilocus phylogenetic analysis coupled with morphological features showed that these isolates were finally identified as six new record species: C. cereale of the C. graminicola species complex, C. fioriniae and C. nymphaeae of the C. acutatum species complex, C. boninense and C. citricola of the C. boninense species complex, and C. nageiae. Pathogenicity tests indicated that all species could induce anthracnose symptoms; of these, C. cereale was more invasive than other species, followed by C. fioriniae, C. nageiae, C. citricola, and C. boninense; C. nymphaeae was the weakest pathogenic species to Italian ryegrass plants (P ≤ 0.05). Fungicide sensitivity assays showed that iprodione, propineb, and oxime·tebuconazole had strong inhibitory effects on the mycelial growth of six Colletotrichum species; in addition, azoxystrobin and fludioxonil also significantly inhibited the mycelial growth of C. nymphaeae and C. fioriniae, respectively. These results provide the basis for the diagnosis and detection in the field, pathogen identification, and management of anthracnose on Italian ryegrass.

意大利黑麦草(Lolium multiflorum L.)营养价值高、适口性好,是世界上广泛种植的重要牧草。由 Colletotrichum 菌引起的炭疽病是意大利黑麦草常见的一种新出现的病害。本研究在 2019 年至 2022 年期间,从中国西南部四川、重庆和贵州省的意大利黑麦草种植区的病叶中收集了 88 株 Colletotrichum 分离物。通过纯培养技术,获得了 15 个具有代表性的单孢分离株,用于进一步研究。多焦点系统进化分析结合形态特征表明,这些分离物最终被鉴定为 6 个新记录种:C.boninense和C. citricola,以及C. nageiae。致病性测试表明,所有物种都能诱发炭疽病症状;其中 C. cereale 的侵染性比其他物种强,其次是 C. fioriniae、C. nageiae、C. citricola 和 C. boninense;C. nymphae 对意大利黑麦草植物的致病性最弱(P ≤ 0.05)。杀菌剂敏感性试验表明,异丙苯啶、丙环唑和肟菌酯对六种 Colletotrichum 的菌丝生长有很强的抑制作用;此外,唑螨酯和氟啶虫酰胺也分别对 C. nymphaeae 和 C. fioriniae 的菌丝生长有显著的抑制作用。这些结果为意大利黑麦草炭疽病的田间诊断和检测、病原体鉴定和管理提供了依据。
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引用次数: 0
First report of paprika as a natural host plant for tomato chlorosis virus in Korea. 在韩国首次报告辣椒是番茄萎黄病病毒的天然宿主植物。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-14 DOI: 10.1094/PDIS-08-24-1697-PDN
Hae-Ryun Kwak, Dong-Wan Kang, Hee-Seong Byun, Bongchoon Lee, Inyoung Han
<p><p>Paprika (<i>Capsicum annuum</i> var. grossum (L.) Sendtn.), also called bell pepper or sweet pepper, is a valuable greenhouse crop that is mostly consumed as fresh fruit in Korea. In 2022, it was cultivated on 726 ha in Korea and 82,042 tons were produced (KOSIS, 2023). In April 2023, interveinal chlorosis and yellowing were observed on the lower and middle leaves of greenhouse paprika in Jinju and Haman, South Korea, and many whiteflies (<i>Bemisia tabaci</i>) were present. The disease incidence was 30-40% on five farms (approximately 4.5 ha) in the two areas. The typical symptoms and presence of whiteflies in greenhouses indicated potential infection by the whitefly-transmitted crinivirus, tomato chlorosis virus (ToCV). Six samples collected from symptomatic plants were examined by leaf dip using transmission electron microscopy (TEM) and revealed filamentous virus particles about 800 nm long. To confirm the TEM results, six symptomatic paprika leaf samples were subject to RT-PCR using the specific primers ToCV-M-4F and ToCV-M-4R to detect ToCV (Choi, 2023). All tested samples were positive for ToCV. To confirm the presence of other viruses and obtain the complete genome sequences, one of the six ToCV-positive samples (ToCV-PAP-JJ6) was subject to high-throughput sequencing (HTS). Total RNA was extracted from symptomatic leaves using the RNeasy Plant Mini Kit (QIAGEN, Germany) and a transcriptome library was generated using the TruSeq Stranded Total RNA LT Sample Prep Kit (Illumina, San Diego, CA, USA) according to standard protocols. HTS was performed on an Illumina NovaSeq 6000 system (Macrogen, Korea). De novo transcriptome assembly of the 85,668,854 reads with Trinity software (r20140717) yielded 817,303 contigs of 201 to 38,987 nucleotides (nt). BLASTn and BLASTx analysis of the contigs against the NCBI viral reference database showed that three large contigs were virus sequences, two of which were homologous to ToCV-RNA1 and -RNA2 and one to bell pepper alphaendornavirus (BPEV). The ToCV-RNA1 (genome 8,594 nt, 11,212 mapped reads, mean read coverage 196.2 times) and ToCV-RNA2 (genome 8,242 nt, 40,122 mapped reads, mean read coverage 734.9 times) contigs showed 100% coverage and 99% base pair matching (8,583/8,594 and 8,233/8,242, respectively) with known genome sequences of ToCV-RNA1 (MG813908) and ToCV-RNA2 (KP114534) isolated from tomato in Korea (Lee et al. 2018). The sequences were deposited in GenBank as isolates ToCV-PAP-JJ6 RNA1 (OR865222) and ToCV-PAP-JJ6 RNA2 (OR865223). The BPEV contig (genome 14,728 nt, 2,928,519 mapped reads, mean read coverage 15,012.4 times) was detected at very high read depth and RT-PCR for BPEV confirmed that all six samples were coinfected with ToCV. BPEV was recently reported in paprika and red pepper in Korea (Jo et al., 2022). The BPEV-infected pepper cultivars have been reported not to produce any symptoms (Escalante & Valverde 2019). ToCV was first reported in tomato greenhouses in Korea in 20
辣椒(Capsicum annuum var.2022 年,韩国的种植面积为 726 公顷,产量为 82 042 吨(KOSIS,2023 年)。2023 年 4 月,在韩国晋州和咸安,温室红辣椒的中下部叶片出现叶脉间枯萎和黄化现象,并有许多粉虱(Bemisia tabaci)。这两个地区的五个农场(约 4.5 公顷)的发病率为 30-40%。温室中的典型症状和粉虱的存在表明可能感染了由粉虱传播的番茄萎黄病病毒(ToCV)。利用透射电子显微镜(TEM)对从有症状植株上采集的六个样本进行了叶片浸渍检查,发现了长约 800 nm 的丝状病毒颗粒。为了证实透射电子显微镜的结果,使用特异引物 ToCV-M-4F 和 ToCV-M-4R 对六个有症状的辣椒叶片样本进行了 RT-PCR 检测(Choi,2023 年)。所有检测样本对 ToCV 均呈阳性。为了确认是否存在其他病毒并获得完整的基因组序列,对六个 ToCV 阳性样本中的一个(ToCV-PAP-JJ6)进行了高通量测序(HTS)。使用 RNeasy Plant Mini Kit(QIAGEN,德国)从有症状的叶片中提取总 RNA,并按照标准协议使用 TruSeq Stranded Total RNA LT Sample Prep Kit(Illumina,San Diego, CA, USA)生成转录组文库。HTS 在 Illumina NovaSeq 6000 系统(Macrogen,韩国)上进行。使用 Trinity 软件(r20140717)对 85,668,854 个读数进行从头转录组组装,得到 817,303 个 201 至 38,987 个核苷酸(nt)的等位组。对照 NCBI 病毒参考数据库对等位基因进行的 BLASTn 和 BLASTx 分析表明,有三个大的等位基因是病毒序列,其中两个与 ToCV-RNA1 和 -RNA2 同源,一个与甜椒α-内多角体病毒(BPEV)同源。ToCV-RNA1 (基因组 8,594 nt,11,212 个映射读数,平均读数覆盖率 196.2 倍)和 ToCV-RNA2 (基因组 8,242 nt,40,122 个映射读数,平均读数覆盖率 734.9倍)等位基因与已知的从韩国番茄中分离出来的ToCV-RNA1(MG813908)和ToCV-RNA2(KP114534)的基因组序列(Lee等人,2018年)显示出100%的覆盖率和99%的碱基对匹配率(分别为8,583/8,594和8,233/8,242)。这些序列作为分离物 ToCV-PAP-JJ6 RNA1 (OR865222) 和 ToCV-PAP-JJ6 RNA2 (OR865223) 保存在 GenBank 中。BPEV contig(基因组 14,728 nt,2,928,519 个映射读数,平均读数覆盖率 15,012.4 倍)以极高的读数深度被检测到,BPEV 的 RT-PCR 检测证实所有六个样本都同时感染了 ToCV。最近在韩国的辣椒和红辣椒中发现了 BPEV(Jo 等人,2022 年)。据报道,受 BPEV 感染的辣椒品种不会产生任何症状(Escalante & Valverde,2019 年)。ToCV 于 2013 年首次在韩国的番茄温室中被报道,与番茄黄叶卷曲病毒一起成为番茄作物中的问题病毒(Jo 等,2023 年;Lee 等,2018 年)。由于番茄温室中持续存在 ToCV、粉虱和天然杂草宿主(Kil 等,2015 年),ToCV 已扩散到其他作物,包括辣椒。据我们所知,这是韩国首次报道 ToCV 感染辣椒。需要在全国范围内开展研究,以防止 ToCV 的传播,这是对辣椒农场的一个新威胁。
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引用次数: 0
Virulence Determinants Are Unevenly Distributed Within Streptomyces Species and Strains Causing Potato Common Scab in the Province of Quebec, Canada. 在加拿大魁北克省,引起马铃薯普通疮痂病的链霉菌种类和菌株中,病毒决定因子分布不均。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-12 DOI: 10.1094/PDIS-04-24-0744-RE
Adrien Biessy, Mélanie Cadieux, Marie Ciotola, Renée St-Onge, Jochen Blom, Martin Filion

Potato common scab is an important bacterial disease afflicting potatoes around the world. Better knowledge of the local Streptomyces spp. populations causing this disease is key to developing durable control strategies. In this study, we isolated 230 Streptomyces strains from scab-infected potato tubers harvested from commercial potato fields located across the province of Quebec, Canada. The genetic diversity of this collection was first studied using repetitive element-based PCR fingerprinting, and the genomes of 36 representative strains were sequenced using PacBio's sequencing technology. This enabled us to identify the strains to the species level, to study the distribution of previously characterized virulence-associated genes and clusters, and to explore the repertoires of putative plant cell wall-degrading enzymes. In parallel, the virulence of the 36 strains was evaluated using a potato tuber slice assay. The diversity was higher than previously reported, as 11 phytopathogenic species were found across the province. Among them, S. scabiei and S. acidiscabies were the most abundant as well as the most virulent. Strains belonging to these two species harbored numerous virulence determinants, including the thaxtomin biosynthetic gene cluster. By contrast, most weakly virulent strains lacked this cluster but harbored at least one known virulence determinant. The results obtained suggest that a higher number of virulence-associated genes and clusters in the genome of phytopathogenic Streptomyces spp. are associated with greater virulence. This study contributes to increasing the publicly available genomic resources of scab-causing Streptomyces spp. and expands our knowledge on the diversity and virulence of this important bacterial pathogen.

马铃薯普通疮痂病是危害世界各地马铃薯的一种重要细菌性病害。更好地了解引起这种病害的当地链霉菌种群是制定持久控制策略的关键。在这项研究中,我们从加拿大魁北克省商业马铃薯田中收获的受疮痂病感染的马铃薯块茎中分离出 230 株链霉菌。我们首先利用基于重复元件的 PCR 指纹图谱研究了这些菌株的遗传多样性,然后利用 PacBio 测序技术对 36 个代表性菌株的基因组进行了测序。这使我们能够对菌株进行物种水平的鉴定,研究先前表征的毒力相关基因和基因簇的分布,并探索假定的植物细胞壁降解酶的复合物。同时,利用马铃薯块茎切片试验评估了 36 株菌株的毒力。与之前的报告相比,该省的植物病原菌多样性更高,在全省范围内发现了 11 种植物病原菌。其中,疥螨属(S. scabiei)和酸螨属(S. acidiscabies)数量最多,毒性最强。属于这两个物种的菌株含有大量毒力决定因子,包括噻吨胺生物合成基因簇。相比之下,大多数弱毒力菌株缺乏这种基因簇,但至少含有一种已知的毒力决定因子。研究结果表明,植物病原链霉菌属基因组中毒力相关基因和基因簇的数量越多,毒力越强。这项研究有助于增加可公开获得的致疮痂病链霉菌属基因组资源,扩大我们对这种重要细菌病原体的多样性和毒力的了解。
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引用次数: 0
Comparison of rDNA-ITS Sequences, Vegetative Compatibility, Biological Characteristics, and Thifluzamide Sensitivity of Rhizoctonia solani AG-3 PT, TM, and TB Subgroup Isolates Collected from Potato and Tobacco in China. 从中国马铃薯和烟草中采集的根瘤菌 AG-3 PT、TM 和 TB 亚群分离株的 rDNA-ITS 序列、植物相容性、生物学特征和对氟唑胺敏感性的比较。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-11 DOI: 10.1094/PDIS-07-24-1407-RE
Mengxiao Liu, Tianbo Liu, Yu Yang, Qian Li, Haiyan Mao, Xinchun Li, Fenglu Wang, Jingtao Zhao, Yuanhua Wu, Chong Zhang
<p><p>Potato black scurf and tobacco target spot are significant agricultural diseases primarily caused by <i>Rhizoctonia solani</i> AG-3, which is further classified into PT (potato type), TM (tomato type), and TB (tobacco type) subgroups, with the naming of these subgroups originally indicating their respective infection hosts potato, tomato, and tobacco. This study determined the subgroup classification of 129 <i>R. solani</i> AG-3 isolates recovered from tobacco leaves exhibiting target spot disease in major tobacco-cultivating regions of China, as well as 80 <i>R. solani</i> AG-3 isolates obtained from potato stem cankers and tuber sclerotia in Liaoning, Jilin, and Heilongjiang provinces, and Inner Mongolia autonomous region of China. Sequence analysis of the internal transcribed spacer (ITS) regions revealed that all potato isolates belonged to the <i>R. solani</i> AG-3 PT subgroup, whereas the tobacco isolates were classified into TM and TB subgroups. Specifically, 51 strains from the northeast (Liaoning, Jilin, and Heilongjiang provinces) belonged to the TM subgroup, while the others belonged to the TB subgroup. Phylogenetic analysis indicated a closer relationship between AG-3 TM and AG-3 PT compared to AG-3 TB. Population genetic analysis highlighted distinct genetic variations among the subgroups, with the AG-3 TB subgroup isolates exhibiting fewer ITS1 variable sites and the AG-3 PT subgroup displaying a higher number of ITS2 variable sites. Furthermore, comparisons were made regarding macroscopic vegetative interactions, biological characteristics, pathogenicity to potato and tobacco, and susceptibility to thifluzamide among the three subgroups. The results indicated no macroscopic somatic interactions among the subgroups, with the highest frequency of somatic incompatibility observed within AG-3 PT, followed by AG-3 TM, and AG-3 TB. Additionally, the three subgroups differed in colony color, sclerotium morphology, production time, and distribution location on potato-dextrose-agar (PDA) medium, with AG-3 TM being the least likely to produce sclerotia. Regarding temperature, optimal growth conditions varied among the subgroups. Regarding pathogenicity, the AG-3 PT subgroup strains were more pathogenic on tobacco and potato stem bases compared to the AG-3 TB subgroup, whereas the AG-3 TB and TM subgroup strains were more pathogenic on leaves. Notably, AG-3 TM exhibited remarkable virulence towards both the stem base and leaves. The AG-3 PT subgroup strains exhibited the highest susceptibility to thifluzamide, with a susceptibility baseline of 0.037±0.013 mg·L<sup>-1</sup>, followed by the AG-3 TM subgroup strains with a baseline of 0.111±0.034 mg·L<sup>-1</sup>, and the AG-3 TB subgroup strains with a baseline of 0.137±0.035 mg·L<sup>-1</sup>. This study contributes to a deeper understanding of the biology and etiology of the three subgroups of <i>R. solani</i> AG-3, providing a valuable theoretical basis for the scientific control and
马铃薯黑斑病和烟草靶斑病是主要由Rhizoctonia solani AG-3引起的重要农业病害,Rhizoctonia solani AG-3又分为PT(马铃薯型)亚群、TM(番茄型)亚群和TB(烟草型)亚群,这些亚群的命名最初是为了表示它们分别感染的寄主马铃薯、番茄和烟草。本研究确定了从中国主要烟草种植区表现靶斑病的烟叶中分离的 129 株 R. solani AG-3 以及从中国辽宁省、吉林省、黑龙江省和内蒙古自治区的马铃薯茎腐病和块茎硬皮病中分离的 80 株 R. solani AG-3 的亚群分类。内部转录间隔区(ITS)序列分析表明,所有马铃薯分离物均属于R. solani AG-3 PT亚群,而烟草分离物则分为TM和TB亚群。具体来说,来自东北(辽宁、吉林和黑龙江省)的 51 株属于 TM 亚群,而其他则属于 TB 亚群。系统发育分析表明,与 AG-3 TB 相比,AG-3 TM 和 AG-3 PT 之间的关系更为密切。群体遗传分析突出显示了亚群之间明显的遗传变异,AG-3 TB 亚群分离物的 ITS1 可变位点较少,而 AG-3 PT 亚群分离物的 ITS2 可变位点较多。此外,还对三个亚群之间的宏观无性繁殖相互作用、生物学特征、对马铃薯和烟草的致病性以及对氟胺的敏感性进行了比较。结果表明,各亚群之间没有宏观的体细胞互作,AG-3 PT 的体细胞不相容频率最高,其次是 AG-3 TM 和 AG-3 TB。此外,三个亚群在菌落颜色、硬菌丝形态、产生时间和在potato-dextrose-agar(PDA)培养基上的分布位置上也有差异,其中AG-3 TM最不容易产生硬菌丝。在温度方面,各亚群的最佳生长条件各不相同。在致病性方面,与 AG-3 TB 亚群相比,AG-3 PT 亚群菌株对烟草和马铃薯茎基部的致病性更强,而 AG-3 TB 和 TM 亚群菌株对叶片的致病性更强。值得注意的是,AG-3 TM 对茎基部和叶片都有显著的致病性。AG-3 PT 亚群菌株对氟虫酰胺的敏感性最高,其敏感性基线为 0.037±0.013 mg-L-1,其次是 AG-3 TM 亚群菌株,其敏感性基线为 0.111±0.034 mg-L-1,AG-3 TB 亚群菌株的敏感性基线为 0.137±0.035 mg-L-1。该研究有助于加深对R. solani AG-3三个亚群的生物学和病原学的认识,为我国烟草靶斑病和马铃薯黑斑病的科学防治提供有价值的理论依据。
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引用次数: 0
Etiology of Pine Ghost Canker in Southern California Urban Forests. 南加州城市森林中松树幽灵腐烂病的病因。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-11 DOI: 10.1094/PDIS-08-24-1718-SR
Marcelo I Bustamante, Karina Elfar, Carlos Carachure, Adam Adaskaveg, John N Kabashima, Christopher Shogren, Akif Eskalen, Shannon Colleen Lynch

Pine ghost canker is a recently described disease affecting multiple pine species in urban forests of Southern California. Symptoms include wedged cankers with irregular margins and cryptic discoloration on cross-sections of branches, which can lead to severe dieback and potentially tree death. In this study, we identified and characterized five Neofusicoccum species (N. luteum, N. mediterraneum, N. parvum, N. stellenboschianum, and N. vitifusiforme) as the primary etiological agents of pine ghost canker. These pathogens were consistently isolated from multiple symptomatic pine samples (n = 41) and identified by morphology and phylogenetic analyses using four DNA barcodes (rDNA ITS, tef1, tub2, and rpb2). Pathogenicity was confirmed on healthy branches of 15-year-old Monterey pines, where the five Neofusicoccum species, caused vascular lesions that were not significantly different in length. Secondary fungi (Diaporthe, Diplodia, Neopestalotiopsis, and Pestalotiopsis spp.) were also recovered from symptomatic tissues but did not cause vascular lesions in pathogenicity tests. The optimal temperature for mycelial growth of N. luteum and N. parvum was 30 °C, whereas for N. mediterraneum, N. stellenboschianum and N. vitifusiforme, it was 25 °C. All five species were able to resume growth at room temperature (20 °C) after showing no growth during a 7-day exposure to 5 °C and 40 °C. This study constitutes the first report of N. luteum, N. stellenboschianum, and N. vitifusiforme causing pine ghost canker in California. Environmental factors such as warmer temperatures, irrigation, and pest infestations are discussed as drivers of disease expression in pine trees. Management practices are also proposed.

松树幽灵腐烂病是最近描述的一种影响南加州城市森林中多个松树品种的疾病。症状包括边缘不规则的楔形溃疡和枝条横截面上的隐性变色,可导致严重枯死,甚至可能造成树木死亡。在这项研究中,我们确定并描述了五种 Neofusicoccum(N. luteum、N. mediterraneum、N. parvum、N. stellenboschianum 和 N. vitifusiforme)为松树鬼皮病的主要病原体。从多个有症状的松树样本(n = 41)中持续分离出了这些病原体,并通过形态学和使用四种 DNA 条形码(rDNA ITS、tef1、tub2 和 rpb2)进行的系统发育分析进行了鉴定。在 15 年树龄的蒙特雷松的健康枝条上证实了致病性,五种 Neofusicoccum 引起的维管病变在长度上没有明显差异。次生真菌(Diaporthe、Diplodia、Neopestalotiopsis 和 Pestalotiopsis spp.)也在有症状的组织中被发现,但在致病性试验中没有引起维管病变。N. luteum 和 N. parvum 菌丝生长的最适温度为 30 °C,而 N. mediterraneum、N. stellenboschianum 和 N. vitifusiforme 的最适温度为 25 °C。在 5 °C 和 40 °C 下暴露 7 天后,所有五个物种都能在室温(20 °C )下恢复生长。这项研究是首次报道 N. luteum、N. stellenboschianum 和 N. vitifusiforme 在加利福尼亚州引起松树鬼腐病。研究讨论了温度升高、灌溉和虫害等环境因素对松树病害表现的影响。此外还提出了一些管理措施。
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引用次数: 0
Pathogen Composition and Fungicide Sensitivity in Rice Spikelet Rot Disease. 水稻小穗腐烂病的病原体组成和杀菌剂敏感性
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-11 DOI: 10.1094/PDIS-09-24-1807-RE
Jiaqi Chen, Xiping Cao, Xiaolong Fu, Mingguo Zhou, Changjun Chen, Xiu-Shi Song

Rice spikelet rot disease (RSRD) is an emerging threat to rice crops, with sporadic but severe outbreaks in China in recent years. The composition of its pathogenic fungi has not been consistently identified, and chemical control methods remain unclear. This study aims to characterize the pathogen species responsible for RSRD and develop effective control strategies. Regional variations in disease symptoms and pathogen compositions were analyzed, which led to the identification of six novel pathogens, including Fusarium tanahbumbuense, Alternaria gaisen, Curvularia verruculosa, Curvularia brachyspora, Curvularia muehlenbeckiae, and Curvularia hominis. The pathogenic composition of RSRD exhibited considerable variation across different latitudes within China. Specifically, Alternaria spp. predominated in the Heilongjiang and Liaoning Provinces. Whereas Fusarium spp. and Curvularia spp. were more prevalent in the Hainan and Fujian Provinces. In contrast, Fusarium spp. and Alternaria spp. were the dominant pathogens in the Anhui and Jiangsu Provinces. Furthermore, an assessment of the sensitivity of these predominant pathogens to four chemical compounds was conducted, which led to the identification of potential fungicides for effective disease control. This research provides valuable insights into the pathogenic profile of RSRD across different regions and offers strategic recommendations for fungicide-based management of the disease.

水稻小穗腐烂病(RSRD)是一种新出现的威胁水稻作物的病害,近年来在中国零星但严重地爆发。其病原真菌的组成尚未得到一致确认,化学防治方法也尚不明确。本研究旨在确定导致 RSRD 的病原菌种类,并制定有效的防治策略。通过分析病害症状和病原菌组成的地区差异,确定了六种新型病原菌,包括 Fusarium tanahbumbuense、Alternaria gaisen、Curvularia verruculosa、Curvularia brachyspora、Curvularia muehlenbeckiae 和 Curvularia hominis。在中国不同纬度地区,RSRD 的病原组成有很大差异。具体来说,黑龙江省和辽宁省主要是交链孢属。而镰刀菌属和卷曲镰刀菌属在海南省和福建省更为普遍。相比之下,安徽省和江苏省的主要病原菌是镰刀菌属和交替孢属。此外,还评估了这些主要病原菌对四种化合物的敏感性,从而确定了有效控制病害的潜在杀菌剂。这项研究为了解不同地区 RSRD 的病原特征提供了宝贵的见解,并为基于杀菌剂的病害管理提供了战略建议。
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Plant disease
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