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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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引用次数: 0
First detection of alfalfa mosaic virus in Ficus benjamina in Chile. 智利首次在榕属植物中发现苜蓿花叶病毒。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-11 DOI: 10.1094/PDIS-12-23-2655-PDN
Elizabeth Peña, Ursula Steinfort, Belén Concha, Inés Marlene Rosales

Ficus benjamina, commonly known as ficus or weeping fig, is an ornamental tree from Southeast Asia in the Moraceae family. It is an evergreen species grown as an interior plant worldwide, prized for its glossy green leaves, gray stem, and multiple branches. F. benjamina can be pruned and kept as a small shrub when grown indoors, with most commercially available cultivars in the ornamental industry derived from sports and propagated cuttings (Chen et al., 2010). In October 2022, symptoms indicative of alfalfa mosaic virus (AMV) infection, such as intense bright yellow spots, mottling and chlorosis were observed in leaves of F. benjamina growing in office environments in Santiago, Chile (supplementary Fig.1). Seven symptomatic and three asymptomatic plants were sampled and traced back to the same plant nursery, and only leaf tissue samples from the symptomatic plants tested positive for the presence of the AMV using enzyme-linked immunosorbent assay (ELISA) (Agdia Inc., Elkhart, IN). These results were verified using reverse transcription (RT)-PCR with specific primers AMV-F and AMV-R, targeting a region of the AMV CP gene (Xu and Nie, 2006). AMV was detected in all symptomatic leaf samples, producing 351-bp amplicons in the RT-PCR assay for all samples that were positive to AMV by ELISA. No amplification product was observed when seronegative samples or non-template control was used as templates in the RT-PCR assays. Three RT-PCR amplicons were directly sequenced in both directions. BLAST analysis of these sequences showed 100% nucleotide sequence identity to an AMV isolate previously reported (GenBank Accession No. KX458469), confirming the detection of AMV in F. benjamina. This pathogen causes disease in several species, including ornamentals plant such as Viburnum tinus L. (Peña et al., 2011), peony (Belardi and Rubies, 2003), phlox (Holcomb et al., 2006), and weeds such as Cayratia japonica (Thunb.) Gagnep, Justicia procumbens L. and Veronica persica Poir. (Gao et al., 2020). F. benjamina is among the most popular ornamental indoor plants in Chile, so nurseries should ensure AMV-free plants to restrict disease incidence and prevent the spread of this virus. However, Chile lacks specific regulations for ornamental nurseries, making adherence to pest and disease recommendations voluntary, and only imported plant material requires a sanitary certificate.

无花果(Ficus benjamina),俗称榕树或垂枝无花果,是一种来自东南亚的桑科观赏树种。它是一种常绿树种,世界各地都将其作为室内植物种植,因其有光泽的绿叶、灰色茎干和多枝而备受青睐。在室内种植时,F. benjamina 可以被修剪成小灌木,观赏植物行业中的大多数商业栽培品种都来自运动和扦插繁殖(Chen 等人,2010 年)。2022 年 10 月,在智利圣地亚哥办公室环境中生长的 F. benjamina 的叶片上观察到了苜蓿花叶病毒(AMV)感染的症状,如强烈的亮黄色斑点、斑驳和萎黄病(补充图 1)。采用酶联免疫吸附测定法(ELISA)(Agdia Inc.)使用针对 AMV CP 基因区域的特异引物 AMV-F 和 AMV-R 进行反转录 (RT)-PCR 检测,验证了这些结果(Xu 和 Nie,2006 年)。在所有有症状的叶片样本中都检测到了 AMV,在 RT-PCR 检测中,所有经 ELISA 检测对 AMV 呈阳性的样本都产生了 351-bp 的扩增子。当血清阴性样本或非模板对照被用作 RT-PCR 检测的模板时,没有观察到扩增产物。对三个 RT-PCR 扩增子进行了双向直接测序。对这些序列的 BLAST 分析表明,它们与之前报道的一个 AMV 分离物(GenBank 编号 KX458469)的核苷酸序列相同度为 100%,这证实在 F. benjamina 中检测到了 AMV。这种病原体会导致多个物种发病,包括观赏植物,如 Viburnum tinus L.(Pena 等人,2011 年)、牡丹(Belardi 和 Rubies,2003 年)、凤仙花(Holcomb 等人,2006 年),以及杂草,如 Cayratia japonica (Thunb.) Gagnep、Justicia procumbens L.和 Veronica persica Poir.(Gao 等人,2020 年)。F. benjamina 是智利最受欢迎的室内观赏植物之一,因此苗圃应确保植物不含 AMV,以限制疾病的发生并防止病毒的传播。然而,智利缺乏针对观赏苗圃的具体法规,因此遵守病虫害建议属于自愿行为,只有进口植物材料才需要卫生证书。
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引用次数: 0
Virulence and Diversity of Puccinia striiformis f. sp. tritici Populations in Northwestern and Southwestern Oversummering Regions of China. 中国西北和西南越夏地区条斑赤霉病菌(Puccinia striiformis f. sp. tritici)种群的致病力和多样性。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-11 DOI: 10.1094/PDIS-09-24-1901-RE
Jie Wang, Xinyun Liu, Mengjie Ma, Yongjin Jin, Liyao Fu, Mingju Li, Lili Huang, Zhensheng Kang, Gangming Zhan

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is the most devastating fungal disease affecting wheat in China, particularly in the northwestern (Qinghai, Gansu, and Ningxia) and southwestern (Sichuan, Yunnan, and Guizhou) oversummering regions. In this study, 302 isolates collected from these six provinces were analyzed using virulence testing and single-nucleotide polymorphism (SNP) markers. Virulence testing on 19 Chinese differentials identified 38 previously reported and 49 novel races. CYR34 was the most prevalent race in both the northwestern and southwestern oversummering regions, with frequencies of 24.00% and 21.05%, respectively. In the northwestern oversummering region, the prevalence of CYR32 (10.70%) and G22-14 (10.67%) was nearly identical, whereas in the southwestern oversummering region, CYR32 (13.20%) and G22-14 (9.87%) exhibited significant differences. Additionally, 112 races were identified using 18 Yr single-gene differentials, although none of the isolates showed virulence to either Yr5 or Yr15. The virulence diversity of Pst populations was greater in the southwestern than in the northwestern oversummering region, as indicated by the differential sets. Further molecular analysis with 20 pairs of polymorphic KASP-SNP markers confirmed that the southwestern Pst population exhibited higher genetic diversity than the northwestern population. Both virulence phenotypes and genotype data demonstrated that the Pst population was more diverse in the southwestern oversummering region, despite presence of the exchange of Pst sources between regions. In addition, mutant isolates indicating resistance to tiadimefon fungicide were detected in both regions. This study provides a systematic comparison of Pst populations in the northwestern and southwestern oversummering regions based on virulence phenotyping and SNP marker genotyping, and the findings are crucial for the integrated management of wheat stripe rust in these regions and across the country.

由条锈病菌(Puccinia striiformis f. sp. tritici,Pst)引起的条锈病是影响中国小麦的最具毁灭性的真菌病害,尤其是在西北(青海、甘肃和宁夏)和西南(四川、云南和贵州)越夏地区。本研究利用毒力测试和单核苷酸多态性(SNP)标记对从这六个省份收集的 302 株分离株进行了分析。通过对 19 个中国差异株进行毒力测试,确定了 38 个以前报道过的菌种和 49 个新菌种。在西北和西南越夏地区,CYR34 是最流行的种族,频率分别为 24.00% 和 21.05%。在西北越夏区,CYR32(10.70%)和 G22-14(10.67%)的流行率几乎相同,而在西南越夏区,CYR32(13.20%)和 G22-14(9.87%)则表现出显著差异。此外,利用 18 个 Yr 单基因差异鉴定出 112 个品系,但没有一个分离物对 Yr5 或 Yr15 有毒力。如差异集所示,西南越夏区 Pst 群体的毒力多样性大于西北越夏区。利用 20 对多态 KASP-SNP 标记进行的进一步分子分析证实,西南部 Pst 群体的遗传多样性高于西北部群体。毒力表型和基因型数据都表明,尽管不同地区之间存在 Pst 来源交换,但西南越夏地区的 Pst 群体更具多样性。此外,在这两个地区都发现了对噻螨酮杀菌剂具有抗性的突变分离株。这项研究基于毒力表型和SNP标记基因分型对西北和西南越夏区的Pst种群进行了系统比较,研究结果对这些地区乃至全国的小麦条锈病综合治理至关重要。
{"title":"Virulence and Diversity of <i>Puccinia striiformis</i> f. sp. <i>tritici</i> Populations in Northwestern and Southwestern Oversummering Regions of China.","authors":"Jie Wang, Xinyun Liu, Mengjie Ma, Yongjin Jin, Liyao Fu, Mingju Li, Lili Huang, Zhensheng Kang, Gangming Zhan","doi":"10.1094/PDIS-09-24-1901-RE","DOIUrl":"https://doi.org/10.1094/PDIS-09-24-1901-RE","url":null,"abstract":"<p><p>Stripe rust, caused by <i>Puccinia striiformis</i> f. sp. <i>tritici</i> (<i>Pst</i>), is the most devastating fungal disease affecting wheat in China, particularly in the northwestern (Qinghai, Gansu, and Ningxia) and southwestern (Sichuan, Yunnan, and Guizhou) oversummering regions. In this study, 302 isolates collected from these six provinces were analyzed using virulence testing and single-nucleotide polymorphism (SNP) markers. Virulence testing on 19 Chinese differentials identified 38 previously reported and 49 novel races. CYR34 was the most prevalent race in both the northwestern and southwestern oversummering regions, with frequencies of 24.00% and 21.05%, respectively. In the northwestern oversummering region, the prevalence of CYR32 (10.70%) and G22-14 (10.67%) was nearly identical, whereas in the southwestern oversummering region, CYR32 (13.20%) and G22-14 (9.87%) exhibited significant differences. Additionally, 112 races were identified using 18 <i>Yr</i> single-gene differentials, although none of the isolates showed virulence to either <i>Yr5</i> or <i>Yr15</i>. The virulence diversity of <i>Pst</i> populations was greater in the southwestern than in the northwestern oversummering region, as indicated by the differential sets. Further molecular analysis with 20 pairs of polymorphic KASP-SNP markers confirmed that the southwestern <i>Pst</i> population exhibited higher genetic diversity than the northwestern population. Both virulence phenotypes and genotype data demonstrated that the <i>Pst</i> population was more diverse in the southwestern oversummering region, despite presence of the exchange of <i>Pst</i> sources between regions. In addition, mutant isolates indicating resistance to tiadimefon fungicide were detected in both regions. This study provides a systematic comparison of <i>Pst</i> populations in the northwestern and southwestern oversummering regions based on virulence phenotyping and SNP marker genotyping, and the findings are crucial for the integrated management of wheat stripe rust in these regions and across the country.</p>","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2024-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142625755","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
Characterization of the species of Pythium associated with corn seedlings in Delaware. 特拉华州与玉米幼苗相关的 Pythium 菌种特征。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-11 DOI: 10.1094/PDIS-04-24-0876-RE
John Bickel, Madeline Grace Henrickson, Alyssa K Betts

Corn (Zea mays L.) is the top grain crop by hectares grown in Delaware (DE). Increased pre- and post-emergence damping-off in corn caused by Pythium species have been observed in recent seasons. To date, no characterization studies have been performed to understand Pythium species diversity in DE. Symptomatic corn seedlings were collected from 33 fields across DE in 2019 and 2020. Isolates were obtained from infected root tissues and identified by molecular sequencing. In total, 14 species were recovered: P. aristosporum, P. arrhenomanes, P. attrantheridium, P. catenulatum, P. dissotocum, P. graminicola, P. inflatum, P. irregulare, P. oligandrum, P. periplocum, P.spinosum, P. sylvaticum, P. tardicrescens, and P. torulosum. The dominant species recovered was P. graminicola, accounting for 66% of the isolates. Five species were screened for fungicide sensitivity to calculate the EC50 of mefenoxam, ethaboxam, and picarbutrazox which are the most commonly used active ingredients to manage species of Pythium in corn seed treatment packages. All species tested were sensitive to mefenoxam and picarbutrazox. For ethaboxam, P. torulosum was the only species with an EC50 value > 10 µg ml 1. Ten species were screened for fungicide sensitivity to mefenoxam, ethaboxam, or picarbutrazox at 18, 21, and 25ºC. As temperatures increased, percent inhibition was reduced in 40, 70, and 80% of the species screened for sensitivity to ethaboxam, picarbutrazox, and mefenoxam, respectively. Nine of these species were screened for isolate aggressiveness in greenhouse panels to examine impact on emergence, shoot height, shoot weight, and root weight. While all species were recovered from inoculated roots, P. arrhenomanes, P. attrantheridium, P. spinosum, and P. sylvaticum had the greatest root weight reduction.

玉米(Zea mays L.)是特拉华州(DE)种植面积最大的粮食作物。最近几季观察到,由 Pythium 物种引起的玉米萌发前和萌发后脱粒现象有所增加。迄今为止,还没有进行过特征研究来了解特拉华州的 Pythium 种类多样性。2019 年和 2020 年,我们从德克萨斯州的 33 块田地中采集了有症状的玉米幼苗。从受感染的根部组织中获得分离株,并通过分子测序进行鉴定。共发现 14 个物种:P. aristosporum、P. arrhenomanes、P. attrantheridium、P. catenulatum、P. dissotocum、P. graminicola、P. inflatum、P. irregulare、P. oligandrum、P. periplocum、P.spinosum、P. sylvaticum、P. tardicrescens 和 P. torulosum。回收的主要菌种是禾本科菌,占分离菌种的 66%。为了计算甲霜灵、乙霜灵和吡蚜酮的半数致死浓度(EC50),对五种菌种进行了杀菌剂敏感性筛选。所有测试物种都对甲螨嗪和吡蚜酮敏感。对于乙草胺,P. torulosum 是唯一 EC50 值大于 10 µg ml 1 的物种。在 18、21 和 25ºC 温度条件下,对 10 个物种进行了杀菌剂敏感性筛选。随着温度的升高,分别有 40%、70% 和 80% 的物种对乙草胺、吡蚜酮和甲霜灵的抑制率降低。在温室面板中对其中九个物种的分离侵染性进行了筛选,以检查其对出苗、芽高、芽重和根重的影响。虽然所有菌种都能从接种的根部恢复,但 P. arrhenomanes、P. attrantheridium、P. spinosum 和 P. sylvaticum 的根重降低幅度最大。
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引用次数: 0
First report of Curvularia chiangmaiensis causing leaf spot in elephant grass in Brazil. 首次报告 Curvularia chiangmaiensis 在巴西引起象草叶斑病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-11 DOI: 10.1094/PDIS-05-24-0960-PDN
Flávia Fernandes Ribeiro Miranda, Marcio Akio Ootani, Raynne Barbosa Santos Rabelo, Eliane Regina Archangelo, Jonahtan Chaves Melo, Raimundo Wagner de Souza Aguiar, Eugenio Eduardo Oliveira, Gil R Santos

Elephant grass, Cenchrus purpureus, exhibits high efficiency in sequesting atmospheric CO2 during photosynthesis, leading to significant biomass production. As a C4 plant, it holds immense potential for alternative biodiesel production and serves as fresh feed for cattle (Negawo et al. 2018). Field observations in commercial pastures planted with elephant grass, located in the Brazilian cerrado, revealed necrotic leaf spots with a brownish center and a yellowish halo, reaching a severity of 30 to 50% between 2022 and 2023 in Palmas (10°24'08"S 48°21'45" W) and Gurupi (11°46'21"S 49°02'08" W), municipalities located in the state of Tocantins, Brazil. In both years, the field incidence of the disease on young and mature leaves approached 100% during the rainy season. Affected leaves were collected, packed in a thermal box, and taken to the phytopathology laboratory. The 198 leaves were sanitised in 1% hypochlorite for 30 seconds and washed thrice in sterile water. Upon sanitisation, leaf tissues were cut and cultured on potato dextrose agar (PDA) medium and incubated in a controlled chamber (BOD) at 28°C with a 12-hour photoperiod. Colonies exhibited a greyish colour with whitish edges, and the top plate was brownish, growing to a diameter of 5 cm over seven days. The colonies obtained from monosporic cultures formed curved conidia, predominantly with three septa, of a brownish colour, with lengths ranging from (7 µm to 14 µm and widths from 5 µm to 7 µm. (N=-50, Fig 1) (Yasanthika et al. 2023). The UFTCC01 isolate's DNA was extracted using the extraction kit (PROMEGA ®), amplified for genes (ITS, Gapdh and Tefα1), sequenced (OR345316.1, OR344427.1 and OR344428.1) and compared in BLASTn at Gen Bank deposited sequences. Phylogenetic analysis using the parsimony method in MEGA X identified the isolate as Curvularia chiangmaiensis, showing 99.5% identity with reference sequences (MN215651.1, MN264085.1, and MN263942.1). Concatenated region comparison revealed 94% similarity to sequences (OP564987.1 and MF490814.1) in the phylogenetic tree, confirming the species. The pathogenicity test used 60 seedlings of 50-day-old elephant grass, 30 of which were inoculated with a spore suspension (3.0 × 106 spores/mL) and 30 with sterile water. The solutions were sprayed twice. Only plants inoculated with the spore suspension exhibited symptoms akin to those observed in the field. Then, the fungus was reisolated from the lesions and Koch's postulates were confirmed twice.Previous studies have documented the C. chiangmaiensis pathogenicity in other grasses, such as maise in Thailand (Marin et al. 2017). Our studies constitute the first report of C. chiangmaiensis causing leaf spots on elephant grass in the Brazil.

象草(Cenchrus purpureus)在光合作用期间可高效封存大气中的二氧化碳,从而产生大量生物质。作为一种 C4 植物,它在替代生物柴油生产方面具有巨大潜力,并可作为牛的新鲜饲料(Negawo 等人,2018 年)。对巴西塞拉多地区种植大象草的商业牧场进行的实地观察显示,在巴西托坎廷斯州的帕尔马斯(西经 48°21'45",南纬 10°24'08")和古鲁皮(西经 49°02'08",南纬 11°46'21"),2022 年至 2023 年期间,坏死叶斑的严重程度达到 30%至 50%。在这两年的雨季,嫩叶和成熟叶片的实地发病率均接近 100%。收集受病害影响的叶片,装入保温箱后送往植物病理学实验室。198 片叶子在 1%的次氯酸盐中消毒 30 秒,然后用无菌水清洗三次。消毒后,切下叶片组织,在马铃薯葡萄糖琼脂(PDA)培养基上培养,并在 28°C 的受控箱(BOD)中培养,光周期为 12 小时。菌落呈灰白色,边缘发白,顶板呈褐色,七天后长到直径 5 厘米。从单孢培养物中获得的菌落形成弯曲的分生孢子,主要有三个隔膜,呈褐色,长度从 7 微米到 14 微米不等,宽度从 5 微米到 7 微米不等。(N=-50, 图 1)(Yasanthika 等人,2023 年)。使用提取试剂盒(PROMEGA ®)提取 UFTCC01 分离物的 DNA,扩增基因(ITS、Gapdh 和 Tefα1),测序(OR345316.1、OR344427.1 和 OR344428.1),并与 Gen Bank 保存的序列进行 BLASTn 比较。利用 MEGA X 中的解析法进行的系统发生分析表明,该分离物为 Curvularia chiangmaiensis,与参考序列(MN215651.1、MN264085.1 和 MN263942.1)的一致性为 99.5%。同源区比较结果显示,该物种与系统发生树中的序列(OP564987.1 和 MF490814.1)相似度为 94%,从而确定了该物种。致病性试验使用了 60 株 50 天大的象草幼苗,其中 30 株接种了孢子悬浮液(3.0 × 106 孢子/毫升),30 株接种了无菌水。溶液喷洒两次。只有接种了孢子悬浮液的植物才会出现与田间观察到的类似症状。之前的研究已经证明了 C. chiangmaiensis 在泰国玉米等其他禾本科植物中的致病性(Marin 等,2017 年)。我们的研究是首次报道 C. chiangmaiensis 在巴西引起象草叶斑病。
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引用次数: 0
First Report in China of Leaf Blight on Spatholobus suberectus Dunn Caused by Diaporthe tulliensis. 中国首次报告由 Diaporthe tulliensis 引起的 Dunn Spatholobus suberectus 叶枯病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-11 DOI: 10.1094/PDIS-09-24-1883-PDN
Xinjie Zhan, Zhuoqiu Qiu, Qi Huang, Shuangshuang Qin, Zhanjiang Zhang, Shugen Wei, Ni Jiang, Lisha Song
<p><p>Spatholobus suberectus Dunn is an important Chinese medicinal plant, which is the only officially recognized authentic source of the Chinese herbal medicine Spatholobi caulis in the China Pharmacopeia (Qin et al. 2024). In November 2023, a serious leaf disease was observed in the S. suberectus planting base in Nanning city (108.22E, 22.51N), Guangxi, China, with an incidence rate of 80%. Initially the symptoms appeared in the diseased leaves tips and was irregular in shape. The necrosis displayed discoloration of the gray color with a deep brown margin and this gradually expanded through into the lamina. To isolate the causal agent, 10 symptomatic leaves were collected and small segments (5×5 mm) of infected tissues were cut, and then they were surface sterilized (30s in 75% EtOH, 2 min in 2.5% NaOCl, and rinsed three times in sterilized distilled water), placed on potato dextrose agar (PDA) and incubated at 28℃ for 5 days. The hyphal tips were transferred to PDA to obtain pure cultures. 36 isolates were obtained, 23 of them had similar morphology, which were isolated from all the symptomatic leaves and were considered pathogens, and named YY-3-S. After 7 d, the colonies were white, fluffy, with white aerial mycelium. Sporulation was induced using pine needle medium (Su et al. 2012). After 30 days, the globose pycnidia developed, which were superficial to semi-immersed on the pine needle medium, and they appeared to be orange to black in color, with pale yellow conidial droplets. The alpha conidia spores were 3.53 to 6.85×1.75 to 3.70 μm (n = 50). They were unicellular, hyaline, aseptate and ellipsoidal to fusiform. These morphological features were consistent with Diaporthe sp (Sun et al. 2021). For further molecular identification, isolate YY-3-S underwent sequencing of the internal transcribed spacer (ITS), beta-tubulin (TUB2), and translation elongation factor 1-alpha (TEF1) sections (Huang et al. 2021). Obtained sequences of ITS, TUB2 and TEF1 (Genebank accessions nos. PP838571, PP865050 and PP865051) displayed a 99% similarity to Diaporthe tulliensis (Genebank accessions nos. OP163562, KR936132, OR662168, respectively). Phylogenetic analysis was based on a combination of ITS, TUB2 and TEF1 sequence data using MEGA 11 software to construct a phylogenetic tree with Neighbor-Joining. In the phylogenetic tree, the combined sequences attributed YY-3-S to D. tulliensis. The pathogenicity test was conducted on 1-year-old S. suberectus seedlings. We sprayed the mixed suspension of spores and mycelia on the unwounded leaves, while control seedlings received sterile water. 3 leaves per plant and 5 plants per treatment were selected for assessment. All seedings were placed in pots in a moist chamber at 28°C and 90% humidity. After 5 d, browning was observed on all the inoculated leaves tips. 14 d after inoculation, necrotic lesions expanded outward and their symptoms like those in the field, while control plants remained healthy. D. tulliensis w
邓楠(Spatholobus suberectus Dunn)是一种重要的中药植物,是《中国药典》中唯一官方认可的中药材刺五加的正品来源(覃等,2024)。2023 年 11 月,中国广西南宁市(东经 108.22 度,北纬 22.51 度)的裸冠菊种植基地出现了严重的叶部病害,发病率高达 80%。症状最初出现在病叶顶端,形状不规则。坏死部位呈灰色,边缘呈深褐色,并逐渐向叶片扩展。为了分离病原菌,采集了 10 片有症状的叶片,切下受感染组织的小片(5×5 毫米),然后进行表面消毒(在 75% 的乙醇中消毒 30 秒,在 2.5% 的 NaOCl 中消毒 2 分钟,再用消毒蒸馏水冲洗 3 次),放在马铃薯葡萄糖琼脂(PDA)上,在 28℃下培养 5 天。将菌丝顶端转移到 PDA 上以获得纯培养物。得到 36 个分离物,其中 23 个形态相似,从所有有症状的叶片中分离出来,被认为是病原体,命名为 YY-3-S。7 d 后,菌落呈白色,绒毛状,有白色气生菌丝。使用松针培养基诱导孢子的产生(Su 等,2012 年)。30 天后,球状分生孢子发育成熟,在松针培养基上呈表层至半浸润状,呈橙色至黑色,有淡黄色的分生孢子液滴。α分生孢子大小为 3.53 至 6.85×1.75 至 3.70 μm(n = 50)。它们呈单细胞、透明、无菌、椭圆形至纺锤形。这些形态特征与 Diaporthe sp 相符(Sun 等,2021 年)。为进一步进行分子鉴定,对分离物 YY-3-S 的内部转录间隔(ITS)、β-微管蛋白(TUB2)和翻译伸长因子 1-α(TEF1)部分进行了测序(Huang 等,2021 年)。获得的 ITS、TUB2 和 TEF1 序列(Genebank 编号:PP838571、PP865050 和 PP865051)与 Diaporthe tulliensis(Genebank 编号:OP163562、KR936132 和 OR662168)的相似度为 99%。利用 MEGA 11 软件结合 ITS、TUB2 和 TEF1 序列数据进行了系统发生分析,构建了一棵邻接系统发生树。在系统进化树中,综合序列将 YY-3-S 定义为 D. tulliensis。致病性试验在 1 年生的 S. suberectus幼苗上进行。我们将孢子和菌丝体的混合悬浮液喷洒在未受伤的叶片上,而对照组幼苗则接受无菌水。每株选取 3 片叶子,每个处理选取 5 株进行评估。所有幼苗都被放入盆中,置于 28°C 和 90% 湿度的潮湿室内。5 d 后,所有接种的叶尖都出现褐变。接种 14 d 后,坏死病斑向外扩展,症状与田间相同,而对照植物则保持健康。从接种的叶片中重新分离出了 D. tulliensis,符合科赫假设。红毛丹坏死斑和叶枯病(Serrato-Diaz 等人,2023 年)、菩提树叶斑病(Li 等人,2022 年)、茉莉茎腐病(Hsu 等人,2023 年)等病害都曾被报道过。据我们所知,这是中国首次报告 D. tulliensis 对亚圆锥花序(S. suberectus)造成叶枯病。确保及时防治这种新出现的病害非常重要。
{"title":"First Report in China of Leaf Blight on <i>Spatholobus suberectus</i> Dunn Caused by <i>Diaporthe tulliensis</i>.","authors":"Xinjie Zhan, Zhuoqiu Qiu, Qi Huang, Shuangshuang Qin, Zhanjiang Zhang, Shugen Wei, Ni Jiang, Lisha Song","doi":"10.1094/PDIS-09-24-1883-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-09-24-1883-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Spatholobus suberectus Dunn is an important Chinese medicinal plant, which is the only officially recognized authentic source of the Chinese herbal medicine Spatholobi caulis in the China Pharmacopeia (Qin et al. 2024). In November 2023, a serious leaf disease was observed in the S. suberectus planting base in Nanning city (108.22E, 22.51N), Guangxi, China, with an incidence rate of 80%. Initially the symptoms appeared in the diseased leaves tips and was irregular in shape. The necrosis displayed discoloration of the gray color with a deep brown margin and this gradually expanded through into the lamina. To isolate the causal agent, 10 symptomatic leaves were collected and small segments (5×5 mm) of infected tissues were cut, and then they were surface sterilized (30s in 75% EtOH, 2 min in 2.5% NaOCl, and rinsed three times in sterilized distilled water), placed on potato dextrose agar (PDA) and incubated at 28℃ for 5 days. The hyphal tips were transferred to PDA to obtain pure cultures. 36 isolates were obtained, 23 of them had similar morphology, which were isolated from all the symptomatic leaves and were considered pathogens, and named YY-3-S. After 7 d, the colonies were white, fluffy, with white aerial mycelium. Sporulation was induced using pine needle medium (Su et al. 2012). After 30 days, the globose pycnidia developed, which were superficial to semi-immersed on the pine needle medium, and they appeared to be orange to black in color, with pale yellow conidial droplets. The alpha conidia spores were 3.53 to 6.85×1.75 to 3.70 μm (n = 50). They were unicellular, hyaline, aseptate and ellipsoidal to fusiform. These morphological features were consistent with Diaporthe sp (Sun et al. 2021). For further molecular identification, isolate YY-3-S underwent sequencing of the internal transcribed spacer (ITS), beta-tubulin (TUB2), and translation elongation factor 1-alpha (TEF1) sections (Huang et al. 2021). Obtained sequences of ITS, TUB2 and TEF1 (Genebank accessions nos. PP838571, PP865050 and PP865051) displayed a 99% similarity to Diaporthe tulliensis (Genebank accessions nos. OP163562, KR936132, OR662168, respectively). Phylogenetic analysis was based on a combination of ITS, TUB2 and TEF1 sequence data using MEGA 11 software to construct a phylogenetic tree with Neighbor-Joining. In the phylogenetic tree, the combined sequences attributed YY-3-S to D. tulliensis. The pathogenicity test was conducted on 1-year-old S. suberectus seedlings. We sprayed the mixed suspension of spores and mycelia on the unwounded leaves, while control seedlings received sterile water. 3 leaves per plant and 5 plants per treatment were selected for assessment. All seedings were placed in pots in a moist chamber at 28°C and 90% humidity. After 5 d, browning was observed on all the inoculated leaves tips. 14 d after inoculation, necrotic lesions expanded outward and their symptoms like those in the field, while control plants remained healthy. D. tulliensis w","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2024-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142625747","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
First report of Pectobacterium peruviense as the causal pathogen of blackleg and soft rot in Solanum tuberosum cv. Superchola in Cañar, Ecuador. 首次报告 Pectobacterium peruviense 是厄瓜多尔卡尼亚尔地区 Solanum tuberosum cv. Superchola 黑腿病和软腐病的病原菌。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-11 DOI: 10.1094/PDIS-08-24-1759-PDN
Estefania Peña-Zuñiga, Noelia Barriga-Medina, Dario Ramirez-Villacis, Madelein Miranda-Guerrero, Corné Pieterse, Jos M Raaijmakers, Antonio Leon-Reyes
<p><p>Potato (Solanum tuberosum) is grown in Ecuador's Andean region at altitudes from 2,200 to 3,600 m.a.s.l., and the most commercially significant cultivar is Superchola, which constitutes 60% of total production, covering around 11,000 hectares (MAG, s.f. 2022). From November 2022 to January 2023, dark, water-soaked lesions progressing upwards from the base of the stems were observed on Superchola potato plants in Cañar, Ecuador (2°23'56.4''S 78°59'13.2''W). Approximately 20 plants in a hectare showed symptoms. Symptomatic stems from five plants were collected, surface-disinfected with 2% sodium hypochlorite, followed by 70% ethanol, and thoroughly rinsed with sterile distilled water. Plant tissue sections (1 cm²) were homogenized in 10 mM MgCl₂, serially diluted, and plated on Tryptic Soy Agar (TSA). After 48 hours of incubation at 28°C, creamy-white, yellowish, and white round colonies appeared. Streak plating was performed, yielding 14 purified bacterial isolates named M1 to M14. For the pathogenicity tests, each purified isolate was inoculated into healthy potato tubers and stem pieces (5 cm) using the prick inoculation method described by Ma et al. (2018) with modifications. Surface-sterilized tubers and stems were wounded using autoclaved toothpicks, into which 5µl of a bacterial suspension, carrying ~1 x 106 CFU/ml of bacteria per strain, was deposited into a 5 mm deep wound. Three independent replicates were conducted. Only one isolate, designated as M3, presented maceration on potato tubers, and dark, watered-soaked lesions appeared on the stem 48 h post-inoculation with strain. Other bacterial isolates showed no symptoms in either stems or tubers. Pathogenicity tests were confirmed on healthy, 4-week-old potato plants grown in a 50% perlite and 50% peat moss substrate. Inoculations were done using two methods: (1) toothpick piercing, where 5 µl of saline solution containing ~1 x 106 CFU/ml of bacteria was applied to a wound approximately 5mm deep (Ma et al. 2018), and (2) immersion of the plant foliage in a bacterial suspension of 1x 108 CFU/ml in saline solution for 10 min Controls used bacteria-free toothpicks (wounded) and sterile 10 mM MgCl₂ solution (unwounded). Plants were enclosed in plastic containers to maintain high humidity and grown at 15/25°C (day/night) with a 12 h photoperiod. Five days post-inoculation, blackleg symptoms were observed in the stems of the infected area, which turned black and rotten (wounded and unwounded). In all cases, negative controls remained symptomless. To complete Koch's postulates, bacteria were reisolated from symptomatic potato stems and showed the same morphology. To identify the isolate M3, whole genome sequencing using Oxford Nanopore technology was performed. Three marker genes were extracted, and a 5,734-bp sequence was concatenated from dnaX (2073 bp), recA (1,074bp), and leuS (2,583bp) (GenBank accession nos. PQ177849, PQ177847 and PQ177848, respectively). The concatenated sequences
马铃薯(Solanum tuberosum)种植于海拔 2,200 至 3,600 米的厄瓜多尔安第斯地区,最具商业价值的栽培品种是 Superchola,占总产量的 60%,种植面积约 11,000 公顷(农业咨询小组,2022 年)。从 2022 年 11 月到 2023 年 1 月,在厄瓜多尔卡尼亚尔(南纬 2°23'56.4''S - 西经 78°59'13.2'')的 Superchola 马铃薯植株上观察到从茎基部向上发展的深色水渍状病变。每公顷约有 20 株出现症状。采集 5 株出现症状的茎秆,用 2% 次氯酸钠进行表面消毒,然后用 70% 乙醇消毒,最后用无菌蒸馏水彻底冲洗。植物组织切片(1 cm²)在 10 mM MgCl₂中匀浆,按比例稀释,然后培养在胰蛋白酶大豆琼脂(TSA)上。28°C 培养 48 小时后,出现乳白色、淡黄色和白色圆形菌落。进行菌株培养后,得到了 14 个纯化的细菌分离株,分别命名为 M1 至 M14。在致病性试验中,采用 Ma 等人(2018 年)描述并修改过的刺穿接种法,将每个纯化的分离株接种到健康的马铃薯块茎和茎(5 厘米)中。使用高压灭菌的牙签刺伤表面消毒的块茎和茎,然后将 5µl 的细菌悬浮液(每个菌株携带 ~1 x 106 CFU/ml 的细菌)滴入 5 mm 深的伤口中。进行了三次独立重复。只有一种被命名为 M3 的分离菌株在马铃薯块茎上出现浸渍现象,在接种菌株 48 小时后,茎干上出现深色、浸水的病变。其他细菌分离物在茎秆或块茎上均未见症状。在 50%珍珠岩和 50%泥炭藓基质中生长的 4 周大的健康马铃薯植株上进行了致病性试验。接种采用两种方法:(1) 牙签穿刺,将 5 µl 含有 ~1 x 106 CFU/ml 细菌的生理盐水涂抹在约 5mm 深的伤口上(Ma 等人,2018 年);(2) 将植物叶片浸泡在生理盐水中 1x 108 CFU/ml 的细菌悬浮液中 10 分钟,对照组使用无菌牙签(有伤口)和无菌 10 mM MgCl₂溶液(无伤口)。将植物置于塑料容器中以保持高湿度,并在 15/25°C (昼/夜)和 12 小时光周期下生长。接种后五天,受感染区域的茎部出现黑腿症状,变黑腐烂(受伤和未受伤)。在所有情况下,阴性对照均无症状。为了完成科赫假设,从有症状的马铃薯茎上重新分离出细菌,并显示出相同的形态。为了确定分离物 M3,使用牛津纳米孔技术进行了全基因组测序。提取了三个标记基因,并从 dnaX(2073bp)、recA(1074bp)和 leuS(2583bp)(GenBank 编号分别为 PQ177849、PQ177847 和 PQ177848)中连接出 5734bp 的序列。使用 BEAST 1.8.4 版构建了 M3 株系和模式株系的多焦点贝叶斯推断系统发生树。M3 株系与 Pectobacterium peruviense 同源,与模式菌株 P. peruviense IFB5232 的序列同一性达到 100%(Drummond 等,2012 年;Portier 等,2019 年;Toth 等,2021 年;Waleron 等,2018 年)。据我们所知,这是首次报道 P. peruviense 在厄瓜多尔马铃薯植株中引起黑腿病和软腐病。进一步的研究至关重要,但这些信息有助于监测病原体的传播并制定病害管理策略。
{"title":"First report of <i>Pectobacterium peruviense</i> as the causal pathogen of blackleg and soft rot in <i>Solanum tuberosum</i> cv. Superchola in Cañar, Ecuador.","authors":"Estefania Peña-Zuñiga, Noelia Barriga-Medina, Dario Ramirez-Villacis, Madelein Miranda-Guerrero, Corné Pieterse, Jos M Raaijmakers, Antonio Leon-Reyes","doi":"10.1094/PDIS-08-24-1759-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-08-24-1759-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Potato (Solanum tuberosum) is grown in Ecuador's Andean region at altitudes from 2,200 to 3,600 m.a.s.l., and the most commercially significant cultivar is Superchola, which constitutes 60% of total production, covering around 11,000 hectares (MAG, s.f. 2022). From November 2022 to January 2023, dark, water-soaked lesions progressing upwards from the base of the stems were observed on Superchola potato plants in Cañar, Ecuador (2°23'56.4''S 78°59'13.2''W). Approximately 20 plants in a hectare showed symptoms. Symptomatic stems from five plants were collected, surface-disinfected with 2% sodium hypochlorite, followed by 70% ethanol, and thoroughly rinsed with sterile distilled water. Plant tissue sections (1 cm²) were homogenized in 10 mM MgCl₂, serially diluted, and plated on Tryptic Soy Agar (TSA). After 48 hours of incubation at 28°C, creamy-white, yellowish, and white round colonies appeared. Streak plating was performed, yielding 14 purified bacterial isolates named M1 to M14. For the pathogenicity tests, each purified isolate was inoculated into healthy potato tubers and stem pieces (5 cm) using the prick inoculation method described by Ma et al. (2018) with modifications. Surface-sterilized tubers and stems were wounded using autoclaved toothpicks, into which 5µl of a bacterial suspension, carrying ~1 x 106 CFU/ml of bacteria per strain, was deposited into a 5 mm deep wound. Three independent replicates were conducted. Only one isolate, designated as M3, presented maceration on potato tubers, and dark, watered-soaked lesions appeared on the stem 48 h post-inoculation with strain. Other bacterial isolates showed no symptoms in either stems or tubers. Pathogenicity tests were confirmed on healthy, 4-week-old potato plants grown in a 50% perlite and 50% peat moss substrate. Inoculations were done using two methods: (1) toothpick piercing, where 5 µl of saline solution containing ~1 x 106 CFU/ml of bacteria was applied to a wound approximately 5mm deep (Ma et al. 2018), and (2) immersion of the plant foliage in a bacterial suspension of 1x 108 CFU/ml in saline solution for 10 min Controls used bacteria-free toothpicks (wounded) and sterile 10 mM MgCl₂ solution (unwounded). Plants were enclosed in plastic containers to maintain high humidity and grown at 15/25°C (day/night) with a 12 h photoperiod. Five days post-inoculation, blackleg symptoms were observed in the stems of the infected area, which turned black and rotten (wounded and unwounded). In all cases, negative controls remained symptomless. To complete Koch's postulates, bacteria were reisolated from symptomatic potato stems and showed the same morphology. To identify the isolate M3, whole genome sequencing using Oxford Nanopore technology was performed. Three marker genes were extracted, and a 5,734-bp sequence was concatenated from dnaX (2073 bp), recA (1,074bp), and leuS (2,583bp) (GenBank accession nos. PQ177849, PQ177847 and PQ177848, respectively). The concatenated sequences ","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2024-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142625831","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
First Report of Pepper Chlorosis-Associated Virus in Burdock (Arctium lappa) in Korea. 韩国首次报告牛蒡(Arctium lappa)中的辣椒萎黄病相关病毒。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-11 DOI: 10.1094/PDIS-08-24-1639-PDN
Sangmin Bak, Daeun Kim, Hyewon Jeong, Minki Kim, Ilkwon Yeon, Seunghan Kim, Seungbin Baek, Su-Heon Lee, Wonkwon Jung
<p><p>Burdock (<i>Arctium lappa</i>) is a fiber-rich root vegetable widely used in Asian cuisine with many health benefits. In July 2023, two burdock leaf samples showing viral symptoms, such as yellowing and interveinal chlorosis, were collected from a field in Daegu, South Korea, with an incidence rate of about 10%. The samples were pooled, ground with liquid nitrogen, and total RNA was extracted using a Maxwell® 16 LEV Plant RNA Kit (Promega, Madison, USA). A cDNA library was constructed using TruSeq Standard Total RNA with Ribo-Zero (Illumina, San Diego, USA) and sequenced on an Illumina NovaSeq 6000 (BIOTO, Daejeon, Korea) with 100 bp paired-end reads. Of the approximately 84 million reads generated, about 61 million clean reads were identified using Trimmomatic 0.39 with default parameters. Subsequently, Trinity 2.13.0 was used for <i>de novo</i> assembly, resulting in 179,910 contigs, which, when annotated as previously described (Bak et al., 2024), revealed the presence of tomato spotted wilt virus (TSWV) and pepper chlorosis-associated virus (PepCaV). Three PepCaV-related contigs, representing a near-complete genome, were submitted to NCBI GenBank (PP502423-PP502425). BLASTn analysis showed 95.21% (7275/7641 bp), 95.89% (1680/1752 bp), and 96.18% (1461/1519 bp) identities with the L, M, and S segments of the PepCaV isolate Higashitsuno_2021 (LC719619-LC719621), respectively. RT-PCR was performed to confirm the presence of viruses using specific primers for PepCaV (5'-CACCTTGATTATAGACATGACAG/GTTCTCAAATCTTCCCAATCG-3') targeting the coat protein region, and TSWV (5'-GCAACAACTTGCAAGAAGATG/CATGACCTTCAGAAGGCTTG-3'; Kim et al., 2021). Initial results showed both samples were positive for TSWV, and one was positive for PepCaV. Additionally, 26 symptomatic and 8 asymptomatic samples were collected from the same field. PepCaV was positive in 3 samples, TSWV in 12, and TSWV+PepCaV in 11 by RT-PCR. All asymptomatic samples were negative. Leaves infected with PepCaV, whether solely or co-infected with TSWV, initially exhibited interveinal chlorosis and vein contraction on one side of the main vein (Fig. S1). In plants infected with PepCaV alone, these symptoms were mild or less noticeable in the initial phases. The majority of PepCaV-infected samples were co-infected with TSWV, during which symptoms appeared more severe. Following this, an amplicon, derived from the sample that initially tested positive for PepCaV in the first RT-PCR was cloned using the All in OneTM Cloning Kit (BioFact, Daejeon, Korea) and sequenced by Bionner (Daejeon, Korea). The resulting sequence, submitted to NCBI GenBank (PP928956), demonstrated 96.87% (805/831 bp) identity with the PepCaV isolate Higashitsuno_2021 (LC719600). Moreover, sap inoculation was performed on <i>A. lappa</i>, <i>Nicotiana benthamiana</i>, <i>N. clevelandii</i>, <i>N. debneyi</i>, <i>N. rustica</i>, <i>N. tabacum</i>, <i>Chenopodium amaranticolor</i>, and <i>C. quinoa</i>, but no transmission was ide
牛蒡(Arctium lappa)是一种富含纤维的根茎类蔬菜,被广泛用于亚洲菜肴,对健康有诸多益处。2023 年 7 月,我们从韩国大邱的一块田地里采集了两份牛蒡叶片样本,这些样本出现了黄化和叶脉间枯萎等病毒症状,发病率约为 10%。样本集中后用液氮研磨,并使用 Maxwell® 16 LEV 植物 RNA 试剂盒(Promega,美国麦迪逊)提取总 RNA。使用 TruSeq Standard Total RNA with Ribo-Zero (Illumina,San Diego,USA)构建 cDNA 文库,并在 Illumina NovaSeq 6000(BIOTO,Daejeon,Korea)上用 100 bp 成对端读数进行测序。在生成的约 8,400 万个读数中,使用 Trimmomatic 0.39(默认参数)鉴定了约 6,100 万个纯净读数。随后,使用 Trinity 2.13.0 进行了从头组装,得到了 179,910 个等位基因,按照之前的描述(Bak 等人,2024 年)进行注释后,发现了番茄斑萎病毒(TSWV)和辣椒萎黄病相关病毒(PepCaV)的存在。三个与 PepCaV 相关的等位基因代表了一个接近完整的基因组,已提交给 NCBI GenBank(PP502423-PP502425)。BLASTn 分析表明,它们与 PepCaV 分离物 Higashitsuno_2021 的 L、M 和 S 片段(LC719619-LC719621)的相同度分别为 95.21%(7275/7641 bp)、95.89%(1680/1752 bp)和 96.18%(1461/1519 bp)。为了确认病毒的存在,使用针对衣壳蛋白区的 PepCaV(5'-CACCTTGATTATAGACATGACAG/GTTCTCAAATCTTCCCAATCG-3')和 TSWV(5'-GCAACAACTTGCAAGAAGATG/CATGACCTTCAGAAGGCTTG-3';Kim 等人,2021 年)的特异引物进行了 RT-PCR 检测。初步结果显示,两份样本的 TSWV 均呈阳性,一份样本的 PepCaV 呈阳性。此外,还从同一田块采集了 26 份无症状样本和 8 份无症状样本。通过 RT-PCR 检测,3 份样本中 PepCaV 呈阳性,12 份样本中 TSWV 呈阳性,11 份样本中 TSWV+PepCaV 呈阳性。所有无症状样本均为阴性。感染了 PepCaV 的叶片,无论是单独感染还是与 TSWV 共同感染,最初都会表现出叶脉间萎黄和主脉一侧的叶脉收缩(图 S1)。在单独感染 PepCaV 的植株中,这些症状在初期比较轻微或不太明显。大多数感染 PepCaV 的样本都同时感染了 TSWV,在此期间症状更加严重。随后,使用 All in OneTM 克隆试剂盒(BioFact,韩国大田)克隆了在第一次 RT-PCR 中检测出 PepCaV 呈阳性的样本的扩增片段,并由 Bionner(韩国大田)进行了测序。测得的序列已提交给 NCBI GenBank(PP928956),显示与 PepCaV 分离物 Higashitsuno_2021 (LC719600) 的同一性为 96.87% (805/831 bp)。此外,还对 A. lappa、Nicotiana benthamiana、N. clevelandii、N. debneyi、N. rustica、N. tabacum、Chenopodium amaranticolor 和 C. quinoa 进行了汁液接种,但均未发现传播。最近发现的 PepCaV 很可能属于蛇形病毒科蛇形病毒属,已知可感染辣椒和烟草(Shimomoto 等人,2023;Wang 等人,2024)。这是 PepCaV 在韩国的首次报道,也是牛蒡作为天然宿主的首次鉴定。虽然缺乏对其传播媒介和致病性的研究,但 PepCaV 的存在提示了潜在的风险。鉴于牛蒡在韩国的广泛种植,PepCaV 可能会影响作物产量和健康,从而对当地农业产生重大影响。
{"title":"First Report of Pepper Chlorosis-Associated Virus in Burdock (<i>Arctium lappa</i>) in Korea.","authors":"Sangmin Bak, Daeun Kim, Hyewon Jeong, Minki Kim, Ilkwon Yeon, Seunghan Kim, Seungbin Baek, Su-Heon Lee, Wonkwon Jung","doi":"10.1094/PDIS-08-24-1639-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-08-24-1639-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Burdock (&lt;i&gt;Arctium lappa&lt;/i&gt;) is a fiber-rich root vegetable widely used in Asian cuisine with many health benefits. In July 2023, two burdock leaf samples showing viral symptoms, such as yellowing and interveinal chlorosis, were collected from a field in Daegu, South Korea, with an incidence rate of about 10%. The samples were pooled, ground with liquid nitrogen, and total RNA was extracted using a Maxwell® 16 LEV Plant RNA Kit (Promega, Madison, USA). A cDNA library was constructed using TruSeq Standard Total RNA with Ribo-Zero (Illumina, San Diego, USA) and sequenced on an Illumina NovaSeq 6000 (BIOTO, Daejeon, Korea) with 100 bp paired-end reads. Of the approximately 84 million reads generated, about 61 million clean reads were identified using Trimmomatic 0.39 with default parameters. Subsequently, Trinity 2.13.0 was used for &lt;i&gt;de novo&lt;/i&gt; assembly, resulting in 179,910 contigs, which, when annotated as previously described (Bak et al., 2024), revealed the presence of tomato spotted wilt virus (TSWV) and pepper chlorosis-associated virus (PepCaV). Three PepCaV-related contigs, representing a near-complete genome, were submitted to NCBI GenBank (PP502423-PP502425). BLASTn analysis showed 95.21% (7275/7641 bp), 95.89% (1680/1752 bp), and 96.18% (1461/1519 bp) identities with the L, M, and S segments of the PepCaV isolate Higashitsuno_2021 (LC719619-LC719621), respectively. RT-PCR was performed to confirm the presence of viruses using specific primers for PepCaV (5'-CACCTTGATTATAGACATGACAG/GTTCTCAAATCTTCCCAATCG-3') targeting the coat protein region, and TSWV (5'-GCAACAACTTGCAAGAAGATG/CATGACCTTCAGAAGGCTTG-3'; Kim et al., 2021). Initial results showed both samples were positive for TSWV, and one was positive for PepCaV. Additionally, 26 symptomatic and 8 asymptomatic samples were collected from the same field. PepCaV was positive in 3 samples, TSWV in 12, and TSWV+PepCaV in 11 by RT-PCR. All asymptomatic samples were negative. Leaves infected with PepCaV, whether solely or co-infected with TSWV, initially exhibited interveinal chlorosis and vein contraction on one side of the main vein (Fig. S1). In plants infected with PepCaV alone, these symptoms were mild or less noticeable in the initial phases. The majority of PepCaV-infected samples were co-infected with TSWV, during which symptoms appeared more severe. Following this, an amplicon, derived from the sample that initially tested positive for PepCaV in the first RT-PCR was cloned using the All in OneTM Cloning Kit (BioFact, Daejeon, Korea) and sequenced by Bionner (Daejeon, Korea). The resulting sequence, submitted to NCBI GenBank (PP928956), demonstrated 96.87% (805/831 bp) identity with the PepCaV isolate Higashitsuno_2021 (LC719600). Moreover, sap inoculation was performed on &lt;i&gt;A. lappa&lt;/i&gt;, &lt;i&gt;Nicotiana benthamiana&lt;/i&gt;, &lt;i&gt;N. clevelandii&lt;/i&gt;, &lt;i&gt;N. debneyi&lt;/i&gt;, &lt;i&gt;N. rustica&lt;/i&gt;, &lt;i&gt;N. tabacum&lt;/i&gt;, &lt;i&gt;Chenopodium amaranticolor&lt;/i&gt;, and &lt;i&gt;C. quinoa&lt;/i&gt;, but no transmission was ide","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2024-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142625838","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
First Report of Root Rot Caused by Fusarium equiseti on Syrian rue (Peganum harmala) in China. 中国首次报告叙利亚芸香(Peganum harmala)上由马镰刀菌(Fusarium equiseti)引起的根腐病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-11-11 DOI: 10.1094/PDIS-06-24-1297-PDN
Ruifang Jia, Jianfeng Yang, Jianjun Hao, Shengze Wang, Jie Wu, Kejian Lin, Zhengqiang Chen, Yuanyuan Zhang
<p><p>Syrian rue (Peganum harmala L) belongs to the family Zygophyllaceae and is mainly distributed in arid and semi-arid areas of Xinjiang, Gansu, Ningxia, Qinghai, and Inner Mongolia in China. It serves as a pioneer species in soil and water conservation, as well as in reclamation of wastelands, playing a crucial role in soil preservation and stabilization against sand encroachment. In 2023, a disease resembling root rot and wilt affected the quality and yield of Syrian rue in Xilingol (122.67°N, 42.78°E), Inner Mongolia, China. Symptoms ranged from yellowing to wilting, accompanied by darkening of the xylem in the roots. This disease was observed in approximately 50% of Syrian rue plants in desertification-prone grassland. A total number of 45 symptomatic root samples were collected from Syrian rue plants to isolate the pahogen. Small pieces of diseased tissue were surface sterilized with 75% alcohol for five seconds, rinsed twice with sterilized water, dried, and then placed on water agar and incubated for 72 h at 25℃ in darkness. A total of 45 isolates were obtained, showing identical morphology and sequences of the internal transcribed spacer region (ITS), translation elongation factor 1-alpha (EF1-α) and RNA polymerase II second largest subunit (RPB2) genes. Therefore, the isolate LTP-5 was used as the representative for further study. On potato dextrose agar (PDA), fungal colonies of LTP-5 exhibited yellow to light brown colore with white aerial mycelia and irregular growth after 7 days of incubation at 25℃. Conidia and chlamydospre development were observed in carboxymethylcellulose sodium medium. Chlamydospores were abundant, terminal or intergrown between hyphae, rough brown walls of 5.9 to 19.82 μm in diameter (n = 75) were observed in the culture. Macroconidia had 5 to 7 septa, thick in the middle and thin at both ends, measured 28.17 to 62.81 μm in length and 1.50 to 4.19 μm in width (n = 128). Microconidia were absent. The morphological characteristics were consistent with the descriptions of Fusarium equiseti (Nelson et al., 1983). The pathogen was further confirmed to be F. equiseti by sequence analysis of the ITS, EF1-α and RPB2 genes amplified using polymerase chain reaction (PCR) with the primers ITS4/ITS5 (White et al., 1990), EF-1/EF-2 (O'Donnell et al., 1998), and RPB2-5F2 /fRPB2-7cR (Zhen et al., 2017). The sequences showed 100% similarity to the F. equiseti strain in the NCBI GenBank with accession numbers MH054914.1, KJ396323.1, and KT213286.1 for ITS, EF1-α, and RPB2, respectively. The sequences of PCR products were submitted to GenBank with accession numbers PP814863.1 (ITS), PP831628.1 (EF1-α) and PP826937.1 (RPB2). Pathogenicity tests were conducted through a pot assay. Five Syrian rue seedlings were inoculated by immersing plant roots in a suspension of 1×107 spores/mL for 30 min and transplanting to pots containing autoclaved soil. Control plants were immersed in sterile water. The inoculated plants showed yellow a
叙利亚芸香(Peganum harmala L)属于芸香科,主要分布在中国新疆、甘肃、宁夏、青海和内蒙古的干旱和半干旱地区。它是水土保持和荒地开垦的先锋物种,在保持和稳定土壤、抵御风沙侵蚀方面发挥着重要作用。2023 年,一种类似根腐病和枯萎病的病害影响了中国内蒙古锡林郭勒(122.67°N,42.78°E)叙利亚芸香的质量和产量。症状从发黄到枯萎不等,同时根部木质部变黑。在易受荒漠化影响的草地上,约 50%的叙利亚芸香属植物都出现了这种病害。从叙利亚芸香科植物上共采集了 45 份有症状的根部样本,以分离出病原体。将小块病组织用 75% 的酒精表面消毒 5 秒钟,再用消毒水冲洗两次,晾干,然后放在水琼脂上,在 25℃ 黑暗条件下培养 72 小时。共获得 45 个分离株,其形态和内部转录间隔区(ITS)、翻译延伸因子 1-α(EF1-α)和 RNA 聚合酶 II 第二大亚基(RPB2)基因的序列完全相同。因此,分离物 LTP-5 被用作进一步研究的代表。在马铃薯葡萄糖琼脂(PDA)上,25℃培养 7 天后,LTP-5 真菌菌落呈黄色至浅棕色,气生菌丝呈白色,生长不规则。在羧甲基纤维素钠培养基中观察到分生孢子和衣壳孢子的发育。培养物中观察到大量衣孢子,顶生或夹生在菌丝之间,直径为 5.9 至 19.82 微米(n = 75)的粗糙棕色壁。大锥体有 5 至 7 个隔膜,中间厚,两端薄,长 28.17 至 62.81 微米,宽 1.50 至 4.19 微米(n = 128)。没有微孢子囊。形态特征与 Fusarium equiseti 的描述一致(Nelson 等人,1983 年)。通过使用引物 ITS4/ITS5 (White 等人,1990 年)、EF-1/EF-2 (O'Donnell 等人,1998 年)和 RPB2-5F2 /fRPB2-7cR (Zhen 等人,2017 年)进行聚合酶链式反应(PCR)扩增 ITS、EF1-α 和 RPB2 基因的序列分析,进一步确认该病原体为马镰刀菌。这些序列与 NCBI GenBank 中的 F. equiseti 菌株相似度达 100%,ITS、EF1-α 和 RPB2 的登录号分别为 MH054914.1、KJ396323.1 和 KT213286.1。PCR 产物的序列已提交至 GenBank,登录号分别为 PP814863.1(ITS)、PP831628.1(EF1-α)和 PP826937.1(RPB2)。致病性试验通过盆栽试验进行。将 5 株叙利亚芸苔幼苗的根部浸泡在 1×107 个孢子/毫升的悬浮液中 30 分钟,然后移栽到装有高压灭菌土壤的花盆中。对照植物浸泡在无菌水中。接种的植株出现黄腐和根腐,但对照植株未出现任何症状。根据形态特征和序列分析,成功地从受感染的根部组织中重新分离并鉴定出了同一种真菌,但未接种的植株上却没有发现。此外,有报告称 F. equiseti 可导致甜菜(Khan 等人,2021 年)、西瓜(Rahman 等人,2021 年)、马铃薯(Cui 等人,2021 年)和其他植物发病。这是第一份证实 F. equiseti 是中国叙利亚芸苔枯萎病和根腐病病原菌的报告。
{"title":"First Report of Root Rot Caused by <i>Fusarium equiseti</i> on Syrian rue (<i>Peganum harmala</i>) in China.","authors":"Ruifang Jia, Jianfeng Yang, Jianjun Hao, Shengze Wang, Jie Wu, Kejian Lin, Zhengqiang Chen, Yuanyuan Zhang","doi":"10.1094/PDIS-06-24-1297-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-06-24-1297-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Syrian rue (Peganum harmala L) belongs to the family Zygophyllaceae and is mainly distributed in arid and semi-arid areas of Xinjiang, Gansu, Ningxia, Qinghai, and Inner Mongolia in China. It serves as a pioneer species in soil and water conservation, as well as in reclamation of wastelands, playing a crucial role in soil preservation and stabilization against sand encroachment. In 2023, a disease resembling root rot and wilt affected the quality and yield of Syrian rue in Xilingol (122.67°N, 42.78°E), Inner Mongolia, China. Symptoms ranged from yellowing to wilting, accompanied by darkening of the xylem in the roots. This disease was observed in approximately 50% of Syrian rue plants in desertification-prone grassland. A total number of 45 symptomatic root samples were collected from Syrian rue plants to isolate the pahogen. Small pieces of diseased tissue were surface sterilized with 75% alcohol for five seconds, rinsed twice with sterilized water, dried, and then placed on water agar and incubated for 72 h at 25℃ in darkness. A total of 45 isolates were obtained, showing identical morphology and sequences of the internal transcribed spacer region (ITS), translation elongation factor 1-alpha (EF1-α) and RNA polymerase II second largest subunit (RPB2) genes. Therefore, the isolate LTP-5 was used as the representative for further study. On potato dextrose agar (PDA), fungal colonies of LTP-5 exhibited yellow to light brown colore with white aerial mycelia and irregular growth after 7 days of incubation at 25℃. Conidia and chlamydospre development were observed in carboxymethylcellulose sodium medium. Chlamydospores were abundant, terminal or intergrown between hyphae, rough brown walls of 5.9 to 19.82 μm in diameter (n = 75) were observed in the culture. Macroconidia had 5 to 7 septa, thick in the middle and thin at both ends, measured 28.17 to 62.81 μm in length and 1.50 to 4.19 μm in width (n = 128). Microconidia were absent. The morphological characteristics were consistent with the descriptions of Fusarium equiseti (Nelson et al., 1983). The pathogen was further confirmed to be F. equiseti by sequence analysis of the ITS, EF1-α and RPB2 genes amplified using polymerase chain reaction (PCR) with the primers ITS4/ITS5 (White et al., 1990), EF-1/EF-2 (O'Donnell et al., 1998), and RPB2-5F2 /fRPB2-7cR (Zhen et al., 2017). The sequences showed 100% similarity to the F. equiseti strain in the NCBI GenBank with accession numbers MH054914.1, KJ396323.1, and KT213286.1 for ITS, EF1-α, and RPB2, respectively. The sequences of PCR products were submitted to GenBank with accession numbers PP814863.1 (ITS), PP831628.1 (EF1-α) and PP826937.1 (RPB2). Pathogenicity tests were conducted through a pot assay. Five Syrian rue seedlings were inoculated by immersing plant roots in a suspension of 1×107 spores/mL for 30 min and transplanting to pots containing autoclaved soil. Control plants were immersed in sterile water. The inoculated plants showed yellow a","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2024-11-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142625750","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}
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