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First Report of Leaf Blight on Paris polyphylla Caused by Ceratobasidium ramicola in China. 中国首次报告由 Ceratobasidium ramicola 引起的巴黎多花植物叶枯病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-23 DOI: 10.1094/PDIS-05-24-1066-PDN
Feiyan Xie, Yuhang Zhong, Xianjing Lin, Yuxi Chen, Li Yang, Ting Tang
<p><p><i>Paris polyphylla</i> var. <i>yunnanensis</i>is a perennial herb with significant medicinal properties in anticancer, anti-inflammatory, antibacterial immunomodulatory and antispasmodic activities (Duan et al. 2018). In April 2022, leaf blight disease emerged in Xiangtan City (Hunan), affecting <i>P</i>. <i>polyphylla</i> plantings over an area of 3×104 m<sup>2</sup> (27.904°N, 112.918°E). The disease incidence reached an average of 22% of the plants in the field, with infected plants initially displaying water-soaked chlorosis, followed by dry yellow shrinkage that gradually spread from the leaf tips to the entire plant. To identify the causal agent, 20 leaf lesions (4 mm2) collected from 20 plants were surface sterilized with 75% ethanol for 10 s, 5% NaOCl for 30 s, rinsed in sterile distilled water three times, transferred to potato dextrose agar (PDA) plates with lactic acid (0.125%) , and incubated at 28 °C in the dark. Four isolates (PP21 to PP24) with similar morphologies were obtained and purified by the hyphal-tip method. Colonies on PDA initially appeared white with cottony mycelium, later turning light-yellow on the underside. Septate hyphae were branched at right angles with a small constriction at the departure of the branch point and measured 4.16 to 7.93 µm in diameter. Binucleate cells within the septate hyphae were visualized using Giemsa staining (Servicebio, China). For molecular identification, the rDNA internal transcribed spacer (ITS), the second largest subunit of nuclear DNA-directed RNA polymerase II (RPB2), and ATP synthase subunit 6 (ATP6) were amplified from genomic DNA of the isolates extracted by Fungus Genomic DNA Extraction Kit (Bioflux, China) using primers ITS1/ITS4 (White et al. 1990), bRPB2-6F/bRPB2-7.1R (Matheny 2005; Reeb et al. 2004) and ATP61/ATP62 (Kretzer and Bruns 1999), respectively. The ITS, RBP2 and ATP6 of the four isolates were sequenced and deposited in GenBank. BLASTn search of sequenced ITS (PQ187050, PQ187051, PP728052, PQ187052), RBP2 (PQ202833, PQ202834, PP735921, PQ202835) and ATP6 (PQ202837, PQ202838, PP735922, PQ202839) revealed a >99% identity with the type strain of <i>Ceratobasidium ramicola</i> CBS133.82 (NR138368, DQ301708, and DQ301577). For phylogenetic analysis, concatenated sequences of ITS, RBP2, and ATP6 were employed using the maximum-likelihood method in MEGA-X. Based on the morphological and molecular analyses, the isolates were classified into the <i>C</i>. <i>ramicola</i> clade (Bandoni 1979; Samuels et al. 2012; Jeong et al. 2023). To test the pathogenicity of the isolate PP23, mycelial plugs (5 mm in diameter) were placed directly on 15 healthy leaves from 15 three-year-old plants after puncturing with sterile needles. Sterile PDA plugs served as controls. All plants were kept in a greenhouse with conditions of 25°C, 80% relative humidity and a photoperiod of 12 h. After 5 days, all infected plants developed leaf blight symptoms similar to those described above, whe
云南白药(Paris polyphylla var. yunnanensis)是一种多年生草本植物,在抗癌、抗炎、抗菌、免疫调节和解痉等方面具有显著的药用价值(Duan等,2018)。2022 年 4 月,湘潭市(湖南)出现了叶枯病,波叶菊种植面积达 3×104 m2(27.904°N,112.918°E)。田间平均发病率为 22%,受感染植株最初表现为水渍状萎黄,随后干枯萎缩,并逐渐从叶尖扩展到整个植株。为鉴定病原菌,从 20 株植物上采集了 20 个叶片病斑(4 平方毫米),用 75% 的乙醇表面消毒 10 秒,5% NaOCl 消毒 30 秒,用无菌蒸馏水冲洗三次,转移到含乳酸(0.125%)的马铃薯葡萄糖琼脂(PDA)平板上,在 28 °C 黑暗中培养。获得了四个形态相似的分离物(PP21 至 PP24),并通过吸头法进行了纯化。PDA 上的菌落最初呈白色,带有棉状菌丝,后来底部变成浅黄色。有节菌丝呈直角分枝,在分枝点的离开处有一个小收缩,直径为 4.16 至 7.93 微米。使用 Giemsa 染色法(中国 Servicebio)可观察到隔膜菌丝内的双核细胞。为了进行分子鉴定,使用 ITS1/ITS4 引物(White et al.1990)、bRPB2-6F/bRPB2-7.1R(Matheny,2005;Reeb 等,2004)和 ATP61/ATP62 (Kretzer 和 Bruns,1999)引物。对四个分离株的 ITS、RBP2 和 ATP6 进行了测序,并存入 GenBank。对已测序的 ITS(PQ187050、PQ187051、PP728052、PQ187052)、RBP2(PQ202833、PQ202834、PP735921、PQ202835)和 ATP6(PQ202837、PQ202838、PP735922、PQ202839)进行 BLASTn 搜索,发现它们与 Ceratobasidium ramicola CBS133.82(NR138368、DQ301708 和 DQ301577)。在系统进化分析中,使用 MEGA-X 中的最大似然法对 ITS、RBP2 和 ATP6 的序列进行了合并分析。根据形态学和分子分析,分离物被归入 C. ramicola 支系(Bandoni,1979 年;Samuels 等,2012 年;Jeong 等,2023 年)。为了测试分离株 PP23 的致病性,用无菌针刺穿菌丝体后,将菌丝体插头(直径 5 毫米)直接插在 15 株三年生植物的 15 片健康叶片上。无菌 PDA 插条作为对照。5 天后,所有受感染的植株都出现了类似上述的叶枯病症状,而对照植株仍无症状。致病性试验进行了三次。从所有有症状的叶片中重新分离出了 C. ramicola,并根据形态和 ITS、RBP2 和 ATP6 的核苷酸序列确认其与原始分离株相同。据报道,从患病可可中分离出的 C. ramicola 很常见,但其对可可的致病性仍不清楚(Samuels 等,2012 年)。据我们所知,这是首次报道 C. ramicola 对 P. polyphylla(一种在中国具有重要经济价值的药材)造成叶枯病。
{"title":"First Report of Leaf Blight on <i>Paris polyphylla</i> Caused by <i>Ceratobasidium ramicola</i> in China.","authors":"Feiyan Xie, Yuhang Zhong, Xianjing Lin, Yuxi Chen, Li Yang, Ting Tang","doi":"10.1094/PDIS-05-24-1066-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-05-24-1066-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;&lt;i&gt;Paris polyphylla&lt;/i&gt; var. &lt;i&gt;yunnanensis&lt;/i&gt;is a perennial herb with significant medicinal properties in anticancer, anti-inflammatory, antibacterial immunomodulatory and antispasmodic activities (Duan et al. 2018). In April 2022, leaf blight disease emerged in Xiangtan City (Hunan), affecting &lt;i&gt;P&lt;/i&gt;. &lt;i&gt;polyphylla&lt;/i&gt; plantings over an area of 3×104 m&lt;sup&gt;2&lt;/sup&gt; (27.904°N, 112.918°E). The disease incidence reached an average of 22% of the plants in the field, with infected plants initially displaying water-soaked chlorosis, followed by dry yellow shrinkage that gradually spread from the leaf tips to the entire plant. To identify the causal agent, 20 leaf lesions (4 mm2) collected from 20 plants were surface sterilized with 75% ethanol for 10 s, 5% NaOCl for 30 s, rinsed in sterile distilled water three times, transferred to potato dextrose agar (PDA) plates with lactic acid (0.125%) , and incubated at 28 °C in the dark. Four isolates (PP21 to PP24) with similar morphologies were obtained and purified by the hyphal-tip method. Colonies on PDA initially appeared white with cottony mycelium, later turning light-yellow on the underside. Septate hyphae were branched at right angles with a small constriction at the departure of the branch point and measured 4.16 to 7.93 µm in diameter. Binucleate cells within the septate hyphae were visualized using Giemsa staining (Servicebio, China). For molecular identification, the rDNA internal transcribed spacer (ITS), the second largest subunit of nuclear DNA-directed RNA polymerase II (RPB2), and ATP synthase subunit 6 (ATP6) were amplified from genomic DNA of the isolates extracted by Fungus Genomic DNA Extraction Kit (Bioflux, China) using primers ITS1/ITS4 (White et al. 1990), bRPB2-6F/bRPB2-7.1R (Matheny 2005; Reeb et al. 2004) and ATP61/ATP62 (Kretzer and Bruns 1999), respectively. The ITS, RBP2 and ATP6 of the four isolates were sequenced and deposited in GenBank. BLASTn search of sequenced ITS (PQ187050, PQ187051, PP728052, PQ187052), RBP2 (PQ202833, PQ202834, PP735921, PQ202835) and ATP6 (PQ202837, PQ202838, PP735922, PQ202839) revealed a &gt;99% identity with the type strain of &lt;i&gt;Ceratobasidium ramicola&lt;/i&gt; CBS133.82 (NR138368, DQ301708, and DQ301577). For phylogenetic analysis, concatenated sequences of ITS, RBP2, and ATP6 were employed using the maximum-likelihood method in MEGA-X. Based on the morphological and molecular analyses, the isolates were classified into the &lt;i&gt;C&lt;/i&gt;. &lt;i&gt;ramicola&lt;/i&gt; clade (Bandoni 1979; Samuels et al. 2012; Jeong et al. 2023). To test the pathogenicity of the isolate PP23, mycelial plugs (5 mm in diameter) were placed directly on 15 healthy leaves from 15 three-year-old plants after puncturing with sterile needles. Sterile PDA plugs served as controls. All plants were kept in a greenhouse with conditions of 25°C, 80% relative humidity and a photoperiod of 12 h. After 5 days, all infected plants developed leaf blight symptoms similar to those described above, whe","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2024-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142505824","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
Field Evaluation of Fluazinam Fungicide in Dollar Spot Populations Confirmed In Vitro Insensitivity. 对氟啶虫酰胺杀菌剂在美元斑病种群中的实地评估证实了其体外不敏感性。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-22 DOI: 10.1094/PDIS-12-23-2639-RE
Xiaojing Shi, Soonhong Min, Geunhwa Jung

Fluazinam, a fungicide widely used in agriculture and turf management, was traditionally thought to pose a low risk of resistance. However, our in vitro sensitivity test conducted in 2021 revealed reduced sensitivity to fluazinam in dollar spot, highlighting the need for more vigilant field monitoring. In 2022 and 2023, we evaluated the field responses of four Clarireedia jacksonii isolates with varying in vitro sensitivity to fluazinam. Fluazinam was used at both a full labeled rate (0.5 oz/1,000 ft2) and a half-rate (0.25 oz/1,000 ft2) to evaluate the effectiveness in isolate-inoculated plots in the field. In 2022, both natural and sensitive isolates showed significantly better control compared to insensitive isolates under both half- and full-rate treatments. However, in 2023, half-rate fluazinam demonstrated limited control under high disease pressure, providing relative disease control of dollar spot less than 45% across all treatments. In contrast, full-rate fluazinam maintained significantly better control of natural and sensitive isolates compared with insensitive isolates. Our results, showing that in vitro insensitivity leads to field insensitivity under inoculated conditions, suggest the development of fluazinam insensitivity in the C. jacksonii population. This highlights the need for judicious use of fluazinam and the establishment of continuous resistance monitoring. Furthermore, the loss of control observed when fluazinam was applied at half-rates under high disease pressure highlights the importance of careful fungicide use.

氟啶胺是一种广泛用于农业和草坪管理的杀菌剂,传统上被认为产生抗药性的风险较低。然而,我们在 2021 年进行的体外敏感性测试表明,氟啶胺对美元斑的敏感性降低,这凸显了进行更仔细的田间监测的必要性。2022 年和 2023 年,我们评估了四种对氟啶虫酰胺具有不同体外敏感性的 Clarireedia jacksonii 分离物的田间反应。氟啶虫酰胺以全标剂量(0.5 盎司/1,000 平方英尺)和半标剂量(0.25 盎司/1,000 平方英尺)使用,以评估分离株接种地块在田间的效果。2022 年,在半量和全量处理中,天然和敏感分离株的防治效果明显优于不敏感分离株。然而,在 2023 年,半量氟啶脲在高病害压力下的防治效果有限,在所有处理中对美元斑的相对病害防治效果均低于 45%。相比之下,与不敏感的分离物相比,全剂量氟啶胺对天然和敏感分离物的控制效果明显更好。我们的结果表明,体外不敏感会导致接种田间条件下的田间不敏感,这表明杰克逊蓟马(C. jacksonii)种群对氟啶胺不敏感。这凸显了明智使用杀菌剂氟啶胺和建立持续抗性监测的必要性。此外,在高病害压力下以半量施用时观察到的防治效果下降也突出了谨慎使用杀菌剂的重要性。
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引用次数: 0
Development and Evaluation of Real-Time Quantitative PCR Assays for Detection of Phellinus noxius Causing Brown Root Rot Disease. 开发和评估实时定量 PCR 检测方法,用于检测引起褐根病的黄粉虫。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-22 DOI: 10.1094/PDIS-01-24-0238-RE
Tse-Yen Liu, Chao-Han Chen, Yi-Chun Ko, Zong-Chi Wu, Ting-Zhi Liao, Hsin-Han Lee, Isheng Jason Tsai, Tun-Tschu Chang, Meng-Ling Wu, Jyh-Nong Tsai, Ned B Klopfenstein, Mee-Sook Kim, Jane E Stewart, Ndeme Atibalentja, Fred E Brooks, Philip G Cannon, A Mohd Farid, Tsutomu Hattori, Hoi-Shan Kwan, Regent Yau Ching Lam, Yuko Ota, Norio Sahashi, Robert L Schlub, Louise S Shuey, Alvin M C Tang, Chia-Lin Chung

Brown root rot disease (BRRD) is a highly destructive tree disease. Early diagnosis of BRRD has been challenging because the first symptoms and signs are often observed after extensive tissue colonization. Existing molecular detection methods, all based on the internal transcribed spacer (ITS) region, were developed without testing against global Phellinus noxius isolates, other wood-decay fungi, or host plant tissues. This study aimed to develop SYBR Green real-time quantitative PCR (qPCR) assays for P. noxius. The primer pair Pn_ITS_F/Pn_ITS_R targets the ITS, and the primer pair Pn_NLR_F/Pn_NLR_R targets a P. noxius-unique group of homologous genes identified through a comparative genomics analysis. The homologous genes belong to the nucleotide-binding-oligomerization-domain-like receptor (NLR) superfamily. The new primer pairs and a previous primer pair G1F/G1R were optimized for qPCR conditions and tested for specificity using 61 global P. noxius isolates, 5 other Phellinus species, and 22 non-Phellinus wood-decay fungal species. Although all three primer pairs could detect as little as 100 fg (approximately 2.99 copies) of P. noxius genomic DNA, G1F/G1R had the highest specificity and Pn_NLR_F/Pn_NLR_R had the highest efficiency. To avoid false positives, the cutoff quantification cycle values were determined as 34 for G1F/G1R, 29 for Pn_ITS_F/Pn_ITS_R, and 32 for Pn_NLR_F/Pn_NLR_R. We further validated these qPCR assays using Ficus benjamina seedlings artificially inoculated with P. noxius, six tree species naturally infected by P. noxius, rhizosphere soil, and bulk soil. The newly developed qPCR assays provide sensitive detection and quantification of P. noxius, which is useful for long-term monitoring of BRRD status.

褐根病(BRRD)是一种破坏性很强的树木病害。由于褐根病的最初症状和体征往往是在大面积组织定植后才被观察到的,因此早期诊断褐根病一直是个难题。现有的分子检测方法都是基于内部转录间隔区(ITS)开发的,没有针对全球黄柏分离物、其他木材腐朽真菌或寄主植物组织进行测试。本研究开发了针对黑黄柏的 SYBR Green 实时定量 PCR (qPCR) 检测方法。引物对 Pn_ITS_F/Pn_ITS_R 针对 ITS,引物对 Pn_NLR_F/Pn_NLR_R 针对通过比较基因组学分析确定的 P. noxius 独有的同源基因组。这些同源基因属于核苷酸结合-同源异构化-类受体(NLR)超家族。对新引物对和以前的引物对 G1F/G1R 的 qPCR 条件进行了优化,并使用 61 个全球野黄柏分离物、5 个其他黄柏种类和 22 个非黄柏木材腐朽真菌种类对其特异性进行了测试。虽然所有三对引物都能检测到低至 100 fg(约 2.99 个拷贝)的 P. noxius 基因组 DNA,但 G1F/G1R 的特异性最高,Pn_NLR_F/Pn_NLR_R 的效率最高。为避免假阳性,我们确定 G1F/G1R 的 Cq 临界值为 34,Pn_ITS_F/Pn_ITS_R 为 29,Pn_NLR_F/Pn_NLR_R 为 32。我们还利用人工接种了鹅掌楸的榕树幼苗、六种自然感染鹅掌楸的树种、根瘤土壤和大块土壤进一步验证了这些 qPCR 检测方法。新开发的 qPCR 检测方法能灵敏地检测和定量 Noxius,有助于长期监测 BRRD 的状况。
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引用次数: 0
Evidence of the involvement of Xylella fastidiosa in the occurrence of walnut leaf scorch in Xinjiang, China. Xylella fastidiosa 参与中国新疆核桃叶焦枯病发生的证据。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-07-23-1430-PDN
Tong Guo, Shiwei Wang, Cunde Pan, Adil Sattar, Changjie Xing, Honglong Hao, Cuifang Zhang
<p><p>Walnut (Juglans regia Linn.) leaf scorch (WLS) was first reported in 2012 in Hotan, Xinjiang, China (Zhang et al., 2012). Initially, brown spots appear at the apex of the leaflets with slight shrinking. These spots spread inward along the leaf margins in a flame-like pattern. The scorched areas curl inward with a yellow halo. No powdery or bacterial signs were observed on the leaf surface. In severe cases, the leaves dried up and shrank, affecting the entire tree. However, new leaves did not show any signs of scorching. We collected 300 symptomatic leaf samples from 10-12 year-old trees of the susceptible WLS species Wen185, located in Daryaboyi (40°72'N, 80°49'E), Xakal (40°69'N, 80°52'E), and Karatal (40°73'N, 80°39'E) for X. fastidiosa PCR detection analysis. X. fastidiosa was detected in asymptomatic leaves of trees with severe WLS, as well as in asymptomatic leaves of trees exhibiting mild WLS symptoms, and it was even found in asymptomatic leaves of trees without any WLS symptoms.To isolate X. fastidiosa, walnut leaves with petioles were disinfected with 3% bleach for 10 minutes, followed by four washes in autoclaved deionized water. The midrib and petiole of the leaf were cut off with a sterile blade, and a 2 mm to 3 mm section was excised. Then, the cut ends were squeezed with a pair of pliers, and the sap was blotted onto Periwinkle Wilt (PW) plates (Davis et al. 1983). The plates were sealed with parafilm and incubated at 28°C in the dark, with daily observations for the development of individual colonies. The sap collected from 5 out of 20 leaf samples with scorch symptoms and from 20 leaves without symptoms never showed bacterial growth in PW.. Six colonies were tested and confirmed positive for X. fastidiosa using the RST31 and RST33 primer pair (Minsavage et al., 1994). The uploaded sequence accession numbers are PP871340-PP871342. Blast analysis of Multilocus Sequence Typing (MLST) sequences from the isolated strain using the X. fastidiosa MLST Database (http://pubmlst.org/xfastidiosa) revealed a perfect match with sequences of alleles leuA_3, petC_3, malF_, cysG_3, holC_4, nuoL_3, and gltT_3 (PP871343-PP871349). Using MEGA software to concatenate the MLST single gene fragments and construct a ML multi-gene fragment phylogenetic tree through Maximum Likelihood (ML) analysis, the bacterium was identified as X. fastidiosa subsp. multiplex To fulfill Koch's postulates, a purified colony from the PW plate was suspended in 500 μL of deionized water at 1×10^8 CFU·mL-1. A sterilized 5 ml medical needle was used to prick a small hole on the main branch, 5 cm from the base, of three-year-old walnut seedlings, and the bacterial solution was injected into the small hole. Three biological replicates were performed for each treatment (5 μL and 10 μL of bacterial solution, deionized water), and the experiment was repeated three times. Scorching symptoms were observed three months after inoculation in 11 out of the 15 seedlings inoculated w
核桃(Juglans regia Linn.)叶焦枯病(WLS)于 2012 年首次在中国新疆和田地区被报道(Zhang 等人,2012 年)。最初,小叶先端出现褐色斑点,并伴有轻微萎缩。这些斑点沿叶缘向内蔓延,呈火焰状。焦枯区域向内卷曲,并带有黄色光晕。叶片表面没有白粉或细菌迹象。严重时,叶片干枯萎缩,影响整棵树。不过,新叶没有任何焦枯迹象。我们从位于 Daryaboyi(北纬 40°72',东经 80°49')、Xakal(北纬 40°69',东经 80°52')和 Karatal(北纬 40°73',东经 80°39')的易感 WLS 品种 Wen185 的 10-12 年树龄树上采集了 300 份有症状的叶片样本,进行 X. fastidiosa PCR 检测分析。为了分离 X. fastidiosa,核桃叶柄用 3% 的漂白剂消毒 10 分钟,然后在高压去离子水中清洗四次。用无菌刀片切下叶片的中脉和叶柄,并切除 2 至 3 毫米的部分。然后用钳子挤压切口,将汁液吸到长春花枯萎病(PW)平板上(Davis 等,1983 年)。平板用保鲜膜密封,28°C 黑暗培养,每天观察单个菌落的生长情况。从 20 个有焦枯症状的叶片样本中的 5 个样本和 20 个无症状的叶片样本中采集的汁液在 PW 中从未出现细菌生长。使用 RST31 和 RST33 引物对 6 个菌落进行了检测,确认其对 X. fastidiosa 呈阳性(Minsavage 等人,1994 年)。上传的序列登录号为 PP871340-PP871342。利用 X. fastidiosa MLST 数据库(http://pubmlst.org/xfastidiosa)对分离菌株的多焦点序列分型(MLST)序列进行的快速分析表明,等位基因 leuA_3、petC_3、malF_、cysG_3、holC_4、nuoL_3 和 gltT_3 的序列完全匹配(PP871343-PP871349)。使用 MEGA 软件连接 MLST 单基因片段,并通过最大似然法(ML)分析构建 ML 多基因片段系统发生树,确定该细菌为 X. fastidiosa subsp. 为了满足科赫假设,将 PW 平板上纯化的菌落悬浮在 500 μL 去离子水中,浓度为 1×10^8 CFU-mL-1。用消毒过的 5 ml 医用针头在三年生核桃树苗主枝上距基部 5 cm 处扎一个小孔,然后将菌液注入小孔中。每个处理进行三个生物重复(5 μL 和 10 μL 菌液、去离子水),实验重复三次。接种 10 μL 菌液的 15 棵树苗中,有 11 棵在接种三个月后出现灼烧症状,其中 5 棵树苗的叶片样本中分离出了 X. fastidiosa,验证了科赫假设。用水浇灌的核桃树苗没有症状,PCR阴性。据我们所知,X. fastidiosa 已在中国陕西的葡萄(Chu,2001 年)和中国台湾的梨(Su 等人,2016 年)中被发现感染。这是中国首次报道 X. fastidiosa 感染 WLS。对该病发生情况的进一步研究将有助于防止该病的传播。
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引用次数: 0
First Report of Bacterial Soft Rot of Potato Caused by Pectobacterium aroidearum strains in China. 中国首次报道由果胶杆菌菌株引起的马铃薯细菌性软腐病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-07-24-1524-PDN
Guoling Chen, Huifang Shen, Xiaoming Pu, Dayuan Sun, Pingping Liu, Qiyun Yang, Shaohua Chen, Jingxin Zhang
<p><p>Potato (<i>Solanum tuberosum</i> L.) is a globally important staple crop, and China has contributed more than 20% of the world's production. In Guangdong Province, potato has become one of the most important winter crops, increased the farmer income. However, the potato were seriously affected by soft rot disease caused by <i>Pectobacterium</i> spp. in the past decade. In February 2024, typical symptoms of potato soft rot were observed on the plants of cultivar "Daxiyang" in Enping City, Guangdong Province, where the disease incidences ranged 3%-5% in the investigated fields. The stems adjacent to the tubes showed typical inky black symptoms, and the plants appeared yellow. The interior of the tubes appeared water-soaked and had developed to overall decay, with an evident smell. Eleven symptomatic plants were collected and used to isolate the causal agents. Pieces of tube tissue (about 5×5 mm) were removed between the rot-symptomatic and non-symptomatic margins, and surface sterilized using 75% ethanol for 30 s and 2% NaClO for 1 min. The pieces were rinsed in sterile water for three times, and then plated on Luria-Bertani (LB) medium agar for 36 h at 30°C. The produced colonies were selected for hypersensitive test on tobacco, and the colonies with a positive reaction were subcultured three times. Two representative strains, EP51-2 and EP51-3, were processed for gene sequencing, including house-keeping genes, <i>danX</i>, <i>leuS</i>, and <i>recA</i> (Portier et al. 2019). The gene sequences (GenBank accessions no. PP870934 to PP870939) of these two representative strains were matched with <i>Pectobacterium aroidearum</i> strain NCPPB 929 (CP166097.1) with identities ranging from 97.15 to 100% and the coverage of 100%. The sequences of three genes were concatenated and used for phylogenetic analyses, along with representative strains from 19 other <i>Pectobacterium</i> species. Phylogenetic analyses supported that EP51-2 and EP51-3 were grouped into <i>P. aroidearum</i> with the representative strains. Koch's postulates were applied to determine the pathogenicity of <i>P. aroidearum</i> strain EP51-2. According to the external inoculation method on potato stems (Czajkowski et al. 2010), each of six pots of healthy potato plants was inoculated at the basal stem with 100 µl of bacterial suspension (10<sup>8</sup> CFU/ml), while six pots inoculated with LB served as controls. The plants were then kept at 30°C and maintained at 95% humidity. After 2 days, all six plants inoculated with the bacteria exhibited typical symptoms on the stems, which progressed to the collapse of the whole plant; the controls remained symptom-free during the observation. Single colonies reisolated from the symptomatic stem of the inoculated plants were confirmed using PCR and sequencing with <i>recA</i> primers as described above, and the causal agent was identified as <i>P. aroidearum</i> strains, fulfilling Koch's postulates. <i>P. aroidearum</i> has been reported
马铃薯(Solanum tuberosum L.)是全球重要的主粮作物,中国的产量占世界总产量的 20% 以上。在广东省,马铃薯已成为最重要的冬季作物之一,增加了农民收入。然而,近十年来,马铃薯受到由果胶杆菌引起的软腐病的严重影响。2024 年 2 月,广东省恩平市栽培品种 "大溪洋 "的植株上出现了典型的马铃薯软腐病症状,调查田块的发病率为 3%-5%。邻近试管的茎部出现典型的墨黑色症状,植株呈黄色。试管内部呈水渍状,整体腐烂,气味明显。收集了 11 株出现症状的植株,用于分离病原体。在有腐烂症状和无症状的边缘之间取下一小块试管组织(约 5×5 毫米),用 75% 的乙醇消毒 30 秒,再用 2% 的 NaClO 消毒 1 分钟。用无菌水冲洗 3 次,然后在 30°C 下在 Luria-Bertani(LB)培养基琼脂上培养 36 小时。选出产生的菌落在烟草上进行高敏试验,阳性反应的菌落再培养三次。对两个代表性菌株 EP51-2 和 EP51-3 进行了基因测序,包括看家基因、danX、leuS 和 recA(Portier 等,2019 年)。这两个代表性菌株的基因序列(GenBank登录号:PP870934 至 PP870939)与果胶杆菌菌株 NCPPB 929(CP166097.1)进行了比对,同一性为 97.15%至 100%,覆盖率为 100%。三个基因的序列被连接起来,与其他 19 种果胶杆菌的代表性菌株一起用于系统发育分析。系统进化分析结果表明,EP51-2 和 EP51-3 与代表菌株被归入 P. aroidearum。应用科赫推定来确定甲壳菌 EP51-2 株的致病性。根据马铃薯茎部外部接种法(Czajkowski 等人,2010 年),在六盆健康马铃薯植株的基部茎部各接种 100 µl 的细菌悬浮液(108 CFU/ml),六盆接种 LB 的植株作为对照。然后将植株放在 30°C 的温度下,保持 95% 的湿度。2 天后,接种了细菌的所有 6 盆植物的茎部都出现了典型症状,随后发展到整株植物倒伏;而对照组在观察期间一直没有出现症状。如上所述,使用 recA 引物进行 PCR 和测序,从接种植株有症状的茎上重新分离出的单菌落得到了确认,病原菌被确定为甲型肝炎嗜血杆菌菌株,符合科赫假说。在中国,大白菜(Xie 等,2018 年)和叶芥(Chu 等,2024 年)以及魔芋(Wei 等,2021 年)、芋头(Zhou 等,2022 年)和半夏(Du 等,2024 年)等天南星科作物上都曾报道过 P. aroidearum。据我们所知,这是 P. aroidearum 在中国引起马铃薯软腐病的首次报道。这种病原菌在广东省的芋头田里很普遍,造成了严重损失。本报告强调了该病原体寄主范围的扩大。应重点关注这种新出现的影响马铃薯的病原体,并立即采取措施控制其传播。
{"title":"First Report of Bacterial Soft Rot of Potato Caused by <i>Pectobacterium aroidearum</i> strains in China.","authors":"Guoling Chen, Huifang Shen, Xiaoming Pu, Dayuan Sun, Pingping Liu, Qiyun Yang, Shaohua Chen, Jingxin Zhang","doi":"10.1094/PDIS-07-24-1524-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-07-24-1524-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Potato (&lt;i&gt;Solanum tuberosum&lt;/i&gt; L.) is a globally important staple crop, and China has contributed more than 20% of the world's production. In Guangdong Province, potato has become one of the most important winter crops, increased the farmer income. However, the potato were seriously affected by soft rot disease caused by &lt;i&gt;Pectobacterium&lt;/i&gt; spp. in the past decade. In February 2024, typical symptoms of potato soft rot were observed on the plants of cultivar \"Daxiyang\" in Enping City, Guangdong Province, where the disease incidences ranged 3%-5% in the investigated fields. The stems adjacent to the tubes showed typical inky black symptoms, and the plants appeared yellow. The interior of the tubes appeared water-soaked and had developed to overall decay, with an evident smell. Eleven symptomatic plants were collected and used to isolate the causal agents. Pieces of tube tissue (about 5×5 mm) were removed between the rot-symptomatic and non-symptomatic margins, and surface sterilized using 75% ethanol for 30 s and 2% NaClO for 1 min. The pieces were rinsed in sterile water for three times, and then plated on Luria-Bertani (LB) medium agar for 36 h at 30°C. The produced colonies were selected for hypersensitive test on tobacco, and the colonies with a positive reaction were subcultured three times. Two representative strains, EP51-2 and EP51-3, were processed for gene sequencing, including house-keeping genes, &lt;i&gt;danX&lt;/i&gt;, &lt;i&gt;leuS&lt;/i&gt;, and &lt;i&gt;recA&lt;/i&gt; (Portier et al. 2019). The gene sequences (GenBank accessions no. PP870934 to PP870939) of these two representative strains were matched with &lt;i&gt;Pectobacterium aroidearum&lt;/i&gt; strain NCPPB 929 (CP166097.1) with identities ranging from 97.15 to 100% and the coverage of 100%. The sequences of three genes were concatenated and used for phylogenetic analyses, along with representative strains from 19 other &lt;i&gt;Pectobacterium&lt;/i&gt; species. Phylogenetic analyses supported that EP51-2 and EP51-3 were grouped into &lt;i&gt;P. aroidearum&lt;/i&gt; with the representative strains. Koch's postulates were applied to determine the pathogenicity of &lt;i&gt;P. aroidearum&lt;/i&gt; strain EP51-2. According to the external inoculation method on potato stems (Czajkowski et al. 2010), each of six pots of healthy potato plants was inoculated at the basal stem with 100 µl of bacterial suspension (10&lt;sup&gt;8&lt;/sup&gt; CFU/ml), while six pots inoculated with LB served as controls. The plants were then kept at 30°C and maintained at 95% humidity. After 2 days, all six plants inoculated with the bacteria exhibited typical symptoms on the stems, which progressed to the collapse of the whole plant; the controls remained symptom-free during the observation. Single colonies reisolated from the symptomatic stem of the inoculated plants were confirmed using PCR and sequencing with &lt;i&gt;recA&lt;/i&gt; primers as described above, and the causal agent was identified as &lt;i&gt;P. aroidearum&lt;/i&gt; strains, fulfilling Koch's postulates. &lt;i&gt;P. aroidearum&lt;/i&gt; has been reported","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2024-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142472376","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 Fire Blight Caused by Erwinia amylovora on Pear in Saudi Arabia. 沙特阿拉伯首次报告由 Erwinia amylovora 引起的梨火疫病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-03-24-0675-PDN
Yasser Eid Eid Ibrahim, Abdur Rehman, Mohammed Ali Al Saleh
<p><p>In the spring of 2020 and 2021, pear trees (Pyrus communis cv. Williams) in orchards (27°46'36.0"N 42°29'44.7"E, 30°00'00.2"N 40°15'11.8"E, and 28°44'52.9"N 36°18'47.8" E) in Hail, Al Jouf, and Tabuk regions exhibited fire blight symptoms. After removing bark, the affected trees showed shoot blight, brown-blighted shoot tips and blossom blight, dead flowers on the stems, and reddish-colored cankers. The disease incidence varied from 10% to 25%. Pathogen was isolated from 21 symptomatic samples including fruits, flowers, and shoots. Bacteria were isolated from washed tissues on King's B (KB) and semi-selective CCT media (Ishimaru and Klos, 1984). After 48 hours, colonies resembling Erwinia amylovora on KB media were 1.5-2 mm in diameter, white, circular, slightly convex, with a smooth surface, and exhibited no fluorescence under ultraviolet light. Colonies on CCT after 72 h were 3-4 mm in diameter, mucoid with shiny surfaces, semitransparent, speckled with craters, and slightly violet. All isolates were purified by subculturing on KB. All isolates were Gram-negative and rod-shaped, fermentative of glucose, positive for catalase and negative for oxidase, and potato rot. They induced a hypersensitive reaction when infiltrated in tobacco leaves (cv. Xanthi). Based on morphology and biochemical tests (EPPO, 2004), the strains were identified as Erwinia sp. Twenty-six strains from Saudi Arabia (SA) and the reference strain (NCPPB 683T) hydrolyzed gelatin and formed white, highly mucous colonies on the levan medium. These strains could not reduce nitrate to nitrite and tested negative for urease and indole production. All the isolates and the reference strain were confirmed to be E. amylovora based on a PCR 0.9-kb DNA fragment amplification with a species-specific primer set, A/B targets pEA29 (Bereswill et al. 1992). 16S-rDNA fragments from Saudi isolates were amplified with 27F and 1492R primers (Lane, 1991). Purified amplicons from PCR were sequenced (OR717505 and OR743536-OR743560), and a BLAST search of the GenBank database revealed 100% (927/927) homology with E. amylovora strain CP066796.1. The housekeeping gene rpoB was PCR amplified with primers CM7-F and CM31b-R (Rezzonico et al. 2012), and the products were sequenced (PP465516-PP465541). BLAST analysis showed 100% (944/944 nt) and 96.19% (908/944 nt) identities with the sequences of E. amylovora ATCC 15580 CP066796.1 and E. pyrifoliae CP201486 CP103445.1, respectively. To fulfill Koch's postulates, SA strains were inoculated on five healthy 3-month-old clones of apple (Malus domestica cv. Gala) per strain with a 10-μl bacterial suspension containing 107 colony-forming units per milliliter by injecting directly in the veins of the upper second leaf plus 5 healthy plants injected with sterile distilled water as control. Plants were incubated at 28°C for six days under a 12-hour light regime. Observed symptoms were similar to the ones observed in the field. The experiment was replicated tw
2020 年和 2021 年春季,海尔、Al Jouf 和塔布克地区果园(北纬 27°46'36.0",东经 42°29'44.7",北纬 30°00'00.2",东经 40°15'11.8",北纬 28°44'52.9",东经 36°18'47.8")中的梨树(Pyrus communis cv. Williams)出现了火疫病症状。除去树皮后,受影响的树木出现嫩枝枯萎病、嫩枝顶端褐色枯萎病和花枯萎病、茎上有枯萎的花朵以及红颜色的溃疡。发病率从 10% 到 25% 不等。从 21 个有症状的样本(包括果实、花和嫩枝)中分离出了病原体。在 King's B(KB)和半选择性 CCT 培养基(Ishimaru 和 Klos,1984 年)上从洗净的组织中分离细菌。48 小时后,KB 培养基上类似于 Erwinia amylovora 的菌落直径为 1.5-2 毫米,白色,圆形,微凸,表面光滑,在紫外线下无荧光。72 小时后,CCT 培养基上的菌落直径为 3-4 毫米,呈粘液状,表面发亮,半透明,有斑点状凹坑,略带紫色。所有分离物均在 KB 上进行亚培养纯化。所有分离物均为革兰氏阴性,杆状,葡萄糖发酵,过氧化氢酶阳性,氧化酶阴性,马铃薯腐烂。当它们渗入烟草叶片(Xanthi 栽培品种)时,会诱发超敏反应。来自沙特阿拉伯(SA)的 26 株菌株和参考菌株(NCPPB 683T)可水解明胶,并在 levan 培养基上形成白色高粘液菌落。这些菌株不能将硝酸盐还原成亚硝酸盐,脲酶和吲哚生产检测呈阴性。根据使用物种特异性引物集 A/B 目标 pEA29(Bereswill 等人,1992 年)进行 PCR 0.9-kb DNA 片段扩增的结果,确认所有分离株和参考菌株均为淀粉伊蚊(E. amylovora)。沙特分离物的 16S-rDNA 片段用 27F 和 1492R 引物扩增(Lane,1991 年)。从 PCR 中纯化的扩增子被测序(OR717505 和 OR743536-OR743560),GenBank 数据库的 BLAST 搜索显示与 E. amylovora 菌株 CP066796.1 的同源性为 100%(927/927)。用引物 CM7-F 和 CM31b-R 对看家基因 rpoB 进行了 PCR 扩增(Rezzonico 等人,2012 年),并对扩增产物进行了测序(PP465516-PP465541)。BLAST 分析表明,与 E. amylovora ATCC 15580 CP066796.1 和 E. pyrifoliae CP201486 CP103445.1 的序列相同度分别为 100%(944/944 nt)和 96.19%(908/944 nt)。为了实现科赫假设,将 SA 菌株接种到 5 株 3 个月大的健康苹果(Malus domestica cv. Gala)克隆上,每株接种 10 微升的细菌悬浮液,每毫升含 107 个菌落形成单位,直接注射到第二片叶子上部的叶脉中,另外 5 株健康植株注射无菌蒸馏水作为对照。植物在 28℃、12 小时光照条件下培养 6 天。观察到的症状与田间观察到的症状相似。实验重复两次。从接种的苹果砧木上重新分离出 CCT 培养基上的细菌菌落,并通过 A/B 引物组进行确认。据我们所知,这是自 2013 年在南澳大利亚州观察到类似火疫病症状以来(Alhudaib,2013 年),南澳大利亚州首次出现 E. amylovora 的同行评审报告。进一步的研究将为南澳大利亚确定火疫病病原体的新寄主植物,这一点非常重要,因为南澳大利亚从邻近的约旦进口梨果幼苗(榅桲、苹果和梨),而约旦曾报道过 E. aylovora(Tehabsim 等人,1992 年)。
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引用次数: 0
Introgression and Mapping of Cotton Leaf Curl Disease (CLCuD) Resistance from Wild Gossypium armourianum Kearney into Upland Cotton (G. hirsutum L.). 棉花卷叶病 (CLCuD) 抗性从野生 Gossypium armourianum Kearney 向陆地棉 (G. hirsutum L.) 的导入和图谱绘制。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-08-24-1645-SC
Dharminder Pathak, Pankaj Rathore, Harpreet Kaur, Bhupinder Singh, Harish Kumar, Akhtar Ali, Sunayana Punia, Parvinder Singh Sekhon, Kuldeep Singh

Cotton leaf curl disease (CLCuD), caused by the whitefly transmitted geminivirus complex (Cotton leaf curl virus - CLCuV and their satellite molecules), is a serious threat to successful upland cotton production in northwest India, Pakistan, and China. The disease causes significant losses in fibre yield and the quality of cotton. Owing to the regular emergence of resistance breaking strains of CLCuV, all the previously available CLCuD resistant germplasms of upland cotton have become compromised and none of the extant upland cotton cultivars is resistant to this disease. Therefore, alternate sources of CLCuD resistance need to be explored, as genetic resistance is the only pragmatic and tenable management strategy to combat this malady. Here, we report for the first time the introgression and mapping of CLCuD resistance from a related non-progenitor wild diploid D-genome cotton species, G. armourianum into upland cotton. A backcross population (G. hirsutum/G. armourianum/G. hirsutum) was developed for this purpose. A single major QTL was found to be associated with resistance to CLCuD and was located on chromosome D01 through the genotyping-by-sequencing technique.

棉花卷叶病(CLCuD)由粉虱传播的 geminivirus 复合体(棉花卷叶病毒 - CLCuV 及其卫星分子)引起,严重威胁印度西北部、巴基斯坦和中国的陆地棉生产。该疾病对棉花的纤维产量和质量造成重大损失。由于 CLCuV 抗性株系的不断出现,所有以前可用的抗 CLCuD 的陆地棉种质都受到了影响,现存的陆地棉栽培品种中没有一个对该病害具有抗性。因此,需要探索 CLCuD 抗性的替代来源,因为遗传抗性是防治该病害的唯一实用且可行的管理策略。在此,我们首次报告了从相关的非祖先野生二倍体 D 基因组棉花物种 G. armourianum 向陆地棉中导入 CLCuD 抗性并绘制抗性图谱的情况。为此开发了一个回交群体(G. hirsutum/G. armourianum/G.hirsutum)。通过基因分型测序技术,发现一个主要的 QTL 与 CLCuD 的抗性有关,并位于 D01 染色体上。
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引用次数: 0
Eight novel diagnostic markers differentiate lineages of the highly invasive myrtle rust pathogen Austropuccinia psidii. 八种新型诊断标记可区分高侵染性桃金娘锈病病原体Austropuccinia psidii的品系。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-10-24-2111-SR
Zhenyan Luo, Jinghang Feng, Austin Bird, Mareike Moeller, Rita Tam, Luc Shepherd, Lydia Murphy, Lavi Singh, Abigail Graetz, Lilian Amorim, Nelson Sidnei Massola Júnior, M Asaduzzaman Prodhan, Louise S Shuey, Douglas Beattie, Alejandro Trujillo Gonzalez, Peri Tobias, Amanda Padovan, Rohan Benjamin Essex Kimber, A R McTaggart, Monica Kehoe, Benjamin Schwessinger, Thais Regina Boufleur

Austropuccinia psidii is the causal agent of myrtle rust in over 480 species within the family Myrtaceae. Lineages of A. psidii are structured by their hosts in the native range, and some have success in infecting newly encountered hosts. For example, the pandemic biotype has spread beyond South America, and proliferation of other lineages is an additional risk to biodiversity and industries. Efforts to manage A. psidii incursions, including lineage differentiation, relies on variable microsatellite markers. Testing these markers is time-consuming, complex, and requires reference material that is not always readily available. We designed a novel diagnostic approach targeting eight selectively chosen loci including the fungal mating-type HD (homeodomain) transcription factor locus. The HD locus (bW1/2-HD1 and bE1/2-HD2) is highly polymorphic, facilitating clear biological predictions about its inheritance from founding populations. To be considered as potentially derived from the same lineage, all four HD alleles must be identical. If all four HD alleles are identical six additional markers can further differentiate lineage identity. Our lineage diagnostics relies on PCR amplification of eight loci in different genotypes of A. psidii followed by amplicon sequencing using Oxford Nanopore Technologies (ONT) and comparative analysis. The lineage-specific assay was validated on four isolates with existing genomes, uncharacterized isolates, and directly from infected leaf material. We reconstructed alleles from amplicons and confirmed their sequence identity relative to their reference. Genealogies of alleles confirmed the variations at the loci among lineages/isolates. Our study establishes a robust diagnostic tool for differentiating known lineages of A. psidii based on biological predictions and available nucleotide sequences. This tool is suited to detecting the origin of new pathogen incursions.

桃金娘锈菌(Austropuccinia psidii)是桃金娘科 480 多个物种桃金娘锈病的病原菌。A.psidii的品系由其在原产地的寄主构成,有些品系能成功感染新遇到的寄主。例如,大流行生物型已扩散到南美洲以外,其他品系的扩散对生物多样性和产业造成了额外的风险。管理 A. psidii 入侵(包括品系区分)的工作依赖于可变的微卫星标记。对这些标记进行测试既费时又复杂,而且需要参考材料,而这些材料并不总能轻易获得。我们设计了一种新的诊断方法,针对八个选择性位点,包括真菌交配型 HD(homeodomain)转录因子位点。HD 基因座(bW1/2-HD1 和 bE1/2-HD2)具有高度多态性,有助于从生物学角度明确预测其在原生种群中的遗传性。要想被视为可能来自同一血统,所有四个 HD 等位基因必须完全相同。如果所有四个 HD 等位基因都相同,则六个附加标记可进一步区分血统身份。我们的品系诊断依赖于对不同基因型 A. psidii 的八个位点进行 PCR 扩增,然后使用牛津纳米孔技术(ONT)进行扩增片段测序和比较分析。我们在四个已有基因组的分离株、未定性的分离株以及直接从感染叶片材料中提取的分离株上验证了该品系特异性检测方法。我们从扩增子中重建了等位基因,并确认了它们相对于参照物的序列同一性。等位基因谱系证实了不同品系/分离株之间基因位点的变异。我们的研究为根据生物学预测和现有核苷酸序列区分 A. psidii 的已知品系提供了一种可靠的诊断工具。该工具适用于检测新病原体入侵的起源。
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引用次数: 0
Comparative Fitness of Monilinia fructicola Isolates with Multiple Fungicide-Resistant Phenotypes. 具有多种抗杀菌剂表型的果蝇科莫尼林菌分离株的适应性比较。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-12-23-2549-RE
Pamela S S Dutra, Thiago A Carraro, Cristiano N Nesi, Lilian Amorim, Louise L May De Mio

This study characterized 52 isolates of Monilinia fructicola from peach and nectarine orchards for their multiresistance patterns to thiophanate-methyl (TF), tebuconazole (TEB), and azoxystrobin (AZO) using in vitro sensitivity assays and molecular analysis. The radial growth of M. fructicola isolates was measured on media amended with a single discriminatory dose of 1 μg/ml for TF and AZO and 0.3 μg/ml for TEB. Cyt b, CYP51, and β-tubulin were tested for point mutations that confer resistance to quinone outside inhibitors (QoIs), demethylation inhibitors (DMIs), and methyl benzimidazole carbamates (MBCs), respectively. Eight phenotypes were identified, including isolates with single, double, and triple in vitro resistance to QoI, MBC, and DMI fungicides. All resistant phenotypes to TF and TEB presented the H6Y mutation in β-tubulin and the G641S mutation in CYP51. None of the point mutations typically linked to QoI resistance were present in the Monilinia isolates examined. Moreover, fitness of the M. fructicola phenotypes was examined in vitro and in detached fruit assays. Phenotypes with single resistance displayed equal fitness in vitro and in fruit assays compared with the wild type. In contrast, the dual- and triple-resistance phenotypes suffered fitness penalties based on osmotic sensitivity and aggressiveness on peach fruit. In this study, multiple resistance to MBC, DMI, and QoI fungicide groups was confirmed in M. fructicola. Results suggest that Monilinia populations with multiple resistance phenotypes are likely to be less competitive in the field than those with single resistance, thereby impeding their establishment over time and facilitating disease management.

本研究通过体外敏感性测定和分子分析,确定了从桃园和油桃园中分离出的 52 株果孢霉对甲基硫菌灵(TF)、戊唑醇(TEB)和唑菌胺(AZO)的多重抗性模式。在使用单次鉴别剂量(TF 和 AZO 为 1 µg/ml,TEB 为 0.3 µg/ml)的培养基上测量了果蝇分离菌的径向生长。对 Cyt b、CYP51 和 ß-tubulin进行了点突变检测,这些点突变分别赋予了它们对醌外抑制剂(QoIs)、去甲基化抑制剂(DMIs)和甲基苯并咪唑氨基甲酸酯(MBCs)的抗性。鉴定出了八种表型,包括对 QoI、MBC 和 DMI 杀菌剂具有单一、双重和三重体外抗性的分离物。所有对 TF 和 TEB 产生抗性的表型都出现了 ß-微管蛋白中的 H6Y 突变和 CYP51 中的 G641S 突变。在所检测的莫尼林菌分离物中,没有发现与 QoI 抗性相关的典型点突变。此外,还在体外和分离果实试验中检验了果蝇表型的适应性。与野生型相比,具有单一抗性的表型在体外和果实试验中显示出相同的适应性。与此相反,双抗和三抗表型在渗透敏感性和对桃果的侵袭性方面的适应性受到了影响。在这项研究中,证实了果孢霉对 MBC、DMI 和 QoI 杀菌剂组的多重抗性。研究结果表明,具有多重抗性表型的莫尼林病菌种群在田间的竞争力可能低于具有单一抗性的种群,从而阻碍了它们在田间的长期繁殖,有利于病害的防治。
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引用次数: 0
Construction of a comprehensive meteorological risk index for wheat Fusarium head blight prediction based on more than a half-century of monitoring data. 基于半个多世纪的监测数据,构建用于小麦镰刀菌头枯病预测的综合气象风险指数。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-05-24-1095-RE
Min Xu, Yifei Xu, Jingwei Xu, Meng Xu, Rongming Yang, Xin Liu

Fusarium head blight (FHB) represents a critical threat to wheat production globally, not only reducing yields but also contaminating crops with harmful mycotoxins. This study aimed to elucidate new spatiotemporal patterns of FHB incidence and to develop a comprehensive meteorological risk index to enhance scientific prevention and control of the disease. Through the analysis of annual and decadal variations from 1965 to 2023, the study assessed FHB trends across four agricultural regions (I, II, III, and IV) in Jiangsu Province, located in the middle and lower reaches of the Yangtze River-a hotspot for FHB in China. Key findings include: Since 1965, Regions I and III consistently exhibited higher FHB incidence rates compared to Regions II and IV. Post-2000, there was a notable increase in years with high incidence rates, with Region III overtaking Region I as the region with the highest incidence. Since 2010, occurrences of FHB reaching the most severe grade (Grade 5) have surpassed those in previous decades across all regions. The study also revealed a stronger correlation between meteorological factors (cumulative precipitation, number of days with rainfall ≥ 0.1 mm, total rainy days with ≥ 2 and ≥3 consecutive days of rain, total rainy days with both average daily air temperature ≥ 15 °C and daily rainfall ≥ 0.1 mm, days with average daily relative humidity ≥ 85%, cumulative sunshine hours, and cumulative cloudy days) and the FHB incidence rates during the heading-flowering-grain filling period in Regions I, II, and III, compared to the heading-flowering period alone. This indicates that optimal temperature and high humidity during the grain filling stage significantly contribute to the final FHB incidence rates. Despite the less apparent correlation between temperature changes and disease rates, the significant warming trend observed since 2000 has likely fostered conditions conducive to the proliferation of FHB. The comprehensive meteorological risk index, constructed to incorporate key meteorological factors during the heading-flowering-filling period, showed a strong correlation with actual disease incidences. The index demonstrated fitting accuracy rates of 84.7% for Region I, 72.9% for Region II, 83.1% for Region III, and 90.9% for Region IV, underscoring its effectiveness in predicting FHB occurrences. This tool offers both convenience and practicality, providing valuable insights for strategically managing FHB risks based on local weather conditions.

镰刀菌白粉病(FHB)对全球小麦生产构成严重威胁,不仅会降低产量,还会使作物受到有害霉菌毒素的污染。本研究旨在阐明 FHB 发病的新时空模式,并开发综合气象风险指数,以加强对该病的科学防控。该研究通过分析 1965 年至 2023 年的年变化和十年变化,评估了位于长江中下游--中国 FHB 的热点地区--江苏省四个农业区(I、II、III 和 IV)的 FHB 趋势。主要发现包括自 1965 年以来,Ⅰ区和Ⅲ区的 FHB 发病率一直高于Ⅱ区和Ⅳ区。2000 年后,高发病率年份明显增多,Ⅲ区超过Ⅰ区成为发病率最高的地区。自 2010 年以来,所有地区达到最严重等级(5 级)的 FHB 发生率都超过了之前几十年的发生率。研究还显示,气象因素(累计降水量、降雨量≥0.1 毫米的天数、连续降雨≥2 天和≥3 天的总雨天数、日平均气温≥15 °C、日降雨量≥0.1 毫米、日平均相对湿度≥ 85% 的天数、累计日照时数和累计阴天数),以及Ⅰ区、Ⅱ区和Ⅲ区在打尖-开花-谷粒灌浆期的 FHB 发病率与仅打尖-开花期的 FHB 发病率相比。这表明,谷粒灌浆期的最佳温度和高湿度对 FHB 的最终发病率有显著影响。尽管温度变化与病害发生率之间的相关性并不明显,但自 2000 年以来观察到的显著变暖趋势很可能为 FHB 的扩散提供了有利条件。综合气象风险指数的构建包含了打顶-开花-灌浆期的关键气象因素,该指数与实际病害发生率有很强的相关性。该指数的拟合准确率在 I 区为 84.7%,II 区为 72.9%,III 区为 83.1%,IV 区为 90.9%,突出了其预测 FHB 发生的有效性。该工具既方便又实用,为根据当地天气条件对 FHB 风险进行战略管理提供了有价值的见解。
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引用次数: 0
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Plant disease
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