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First report of Botryosphaeria dothidea causing root rot of sugar beet in Serbia. 首次报告 Botryosphaeria dothidea 在塞尔维亚引起甜菜根腐病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-06-24-1275-PDN
Nina Vuckovic, Natasa Duduk, Emil Rekanovic, Bojan Duduk, Ivana Vico
<p><p>Botryosphaeria dothidea (Moug.:Fr.) Ces. & De Not. is predominantly recognized as a pathogen of various woody plants, inducing symptoms of stem canker, dieback, and fruit rot worldwide. However, sporadic reports suggest its impact on field crops, including B. dothidea associated with stem canker in soybean and tobacco (Bian et al. 2015; Chen et al. 2021), as well as B. quercuum on sugar beet (Alfieri et al. 1984). In September 2023, during a survey of root rot pathogens of sugar beet (Beta vulgaris L.) in Rimski Šančevi, Serbia (N 45°19´57″; E 19°49'58″), 3% of collected samples exhibited root rot symptoms. Externally, the lesions exhibited a dark brown coloration. On cross-section, the tissue displayed a gradient of discoloration ranging from light to dark brown throughout the roots. The roots were completely rotted. From these samples, two fungal isolates (SR28/II and SR4/III) were obtained from rotted internal root fragments, after washing, surface disinfection (70% ethanol), and plating on potato dextrose agar (PDA). Colonies on PDA were fluffy with abundant aerial mycelium, surface light to dark grey-brown, reverse black in the centre and grey-brown towards the irregular margin, after 7 days at 25°C in the dark. To induce sporulation, isolates were cultivated on 2% water agar with pine needles and incubated under continuous near ultraviolet (NUV) light at room temperature for 30 days. Conidia were hyaline, aseptate, fusiform, subtruncate at the base, subobtuse at the apex, and measured (19.56-) 24.12 - 26.31 (-28.99) x (5.13-) 5.94 - 6.54 (-7.44) µm (mean 25.06 x 6.26 µm, n=100), consistent with description of B. dothidea (Phillips et al. 2013). For molecular identification, DNA was extracted from mycelium of 7-day-old cultures, and internal transcribed spacer region (ITS), partial translation elongation factor 1-alpha gene (TEF) and partial β-tubulin gene (TUB) were amplified using the primer pairs ITS1/ITS4 (White et al. 1990), EF1-728F/EF1-986R (Carbone and Kohn 1999), and Bt2a/Bt2b (Glass and Donaldson 1995), respectively. Multiple sequence alignment and BLAST analyses showed that our isolates had 100% sequence similarity with reference isolates of B. dothidea, including ex-type isolate CBS 115476 (= CMW 8000) in ITS (AY236949), TEF (AY236898), and TUB (AY236927). Maximum likelihood phylogeny of concatenated sequences confirmed the identity of the isolates as B. dothidea. Sequences of ITS, TEF and TUB of isolate SR4/III were submitted to GenBank under accession numbers PP908658, PP911334, PP911333, respectively. The pathogenicity of obtained isolates was assessed on 3-month-old sugar beet plants grown in sterile substrate in the greenhouse. For inoculation, the upper parts of the roots were wounded (10x3 mm) 1 cm above the substrate, using a sterilized nail, and 2x2 mm mycelial plugs of 7-day-old culture grown on PDA were inserted and sealed with parafilm. Control plants were inoculated with sterile PDA plugs. Six plants were used
Botryosphaeria dothidea (Moug.:Fr.) Ces. & De Not.主要被认为是各种木本植物的病原体,在全球范围内引起茎腐病、枯萎病和果实腐烂等症状。然而,也有零星报道表明它对大田作物有影响,包括与大豆和烟草茎腐病相关的 B. dothidea(Bian 等,2015 年;Chen 等,2021 年),以及甜菜上的 B. quercuum(Alfieri 等,1984 年)。2023 年 9 月,在塞尔维亚 Rimski Šančevi (北纬 45°19´57″;东经 19°49'58″)对甜菜(Beta vulgaris L.)根腐病病原体进行调查期间,采集的样本中有 3% 出现根腐病症状。从外观上看,病害呈深褐色。从横截面上看,整个根部的组织呈现出从浅棕色到深棕色的渐变色。根部完全腐烂。从这些样本中,经过清洗、表面消毒(70% 乙醇)并在马铃薯葡萄糖琼脂(PDA)上培养后,从腐烂的内部根部碎片中获得了两种真菌分离物(SR28/II 和 SR4/III)。在 25°C 黑暗环境中培养 7 天后,PDA 上的菌落呈绒毛状,有丰富的气生菌丝,表面为浅灰褐色至深灰褐色,中部反转为黑色,边缘不规则处为灰褐色。为诱导分生孢子,将分离物放在含松针的 2% 水琼脂上培养,并在室温下持续近紫外线(NUV)光照下培养 30 天。分生孢子呈透明、无菌、纺锤形、基部近截形、先端近钝,尺寸为 (19.56-) 24.12 - 26.31 (-28.99) x (5.13-) 5.94 - 6.54 (-7.44) µm(平均 25.06 x 6.26 µm,n=100),与 B. dothidea 的描述一致(Phillips 等,2013 年)。为进行分子鉴定,从 7 天培养物的菌丝中提取 DNA,并使用引物对 ITS1/ITS4(White 等,1990 年)、EF1-728F/EF1-986R(Carbone 和 Kohn,1999 年)和 Bt2a/Bt2b (Glass 和 Donaldson,1995 年)分别扩增内部转录间隔区(ITS)、部分翻译伸长因子 1-α 基因(TEF)和部分 β-微管蛋白基因(TUB)。多重序列比对和 BLAST 分析表明,我们的分离物在 ITS (AY236949)、TEF (AY236898) 和 TUB (AY236927) 方面与 B. dothidea 的参考分离物(包括前型分离物 CBS 115476(= CMW 8000))具有 100% 的序列相似性。连接序列的最大似然系统发生证实了分离物为 B. dothidea。分离株 SR4/III 的 ITS、TEF 和 TUB 序列已提交至 GenBank,登录号分别为 PP908658、PP911334 和 PP911333。在温室无菌基质中生长 3 个月的甜菜植株上评估了所获分离株的致病性。接种时,用灭菌钉在基质上方 1 厘米处伤根上部(10x3 毫米),插入在 PDA 上培养 7 天的 2x2 毫米菌丝塞,并用保鲜膜密封。对照植株接种无菌 PDA 插条。每种分离物和对照均使用 6 株植物。培养 3 周后,将接种的植株从基质中移出,横切面检查可观察到褐腐症状,而对照组根系仍无症状。病原体被成功地重新分离出来,经形态鉴定为 B. dothidea,符合科赫假设。甜菜是塞尔维亚重要的工业作物,因其含糖量高而受到各种病原体的威胁。据我们所知,这是塞尔维亚乃至世界上第一份关于 B. dothidea 导致甜菜根腐病的报告。鉴于其在其他寄主上的广泛分布和潜在的侵袭性,甜菜中存在的 B. dothidea 不应被忽视。
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引用次数: 0
Characterization and Pathogenicity of Soilborne Pathogens in Gloriosa superba: Effects of Single- and Multiple-Pathogen Coinfection on Disease Responses. Gloriosa superba 中土传病原体的特征和致病性:单病原体和多病原体共感染对疾病反应的影响
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-03-24-0496-RE
Shanmuga Priya Dhanabalan, Iruthayasamy Johnson, Parthiban V Kumaresan, Rajamani Kandasamy, Senthil Natesan, Sambasivam Periyannan, Karthikeyan Muthusamy

Glory lily (Gloriosa superba), an ornamental climbing plant, contains the bioactive compound colchicine, attracting attention from the pharmaceutical industry. However, soilborne pathogens have emerged as a serious threat to the cultivation of glory lily, leading to substantial economic losses in the southern parts of India. Among these, the three major pathogens are Macrophomina phaseolina, Fusarium oxysporum, and Agroathelia rolfsii, causing dry root rot (also referred to as charcoal rot), wilt, and stem rot, respectively. Here, we characterized these pathogens using morphological characteristics and phylogenetic analysis of DNA sequences related to the internal transcribed spacer of ribosomal DNA, calmodulin (CAL), and translation elongation factor 1-alpha (TEF-1α). Furthermore, in the pathogenicity tests, the inoculation of M. phaseolina alone resulted in lesions measuring 7.54 ± 0.01 mm on tubers and 90% seedling mortality. This severity was comparable to the simultaneous inoculation of all three pathogens, indicating the prominence of dry root rot among soilborne diseases. This study marks the first detailed investigation of soilborne pathogens combined infection in G. superba, contributing to the understanding of fungal disease complexity in medicinal plants.

荣光百合(Gloriosa superba)是一种观赏攀援植物,含有生物活性化合物秋水仙碱,受到制药业的关注。然而,土传病原体已成为光辉百合种植的严重威胁,导致印度南部地区蒙受重大经济损失。其中,三种主要病原体是相叶巨霉菌(Macrophomina phaseolina)、氧孢镰刀菌(Fusarium oxysporum)和根腐菌(Agroathelia rolfsii),它们分别导致根部干腐(也称为炭腐)、枯萎和茎腐。在此,我们利用形态特征以及与核糖体 DNA 内部转录间隔序列(ITS)、钙调素(CAL)和翻译伸长因子(TEF)-1α 相关的 DNA 序列的系统发育分析,对这些病原体进行了鉴定。此外,在致病性试验中,单独接种 M. phaseolina 会导致块茎出现 7.54±0.01 毫米的病变,幼苗死亡率达 90%。这一严重程度与同时接种三种病原体的结果相当,表明干根腐病在土传病害中的突出地位。这项研究首次详细调查了土传病原体对 G. superba 的联合感染,有助于了解药用植物真菌病害的复杂性。
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引用次数: 0
A new leaf spot caused by Alternaria alternata on Atractylodes lancea in Jiangsu, China. 中国江苏白术上由交替丝核菌(Alternaria alternata)引起的一种新的叶斑病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-07-24-1349-PDN
Longjiao Hu, Yang Li, Jiping Xuan, Yonghua Gu, Dongling Li, Zhenghai Mo, Panhua Liao
<p><p>Atractylodes lancea is an important Chinese herbal medicine, which is mainly produced in Jiangsu province, China. In June 2022, leaf spots symptom were observed on some A. lancea seedlings growing in a Chinese herbal medicine resource garden of Nanjing Botanical Garden, Jiangsu. Approximately 75% of 100 A. lancea seedlings suffered from the disease. Initially, gray to black spots appeared at the tip of the blade, then spread deep into the petiole, finally causing the blade to wither and fall. To isolate the pathogen, five diseased leaves were collected from five different seedlings. Leaf sections (3 to 4 mm) were excised from the margins between healthy and diseased tissues, surface sterilized in 75% alcohol for 30 s, then in 1.5% NaClO for 90 s, rinsed three times in sterilized distilled water, plated on potato dextrose agar (PDA) and incubated at 25℃in darkness. Pure cultures were obtained by monosporic isolation. Eighteen isolates were obtained, and 77.8% of isolates was identified as Alternaria spp. A representative isolate, CS4-1 was used for further investigation. The colony of CS4-1, growing on PDA was cotton-like and black to brown with gray-white aerial hyphae on their surfaces, and dark gray on the back. The conidia were solitary on conidiophores and were oval to pear-shaped, brown in color, with 1 to 4 transverse septa and 0 to 1 oblique septa, parietal cells extending into the beak, and measured 8.9 to 39.5×6.0 to 13.5 µm (n=35). These characteristics were consistent with the description of Alternaria spp. (Simmons 2007). Six DNA regions, i.e.,internal transcribed spacer (ITS), large subunit ribosomal RNA (LSU), small subunit ribosomal RNA (SSU), anonymous region OPA10-2, Alt a 1 major allergen (Alta1), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and translation elongation factor 1-alpha (TEF1) with the respective GenBank Accession No. OP836052, OP836054, OP836051, OP851487, OP851488, OP851489, and OP851490, were amplified and sequenced with the primer pairs described by White et al. (1990) and Woudenberg et al. (2015). A neighbor-joining phylogenetic tree was generated by combining all sequenced loci in MEGA7, and CS4-1 clustered in the A. alternata clade. To test pathogenicity, 12 leaves, on three one-month-old A. lancea seedlings (four leaves from each seedling) were wounded with a sterile needle and inoculated with 20 μL of conidial suspension (1×106 spores/mL) on the left sides of leaves. The right sides of the leaves were inoculated with 20 µL sterile water and used as the control. All inoculated detached leaves and seedlings were covered with clear polyethylene bags to keep moisture and were incubated in a greenhouse at 25℃, 80% relative humidity, and a 12-h light/dark cycle. The experiment was repeated three times. After 4 days, typical gray to black spots were visible on the left sides of all inoculated leaves and the inoculated seedlings, and the right sides remained asymptomatic. Subsequently, the same fungus was
白术是一种重要的中药材,主产于中国江苏省。2022 年 6 月,江苏南京植物园中药材资源圃的部分白术幼苗出现叶斑病症状。在 100 株长春花幼苗中,约 75% 的幼苗发病。最初,叶片顶端出现灰黑色病斑,然后向叶柄深处扩展,最后导致叶片枯萎和倒伏。为了分离病原体,从五株不同的秧苗上采集了五片病叶。从健康组织和病变组织之间的边缘切除叶片(3 至 4 毫米),在 75% 的酒精中进行表面消毒 30 秒,然后在 1.5% 的 NaClO 中消毒 90 秒,再用消毒蒸馏水冲洗三次,然后将其置于马铃薯葡萄糖琼脂(PDA)上,在 25℃的黑暗环境中培养。通过单孢分离获得纯培养物。其中具有代表性的分离物 CS4-1 被用于进一步研究。CS4-1 生长在 PDA 上的菌落呈棉花状,黑色至棕色,表面有灰白色气生菌丝,背面呈深灰色。分生孢子单生在分生孢子梗上,呈椭圆形至梨形,褐色,有 1 至 4 个横隔膜和 0 至 1 个斜隔膜,顶细胞伸入喙内,大小为 8.9 至 39.5×6.0 至 13.5 µm(n=35)。这些特征与 Alternaria spp.的描述一致(Simmons,2007 年)。六个 DNA 区域,即内转录间隔区(ITS)、大亚基核糖体 RNA(LSU)、小亚基核糖体 RNA(SSU)、匿名区 OPA10-2、Alt a 1 主要过敏原(Alta1)、3-磷酸甘油醛脱氢酶(GAPDH)和翻译延伸因子 1-α(TEF1),其 GenBank Accession No.OP836052、OP836054、OP836051、OP851487、OP851488、OP851489 和 OP851490,用 White 等人(1990 年)和 Woudenberg 等人(2015 年)描述的引物对进行扩增和测序。在 MEGA7 中结合所有测序位点生成了邻接系统发生树,CS4-1 聚类在交替蓟马支系中。为了测试致病性,用无菌针刺伤三株一个月大的 A. lancea 幼苗(每株幼苗四片叶子)的 12 片叶子,然后在叶子左侧接种 20 μL 分生孢子悬浮液(1×106 个孢子/毫升)。叶片右侧接种 20 µL 无菌水作为对照。所有接种的离体叶片和幼苗都用透明聚乙烯袋覆盖以保持湿度,并在 25℃、相对湿度 80% 和 12 小时光照/黑暗循环的温室中培养。实验重复三次。4 天后,所有接种叶片和接种秧苗的左侧都出现了典型的灰黑色斑点,右侧仍无症状。随后,根据形态和分子特征重新分离并鉴定了同一种真菌。交替花叶病毒的寄主范围很广,但在全球范围内从未有关于 A. lancea 的报道(nt.ars-grin.gov)。由于 A. lancea 具有药用价值,因此应进一步研究如何控制这种疾病。
{"title":"A new leaf spot caused by <i>Alternaria alternata</i> on <i>Atractylodes lancea</i> in Jiangsu, China.","authors":"Longjiao Hu, Yang Li, Jiping Xuan, Yonghua Gu, Dongling Li, Zhenghai Mo, Panhua Liao","doi":"10.1094/PDIS-07-24-1349-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-07-24-1349-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Atractylodes lancea is an important Chinese herbal medicine, which is mainly produced in Jiangsu province, China. In June 2022, leaf spots symptom were observed on some A. lancea seedlings growing in a Chinese herbal medicine resource garden of Nanjing Botanical Garden, Jiangsu. Approximately 75% of 100 A. lancea seedlings suffered from the disease. Initially, gray to black spots appeared at the tip of the blade, then spread deep into the petiole, finally causing the blade to wither and fall. To isolate the pathogen, five diseased leaves were collected from five different seedlings. Leaf sections (3 to 4 mm) were excised from the margins between healthy and diseased tissues, surface sterilized in 75% alcohol for 30 s, then in 1.5% NaClO for 90 s, rinsed three times in sterilized distilled water, plated on potato dextrose agar (PDA) and incubated at 25℃in darkness. Pure cultures were obtained by monosporic isolation. Eighteen isolates were obtained, and 77.8% of isolates was identified as Alternaria spp. A representative isolate, CS4-1 was used for further investigation. The colony of CS4-1, growing on PDA was cotton-like and black to brown with gray-white aerial hyphae on their surfaces, and dark gray on the back. The conidia were solitary on conidiophores and were oval to pear-shaped, brown in color, with 1 to 4 transverse septa and 0 to 1 oblique septa, parietal cells extending into the beak, and measured 8.9 to 39.5×6.0 to 13.5 µm (n=35). These characteristics were consistent with the description of Alternaria spp. (Simmons 2007). Six DNA regions, i.e.,internal transcribed spacer (ITS), large subunit ribosomal RNA (LSU), small subunit ribosomal RNA (SSU), anonymous region OPA10-2, Alt a 1 major allergen (Alta1), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and translation elongation factor 1-alpha (TEF1) with the respective GenBank Accession No. OP836052, OP836054, OP836051, OP851487, OP851488, OP851489, and OP851490, were amplified and sequenced with the primer pairs described by White et al. (1990) and Woudenberg et al. (2015). A neighbor-joining phylogenetic tree was generated by combining all sequenced loci in MEGA7, and CS4-1 clustered in the A. alternata clade. To test pathogenicity, 12 leaves, on three one-month-old A. lancea seedlings (four leaves from each seedling) were wounded with a sterile needle and inoculated with 20 μL of conidial suspension (1×106 spores/mL) on the left sides of leaves. The right sides of the leaves were inoculated with 20 µL sterile water and used as the control. All inoculated detached leaves and seedlings were covered with clear polyethylene bags to keep moisture and were incubated in a greenhouse at 25℃, 80% relative humidity, and a 12-h light/dark cycle. The experiment was repeated three times. After 4 days, typical gray to black spots were visible on the left sides of all inoculated leaves and the inoculated seedlings, and the right sides remained asymptomatic. Subsequently, the same fungus was","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":"142472369","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
Fusarium species associated with Bakanae Disease of Rice in Bangladesh. 与孟加拉国水稻 Bakanae 病相关的镰刀菌物种。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-03-24-0655-SR
Asmaul Husna, Md Assaduzzaman Miah, Latiffah Zakaria, Masratul Hawa Mohd, Nur Ain Izzati Mohd Zainudin, Nik Mohd Izham Mohamed Nor

Bakanae disease has become a serious threat for sustainable rice production in Asian countries including Bangladesh. Fusarium species are important seedborne pathogens that cause bakanae disease of rice. Typical bakanae symptomatic samples were collected through a series of sampling conducted in several districts of Bangladesh for 4 consecutive years from 2019 - 2022. The pathogens were confirmed using morphological characteristics, DNA sequences, and phylogenetic analyses of two genes, namely, translation elongation factor 1-alpha (TEF1-α), and RNA polymerase subunit II (RPB2). A total of 121 Fusarium isolates were recovered from diseased rice samples at different geographical locations. From the phylogenetics analyses of TEF1-α and RPB2 gene sequences coupled with morphological characterization revealed that the collected isolates belonged to five species viz. F. fujikuroi (75.2% isolation frequency), F. incarnatum (17.35%), F. commune (4.95%), F. verticillioides (1.65%), and F. proliferatum (0.82%). Phylogenetic analysis also showed that 28 representative strains were attributed to five species. Finally, four Fusarium spp. F. fujikuroi, F. commune, F. verticillioides and F. proliferatum were found to be pathogenic under virulence assays of the isolates. The pathogenicity test results demonstrated that F. fujikuroi caused typical symptoms of bakanae, leaf elongation and chlorosis, whereas F. proliferatum and F. verticillioides only caused stunting of seedlings and F. commune caused wilt and root rot. F. incarnatum was found to be associated with bakanae disease of rice, however their pathogenicity could not be established. This study provides insight into the diversity and pathogenicity of Fusarium populations associated with bakanae disease in Bangladesh, which will help in formulating effective strategies and policies for better control of the bakanae disease.

稻瘟病已成为包括孟加拉国在内的亚洲国家水稻可持续生产的严重威胁。镰刀菌是引起水稻白粉病的重要种子传播病原体。从 2019 年到 2022 年,连续 4 年在孟加拉国的几个地区进行了一系列采样,收集了典型的 bakanae 症状样本。利用形态特征、DNA 序列和两个基因(即翻译延伸因子 1-α(TEF1-α)和 RNA 聚合酶亚基 II(RPB2))的系统发育分析确认了病原体。从不同地理位置的病害水稻样本中共分离出 121 株镰刀菌。通过对 TEF1-α 和 RPB2 基因序列的系统发育分析以及形态特征描述,发现所收集的分离株属于五个种,即 F. fujikuroi(分离频率为 75.2%)、F. incarnatum(17.35%)、F. commune(4.95%)、F. verticillioides(1.65%)和 F. proliferatum(0.82%)。系统发育分析还显示,28 个代表性菌株归属于 5 个种。最后,在对分离物进行致病性检测时发现,富士黑镰刀菌属(F. fujikuroi)、共生镰刀菌属(F. commune)、疣镰刀菌属(F. verticillioides)和增殖镰刀菌属(F. proliferatum)这四种镰刀菌属具有致病性。致病性试验结果表明,F. fujikuroi 会引起典型的包枯病、叶片伸长和萎黄病,而 F. proliferatum 和 F. verticillioides 只引起幼苗发育不良,F. commune 会引起枯萎病和根腐病。发现 F. incarnatum 与水稻白粉病有关,但其致病性尚未确定。这项研究有助于深入了解孟加拉国与稻瘟病相关的镰刀菌种群的多样性和致病性,有助于制定有效的战略和政策,更好地控制稻瘟病。
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引用次数: 0
Pseudomonas chlororaphis Metabolites Reduce MfCYP51 Expression and Yield Synergistic Efficacy in Mixture with Reduced Rates of Propiconazole Against DMI-Resistant Monilinia fructicola Isolates. 氯唑假单胞菌代谢物可减少 MfCYP51 的表达,与丙环唑混合使用可产生协同效应,降低丙环唑对耐 DMI 的果蝇科莫尼林菌分离物的药效。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-04-24-0869-RE
Johanna Wesche, Zhezheng Zeng, Chao-Xi Luo, Guido Schnabel

Brown rot caused by Monilinia fructicola is one of the most important diseases affecting peach production in the southeastern United States. Management often involves the use of demethylation inhibitor (DMI) fungicides, but efficacy can be compromised because of overexpression of the MfCYP51 gene encoding the 14α-demethylase of the ergosterol biosynthesis pathway. This study aimed to investigate the influence of the biorational fungicide Howler EVO containing Pseudomonas chlororaphis ASF009 metabolites on the expression of MfCYP51 in M. fructicola and associated synergy with a DMI fungicide for control of DMI-resistant strains. Mycelia from two DMI-sensitive and three DMI-resistant M. fructicola isolates were exposed or not to propiconazole (0.3 μg/ml), Howler (88.1 μg/ml), or the combination propiconazole + Howler for 6 h prior to RNA extraction. Real-time PCR indicated that Howler reduced the constitutive expression of MfCYP51 in DMI-sensitive and two of three DMI-resistant isolates. Propiconazole-induced expression of the DMI target gene was significantly reduced by Howler and by the mixture of Howler plus propiconazole in all isolates. Detached fruit studies on apple revealed that the combination of Howler plus a reduced label rate of Mentor (50 μg/ml propiconazole) was synergistic against brown rot caused by a DMI-resistant isolate in high and low inoculum spore concentration experiments (synergy values of 40.1 and 4.9, respectively). We hypothesize that the synergistic effects against M. fructicola resistant to DMI fungicides based on MfCYP51 gene overexpression can be attributed to reduced 14α demethylase production due to transcription inhibition, which may necessitate fewer DMI fungicide molecules to arrest fungal growth. The use of Howler/DMI mixtures for brown rot control warrants further investigation because such mixtures could potentially allow for reduced DMI fungicide use rates in the field without compromising yield or increased resistance selection.

由果核菌(Monilinia fructicola)引起的褐腐病是影响美国东南部桃子生产的最重要病害之一。防治方法通常包括使用去甲基化抑制剂(DMI)杀菌剂,但由于编码麦角甾醇生物合成途径 14α- 去甲基化酶的 MfCYP51 基因过度表达,药效可能会受到影响。本研究旨在探究含有绿假单胞菌 ASF009 代谢物的生物杀菌剂 Howler EVO 对果蝇中 MfCYP51 基因表达的影响,以及与 DMI 杀菌剂在控制抗 DMI 菌株方面的协同作用。在提取 RNA 之前,将两种对 DMI 敏感和三种对 DMI 抗性的果蝇菌分离菌丝暴露于或不暴露于丙环唑(0.3 µg/ml)、Howler(78.5 µg/ml)或丙环唑 + Howler 组合药剂 6 小时。实时聚合酶链式反应(Real-time PCR)表明,在对 DMI 敏感的分离株和三个对 DMI 抗性的分离株中的两个中,Howler 可减少 MfCYP51 的组成型表达。Howler 和 Howler 加丙环唑的混合物显著降低了所有分离物中丙环唑诱导的 DMI 目标基因的表达。对苹果进行的离体果实研究表明,在高、低接种孢子浓度实验中,Howler 与标签率降低的 Mentor(50 µg/ml 丙环唑)的组合对抗性 DMI 分离物引起的褐腐病具有协同作用(协同作用值分别为 40.1 和 4.9)。我们推测,基于 MfCYP51 基因的过表达对果蝇抗 DMI 杀菌剂产生的增效作用可归因于转录抑制导致的 14α 去甲基化酶生成减少,这可能需要更少的 DMI 杀菌剂分子来抑制真菌生长。使用 Howler /DMI 混合物控制褐腐病值得进一步研究,因为这种混合物有可能降低田间 DMI 杀真菌剂的使用率,而不会影响产量或增加抗性选择。
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引用次数: 0
Biocontrol Efficacy of Pseudomonas Consortia Against Botrytis Blight in Petunias. 假单胞菌群对矮牵牛灰霉病的生物防治功效
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-06-24-1210-RE
Sachin Naik, Laura J Chapin, Kaylee A South, Michelle Lyn Jones

Botrytis cinerea, a fungal pathogen causing Botrytis blight, significantly impacts greenhouse crop management due to its broad host range and infection capabilities at various growth stages. Traditional control methods, primarily reliant on fungicides, are challenged by environmental concerns and the rise of fungicide-resistant strains. This study investigates the use of beneficial Pseudomonas bacteria as a sustainable alternative. We hypothesized that specific Pseudomonas consortia could provide more effective biocontrol of B. cinerea than individual strains. Our research investigated five Pseudomonas strains (14B11, AP54, 15H3, 94G2, and 89F1) known to reduce Botrytis blight in Petunia × hybrida. Compatibility for bacterial consortia was assessed through biofilm formation and direct bacterial inhibition assays. The biocontrol effects of the bacteria against B. cinerea were investigated in vitro using shared-air space dual culture assays and in planta by inoculating detached petunia flowers. We found strain 14B11 exhibited the highest biofilm formation, with consortia of 14B11 and 89F1 showing significant enhancement compared to individual cultures, while a slight, non-significant increase was observed in 14B11 and AP54 consortia. However, strain 14B11 efficacy was inhibited by strain 15H3. Genomic analyses identified antifungal compound-related gene clusters in 14B11 and AP54, contributing to their biocontrol potential. Trials with detached flowers of Petunia × hybrida 'Carpet Red Bright' confirmed significant disease severity reduction with 14B11, AP54, and their consortia. This research highlights strategic Pseudomonas consortia as promising, eco-friendly alternatives to chemical fungicides, promoting sustainable agriculture by enhancing our understanding of how microbial interactions can be used to manage Botrytis blight.

灰霉病菌是一种导致灰霉病的真菌病原体,由于其寄主范围广泛,在不同生长阶段都有感染能力,因此对温室作物的管理产生了重大影响。传统的控制方法主要依赖杀菌剂,但受到环境问题和抗杀菌剂菌株增加的挑战。本研究调查了有益假单胞菌作为可持续替代品的使用情况。我们假设,与单个菌株相比,特定的假单胞菌群能更有效地生物防治银环蛇属真菌。我们的研究调查了五种假单胞菌菌株(14B11、AP54、15H3、94G2 和 89F1),这些菌株已知能减轻矮牵牛×杂交种的灰霉病。通过生物膜形成和直接细菌抑制试验评估了细菌联合体的兼容性。在体外,我们使用共享空气空间双培养试验研究了细菌对灰霉病菌的生物防治效果;在植物体内,我们接种了分离的矮牵牛花。我们发现,菌株 14B11 的生物膜形成率最高,与单个培养物相比,14B11 和 89F1 的联合体显示出显著的增强效果,而 14B11 和 AP54 联合体的生物膜形成率略有增加,但不显著。不过,菌株 14B11 的功效受到菌株 15H3 的抑制。基因组分析在 14B11 和 AP54 中发现了与抗真菌化合物相关的基因簇,这有助于提高它们的生物防治潜力。对矮牵牛×杂交种'地毯红亮'的脱落花进行的试验证实,14B11、AP54 和它们的联合体能显著减轻病害的严重程度。这项研究强调了战略性假单胞菌菌群作为化学杀真菌剂的有前途的生态友好型替代品,通过加强我们对如何利用微生物相互作用来控制灰霉病的了解,促进可持续农业的发展。
{"title":"Biocontrol Efficacy of <i>Pseudomonas</i> Consortia Against Botrytis Blight in Petunias.","authors":"Sachin Naik, Laura J Chapin, Kaylee A South, Michelle Lyn Jones","doi":"10.1094/PDIS-06-24-1210-RE","DOIUrl":"https://doi.org/10.1094/PDIS-06-24-1210-RE","url":null,"abstract":"<p><p><i>Botrytis cinerea</i>, a fungal pathogen causing Botrytis blight, significantly impacts greenhouse crop management due to its broad host range and infection capabilities at various growth stages. Traditional control methods, primarily reliant on fungicides, are challenged by environmental concerns and the rise of fungicide-resistant strains. This study investigates the use of beneficial <i>Pseudomonas</i> bacteria as a sustainable alternative. We hypothesized that specific <i>Pseudomonas</i> consortia could provide more effective biocontrol of <i>B. cinerea</i> than individual strains. Our research investigated five <i>Pseudomonas</i> strains (14B11, AP54, 15H3, 94G2, and 89F1) known to reduce Botrytis blight in <i>Petunia × hybrida</i>. Compatibility for bacterial consortia was assessed through biofilm formation and direct bacterial inhibition assays. The biocontrol effects of the bacteria against <i>B. cinerea</i> were investigated in vitro using shared-air space dual culture assays and in planta by inoculating detached petunia flowers. We found strain 14B11 exhibited the highest biofilm formation, with consortia of 14B11 and 89F1 showing significant enhancement compared to individual cultures, while a slight, non-significant increase was observed in 14B11 and AP54 consortia. However, strain 14B11 efficacy was inhibited by strain 15H3. Genomic analyses identified antifungal compound-related gene clusters in 14B11 and AP54, contributing to their biocontrol potential. Trials with detached flowers of <i>Petunia × hybrida</i> 'Carpet Red Bright' confirmed significant disease severity reduction with 14B11, AP54, and their consortia. This research highlights strategic <i>Pseudomonas</i> consortia as promising, eco-friendly alternatives to chemical fungicides, promoting sustainable agriculture by enhancing our understanding of how microbial interactions can be used to manage Botrytis blight.</p>","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":"142472370","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 Fusarium falciforme Causing Fusarium Wilt on pepper in Hainan, China. Fusarium falciforme 在中国海南辣椒上引起镰刀菌枯萎病的首次报告。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-16 DOI: 10.1094/PDIS-09-24-1854-PDN
Xingliang Wang, Yue Huang, Na Yang, Xue Wang, Xiaoyu Shen, Lijuan Pei, Ying Wang, Hui Zhang
<p><p>Pepper (<i>Capsicum annuum</i> L.) is a significant vegetable crop, valued for its nutritional and economic importance (Pang et al. 2023). Pepper cultivation in China accounts for about 8-10% of the total vegetable planting area, contributing an output value of approximately 250 billion yuan. This makes pepper the leading vegetable in terms of both planting area and economic value. In December 2023, a total of 70% disease incidence of Fusarium wilt was observed in a 1200 m² pepper seed breeding base in Sanya City, Hainan Province, China (18°38'60″ N, 109°16'51″ E). Symptoms initially appeared as wilting on upper leaves. Subsequently, the base of the stem started to necrosis, browning, and gradually spreading upward along the stem. As the lesions expanded, the whole plant gradually wilted and died. Ten diseased plants were randomly selected from the most severely affected area (667 m²). Diseased tissues (5 mm²) were subsequently removed from the lesion edges of these plants, surface sterilized in 75% ethanol for 30 s, and rinsed with sterile distilled water three times, finally cultured on potato dextrose agar (PDA) at 25 °C. Six fungal isolates were obtained using the single-spore isolation method (HN-01 to HN-06). Colonies produced white aerial mycelia with apricot pigments in the PDA medium. The spore morphology and size were observed and measured using synthetic nutrient-poor agar (SNA) medium. Macroconidia were hyaline, slightly curved in shape with 3 or 4 septa, measuring 28.6 to 41.4 × 3.2 to 6.2 μm (av. = 34.8 ± 3.32 × 4.6 ± 0.85 um, n = 20). Microconidia were elongated, oval with 0 or 1 septum, and measured 11.2 to 16.8 × 2.6 to 5.8 μm (av. = 13.5 ± 1.47 × 4.12 ± 1.03 um, n = 20). Chlamydospores were spherical, terminal or intercalary, solitary or chain-forming, with diameters ranging from 2.8 to 10.5 um (av. = 5.8 ± 2.31 um, n = 20). For molecular identification, genomic DNA from all six isolates was extracted using the cetyl trimethyl ammonium bromide (CTAB) method, and the internal transcribed spacer of rDNA (ITS), translation elongation factor 1-α (EF1-α), and RNA polymerase II beta subunit (RPB2) regions were amplified and sequenced using the primers ITS1/ITS4, EF-1/EF-2, and RPB2-5F/7cR (White et al. 1990; O'Donnell et al. 2010). The sequences were deposited in GenBank (ITS: PP779839, PP779840, PP779841, PP779842, PP779843, PP779844; EF1-α: PP797138, PP797139, PP797140, PP797141, PP797142, PP797143; RPB2: PP797144, PP797145, PP797146, PP797147, PP797148, PP797149). The sequences of all three genes showed 99 to 100% similarity with <i>Fusarium falciforme</i> and other closely related <i>Fusarium</i> species (ITS: PP735125, EF1-α: OP163897 and RPB2: MF467484). A maximum likelihood phylogenetic tree was constructed based on the ITS, EF1-α, and RPB2 sequences of all isolates, along with other closely related <i>Fusarium</i> species. Based on morphological and phylogenetic characteristics, all isolates were identified as <i>F. falc
辣椒(Capsicum annuum L.)是一种重要的蔬菜作物,具有重要的营养和经济价值(Pang 等,2023 年)。中国的辣椒种植面积约占蔬菜种植总面积的 8-10%,产值约为 2500 亿元。这使得辣椒在种植面积和经济价值方面都成为蔬菜中的佼佼者。2023 年 12 月,在中国海南省三亚市(北纬 18°38'60″,东经 109°16'51″)的一个 1200 平方米的辣椒种子繁育基地,发现镰刀菌枯萎病的发病率高达 70%。症状最初表现为上部叶片枯萎。随后,茎基部开始坏死、变褐,并逐渐沿茎向上蔓延。随着病斑扩大,整个植株逐渐枯萎死亡。从受灾最严重的地区(667 平方米)随机抽取了 10 株病株。随后从这些植株的病斑边缘取下病变组织(5 平方毫米),在 75% 的乙醇中表面消毒 30 秒,并用无菌蒸馏水冲洗三次,最后在 25 °C 的马铃薯葡萄糖琼脂(PDA)上培养。采用单孢分离法获得了六种真菌分离物(HN-01 至 HN-06)。菌落在 PDA 培养基中产生白色气生菌丝体,带有杏色素。使用贫养分合成琼脂(SNA)培养基对孢子形态和大小进行了观察和测量。大锥体呈透明状,形状略微弯曲,有 3 或 4 个隔膜,大小为 28.6 至 41.4 × 3.2 至 6.2 μm(平均值 = 34.8 ± 3.32 × 4.6 ± 0.85 um,n = 20)。微囊拉长,椭圆形,有 0 或 1 个隔膜,大小为 11.2-16.8 × 2.6-5.8 μm(平均值 = 13.5 ± 1.47 × 4.12 ± 1.03 um,n = 20)。衣孢子呈球形,顶生或闰生,单生或成链,直径范围为 2.8 至 10.5 微米(平均值 = 5.8 ± 2.31 微米,n = 20)。为了进行分子鉴定,使用十六烷基三甲基溴化铵(CTAB)法提取了所有 6 个分离株的基因组 DNA,并使用引物 ITS1/ITS4、EF-1/EF-2 和 RPB2-5F/7cR 对 rDNA 内部转录间隔区(ITS)、翻译延伸因子 1-α (EF1-α)和 RNA 聚合酶 II beta 亚基(RPB2)区进行了扩增和测序(White et al.1990;O'Donnell 等人,2010)。序列已存入 GenBank(ITS:PP779839, PP779840, PP779841, PP779842, PP779843, PP779844; EF1-α:PP797138、PP797139、PP797140、PP797141、PP797142、PP797143;RPB2:PP797144、PP797145、PP797146、PP797147、PP797148、PP797149)。这三个基因的序列与镰刀菌和其他近缘镰刀菌(ITS:PP735125;EF1-α:OP163897;RPB2:MF467484)的相似度为 99%至 100%。根据所有分离物的 ITS、EF1-α 和 RPB2 序列以及其他近缘镰刀菌种的序列,构建了最大似然系统发生树。根据形态学和系统发育特征,所有分离株都被鉴定为镰刀菌(Xu 等,2023 年;Wang 等,2023 年)。利用三个代表性分离株对 10 个辣椒近交系 A23-41(接种处理 5 个,对照 5 个)进行了致病性测试:HN-01、HN-02 和 HN-03。将浓度为 106 个孢子/毫升的孢子悬浮液共 20 ul 注入茎干土壤表面附近,而对照处理则接种 20 μl 无菌水。接种后,将植物置于相对湿度为 80-90%、温度为 25/20 °C(昼/夜)的恒温室中。实验重复三次。10 天后,接种的植株茎干出现坏死、褐变,而对照组仍无症状。通过 EF1-α 和 RPB2 序列分析,从人工感染的茎秆中重新分离出的真菌被鉴定为镰刀菌,因此符合科赫假说。据报道,镰刀菌(Fusarium falciforme)曾在多个国家的不同寄主上引起多种病害,包括韩国(Kang 等人,2024 年)、马来西亚(Balasubramaniam 等人,2023 年)和墨西哥(Payán-Arzapalo 等人,2024 年)。据我们所知,这是 F. falciforme 在中国引起辣椒镰刀菌枯萎病的首次报道。研究结果可为今后研究该病害的发生、预防和管理提供依据。
{"title":"First Report of <i>Fusarium falciforme</i> Causing Fusarium Wilt on pepper in Hainan, China.","authors":"Xingliang Wang, Yue Huang, Na Yang, Xue Wang, Xiaoyu Shen, Lijuan Pei, Ying Wang, Hui Zhang","doi":"10.1094/PDIS-09-24-1854-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-09-24-1854-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Pepper (&lt;i&gt;Capsicum annuum&lt;/i&gt; L.) is a significant vegetable crop, valued for its nutritional and economic importance (Pang et al. 2023). Pepper cultivation in China accounts for about 8-10% of the total vegetable planting area, contributing an output value of approximately 250 billion yuan. This makes pepper the leading vegetable in terms of both planting area and economic value. In December 2023, a total of 70% disease incidence of Fusarium wilt was observed in a 1200 m² pepper seed breeding base in Sanya City, Hainan Province, China (18°38'60″ N, 109°16'51″ E). Symptoms initially appeared as wilting on upper leaves. Subsequently, the base of the stem started to necrosis, browning, and gradually spreading upward along the stem. As the lesions expanded, the whole plant gradually wilted and died. Ten diseased plants were randomly selected from the most severely affected area (667 m²). Diseased tissues (5 mm²) were subsequently removed from the lesion edges of these plants, surface sterilized in 75% ethanol for 30 s, and rinsed with sterile distilled water three times, finally cultured on potato dextrose agar (PDA) at 25 °C. Six fungal isolates were obtained using the single-spore isolation method (HN-01 to HN-06). Colonies produced white aerial mycelia with apricot pigments in the PDA medium. The spore morphology and size were observed and measured using synthetic nutrient-poor agar (SNA) medium. Macroconidia were hyaline, slightly curved in shape with 3 or 4 septa, measuring 28.6 to 41.4 × 3.2 to 6.2 μm (av. = 34.8 ± 3.32 × 4.6 ± 0.85 um, n = 20). Microconidia were elongated, oval with 0 or 1 septum, and measured 11.2 to 16.8 × 2.6 to 5.8 μm (av. = 13.5 ± 1.47 × 4.12 ± 1.03 um, n = 20). Chlamydospores were spherical, terminal or intercalary, solitary or chain-forming, with diameters ranging from 2.8 to 10.5 um (av. = 5.8 ± 2.31 um, n = 20). For molecular identification, genomic DNA from all six isolates was extracted using the cetyl trimethyl ammonium bromide (CTAB) method, and the internal transcribed spacer of rDNA (ITS), translation elongation factor 1-α (EF1-α), and RNA polymerase II beta subunit (RPB2) regions were amplified and sequenced using the primers ITS1/ITS4, EF-1/EF-2, and RPB2-5F/7cR (White et al. 1990; O'Donnell et al. 2010). The sequences were deposited in GenBank (ITS: PP779839, PP779840, PP779841, PP779842, PP779843, PP779844; EF1-α: PP797138, PP797139, PP797140, PP797141, PP797142, PP797143; RPB2: PP797144, PP797145, PP797146, PP797147, PP797148, PP797149). The sequences of all three genes showed 99 to 100% similarity with &lt;i&gt;Fusarium falciforme&lt;/i&gt; and other closely related &lt;i&gt;Fusarium&lt;/i&gt; species (ITS: PP735125, EF1-α: OP163897 and RPB2: MF467484). A maximum likelihood phylogenetic tree was constructed based on the ITS, EF1-α, and RPB2 sequences of all isolates, along with other closely related &lt;i&gt;Fusarium&lt;/i&gt; species. Based on morphological and phylogenetic characteristics, all isolates were identified as &lt;i&gt;F. falc","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":"142472375","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
Evaluation of Stripe Rust Resistance and Chip Detection Resistance Genes in 286 Xinjiang Wheat Cultivars and Breeding Lines. 对 286 个新疆小麦栽培品种和育种系的抗条锈病基因和抗芯片检测基因进行评估
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-14 DOI: 10.1094/PDIS-04-24-0780-RE
Haohao Yan, Jianing Zhu, Yongjin Jin, Xingxuan Bai, Qingdong Zeng, Haifeng Gao, Jinbiao Ma, Lili Huang, Zhensheng Kang, Gangming Zhan

Wheat stripe rust is a destructive disease worldwide, caused by Puccinia striiformis f. sp. tritici (Pst). Resistance breeding is the most effective method of controlling stripe rust. Xinjiang is a relatively independent epidemic region of wheat stripe rust in China. In recent years, wheat stripe rust in this area has shown an upward trend. Therefore, the purpose of this study was to evaluate the resistance level of wheat cultivars (lines) to the prevalent Pst races and determine the genetic background of stripe rust resistance genes in Xinjiang. Six predominant Pst races in China were used to study resistance of 286 wheat cultivars (lines) at both the seedling stage under controlled conditions and the adult-plant stage under field conditions. In the seedling tests, 175 (61.19%) entries were resistant to the race CYR23, 125 (43.71%) to CYR29, 153 (53.50%) to CYR31, 88 (30.77%) to CYR32, 174 (60.84%) to CYR33, and 98 (34.27%) to CYR34. Among the resistant entries, 23 (8.04%) were resistant to all six races. In the field test, 135 (47.20%) entries were resistant to the tested mixed races. Through comparing the responses in the seedling and adult-plant stages, 109 (38.11%) entries were found to have adult-plant resistance (APR), and 14 (4.90%) entries have all-stage resistance (ASR). The 286 wheat entries were also tested using a wheat breeder chip containing 12 Yr resistance loci. Among these entries, 44 (15.38%) were found to have a single gene, 221 (77.27%) have two or more genes, and 21 (7.34%) have none of the 12 genes, including 144 (50.35%) with Yr30 and 5 (1.75%) with YrSP. Entries with two or more genes have stronger resistance to Pst. Overall, the majority of entries have all-stage and/or adult-plant resistance, but their genes for resistance in addition to the 12 tested Yr genes need to be determined. It is also necessary to introduce more effective resistance genes in the breeding programs to improve stripe rust resistance in wheat cultivars in Xinjiang.

小麦条锈病是一种世界性毁灭性病害,由条锈病菌(Puccinia striiformis f. sp. tritici,Pst)引起。抗性育种是防治条锈病最有效的方法。新疆是我国小麦条锈病相对独立的流行区。近年来,该地区的小麦条锈病呈上升趋势。因此,本研究旨在评估新疆小麦栽培品种(品系)对流行的 Pst 株系的抗性水平,并确定新疆小麦条锈病抗性基因的遗传背景。本研究利用中国的 6 个主要 Pst 株系,研究了 286 个小麦栽培品种(品系)在对照条件下的幼苗期和田间条件下的成株期的抗性。在苗期试验中,175 个品种(61.19%)对 CYR23 感抗,125 个品种(43.71%)对 CYR29 感抗,153 个品种(53.50%)对 CYR31 感抗,88 个品种(30.77%)对 CYR32 感抗,174 个品种(60.84%)对 CYR33 感抗,98 个品种(34.27%)对 CYR34 感抗。在抗性品种中,有 23 个品种(8.04%)对所有六个品系都具有抗性。在田间试验中,135 个条目(47.20%)对测试的混合品系具有抗性。通过比较苗期和成株期的反应,发现 109 个(38.11%)品种具有成株抗性(APR),14 个(4.90%)品种具有全生育期抗性(ASR)。这 286 个小麦品种还使用含有 12 个 Yr 抗性位点的小麦育种芯片进行了测试。在这些条目中,发现 44 个(15.38%)条目具有单基因,221 个(77.27%)条目具有两个或多个基因,21 个(7.34%)条目没有 12 个基因,其中 144 个(50.35%)条目具有 Yr30 基因,5 个(1.75%)条目具有 YrSP 基因。含有两个或更多基因的品种对 Pst 的抗性更强。总体而言,大多数品种具有全生育期和/或成株抗性,但除了 12 个测试的 Yr 基因外,还需要确定它们的抗性基因。在育种计划中引入更多有效的抗性基因以提高新疆小麦品种的条锈病抗性也是必要的。
{"title":"Evaluation of Stripe Rust Resistance and Chip Detection Resistance Genes in 286 Xinjiang Wheat Cultivars and Breeding Lines.","authors":"Haohao Yan, Jianing Zhu, Yongjin Jin, Xingxuan Bai, Qingdong Zeng, Haifeng Gao, Jinbiao Ma, Lili Huang, Zhensheng Kang, Gangming Zhan","doi":"10.1094/PDIS-04-24-0780-RE","DOIUrl":"10.1094/PDIS-04-24-0780-RE","url":null,"abstract":"<p><p>Wheat stripe rust is a destructive disease worldwide, caused by <i>Puccinia striiformis</i> f. sp. <i>tritici</i> (<i>Pst</i>). Resistance breeding is the most effective method of controlling stripe rust. Xinjiang is a relatively independent epidemic region of wheat stripe rust in China. In recent years, wheat stripe rust in this area has shown an upward trend. Therefore, the purpose of this study was to evaluate the resistance level of wheat cultivars (lines) to the prevalent <i>Pst</i> races and determine the genetic background of stripe rust resistance genes in Xinjiang. Six predominant <i>Pst</i> races in China were used to study resistance of 286 wheat cultivars (lines) at both the seedling stage under controlled conditions and the adult-plant stage under field conditions. In the seedling tests, 175 (61.19%) entries were resistant to the race CYR23, 125 (43.71%) to CYR29, 153 (53.50%) to CYR31, 88 (30.77%) to CYR32, 174 (60.84%) to CYR33, and 98 (34.27%) to CYR34. Among the resistant entries, 23 (8.04%) were resistant to all six races. In the field test, 135 (47.20%) entries were resistant to the tested mixed races. Through comparing the responses in the seedling and adult-plant stages, 109 (38.11%) entries were found to have adult-plant resistance (APR), and 14 (4.90%) entries have all-stage resistance (ASR). The 286 wheat entries were also tested using a wheat breeder chip containing 12 <i>Yr</i> resistance loci. Among these entries, 44 (15.38%) were found to have a single gene, 221 (77.27%) have two or more genes, and 21 (7.34%) have none of the 12 genes, including 144 (50.35%) with <i>Yr30</i> and 5 (1.75%) with <i>YrSP</i>. Entries with two or more genes have stronger resistance to <i>Pst</i>. Overall, the majority of entries have all-stage and/or adult-plant resistance, but their genes for resistance in addition to the 12 tested <i>Yr</i> genes need to be determined. It is also necessary to introduce more effective resistance genes in the breeding programs to improve stripe rust resistance in wheat cultivars in Xinjiang.</p>","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":"PDIS04240780RE"},"PeriodicalIF":4.4,"publicationDate":"2024-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141470071","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
Stevia rebaudiana Is a New Host of Stagonosporopsis pogostemonis Associated with Leaf Spot Disease in China. 甜叶菊是中国与叶斑病有关的 Stagonosporopsis pogostemonis 的新寄主
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-11 DOI: 10.1094/PDIS-11-23-2325-PDN
Shun Cao, Amei Xu, Yuankai Chi, Xiaofang Cui, Xueying Zhang, Rende Qi, Wei Zhao
<p><p>Stevia rebaudiana is a promising medicinal and edible plant, widely cultivated in China. In 2022-2023, a new leaf spot disease occurred in S. rebaudiana in Hongxing country (32°34'55″N, 118°2'12″E), Dingyuan city, Anhui province. Symptoms were observed on 10 to 15% of plants in three S. rebaudiana nursery beds (0.1 ha in total). The typical symptoms included dark brown spots on the leaves and foliar wilting, the development of brown stems with dieback of top buds, and occasional plant death (Fig. 1a). To identify the pathogen, twenty diseased leaves were collected, cut into small pieces, surface sterilized with 75% ethanol for 30 s, in 0.5% sodium hypochlorite for 2 min, washed three times in sterile water, placed on PDA, and incubated at 25℃ for 5 days. Pure cultures were prepared by subculturing hyphal tips. Twenty-five Stagonosporopsis-like isolates with similar morphology were obtained. After 7 days growth on PDA, colonies had a regular margin, were cottony, and formed concentric circles on the surface that were gray-green. The reverse side of the culture was dark brown with a creme-orange and white, margin. The growth rate was 9.5 mm/day on PDA. Pycnidia were mostly solitary, globose or subglobose, pale to dark brown, thin-walled, glabrous, ostiolate, 95.735~250.851×90.93~266.32 μm (n=50). Conidia were oblong, cylindrical to ellipsoidal, smooth-walled, aseptate, with rounded ends and two polar guttules and measured 3.41 to 5.83 × 1.78 to 3.07 μm (n = 50) (Fig.1 b-d). For molecular identification, the internal transcribed spacer (ITS) rDNA, large ribosomal subunit (LSU) gene, β-tubulin (TUB2) gene and RNA polymerase II (RPB2) gene sequences of two representative isolates (TYJ-SP1 and TYJ-SP2) were amplified by PCR (Woudenberg et al. 2009; Dong et al. 2021). The sequences were deposited in GenBank (accession nos.: OR506193 and OR506194 for ITS, OR533526 and OR533527 for LSU, OR545221and OR545222 for TUB; OR545223 and OR545224 for RPB2) and showed 99.60% to 99.2% similarity to ITS (502/504 bp and 507/511 bp; MZ156571), 100% similarity to LSU (857/857 bp and 857/857 bp; MZ191532), 98.67% to 99.3% similarity to TUB2 (296/300 bp and 298/300 bp; MZ203132) and 99.78% (888/890 bp and 868/870 bp; MZ203135) of S. pogostemonis strain ZHKUCC 21-0001. A maximum likelihood phylogenetic analysis based on the concatenated sequences of ITS, LSU, TUB2 and RPB2 using MEGA 11.0 showed the strains TYJ-SP1 and TYJ-SP2 formed a clade with S. pogostemonis (Fig. 2). Thus, the strains were identified as S. pogostemonis (Dong et al. 2021). To test pathogenicity, the strain TYJ-SP1 was inoculated onto 30-day-old S. rebaudiana seedlings which were surface sterilized with 70% alcohol and washed 3 times with water and air dried prior to inoculation. Ten seedlings were sprayed with a conidial suspension (105 conidia/mL) and ten seedlings were sprayed with sterile water to serve as the negative control. All seedlings were maintained in a growth chamber (25°C, 90% relat
甜叶菊(Stevia rebaudiana)是一种前景广阔的药用和食用植物,在中国广泛种植。2022-2023 年,安徽省定远市红星乡(32°34'55″N,118°2'12″E)的甜叶菊发生了新的叶斑病。在三个 S. rebaudiana 苗圃(共 0.1 公顷)中,10% 至 15% 的植株出现了病害症状。典型症状包括叶片上出现黑褐色斑点,叶片枯萎,茎干变褐,顶芽枯死,偶尔出现植株死亡(图 1a)。为了鉴定病原体,采集了 20 片病叶,切成小块,用 75% 的乙醇进行表面消毒 30 秒,再用 0.5% 的次氯酸钠消毒 2 分钟,然后用无菌水洗 3 次,放在 PDA 上,在 25℃下培养 5 天。通过对菌丝尖端进行亚培养,制备纯培养物。获得了 25 个形态相似的拟石蒜孢子分离物。菌落在 PDA 上生长 7 天后,边缘规则,呈棉状,表面形成灰绿色的同心圆。培养物的反面呈深褐色,边缘呈橘黄色和白色相间。在 PDA 上的生长速度为 9.5 毫米/天。分生孢子多为单生,球形或近球形,淡褐色至深褐色,薄壁,无毛,具柄,直径 95.735~250.851×90.93~266.32 μm(n=50)。分生孢子长圆形、圆柱形至椭圆形,壁光滑,无菌,末端圆形,有两个极性菌囊,大小为 3.41 至 5.83 × 1.78 至 3.07 μm(n = 50)(图 1 b-d)。为进行分子鉴定,通过 PCR 扩增了两个代表性分离株(TYJ-SP1 和 TYJ-SP2)的内部转录间隔(ITS)rDNA、大核糖体亚基(LSU)基因、β-微管蛋白(TUB2)基因和 RNA 聚合酶 II(RPB2)基因序列(Woudenberg 等,2009 年;Dong 等,2021 年)。序列已存入 GenBank(登录号:OR506193 和 OR506193):ITS为 OR506193 和 OR506194,LSU 为 OR533526 和 OR533527,TUB 为 OR545221 和 OR545222;RPB2 为 OR545223 和 OR545224),与 ITS 的相似度为 99.60% 至 99.2%(502/504 bp 和 507/511 bp;MZ156571),与 LSU 的相似度为 100% (857/857 bp 和 857/857 bp;MZ191532),与 RPB2 的相似度为 98.67%至 99.3% 的相似性(296/300 bp 和 298/300 bp;MZ203132)和 99.78% 的相似性(888/890 bp 和 868/870 bp;MZ203135)。基于 ITS、LSU、TUB2 和 RPB2 序列的最大似然系统进化分析(MEGA 11.0)表明,TYJ-SP1 和 TYJ-SP2 与 S. pogostemonis 形成一个支系(图 2)。因此,这两株菌株被鉴定为 S. pogostemonis(Dong 等,2021 年)。为了测试致病性,将菌株 TYJ-SP1 接种到 30 天大的 S. rebaudiana 秧苗上,秧苗表面用 70% 的酒精消毒,接种前用水冲洗 3 次并晾干。10 株幼苗喷洒分生孢子悬浮液(105 个分生孢子/毫升),10 株幼苗喷洒无菌水作为阴性对照。所有幼苗均在光周期为 16 小时的生长室(25°C,相对湿度 90%)中养护。接种后 48 小时,在接种叶片上首次观察到褐斑;接种后 7 天出现典型症状。所有接种的植株都出现了与苗圃中自然感染植株相似的症状,发病率达到 100%,而对照植株则没有症状(图 1 e-f)。根据形态学和系统发育分析,从接种植物中重新分离出了相同的 Stagonosporopsis 分离物并进行了鉴定。据报道,S. pogostemonis 可导致卡布其林和甘蓝变种叶斑病(Dong 等人,2021 年;Habib 等人,2024 年)。据我们所知,这是中国首次报道 S. pogostemonis 导致 S. rebaudiana 叶斑病。作为一种药用和经济植物,S. rebaudiana 在中国和其他亚洲国家被广泛种植。叶斑病的发生严重影响了其药用和经济价值。因此,建立和实施有效的病害管理方法以减少该病害的影响至关重要。
{"title":"<i>Stevia rebaudiana</i> Is a New Host of <i>Stagonosporopsis pogostemonis</i> Associated with Leaf Spot Disease in China.","authors":"Shun Cao, Amei Xu, Yuankai Chi, Xiaofang Cui, Xueying Zhang, Rende Qi, Wei Zhao","doi":"10.1094/PDIS-11-23-2325-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-11-23-2325-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Stevia rebaudiana is a promising medicinal and edible plant, widely cultivated in China. In 2022-2023, a new leaf spot disease occurred in S. rebaudiana in Hongxing country (32°34'55″N, 118°2'12″E), Dingyuan city, Anhui province. Symptoms were observed on 10 to 15% of plants in three S. rebaudiana nursery beds (0.1 ha in total). The typical symptoms included dark brown spots on the leaves and foliar wilting, the development of brown stems with dieback of top buds, and occasional plant death (Fig. 1a). To identify the pathogen, twenty diseased leaves were collected, cut into small pieces, surface sterilized with 75% ethanol for 30 s, in 0.5% sodium hypochlorite for 2 min, washed three times in sterile water, placed on PDA, and incubated at 25℃ for 5 days. Pure cultures were prepared by subculturing hyphal tips. Twenty-five Stagonosporopsis-like isolates with similar morphology were obtained. After 7 days growth on PDA, colonies had a regular margin, were cottony, and formed concentric circles on the surface that were gray-green. The reverse side of the culture was dark brown with a creme-orange and white, margin. The growth rate was 9.5 mm/day on PDA. Pycnidia were mostly solitary, globose or subglobose, pale to dark brown, thin-walled, glabrous, ostiolate, 95.735~250.851×90.93~266.32 μm (n=50). Conidia were oblong, cylindrical to ellipsoidal, smooth-walled, aseptate, with rounded ends and two polar guttules and measured 3.41 to 5.83 × 1.78 to 3.07 μm (n = 50) (Fig.1 b-d). For molecular identification, the internal transcribed spacer (ITS) rDNA, large ribosomal subunit (LSU) gene, β-tubulin (TUB2) gene and RNA polymerase II (RPB2) gene sequences of two representative isolates (TYJ-SP1 and TYJ-SP2) were amplified by PCR (Woudenberg et al. 2009; Dong et al. 2021). The sequences were deposited in GenBank (accession nos.: OR506193 and OR506194 for ITS, OR533526 and OR533527 for LSU, OR545221and OR545222 for TUB; OR545223 and OR545224 for RPB2) and showed 99.60% to 99.2% similarity to ITS (502/504 bp and 507/511 bp; MZ156571), 100% similarity to LSU (857/857 bp and 857/857 bp; MZ191532), 98.67% to 99.3% similarity to TUB2 (296/300 bp and 298/300 bp; MZ203132) and 99.78% (888/890 bp and 868/870 bp; MZ203135) of S. pogostemonis strain ZHKUCC 21-0001. A maximum likelihood phylogenetic analysis based on the concatenated sequences of ITS, LSU, TUB2 and RPB2 using MEGA 11.0 showed the strains TYJ-SP1 and TYJ-SP2 formed a clade with S. pogostemonis (Fig. 2). Thus, the strains were identified as S. pogostemonis (Dong et al. 2021). To test pathogenicity, the strain TYJ-SP1 was inoculated onto 30-day-old S. rebaudiana seedlings which were surface sterilized with 70% alcohol and washed 3 times with water and air dried prior to inoculation. Ten seedlings were sprayed with a conidial suspension (105 conidia/mL) and ten seedlings were sprayed with sterile water to serve as the negative control. All seedlings were maintained in a growth chamber (25°C, 90% relat","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2024-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142406700","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 lily mottle virus naturally infecting lily (Lilium asiatica) in Texas, USA. 首次报告美国得克萨斯州百合花(Lilium asiatica)自然感染百合斑驳病毒的情况。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-11 DOI: 10.1094/PDIS-07-24-1485-PDN
John Oladeji Oladokun, Isaias Hernandez, Aryed N Perez-Baez, Olufemi Joseph Alabi
<p><p>Lily (Lilium asiatica) is an important plant grown for its range of flower colors and heavy scent. In March 2024, potyvirus-like symptoms consisting of light-yellow mottling and mosaic were noticed on 12/20 lily plants in a private property in Weslaco, Hidalgo County, Texas. Two symptomatic plants (WTX1 and WTX2) were sampled randomly for virus diagnosis. The leaf extracts of both samples were negative for the potyvirus group using Agdia's Poty ImmunoStrip® (Agdia, Inc., Elkhart, IN, USA). However, rub-inoculation of the extracts onto healthy Nicotiana benthamiana and Vigna unguiculata plants (n=4, each) induced mild mottle symptoms on the systemic leaves of both herbaceous test plants 20 to 28 days post inoculation, indicating the presence of a mechanically transmissible agent in the samples. No virus-like symptoms were observed on the mock-inoculated plants (n=1, each) of both species. To test for suspected potyvirus infection, 2-µg of total nucleic acid extracts (Dellaporta et al. 1983) from WTX1 and WTX2 were used for complimentary DNA (cDNA) synthesis with Oligo(dT) primer and the PrimeScript 1st strand cDNA synthesis kit (Takara Bio, USA). One microliter aliquot of each cDNA served as template in 12.5-µl conventional PCR reaction volumes with 5 U Taq polymerase (Roche Diagnostics, Indianapolis, IN), and two pairs of degenerate primers targeting the partial cylindrical inclusion (CI) gene and the helper component protease (HC-Pro) of potyviruses (Ha et al. 2008). The expected ~700-bp DNA product of each gene target was amplified from both samples. The amplicons were excised, gel eluted (Zymoclean™ Gel DNA Recovery kit) and cloned individually into the pJET1.2/Blunt vector (Life Technologies). Recombinant plasmids from two transformed Escherichia coli cells per cloned DNA insert were Sanger sequenced. BLASTn analysis of each gene-specific nucleotide (nt) sequence fragment revealed their identities (83 to 92 % nt) as lily mottle virus (LMoV; genus Potyvirus) at 93 to 98% query coverage. In pairwise comparison, the obtained sequences of LMoV isolates from TX specific to the CI (GenBank accession nos. PQ037260-61) and the HC-Pro (PQ037262-63) shared ~99% nt and 100% amino acid (aa) identities with each other and ~91-92% nt and 98-100% aa identities with the closest isolate, Bate5 (JN127341) from Australia. Based on these results, a pair of LMoV-specific primers (LM_v109-F: 5'-GGCCAGTAATGTGCACAAGC-3' and LM_c527-R: 5'-TCGCTGTAGCTAGCGACGTAC-3') was newly designed to target the partial CI gene, internal to the 700-bp fragment obtained above with the generic primers. The primer pair was used to screen each of the mechanically inoculated test plants by RT-PCR as previously described above. The expected 438-bp fragment of the LMoV CI gene was amplified from all the inoculated plants of both N. benthamiana and V. unguiculata; the results were confirmed by Sanger sequencing as described above. The mock-inoculated plant of each experimental host pl
百合(Lilium asiatica)是一种重要的植物,因其花色繁多、香味浓郁而被广泛种植。2024 年 3 月,在得克萨斯州伊达尔戈县韦斯拉科的一处私人庄园中,12/20 株百合植株上出现了由浅黄色斑驳和马赛克组成的潜在病毒样症状。我们随机抽取了两株出现症状的植物(WTX1 和 WTX2)进行病毒诊断。使用 Agdia 的 Poty ImmunoStrip® (Agdia, Inc.)然而,在接种后 20 至 28 天,将提取物摩擦接种到健康的烟草植物(Nicotiana benthamiana)和木樨(Vigna unguiculata)植物(各 4 株)的系统叶片上会诱发轻微的斑驳症状,这表明样品中存在可机械传播的病原体。在两种草本植物的模拟接种植株(各 1 株)上均未观察到病毒样症状。为了检测是否感染了疑似壶状病毒,使用 Oligo(dT) 引物和 PrimeScript 第一链 cDNA 合成试剂盒(Takara Bio,美国)从 WTX1 和 WTX2 提取 2µg 的总核酸(Dellaporta 等,1983 年)用于合成互补 DNA(cDNA)。用 5 U Taq 聚合酶(罗氏诊断公司,印第安纳波利斯,IN)和两对针对部分圆柱包涵体(CI)基因和壶状病毒辅助成分蛋白酶(HC-Pro)的退化引物(Ha 等人,2008 年),在 12.5µl 常规 PCR 反应体积中以每个 cDNA 的一微升等分量作为模板。从两个样本中扩增了每个目标基因的预期 ~700-bp DNA 产物。扩增子被切除、凝胶洗脱(Zymoclean™ Gel DNA Recovery kit)并分别克隆到 pJET1.2/Blunt 载体(Life Technologies)中。对每个克隆 DNA 插入物的两个转化大肠杆菌细胞中的重组质粒进行了桑格测序。对每个基因特异性核苷酸(nt)序列片段的 BLASTn 分析表明,它们与百合斑驳病病毒(LMoV;Potyvirus 属)的相同度(83%-92% nt)为 93%-98%,查询覆盖率为 93%-98%。在配对比较中,所获得的德克萨斯州百合斑驳病病毒(LMoV)特异分离物序列与CI(GenBank登录号:PQ037260-61)和HC-Pro(PQ037262-63)的序列具有约99%的nt和100%的氨基酸(aa)相同性,与最接近的澳大利亚分离物Bate5(JN127341)的序列具有约91-92%的nt和98-100%的aa相同性。根据这些结果,我们新设计了一对 LMoV 特异引物(LM_v109-F:5'-GGCCAGTAATGTGCACAAGC-3'和 LM_c527-R:5'-TCGCTGTAGCTAGCGACGTAC-3'),以上述通用引物获得的 700-bp 片段内部的部分 CI 基因为靶标。如前所述,该引物对用于通过 RT-PCR 筛选每一株机械接种的试验植株。从 N. benthamiana 和 V. unguiculata 的所有接种植株中都扩增出了预期的 LMoV CI 基因的 438-bp 片段;如上所述,结果通过 Sanger 测序得到了证实。每种实验寄主植物的模拟接种植株对 LMoV 的检测结果均为阴性。LMoV 是一种检疫性害虫,以前在美国(Brierley 和 Smith,1944 年)和世界其他百合生产地区(如荷兰、中国、韩国和巴西)都有报道。据我们所知,这是德克萨斯州首次确诊该病毒,从而扩大了其地理分布范围。为防止 LMoV 通过种植材料进一步传播,在繁殖前对百合基础种群进行检测至关重要。
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