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First Report of Colletotrichum siamense Causing Leaf Anthracnose on Jackfruit in Thailand. 泰国首次报告 Colletotrichum siamense 在菠萝上引起叶炭疽病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-09 DOI: 10.1094/PDIS-06-24-1273-PDN
Surapong Khuna, Jaturong Kumla, Tanapol Thitla, Chanokned Senwanna, Nakarin Suwannarach
<p><p>Jackfruit (Artocarpus heterophyllus Lam.) is commonly grown in Thailand. In June 2023, leaf anthracnose on this plant was observed at a field in Chai Prakan District (19°42'24"N, 99°01'59"E), Chiang Mai Province, Thailand, with ~25% disease incidence in a 1000-m2 plantation area. The initial symptom had brown spots with a yellow halo, enlarged, elongated, 0.2 to 2 cm in diameter, irregular, sunken, brown, with a dark brown halo, and leaves withered and dried. Pale yellow conidiomata developed on the lesions in high humidity. Ten symptomatic leaves were used to isolate the fungal causal agents through a single spore isolation method (Tovar-Pedraza et al. 2020). Four fungal isolates (SDBR-CMU492 to SDBR-CMU495) with similar morphology were obtained. Colonies on potato dextrose agar (PDA) were 70 to 85 mm in diameter, white to grayish white with cottony mycelia, the reverse pale yellow after incubation at 25°C for 1 week. All isolates produced asexual structures. Setae were brown with 1 to 3 septa, 40 to 100 × 2.2 to 4.0 µm, a cylindrical base, and acuminate tip. Conidiophores were hyaline to pale brown, septate, and branched. Conidiogenous cells were hyaline to pale brown, cylindrical to ampulliform, 7.4 to 27.2 × 2.0 to 4.5 µm. Conidia were one celled, hyaline, smooth walled, cylindrical, ends rounded, guttulate, 11.1 to 15.7 × 3.4 to 6.1 µm. Appressoria were dark brown to black, oval to irregular, 8.8 to 24.9 × 3.6 to 10 µm. Morphologically, all isolates resembled the Colletotrichum gloeosporioides species complex (Weir et al. 2012). The internal transcribed spacer (ITS) region, actin (act), β-tubulin (tub2), calmodulin (CAL), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were amplified using primer pairs ITS5/ITS4, ACT-512F/ACT-783R, T1/T22, CL1C/CL2C, and GDF1/GDR1, respectively (White et al. 1990; Weir et al. 2012). Sequences were deposited in GenBank (ITS: PP068858, PP068859, PP446789, PP446790; act: PP079636, PP079637, PP460760, PP460761; tub2: PP079638, PP079639, PP460762, PP460763; CAL: PP079634, PP079635, PP460758, PP460759; GAPDH: PP079632, PP079633, PP460756, PP460757). Maximum likelihood phylogenetic analyses of the concatenated five genes identified all isolates as C. siamense. To pathogenicity test, the mature leaves of a healthy plant were surface disinfested using 0.1% NaClO for 3 min, rinsed three times with sterile water, and wounded. Conidia suspensions (15 µl of 1 × 106 conidia/ml) of each isolate grown on PDA at 25°C for 2 weeks were used to inoculate wounded and unwounded samples by the attached method. Control leaves were mock inoculated with sterile distilled water. Ten replications were conducted for each treatment and repeated twice. Plants were placed in a greenhouse at 25 to 30°C and 80 to 90% relative humidity. After 7 days, all inoculated leaves displayed brown lesions, while control leaves had no symptoms. Colletotrichum siamense was reisolated from inoculated tissues on PDA to complete Koch's post
菠萝蜜(Artocarpus heterophyllus Lam.)2023 年 6 月,在泰国清迈府 Chai Prakan 县(北纬 19°42'24",东经 99°01'59")的一块田地里观察到这种植物的叶炭疽病,在 1000 平方米的种植区中发病率约为 25%。初期症状为带黄晕的褐色斑点,扩大、拉长,直径 0.2 至 2 厘米,不规则,凹陷,褐色,带深褐色晕,叶片枯萎干枯。在高湿度条件下,病斑上长出淡黄色的分生孢子器。通过单孢子分离法(Tovar-Pedraza et al.)获得了四种形态相似的真菌分离物(SDBR-CMU492 至 SDBR-CMU495)。马铃薯葡萄糖琼脂(PDA)上的菌落直径为 70 至 85 毫米,白色至灰白色,带有棉状菌丝,在 25°C 下培养 1 周后,菌落呈淡黄色。所有分离物均产生无性结构。刚毛呈棕色,有 1 至 3 个隔膜,长 40 至 100 × 2.2 至 4.0 µm,基部呈圆柱形,顶端渐尖。分生孢子梗呈透明至淡褐色,有隔膜,有分枝。分生孢子细胞呈透明至淡褐色,圆柱形至瓶形, 7.4 - 27.2 × 2.0 - 4.5 µm。分生孢子单细胞,透明,壁光滑,圆柱形,末端圆形,具沟, 11.1-15.7 × 3.4-6.1 µm。外稃呈深褐色至黑色,椭圆形至不规则形,8.8 至 24.9 × 3.6 至 10 微米。从形态上看,所有分离物都与 Colletotrichum gloeosporioides 种群相似(Weir 等人,2012 年)。使用引物对 ITS5/ITS4、ACT-512F/ACT-783R、T1/T22、CL1C/CL2C 和 GDF1/GDR1 分别扩增了内部转录间隔区(ITS)、肌动蛋白(act)、β-微管蛋白(tub2)、钙调蛋白(CAL)和甘油醛-3-磷酸脱氢酶(GAPDH)基因(White 等,1990 年;Weir 等,2012 年)。序列已存入 GenBank(ITS:PP068858, PP068859, PP446789, PP446790; act:PP079636, PP079637, PP460760, PP460761; tub2: PP079638, PP079639, PP460762, PP460763; CAL:PP079634、PP079635、PP460758、PP460759;GAPDH:PP079632、PP079633、PP460756、PP460757)。通过对五个基因的最大似然系统发生分析,确定所有分离物均为 C. siamense。在致病性试验中,用 0.1% 的 NaClO 对健康植物的成熟叶片进行表面消毒 3 分钟,然后用无菌水冲洗 3 次,并进行伤口处理。在 25°C 的 PDA 上生长 2 周的每种分离株的分生孢子悬浮液(15 µl 含 1 × 106 个分生孢子/ml)被用来按附着法接种受伤和未受伤的样本。对照叶片用无菌蒸馏水模拟接种。每种处理进行 10 次重复,重复两次。将植物置于温度为 25 至 30°C、相对湿度为 80 至 90% 的温室中。7 天后,所有接种的叶片都出现褐色病斑,而对照叶片则没有症状。从 PDA 上的接种组织中重新分离出了暹罗毛霉菌(Colletotrichum siamense),从而完成了科赫假说(Koch's postulates)。在本研究之前,C. fructicola 和 C. gloeosporioides 在全球范围内引起了菠萝的叶炭疽病(Sangchote 等人,2003 年;Chitambar,2016 年)。澳大利亚(James 等人,2014 年)和 Bazil(Borges 等人,2023 年)报道了由 C. siamense 引起的胡柚叶炭疽病。在泰国,Bhunjun 等人(2019 年)报道了 C. artocarpicola 在菠萝中引起的叶炭疽病。因此,这是泰国首次报道 C. siamense 在菠萝上引起叶炭疽病。这一发现将为流行病学调查和未来管理这种疾病的方法提供信息。
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引用次数: 0
First Report of Rhizopus arrhizusi causing rot of Dictyophora rubrovolvata in China. 在中国首次报道了由根瘤菌(Rhizopus arrhizusi)引起的红豆杉(Dictyophora rubrovolvata)腐烂病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-09 DOI: 10.1094/PDIS-08-24-1767-PDN
Shiqi Tu, Feng Zhou, Yin Zhang, YiFan Chen, ShengQian Chao, Beibei Lü, Hailong Yu
<p><p>Dictyophora rubrovolvata, as an edible fungus with high medicinal value, is widely cultivated in several provinces in China (Hang et al. 2012). However, between December 2023 and March 2024, a rot disease occurred in the main production area in Fengxian District, Shanghai, China (N30°93', E121°49'). The disease incidence was 25% in the affected 1.33-ha growing area. High temperatures (>25℃) and poor ventilation provide favorable conditions for the spread of this disease. The disease mainly occurs at the stage of fruiting bodies formation of D. rubrovolvata. When the epidermis is damaged and broken, it becomes infested with mold, which then produces a layer of moldy rot with pus. The infected D. rubrovolvata tissues at the edge of the lesions were isolated, surface sterilized and cultured on potato dextrose agar (PDA) at 30 ℃ under dark conditions. Pure cultures were obtained by single-spore isolation. After 3 days, isolates were transferred to Czapek Yeast agar (CYA) (Samson et al, 2014). On CYA, the fungal colony consisted of white flocculent hyphae. Scanning electron microscopy analysis showed that the mycelium was white, and the internodes of the stolons formed characteristic pseudoroots, from which upwardly clustered erect, unbranched sporocarp peduncles expanded apically to form rounded sporocarp sacs, within which sporocarp spores were produced. (Hariprasath P, 2019). To confirm the identity of the pathogen, the genomic fragments for the internal transcribed spacer (ITS) and intergenic spacer (IGS) gene of the isolate were amplified by PCR (White et al. 1990; Liu XY. 2008). The resulting sequence was deposited in GenBank with accession PP951880 and PQ001670, respectively. PCR results and morphological observations indicated the isolated strain was a pure culture and the strain was designated as DIC01. Comparative results showed that the sequences with accession numbers MT603964.1 and DQ990323.1 showed high homology of 99.15% and 98.96% to the ITS and IGS sequences of Rhizopus arrhizusi DIC01, respectively. Phylogenetic analysis with ITS and IGS genes of the isolated strain and 7 Rhizopus spp. strains were performed using MEGAX with Neighbor-Joining (NJ) method. Based on the results of growth habits, morphological observations, and phylogenetic analysis, the pathogen was identified as R. arrhizusi. A spore suspension of the R. arrhizusi DIC01 (1 x107 conidia/mL) was inoculated back to healthy D. rubrovolvata. Five healthy fruit bodies of D. rubrovolvata were injected, and another five healthy morels were treated with potato dextrose broth (PDB) as controls. D. rubrovolvata was incubated at 25°C and 90% relative humidity without ventilation for 5 days. The pathogen successfully infected the D. rubrovolvata, which developed white moldy lesions similar to those of natural diseases. The controls remained healthy without any symptoms. The pathogen was reisolated from the affected lesions and identified as R. arrhizusi DIC01 based on its mor
红豆杉(Dictyophora rubrovolvata)是一种药用价值极高的食用菌,在中国多个省份广泛种植(Hang 等,2012 年)。然而,在 2023 年 12 月至 2024 年 3 月期间,中国上海市奉贤区(N30°93',E121°49')的主产区发生了腐烂病。发病面积为 1.33 公顷,发病率为 25%。高温(>25℃)和通风不良为该病的传播提供了有利条件。该病主要发生在 D. rubrovvolata 子实体形成阶段。当表皮受损和破损时,就会被霉菌侵染,然后产生一层带有脓液的霉烂层。分离病斑边缘受感染的红铃虫组织,进行表面灭菌,在 30 ℃ 黑暗条件下置于马铃薯葡萄糖琼脂(PDA)上培养。通过单孢分离获得纯培养物。3 天后,将分离物转移到 Czapek 酵母菌琼脂(CYA)上(Samson 等人,2014 年)。在 CYA 上,真菌菌落由白色絮状菌丝组成。扫描电子显微镜分析表明,菌丝是白色的,匍匐茎节间形成特征性的假根,从假根向上簇生直立、不分枝的孢子囊梗,顶部膨大形成圆形的孢子囊,孢子囊内产生孢子。(Hariprasath P,2019 年)。为确认病原体的身份,通过 PCR 扩增了分离株的内部转录间隔(ITS)和基因间间隔(IGS)基因的基因组片段(White 等,1990 年;Liu XY,2008 年)。所得序列分别以 PP951880 和 PQ001670 的登录号存入 GenBank。聚合酶链式反应结果和形态学观察结果表明,分离出的菌株是纯培养物,该菌株被命名为 DIC01。比较结果表明,登录号为 MT603964.1 和 DQ990323.1 的序列与根瘤菌 DIC01 的 ITS 和 IGS 序列的同源性分别为 99.15%和 98.96%。利用 MEGAX 和 Neighbor-Joining (NJ) 方法对分离菌株和 7 株根瘤菌的 ITS 和 IGS 基因进行了系统进化分析。根据生长习性、形态观察和系统发育分析的结果,确定病原体为 R. arrhizusi。将 R. arrhizusi DIC01 的孢子悬浮液(1 x107 分生孢子/毫升)接种回健康的 D. rubrovolvata。注射 5 个健康的 D. rubrovolvata 果体,另用马铃薯葡萄糖肉汤(PDB)处理 5 个健康的羊肚菌作为对照。在 25°C、相对湿度为 90%、不通风的条件下培养 D. rubrovolvata 5 天。病原体成功感染了 D. rubrovolvata,它出现了与自然病害类似的白色霉斑。对照组仍然健康,没有出现任何症状。根据病原体的形态特征和系统发育标记基因,从受害病斑中重新分离出病原体,并确定其为 R. arrhizusi DIC01。据报道,R. arrhizusi 可造成内皮细胞损伤,菌丝侵入血管,导致血栓形成和组织坏死。(Hariprasath P, 2019)。据我们所知,这是第一份关于 R. arrhizusi 导致 D. rubrovolvata 腐烂病的报告。本研究证实,R. arrhizusi 是导致上海奉贤红富士农场红富士腐烂病的病原真菌。
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引用次数: 0
First report of Colletotrichum nymphaeae causing anthracnose on Hass avocados in Brazil. 首次报告 Colletotrichum nymphaeae 在巴西哈斯鳄梨上引起炭疽病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-09 DOI: 10.1094/PDIS-08-24-1674-PDN
Vanessa Caroline de Oliveira, Hiago Antonio Oliveira da Silva, Fabrícia Queiroz Mendes, Eduardo Basílio de Oliveira, Lucas Magalhães de Abreu, Érica Nascif Rufino Vieira
<p><p>The Hass cultivar is one of the most cultivated and distributed avocado varieties throughout the world, having high productivity, nutritional quality, market acceptance and adaptation to different climatic zones (Dreher and Davenport 2013). Anthracnose affects avocado production in tropical and subtropical regions, causing economic losses, especially post-harvest (Fuentes-Aragón et al. 2020). Correct identification of Colletotrichum species causing anthracnose is essential, as different species vary in important phenotypes such as virulence and sensitivity to fungicides (Chen et al. 2016). Twelve samples from batches of avocados with anthracnose were collected in Minas Gerais State, Brazil, in September 2023. The observed symptoms were brown to black depressed circular spots, ranging from a few millimeters to 3 cm in diameter on the epicarp of the fruits, covered in center by mucilaginous layers of pathogen sporulation. Isolation was performed directly from the spore masses and monoconidial isolates were cultured in PDA at 25°C for 7 days for morphological characterization and preserved in sterile water at 4°C. One of the morphotypes commonly recovered from lesions, represented by isolate UCBV 362 (Culture Collection COAD 3843), formed fast-growing colonies having white aerial mycelium and intense salmon-colored sporulation. The cylindrical conidia were 13 to 17.5 μm long and 4.5 to 7 μm wide (average 14.5 x 5.7 μm, N=100), produced on conidiophores dispersed in the aerial mycelium or aggregated on melanized conidiomata formed on the agar. The partial sequence of the second largest subunit of the RNA polymerase II gene (RPB2) from isolate UCBV 362 (GenBank: PQ034617, 1116 nt) showed 99% of coverage and 99.37% of nucleotide identity with the RPB2 sequence of the ex-epitype strain of Colletotrichum nymphaeae ICMP 17918 (=CBS 515.78) (GenBank: JN985506). In a Maximum Likelihood phylogenetic tree composed with RBP2 sequences from reference strains of the Colletotrichum acutatum species complex, the isolate UCBV 362 formed a highly supported clade with the ex-epitype and other reference strains of Colletotrichum nymphaeae, occupying the Clade 2 of the species complex together with C. scovillae and C. simmondsii (Damm et al. 2012). This result shows the reliability of RPB2 for phylogeny and species delimitation within Colletotrichum. To confirm pathogenicity, 10-mm discs from a 7-day-old colony were inoculated at 3 different points on healthy-looking avocado fruits and incubated at 28°C. Uninoculated fruits served as controls. The first symptoms appeared 5 days after inoculation and were similar to those observed in the original samples, while the fruits in the control group remained asymptomatic. The pathogen was reisolated from the lesions and identified morphologically, fulfilling Koch's postulates. Colletotrichum nymphaeae has been associated with avocado anthracnose in Mexico (Fuentes-Aragón et al. 2020). In Brazil, a study based on molecula
哈斯栽培品种是全世界栽培和分布最广的鳄梨品种之一,具有产量高、营养品质好、市场接受度高以及适应不同气候带等特点(Dreher 和 Davenport,2013 年)。炭疽病影响热带和亚热带地区的鳄梨生产,造成经济损失,尤其是收获后的损失(Fuentes-Aragón 等人,2020 年)。正确识别引起炭疽病的 Colletotrichum 菌种至关重要,因为不同菌种在毒力和对杀菌剂的敏感性等重要表型上存在差异(Chen 等,2016 年)。2023 年 9 月,在巴西米纳斯吉拉斯州采集了 12 批患有炭疽病的鳄梨样本。观察到的症状是在果实的外果皮上出现棕色至黑色的凹陷圆斑,直径从几毫米到 3 厘米不等,中间被病原体孢子的粘液层覆盖。直接从孢子团中进行分离,单孢分离物在 25°C 的 PDA 中培养 7 天,以确定形态特征,并在 4°C 的无菌水中保存。以分离株 UCBV 362(培养物保藏中心 COAD 3843)为代表的一种常见形态型是从病变中恢复的,形成快速生长的菌落,具有白色气生菌丝和浓鲑鱼色孢子。圆柱形分生孢子长 13 至 17.5 μm,宽 4.5 至 7 μm(平均 14.5 x 5.7 μm,N=100),产生于分散在气生菌丝中的分生孢子梗上或聚集在琼脂上形成的黑色化分生孢子器上。分离株 UCBV 362(GenBank:PQ034617,1116 nt)的 RNA 聚合酶 II 基因第二大亚基(RPB2)的部分序列与 Colletotrichum nymphaeae ICMP 17918(=CBS 515.78)(GenBank:JN985506)前表皮菌株的 RPB2 序列的覆盖率为 99%,核苷酸同一性为 99.37%。在与来自 acutatum Colletotrichum 种类复合体参考菌株的 RBP2 序列组成的最大似然系统发生树中,分离株 UCBV 362 与 nymphae Colletotrichum 的前表皮菌株和其他参考菌株形成了一个高度支持的支系,与 C. scovillae 和 C. simmondsii 一起占据了种类复合体的支系 2(Damm 等人,2012 年)。这一结果显示了 RPB2 在 Colletotrichum 系统发育和物种划分方面的可靠性。为确认致病性,在外观健康的鳄梨果实上的 3 个不同点接种了 7 天菌落的 10 毫米圆片,并在 28°C 下培养。未接种的果实作为对照。接种 5 天后出现了第一个症状,与在原始样本中观察到的症状相似,而对照组中的果实仍无症状。病原体从病变处重新分离出来,并通过形态学鉴定,符合科赫假说。在墨西哥,Colletotrichum nymphaeae 与鳄梨炭疽病有关(Fuentes-Aragón 等人,2020 年)。在巴西,一项基于分子系统发育的研究确定 Colletotrichum siamense 和 C. karsti 为该病害的病原菌(Soares 等,2021 年)。本报告扩大了导致巴西鳄梨炭疽病的物种范围,并为实施和监测控制方法(尤其是化学防治)提供了病原学信息。
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引用次数: 0
First Report of Ralstonia pseudosolanacearum Causing Bacterial Wilt in Ginger in Peru. 秘鲁首次报告 Ralstonia pseudosolanacearum 导致生姜细菌性枯萎病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-09 DOI: 10.1094/PDIS-09-24-1874-PDN
José Soto-Heredia, Salome Ramos-Tito, Angelica Rodrigues Alves, Luciellen da Costa Ferreira, Luz Leonor Mattos Calderon, Maurício Rossato
<p><p>Around 90% of Peru's ginger (Zingiber officinale) production is concentrated in the Junín region, due to the optimal agroecological conditions for its cultivation. In March 2024, fields with ginger plants (cultivar Criollo) in Junín region, provinces Chanchamayo and Satipo, specifically in the cities of Pichanaqui and Satipo respectively, exhibited approximately 40% of plants with severe symptoms of a disease characterized initially by plant yellowing and rapid progressing to necrosis. Affected rhizomes showed dark vascular bundles with milky white exudates upon cutting, while stems displayed vascular necrosis hindering water and nutrient transport, often resulting in plant death. Fifteen plants were sampled and diseased vascular tissues from rhizomes and stems were cultured on nutrient agar (NA) and incubated at 28°C. After 72 h, all isolations resulted in colonies with typical characteristics of Ralstonia solanacearum species complex (RSSC) were produced, with appearing fluid, irregularly round, and creamy white. Three isolates were selected for the identification steps (UNALM-RP01 to 03) were identified by PCR using primers 759/760 (Opina et al. 1997) confirming as RSSC with a 282 bp amplification product. Additionally, isolates were assigned to biovar 3 based on their ability to metabolize three acid-producing disaccharides (maltose, lactose, cellobiose) and three hexose alcohols (mannitol, sorbitol, dulcitol) (Hayward, 1964). Phylotype I was identified by multiplex PCR (primers Nmult) with a 114 bp amplification product (Fegan and Prior 2005). For the identification of the sequevars of the three isolates, DNA was extracted and PCR with primers ENDO-F/R (Ji et al. 2007) were performed to amplify and sequence the partial gene sequence of egl gene with 681 bp in length. The phylogeny by Neighbor joining with 10,000 bootstraps clustered the UNALM isolates along other sequevar 30 of R. pseudosolanacearum. The sequences were deposited in Genbank under accessions PQ213016, PQ213017 and PQ213018. For pathogenicity tests, bacterial colonies of isolate UNALM-RP01 were scraped from the culture media with a sterile needle and introduced into the stems of three 2-month-old ginger plants (cultivar Gigante). The plants subsequently exhibited yellowing seven days post-inoculation. Additionally, the rhizomes showed internal discoloration and bacterial exudation. Three plants were used as a control, which were pierced with a sterilized needle and showed no symptoms. All tested plants were kept in a greenhouse with controlled temperature (20-40 °C) The pathogen was successfully re-isolated from infected plants on NA medium, presenting typical colonies of RSSC and identified via PCR with primers 759/760, fulfilling Koch's postulates. This represents the first case in Peru of ginger plants infected with a Ralstonia species, specifically R. solanacearum phylotype I, corresponding to R. pseudosolanacearum. This species of RSSC and sequevar is known for causi
秘鲁约 90% 的生姜(Zingiber officinale)产量集中在胡宁地区,因为该地区拥有种植生姜的最佳农业生态条件。2024 年 3 月,胡宁地区、昌查马约省和萨蒂波省(特别是分别位于皮查纳基市和萨蒂波市)种植生姜(栽培品种 Criollo)的田地中,约 40% 的植株出现了严重的病害症状,最初表现为植株发黄,随后迅速发展为坏死。受影响的根茎表现为深色维管束,切开后有乳白色渗出物,而茎则表现为维管束坏死,阻碍水分和养分的运输,往往导致植株死亡。对 15 株植物进行取样,将根茎和茎上的病变维管束组织培养在营养琼脂(NA)上,在 28°C 下培养。72 小时后,所有分离菌株都产生了具有茄属拉氏菌(Ralstonia solanacearum)复合菌种(RSSC)典型特征的菌落,呈液态、不规则圆形和乳白色。通过使用引物 759/760(Opina 等人,1997 年)进行 PCR 鉴定,确认三个分离物(UNALM-RP01 至 03)为 RSSC,扩增产物为 282 bp。此外,根据分离物代谢三种产酸二糖(麦芽糖、乳糖、纤维生物糖)和三种六糖醇(甘露醇、山梨醇、杜冷丁醇)的能力,将其归入生物变种 3(Hayward,1964 年)。系统型 I 通过多重 PCR(引物 Nmult)与 114 bp 的扩增产物进行鉴定(Fegan 和 Prior,2005 年)。为了鉴定这三个分离株的序列,提取了 DNA,并用引物ENDO-F/R(Ji 等人,2007 年)进行 PCR 扩增和测序,得到了长度为 681 bp 的egl 基因部分序列。通过 10,000 次引导的邻接法进行系统发育,将 UNALM 分离物与 R. pseudosolanacearum 的其他 30 个序列聚类。这些序列以编号 PQ213016、PQ213017 和 PQ213018 存入 Genbank。在致病性试验中,用无菌针从培养基中刮取分离物 UNALM-RP01 的细菌菌落,并将其引入三株 2 个月大的生姜植株(栽培品种 Gigante)的茎中。接种后七天,植株出现黄化现象。此外,根茎内部也出现褪色和细菌渗出。三株植物作为对照,用消毒针刺入,未出现任何症状。病原体在 NA 培养基上从受感染的植物中成功地重新分离出来,呈现出 RSSC 的典型菌落,并通过引物 759/760 进行 PCR 鉴定,符合科赫假说。这是秘鲁首例生姜植物感染 Ralstonia 菌种的病例,特别是 R. solanacearum 系统型 I,相当于 R. pseudosolanacearum。众所周知,RSSC 和 sequevar 的这一菌种会导致生姜发病。然而,它在秘鲁的出现可能是病原体引入的结果,因为它的地理起源与亚洲有关(Fegan 和 Prior,2005 年)。据我们所知,这是秘鲁首次报道 R. pseudosolanacearum 引起生姜枯萎病。据估计,2024 年胡宁地区的平均产量将因枯萎病损失 30%,这对种植业构成了重大威胁。为了控制和减少进一步的损失,必须采取紧急和有效的病害管理策略。
{"title":"First Report of <i>Ralstonia pseudosolanacearum</i> Causing Bacterial Wilt in Ginger in Peru.","authors":"José Soto-Heredia, Salome Ramos-Tito, Angelica Rodrigues Alves, Luciellen da Costa Ferreira, Luz Leonor Mattos Calderon, Maurício Rossato","doi":"10.1094/PDIS-09-24-1874-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-09-24-1874-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Around 90% of Peru's ginger (Zingiber officinale) production is concentrated in the Junín region, due to the optimal agroecological conditions for its cultivation. In March 2024, fields with ginger plants (cultivar Criollo) in Junín region, provinces Chanchamayo and Satipo, specifically in the cities of Pichanaqui and Satipo respectively, exhibited approximately 40% of plants with severe symptoms of a disease characterized initially by plant yellowing and rapid progressing to necrosis. Affected rhizomes showed dark vascular bundles with milky white exudates upon cutting, while stems displayed vascular necrosis hindering water and nutrient transport, often resulting in plant death. Fifteen plants were sampled and diseased vascular tissues from rhizomes and stems were cultured on nutrient agar (NA) and incubated at 28°C. After 72 h, all isolations resulted in colonies with typical characteristics of Ralstonia solanacearum species complex (RSSC) were produced, with appearing fluid, irregularly round, and creamy white. Three isolates were selected for the identification steps (UNALM-RP01 to 03) were identified by PCR using primers 759/760 (Opina et al. 1997) confirming as RSSC with a 282 bp amplification product. Additionally, isolates were assigned to biovar 3 based on their ability to metabolize three acid-producing disaccharides (maltose, lactose, cellobiose) and three hexose alcohols (mannitol, sorbitol, dulcitol) (Hayward, 1964). Phylotype I was identified by multiplex PCR (primers Nmult) with a 114 bp amplification product (Fegan and Prior 2005). For the identification of the sequevars of the three isolates, DNA was extracted and PCR with primers ENDO-F/R (Ji et al. 2007) were performed to amplify and sequence the partial gene sequence of egl gene with 681 bp in length. The phylogeny by Neighbor joining with 10,000 bootstraps clustered the UNALM isolates along other sequevar 30 of R. pseudosolanacearum. The sequences were deposited in Genbank under accessions PQ213016, PQ213017 and PQ213018. For pathogenicity tests, bacterial colonies of isolate UNALM-RP01 were scraped from the culture media with a sterile needle and introduced into the stems of three 2-month-old ginger plants (cultivar Gigante). The plants subsequently exhibited yellowing seven days post-inoculation. Additionally, the rhizomes showed internal discoloration and bacterial exudation. Three plants were used as a control, which were pierced with a sterilized needle and showed no symptoms. All tested plants were kept in a greenhouse with controlled temperature (20-40 °C) The pathogen was successfully re-isolated from infected plants on NA medium, presenting typical colonies of RSSC and identified via PCR with primers 759/760, fulfilling Koch's postulates. This represents the first case in Peru of ginger plants infected with a Ralstonia species, specifically R. solanacearum phylotype I, corresponding to R. pseudosolanacearum. This species of RSSC and sequevar is known for causi","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":null,"pages":null},"PeriodicalIF":4.4,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142392448","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 Paprika mild mottle virus Infecting pepper (Capsicum annuum L.) in China. 中国首次报告辣椒轻度斑驳病毒感染辣椒(Capsicum annuum L.)
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-09 DOI: 10.1094/PDIS-01-24-0191-PDN
Zhilei Han, Mingrui Gao, Zhiyong Yan, Hao Hong, Xiao Yin, Shuai Xu, Jiejun Peng, Fei Yan, Xiangdong Li, Shanshan Jiang
<p><p>Paprika mild mottle virus (PaMMV), a tobamovirus from the Virgaviridae family, has been reported to infect pepper (<i>Capsicum annuum</i> L.) in the Netherlands (García-Luque et al., 1993), Japan (Hamada et al., 2003), Bulgaria (Ruíz del Pino et al., 2003), and Israel (Luria et al., 2018). Pepper has become the most widely planted vegetable and the most heavily consumed spicy condiment, with a large planting area (>21,000 km<sup>2</sup>) in China (Zou et al., 2021). However, the crop's productivity is limited by viral diseases. In August 2021, a survey of viral diseases was conducted across five pepper-growing regions in Linyi, Shandong Province, China, revealed that 5% to 10% of plants in most fields exhibited symptoms such as stunting, leaf narrowing, chlorosis, and crinkling of leaf tissue (Fig. S1-a). Subsequently, seven symptomatic samples (Fig. S1-b) were collected from a plot (118° 29' E, 34° 65' N) where thin-skinned peppers were grown. All samples tested positive in a Western blot assay using a polyclonal antibody for tobacco mosaic virus (TMV), indicating the presence of a tobamovirus (Fig. S1-c). To identify the virus, total RNAs were extracted from 10 different symptomatic pepper plant samples, using the RNeasy Plant Mini Kit (QIAGEN, Hilden, Germany) and combined into a single sample in equal amounts (100 ng/μL each). RNA-Seq was performed on this mixture using the Illumina NovaSeq 6000 (Illumina, San Diego, USA). Raw reads (6.92G) in fastq format were processed with in-house perl scripts to yield clean reads (6.68G), which were assembled into 780 contigs (mean length: 2036 bp) using Trinity software (v2.6.6). These contigs were then analysed using the NCBI BLASTX program (http://www.ncbi.nlm.nih.gov/blast) against the viral RefSeq database. The results demonstrated that the contigs exhibited an average genome coverage of 40.16%. Notably, one unique contig (6453 bp) was mapped to the PaMMV genome (reference sequence KX187305.1) with 98.34% identity. To verify the RNA-Seq result, a PaMMV-specific primer pair (forward: 5'-GAGTTCATAGAGGCAGTACC-3'; reverse: 5'-CTTCGATTTAAGTGGAGGGAT-3') was designed for amplifying an 800-nt fragment of the PaMMV coat protein (<i>CP</i>) gene by RT-PCR. The 800-nt fragment was successfully amplified from all the symptomatic samples (Fig. S1-d). Sequenced RT-PCR products (GenBank No. OR365081.1) showed 99.75% nucleotide identity with PaMMV isolates (OQ198318.1, China, goat) according to BLAST analysis. Mechanical transmission of PaMMV from three infected pepper plants collected in Linyi to healthy pepper plants confirmed pathogenicity. Three pepper cultivars (trade name: Qiemen, Tianyu, Haonong11) were tested and showed varying symptoms post-inoculation (Fig. S1-e). All cultivars were confirmed in the lab to be PMMoV-sensitive and exhibited significant viral disease symptoms after PMMoV inoculation (data not shown). Each variety was inoculated with at least 3 seedlings from different field-collected v
据报道,荷兰(García-Luque等人,1993年)、日本(Hamada等人,2003年)、保加利亚(Ruíz del Pino等人,2003年)和以色列(Luria等人,2018年)的辣椒(Capsicum annuum L.)感染了红辣椒轻微斑驳病毒(PaMMV),这是一种病毒科的托布病毒。辣椒已成为种植面积最广的蔬菜和消费量最大的辛辣调味品,在中国的种植面积很大(>21,000 平方公里)(邹等人,2021 年)。然而,该作物的产量受到病毒病的限制。2021 年 8 月,在中国山东省临沂市的五个辣椒种植区进行了一次病毒病调查,结果显示,大多数田块中有 5%至 10%的植株表现出发育不良、叶片变窄、萎蔫和叶片组织皱缩等症状(图 S1-a)。随后,从一块种植薄皮辣椒的地块(东经 118°29',北纬 34°65')采集了 7 个症状样本(图 S1-b)。使用烟草花叶病毒(TMV)多克隆抗体进行 Western 印迹检测,所有样本均呈阳性,表明存在烟草花叶病毒(图 S1-c)。为鉴定病毒,使用 RNeasy Plant Mini Kit(QIAGEN,Hilden,Germany)从 10 个不同症状的辣椒植株样本中提取总 RNA,并等量(每个 100 ng/μL)合并成一个样本。使用 Illumina NovaSeq 6000(Illumina,美国圣地亚哥)对该混合物进行 RNA 序列分析。使用内部 perl 脚本对 fastq 格式的原始读数(6.92G)进行处理,得到干净的读数(6.68G),并使用 Trinity 软件(v2.6.6)将其组装成 780 个等位组(平均长度:2036 bp)。然后使用 NCBI BLASTX 程序(http://www.ncbi.nlm.nih.gov/blast)对照病毒 RefSeq 数据库对这些等位基因进行分析。结果显示,等位基因的平均基因组覆盖率为 40.16%。值得注意的是,一个独特的等位基因(6453 bp)与 PaMMV 基因组(参考序列 KX187305.1)有 98.34% 的一致性。为了验证 RNA-Seq 的结果,研究人员设计了一对 PaMMV 特异引物(正向:5'-GAGTTCATAGAGGCAGTACC-3';反向:5'-CTTCGATTTAAGTGGAGGGAT-3'),用于通过 RT-PCR 扩增 PaMMV 衣壳蛋白(CP)基因的 800-nt 片段。从所有症状样本中成功扩增出了 800-nt 片段(图 S1-d)。根据 BLAST 分析,RT-PCR 产物(GenBank 编号:OR365081.1)与 PaMMV 分离物(OQ198318.1,中国,山羊)的核苷酸一致性为 99.75%。从临沂采集的三株受感染的辣椒植株上将 PaMMV 机械传播给健康的辣椒植株证实了其致病性。三个辣椒栽培品种(商品名:祁门、天宇、昊农11)在接种后出现了不同的症状(图 S1-e)。实验室证实所有栽培品种都对 PMMoV 敏感,接种 PMMoV 后表现出明显的病毒病症状(数据未显示)。每个品种至少接种了 3 株来自不同田间采集毒源的幼苗。祁门(栽培品种:甜椒)表现出严重的坏死斑症状,天宇(栽培品种:朝天椒)出现轻微的叶片皱缩,昊农 11 号(栽培品种:泡椒)出现黄色斑驳和叶片萎缩。通过 RT-PCR 检测,所有受试幼苗均对 PaMMV 呈阳性反应(图 S1-f)。据我们所知,这是中国首次报道辣椒感染PaMMV病毒,这突出表明有必要进行警惕性监测,以保护辣椒产业并防止病毒传播到其他作物。
{"title":"First Report of Paprika mild mottle virus Infecting pepper (<i>Capsicum annuum</i> L.) in China.","authors":"Zhilei Han, Mingrui Gao, Zhiyong Yan, Hao Hong, Xiao Yin, Shuai Xu, Jiejun Peng, Fei Yan, Xiangdong Li, Shanshan Jiang","doi":"10.1094/PDIS-01-24-0191-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-01-24-0191-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Paprika mild mottle virus (PaMMV), a tobamovirus from the Virgaviridae family, has been reported to infect pepper (&lt;i&gt;Capsicum annuum&lt;/i&gt; L.) in the Netherlands (García-Luque et al., 1993), Japan (Hamada et al., 2003), Bulgaria (Ruíz del Pino et al., 2003), and Israel (Luria et al., 2018). Pepper has become the most widely planted vegetable and the most heavily consumed spicy condiment, with a large planting area (&gt;21,000 km&lt;sup&gt;2&lt;/sup&gt;) in China (Zou et al., 2021). However, the crop's productivity is limited by viral diseases. In August 2021, a survey of viral diseases was conducted across five pepper-growing regions in Linyi, Shandong Province, China, revealed that 5% to 10% of plants in most fields exhibited symptoms such as stunting, leaf narrowing, chlorosis, and crinkling of leaf tissue (Fig. S1-a). Subsequently, seven symptomatic samples (Fig. S1-b) were collected from a plot (118° 29' E, 34° 65' N) where thin-skinned peppers were grown. All samples tested positive in a Western blot assay using a polyclonal antibody for tobacco mosaic virus (TMV), indicating the presence of a tobamovirus (Fig. S1-c). To identify the virus, total RNAs were extracted from 10 different symptomatic pepper plant samples, using the RNeasy Plant Mini Kit (QIAGEN, Hilden, Germany) and combined into a single sample in equal amounts (100 ng/μL each). RNA-Seq was performed on this mixture using the Illumina NovaSeq 6000 (Illumina, San Diego, USA). Raw reads (6.92G) in fastq format were processed with in-house perl scripts to yield clean reads (6.68G), which were assembled into 780 contigs (mean length: 2036 bp) using Trinity software (v2.6.6). These contigs were then analysed using the NCBI BLASTX program (http://www.ncbi.nlm.nih.gov/blast) against the viral RefSeq database. The results demonstrated that the contigs exhibited an average genome coverage of 40.16%. Notably, one unique contig (6453 bp) was mapped to the PaMMV genome (reference sequence KX187305.1) with 98.34% identity. To verify the RNA-Seq result, a PaMMV-specific primer pair (forward: 5'-GAGTTCATAGAGGCAGTACC-3'; reverse: 5'-CTTCGATTTAAGTGGAGGGAT-3') was designed for amplifying an 800-nt fragment of the PaMMV coat protein (&lt;i&gt;CP&lt;/i&gt;) gene by RT-PCR. The 800-nt fragment was successfully amplified from all the symptomatic samples (Fig. S1-d). Sequenced RT-PCR products (GenBank No. OR365081.1) showed 99.75% nucleotide identity with PaMMV isolates (OQ198318.1, China, goat) according to BLAST analysis. Mechanical transmission of PaMMV from three infected pepper plants collected in Linyi to healthy pepper plants confirmed pathogenicity. Three pepper cultivars (trade name: Qiemen, Tianyu, Haonong11) were tested and showed varying symptoms post-inoculation (Fig. S1-e). All cultivars were confirmed in the lab to be PMMoV-sensitive and exhibited significant viral disease symptoms after PMMoV inoculation (data not shown). Each variety was inoculated with at least 3 seedlings from different field-collected v","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":null,"pages":null},"PeriodicalIF":4.4,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142392451","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 Exserohilum pedicellatum Causing Root Rot of Wheat (Triticum aestivum L.) in Uzbekistan. 乌兹别克斯坦首次报告 Exserohilum pedicellatum 引起小麦(Triticum aestivum L.)根腐病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-09 DOI: 10.1094/PDIS-08-24-1788-PDN
Anvar Gulmirzaevich Sherimbetov, Bakhtiyor Shukhratovich Adilov, Dilshod Rustam Ogli Ruzmetov
<p><p>Wheat is the major staple food in Uzbekistan, and it occupies the largest harvested area (1,3 million hectares) in the country (USDA 2024). In June 2023, a survey was conducted to investigate root pathogens in wheat growing fields of Kaspi district in the Kashkadarya region of Uzbekistan. A total of 24 symptomatic plants with root rot and dark brown root lesions were collected from focal lesions in 4 different fields. From each plant, roots were excised and surface sterilized with 1% sodium hypochlorite for four minutes, then rinsed three times with sterile distilled water. Following surface sterilization, the excised roots were air dried in a laminar flow on sterile tissue sheets, rinsed twice with sterile distilled water, and then cut into 1 cm lengths segments (5 segments per one plant). The root pieces were cultured at 24°C for 4 days with a 12-hour photoperiod on potato dextrose agar supplemented with streptomycin (0.1 g/liter) and chloramphenicol (0.05 g/liter). From 24 symptomatic plants a 5 dematiaceous hyphomycete monoconidial pure isolates with abundant conidia were isolated. The conidia (n = 60) were mostly fusiform, straight, four to seven distoseptate, olivaceous brown to dark brown, and measured 51 to 88.7 × 17.9 to 25.4 μm (average 69.7 × 21.57 μm). Based on morphological characteristics the fungus was identified as E. pedicellatum according to Sivanesan (1987) and Hernandez-Restrepo et al. (2018). From five isolated monoconidial colonies, one has been chosen for molecular-genetic identification. Total DNA was extracted from it using PureLink™ Genomic DNA Mini Kit (Thermo Fisher Scientific, Waltham, MA, USA). For more informative analysis two loci, he translation elongation factor 1-alpha (tef1) and beta-tubulin (tub) genes were PCR-amplified and sequenced using gene specific primers: EF-1F (5'-CGGTGGTATCGACAAGCGT-3'), EF-2R (5'-AGCATGTTGTCGCCGTTGAAG-3')designed by Primer3web v4.1.0 software (Untergasser et al. 2012), and Bt2a (5'- GGTAACCAAATCGGTGCTGCTTTC, Bt2b (5'-ACCCTCAGTGTAGTGACCCTTGGC -3')described by Glass and Donaldson (1995), respectively. The resulting sequences were deposited in NCBI database under accession number PQ095881 and PQ095882. After BLAST analysis they showed highest similarity with the corresponding sequences of tef1 JQ672389 (100% identity, from 287 bp 287 bp are matching) and tub JQ671941 (100% identity, from 273 bp 273 bp are matching) of BMP 0384 isolate of E. pedicellatum from USA. In the plant inoculations (pathogenicity test), three isolates of E. pedicellatum were evaluated. For the pathogenicity test, conidia were scraped from PDA plate, suspended in water, and mixed with sterile sand to obtain a density of 500 conidia/g. A total of 20 wheat seed (Grom variety) previously disinfected 2 min with 10% NaOCl, were sown in each plastic pot (14 cm x 4 cm, 2 seeds per pot) filled with the inoculated soil (5 pots) and with sterilized soil (5 pots) as a control. Plants were grown in a growth chamber wit
小麦是乌兹别克斯坦的主要主食,其收获面积居全国之首(130 万公顷)(美国农业部,2024 年)。2023 年 6 月,对乌兹别克斯坦卡什卡达里亚地区卡斯皮区小麦种植地的根部病原体进行了调查。在 4 块不同的田地里,从病灶处共采集了 24 株有根腐病和深褐色根部病变症状的植株。从每株植物上切除根部,用 1%次氯酸钠进行表面消毒 4 分钟,然后用无菌蒸馏水冲洗 3 次。表面消毒后,将切除的根在无菌组织板上层流风干,用无菌蒸馏水冲洗两次,然后切成 1 厘米长的小段(每株植物 5 段)。将根块放在添加了链霉素(0.1 克/升)和氯霉素(0.05 克/升)的马铃薯葡萄糖琼脂上,在 24℃、12 小时光周期下培养 4 天。从 24 株有症状的植株中分离出了 5 个具有大量分生孢子的半知菌单胞纯分离株。分生孢子(n = 60)多呈纺锤形,直,4 至 7 个二裂,橄榄褐色至深褐色,大小为 51 至 88.7 × 17.9 至 25.4 μm(平均 69.7 × 21.57 μm)。根据形态特征,按照 Sivanesan(1987 年)和 Hernandez-Restrepo 等人(2018 年)的说法,该真菌被鉴定为 E. pedicellatum。从五个分离的单核菌落中,选择了一个进行分子遗传鉴定。使用 PureLink™ Genomic DNA Mini Kit(赛默飞世尔科技公司,美国马萨诸塞州沃尔瑟姆)从其中提取总 DNA。为了进行更翔实的分析,使用基因特异性引物对翻译伸长因子 1-α(tef1)和 beta-tubulin(tub)基因的两个位点进行了 PCR 扩增和测序:EF-1F (5'-CGGTGGTATCGACAAGCGT-3')、EF-2R (5'-AGCATGTTGTCGCCGTTGAAG-3')由 Primer3web v4.1.0 软件设计(Untergasser et al.2012)和 Bt2a (5'- GGTAACCAAATCGGTGCTGCTTTC,Bt2b (5'-ACCCTCAGTGTAGTGACCCTTGGC -3')分别由 Glass 和 Donaldson(1995)描述。得到的序列以 PQ095881 和 PQ095882 的登录号存入 NCBI 数据库。经过 BLAST 分析,它们与美国 BMP 0384 pedicellatum E. 分离物的 tef1 JQ672389(100% 相同,从 287 bp 起 287 bp 匹配)和 tub JQ671941(100% 相同,从 273 bp 起 273 bp 匹配)的相应序列显示出最高的相似性。在植物接种(致病性试验)中,评估了三种赤霉病菌分离物。在致病性试验中,从 PDA 平板上刮取分生孢子,悬浮于水中,并与无菌沙混合,以获得 500 个分生孢子/克的密度。在每个塑料盆(14 厘米 x 4 厘米,每盆 2 粒种子)中播种 20 粒小麦种子(Grom 品种),每粒种子事先用 10%的 NaOCl 消毒 2 分钟,然后播种接种土壤(5 盆)和消毒土壤(5 盆)作为对照。植物在光周期为 12 小时、温度为 24°C 的生长室中生长 4 周。在接种土壤中生长的植物根部出现了与田间种植植物相似的症状,而对照植物的根部则没有症状。从有症状的根部重新分离出真菌,并从形态学和分子遗传学上确认为 E. pedicellatum,这符合科赫假说。由于 GPEB-70 菌株的系统发育分析表明该菌株与美国的菌株具有聚类关系,同时考虑到农业全球化的加剧、全球种子市场的兴起以及中亚农民对美国和加拿大高产小麦种子需求的增加,我们推测最近可能有 E. pedicellatum 真菌从美国传入乌兹别克斯坦。鉴于小麦是乌兹别克斯坦重要且受欢迎的主食,进一步的工作重点将是制定有效的策略来管理这种根腐病,而制定有效的根腐病管理策略将是未来研究的重点。
{"title":"First Report of <i>Exserohilum pedicellatum</i> Causing Root Rot of Wheat (<i>Triticum aestivum</i> L.) in Uzbekistan.","authors":"Anvar Gulmirzaevich Sherimbetov, Bakhtiyor Shukhratovich Adilov, Dilshod Rustam Ogli Ruzmetov","doi":"10.1094/PDIS-08-24-1788-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-08-24-1788-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Wheat is the major staple food in Uzbekistan, and it occupies the largest harvested area (1,3 million hectares) in the country (USDA 2024). In June 2023, a survey was conducted to investigate root pathogens in wheat growing fields of Kaspi district in the Kashkadarya region of Uzbekistan. A total of 24 symptomatic plants with root rot and dark brown root lesions were collected from focal lesions in 4 different fields. From each plant, roots were excised and surface sterilized with 1% sodium hypochlorite for four minutes, then rinsed three times with sterile distilled water. Following surface sterilization, the excised roots were air dried in a laminar flow on sterile tissue sheets, rinsed twice with sterile distilled water, and then cut into 1 cm lengths segments (5 segments per one plant). The root pieces were cultured at 24°C for 4 days with a 12-hour photoperiod on potato dextrose agar supplemented with streptomycin (0.1 g/liter) and chloramphenicol (0.05 g/liter). From 24 symptomatic plants a 5 dematiaceous hyphomycete monoconidial pure isolates with abundant conidia were isolated. The conidia (n = 60) were mostly fusiform, straight, four to seven distoseptate, olivaceous brown to dark brown, and measured 51 to 88.7 × 17.9 to 25.4 μm (average 69.7 × 21.57 μm). Based on morphological characteristics the fungus was identified as E. pedicellatum according to Sivanesan (1987) and Hernandez-Restrepo et al. (2018). From five isolated monoconidial colonies, one has been chosen for molecular-genetic identification. Total DNA was extracted from it using PureLink™ Genomic DNA Mini Kit (Thermo Fisher Scientific, Waltham, MA, USA). For more informative analysis two loci, he translation elongation factor 1-alpha (tef1) and beta-tubulin (tub) genes were PCR-amplified and sequenced using gene specific primers: EF-1F (5'-CGGTGGTATCGACAAGCGT-3'), EF-2R (5'-AGCATGTTGTCGCCGTTGAAG-3')designed by Primer3web v4.1.0 software (Untergasser et al. 2012), and Bt2a (5'- GGTAACCAAATCGGTGCTGCTTTC, Bt2b (5'-ACCCTCAGTGTAGTGACCCTTGGC -3')described by Glass and Donaldson (1995), respectively. The resulting sequences were deposited in NCBI database under accession number PQ095881 and PQ095882. After BLAST analysis they showed highest similarity with the corresponding sequences of tef1 JQ672389 (100% identity, from 287 bp 287 bp are matching) and tub JQ671941 (100% identity, from 273 bp 273 bp are matching) of BMP 0384 isolate of E. pedicellatum from USA. In the plant inoculations (pathogenicity test), three isolates of E. pedicellatum were evaluated. For the pathogenicity test, conidia were scraped from PDA plate, suspended in water, and mixed with sterile sand to obtain a density of 500 conidia/g. A total of 20 wheat seed (Grom variety) previously disinfected 2 min with 10% NaOCl, were sown in each plastic pot (14 cm x 4 cm, 2 seeds per pot) filled with the inoculated soil (5 pots) and with sterilized soil (5 pots) as a control. Plants were grown in a growth chamber wit","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":null,"pages":null},"PeriodicalIF":4.4,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142392447","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 Bacterial Leaf Spot Disease on Ginger Caused by Enterobacter quasiroggenkampii in China. 中国首次报道由准姜黄色肠杆菌引起的生姜细菌性叶斑病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-09 DOI: 10.1094/PDIS-06-24-1254-PDN
Bo Tao, Xiaoli Li, Shengmao Zhou, Gaoqing Yuan
<p><p>In autumn 2023, an unknown leaf spot disease has occurred on ginger (Zingiber officinale Roscoe) in two fields of approximately 1800 m2 in Yongning District (22°49'N; 108°48'E), Nanning, China, with a incidence of 20-30%. The symptoms began as yellow spots on the leaves, expanding into elliptical to irregular lesions with yellow edges, the middle of the lesion turning grey-white in dry weather. Finally, multiple spots caused necrosis of the whole leaf. Twelve diseased leaves from six plants of two fields were collected, surface disinfected and ground. The ground samples were diluted and plated on nutrient agar (NA) medium at 28 °C for 48-72 h. The purified colonies appeared milky white and round, with smooth edges. Three isolates (GL1, GL2 and GL3) were selected for identification and pathogenic determination. They were gram negative, could utilize sorbitol, mannitol, inositol, raffinose, melibiose, disaccharides, and citrate; negative for methyl red, phenylalanine decarboxylase, hydrogen sulfide, urease; positive for voges-proskauer test and ornithine decarboxylase. These characteristics were consistent with Enterobacter genus (Wu et al., 2020). Genomic DNA was extracted from three isolates. The 16S rDNA region was amplified using 27F/1492R primers (Weisburg et al. 1991) and sequenced (accession no. PP837703-PP837705). Blastn analysis revealed that 16S rDNA sequences for GL1 was 99% identical (1373/1387 nt), GL2 96% (1364/1422 nt) and GL3 95% (1365/1435 nt) to Enterobacter quasiroggenkampii WCHECL1060 (NR_179166). To determine the species, the sequences of gyrB, rpoB and atpD genes were amplified using primers gyrB 01-F/gyrB 02-R, rpoB CM7/rpoB CM31b, and atpD 01-F/atpD 02-R, respectively (Lin et al. 2015; Zhu at al. 2010; Zhang et al. 2013). The GenBank accession numbers for the sequences were PP857680-PP857688. A multilocus phylogenetic tree was constructed with the concatenated sequence of 16S rDNA-gyrB-rpoB-atpD by using the Neighbor-Joining (NJ) method with 1000 bootstrap replicates in MEGA6 software. The three isolates clustered with E. quasiroggenkampii. Fifteen Darou ginger variety plants at the 4-5 leaf stage were tested for pathogenicity. Two to three leaves of each ginger plant were pricked with a syringe needle of 0.36mm in diameter or not and inoculated by spraying the bacterial suspension (108 CFU/mL), sterile water was used as a control. Five plants were inoculated with each isolate and the test was repeated three times. After 3-4 days of inoculation, all wounded leaves and about 10% of the unwounded leaves showed symptoms similar to those observed in the field. Control plants did not develop symptoms. Enterobacter quasiroggenkampii isolates were re-isolated from the inoculated leaves with symptoms, and their identity was confirmed by gyrB sequencing and colony morphology, completing Koch's postulates. Enterobacter quasiroggenkampii is a pathogen of humans that can cause nosocomial infections (Wu et al., 2020). In Guangxi, E
2023 年秋季,中国南宁市邕宁区(22°49'N; 108°48'E)两块约 1800 平方米的生姜(Zingiber officinale Roscoe)发生了一种未知叶斑病,发病率为 20-30%。症状开始是叶片上出现黄色斑点,然后扩大为椭圆形至不规则形病斑,病斑边缘呈黄色,在干燥天气下,病斑中部变为灰白色。最后,多个病斑导致整个叶片坏死。从两块田地的 6 株植物上采集了 12 片病叶,进行表面消毒后磨碎。磨碎的样品稀释后在营养琼脂(NA)培养基上培养 48-72 小时,纯化的菌落呈乳白色,圆形,边缘光滑。选取了三个分离菌株(GL1、GL2 和 GL3)进行鉴定和致病性测定。它们均为革兰氏阴性,能利用山梨醇、甘露醇、肌醇、棉子糖、三聚糖、双糖和柠檬酸盐;甲基红、苯丙氨酸脱羧酶、硫化氢、脲酶均为阴性;伏格-普罗斯考尔试验和鸟氨酸脱羧酶均为阳性。这些特征与肠杆菌属一致(Wu 等人,2020 年)。从三个分离株中提取了基因组 DNA。使用 27F/1492R 引物(Weisburg 等,1991 年)扩增了 16S rDNA 区域并进行了测序(登录号:PP837703-PP837705)。Blastn 分析表明,GL1 的 16S rDNA 序列与肠杆菌 quasiroggenkampii WCHECL1060 (NR_179166) 的相同度分别为 99% (1373/1387 nt)、96% (1364/1422 nt) 和 95% (1365/1435 nt)。为了确定物种,分别使用引物 gyrB 01-F/gyrB 02-R、rpoB CM7/rpoB CM31b 和 atpD 01-F/atpD 02-R 扩增了 gyrB、rpoB 和 atpD 基因的序列(Lin 等,2015 年;Zhu 等,2010 年;Zhang 等,2013 年)。这些序列的GenBank登录号为PP857680-PP857688。在 MEGA6 软件中使用邻接法(NJ)和 1000 次引导重复,用 16S rDNA-gyrB-rpoB-atpD 的序列连接构建了多焦点系统发生树。这三个分离物与 E. quasiroggenkampii 聚类。对 15 株处于 4-5 叶期的 Darou 生姜品种进行了致病性测试。用直径为 0.36 毫米的注射器针头刺破每株生姜的两到三片叶子,然后喷洒细菌悬浮液(108 CFU/mL)进行接种,无菌水作为对照。每种分离物接种 5 株植物,重复试验 3 次。接种 3-4 天后,所有受伤的叶片和大约 10% 未受伤的叶片都出现了与田间观察到的症状相似的症状。对照植物没有出现症状。从出现症状的接种叶片中再次分离出准根康比肠杆菌,并通过 gyrB 测序和菌落形态确认了它们的身份,从而完成了科赫假说。准罗根坎皮氏肠杆菌是一种人类病原体,可引起医院内感染(Wu 等,2020 年)。在广西,E. quasiroggenkampii 被确定为导致桑枯萎病的病原体之一(Jiao,2022 年)。据我们所知,这是首次报道 E. quasiroggenkampii 导致生姜细菌性叶斑病。该研究结果不仅对生姜叶斑病的防治具有现实意义,而且可为研究肠杆菌属的分化和致病机理提供极好的材料,具有重要的学术价值。
{"title":"First Report of Bacterial Leaf Spot Disease on Ginger Caused by <i>Enterobacter quasiroggenkampii</i> in China.","authors":"Bo Tao, Xiaoli Li, Shengmao Zhou, Gaoqing Yuan","doi":"10.1094/PDIS-06-24-1254-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-06-24-1254-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;In autumn 2023, an unknown leaf spot disease has occurred on ginger (Zingiber officinale Roscoe) in two fields of approximately 1800 m2 in Yongning District (22°49'N; 108°48'E), Nanning, China, with a incidence of 20-30%. The symptoms began as yellow spots on the leaves, expanding into elliptical to irregular lesions with yellow edges, the middle of the lesion turning grey-white in dry weather. Finally, multiple spots caused necrosis of the whole leaf. Twelve diseased leaves from six plants of two fields were collected, surface disinfected and ground. The ground samples were diluted and plated on nutrient agar (NA) medium at 28 °C for 48-72 h. The purified colonies appeared milky white and round, with smooth edges. Three isolates (GL1, GL2 and GL3) were selected for identification and pathogenic determination. They were gram negative, could utilize sorbitol, mannitol, inositol, raffinose, melibiose, disaccharides, and citrate; negative for methyl red, phenylalanine decarboxylase, hydrogen sulfide, urease; positive for voges-proskauer test and ornithine decarboxylase. These characteristics were consistent with Enterobacter genus (Wu et al., 2020). Genomic DNA was extracted from three isolates. The 16S rDNA region was amplified using 27F/1492R primers (Weisburg et al. 1991) and sequenced (accession no. PP837703-PP837705). Blastn analysis revealed that 16S rDNA sequences for GL1 was 99% identical (1373/1387 nt), GL2 96% (1364/1422 nt) and GL3 95% (1365/1435 nt) to Enterobacter quasiroggenkampii WCHECL1060 (NR_179166). To determine the species, the sequences of gyrB, rpoB and atpD genes were amplified using primers gyrB 01-F/gyrB 02-R, rpoB CM7/rpoB CM31b, and atpD 01-F/atpD 02-R, respectively (Lin et al. 2015; Zhu at al. 2010; Zhang et al. 2013). The GenBank accession numbers for the sequences were PP857680-PP857688. A multilocus phylogenetic tree was constructed with the concatenated sequence of 16S rDNA-gyrB-rpoB-atpD by using the Neighbor-Joining (NJ) method with 1000 bootstrap replicates in MEGA6 software. The three isolates clustered with E. quasiroggenkampii. Fifteen Darou ginger variety plants at the 4-5 leaf stage were tested for pathogenicity. Two to three leaves of each ginger plant were pricked with a syringe needle of 0.36mm in diameter or not and inoculated by spraying the bacterial suspension (108 CFU/mL), sterile water was used as a control. Five plants were inoculated with each isolate and the test was repeated three times. After 3-4 days of inoculation, all wounded leaves and about 10% of the unwounded leaves showed symptoms similar to those observed in the field. Control plants did not develop symptoms. Enterobacter quasiroggenkampii isolates were re-isolated from the inoculated leaves with symptoms, and their identity was confirmed by gyrB sequencing and colony morphology, completing Koch's postulates. Enterobacter quasiroggenkampii is a pathogen of humans that can cause nosocomial infections (Wu et al., 2020). In Guangxi, E","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":null,"pages":null},"PeriodicalIF":4.4,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142392450","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
The Efficacy of Orange Terpene and Bacillus mycoides Strain BM103 on the Control of Periwinkle Leaf Yellowing Phytoplasma. 橙色萜烯和枯草芽孢杆菌 BM103 菌株对长春花叶片黄化病原菌的防治效果。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-09 DOI: 10.1094/PDIS-07-24-1547-RE
Jui-Chen Hung, Tzu-Pi Huang, Jenn-Wen Huang, Chung Jan Chang, Fuh-Jyh Jan

Phytoplasmas are obligate phytopathogenic bacteria belonging to the class Mollicutes. The pathogens, transmitted by insect vectors, associated with hundreds of plant diseases worldwide. Due to the regulation on banning use of antibiotics and limited efficacy of the traditional disease management manners, an eco-friendly alternative is needed. Given that terpene and probiotics have antibiotic activity and the ability to induce systemic resistance, in this study, the effectiveness of orange terpene and a Bacillus mycoides strain, BM103, was evaluated in periwinkle plants infected with periwinkle leaf yellowing (PLY) phytoplasma derived from a shoot-tip tissue culture system. Weekly drenching of 1,000 ppm diluted orange terpene emulsion or pre-activated strain BM103 liquid culture dilution exhibited the ability to inhibit PLY phytoplasma accumulation. The expression of the genes associated with plant defense response and flower development was upregulated after treatment. Moreover, pre-treatment of orange terpene or strain BM103 delayed PLY infection via cleft-grafting inoculation. While orange terpene did not suppress the symptoms, strain BM103 did result in a milder symptom expression that might partially attribute to its plant growth-promoting characteristics. Additionally, the pre-activation of strain BM103 may contribute to its efficacy. Taken together, this research indicates that orange terpene and B. mycoides BM103, with the ability to rapidly induce plant defense responses, could potentially be developed into biological control materials as preventive agents or biofertilizers.

植物病原菌是属于真菌门的强制性植物病原菌。病原体通过昆虫媒介传播,与全球数百种植物病害有关。由于禁止使用抗生素的规定以及传统病害防治方法的效果有限,因此需要一种生态友好型替代品。鉴于萜烯和益生菌具有抗生素活性和诱导系统抗性的能力,本研究评估了橙色萜烯和枯草芽孢杆菌 BM103 菌株对感染了长春花叶黄病(PLY)植原体的长春花植物的有效性。每周淋洗 1,000 ppm 稀释的橙色萜乳液或预活化菌株 BM103 的液体培养稀释液,均能抑制 PLY 植原体的积累。处理后,与植物防御反应和花发育相关的基因表达上调。此外,橙萜素或菌株 BM103 的预处理可延缓 PLY 通过裂殖接种的感染。虽然橙色萜烯没有抑制症状,但菌株 BM103 确实导致了较轻微的症状表现,这可能部分归因于其促进植物生长的特性。此外,菌株 BM103 的预激活也可能有助于发挥其功效。综上所述,这项研究表明,橙萜和霉菌菌株 BM103 具有快速诱导植物防御反应的能力,有可能被开发成生物防治材料,用作预防剂或生物肥料。
{"title":"The Efficacy of Orange Terpene and <i>Bacillus mycoides</i> Strain BM103 on the Control of Periwinkle Leaf Yellowing Phytoplasma.","authors":"Jui-Chen Hung, Tzu-Pi Huang, Jenn-Wen Huang, Chung Jan Chang, Fuh-Jyh Jan","doi":"10.1094/PDIS-07-24-1547-RE","DOIUrl":"https://doi.org/10.1094/PDIS-07-24-1547-RE","url":null,"abstract":"<p><p>Phytoplasmas are obligate phytopathogenic bacteria belonging to the class Mollicutes. The pathogens, transmitted by insect vectors, associated with hundreds of plant diseases worldwide. Due to the regulation on banning use of antibiotics and limited efficacy of the traditional disease management manners, an eco-friendly alternative is needed. Given that terpene and probiotics have antibiotic activity and the ability to induce systemic resistance, in this study, the effectiveness of orange terpene and a Bacillus mycoides strain, BM103, was evaluated in periwinkle plants infected with periwinkle leaf yellowing (PLY) phytoplasma derived from a shoot-tip tissue culture system. Weekly drenching of 1,000 ppm diluted orange terpene emulsion or pre-activated strain BM103 liquid culture dilution exhibited the ability to inhibit PLY phytoplasma accumulation. The expression of the genes associated with plant defense response and flower development was upregulated after treatment. Moreover, pre-treatment of orange terpene or strain BM103 delayed PLY infection via cleft-grafting inoculation. While orange terpene did not suppress the symptoms, strain BM103 did result in a milder symptom expression that might partially attribute to its plant growth-promoting characteristics. Additionally, the pre-activation of strain BM103 may contribute to its efficacy. Taken together, this research indicates that orange terpene and B. mycoides BM103, with the ability to rapidly induce plant defense responses, could potentially be developed into biological control materials as preventive agents or biofertilizers.</p>","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":null,"pages":null},"PeriodicalIF":4.4,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142392452","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
Pyrrolnitrin in Pseudomonas chlororaphis strain AFS009 metabolites reduces constitutive and DMI-induced MfCYP51 gene expression in Monilinia fructicola. 绿假单胞菌 AFS009 菌株代谢物中的吡咯烷酮可减少果核莫尼菌中构成型和 DMI 诱导型 MfCYP51 基因的表达。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-02 DOI: 10.1094/PDIS-07-24-1470-RE
Johanna Wesche, Peishan Wu, Chaoxi Luo, Guido Schnabel

Brown rot, caused by Monilinia fructicola, is one of the most economically important diseases of peach. Demethylation inhibitor (DMI) fungicides play an important part in managing brown rot in the southeastern U.S., but over the last 20 years, reduced efficacy to DMIs has been reported in field isolates overexpressing the DMI target enzyme encoding MfCYP51 gene. Metabolites of the biocontrol agent (BCA) Pseudomonas chlororaphis strain AFS009 suppressed the MfCYP51 gene in sensitive and resistant M. fructicola isolates previousely, but it is not known what molecule was responsible. The goals of this study were to determine the presence and role of pyrrolnitrin (PRN), a common metabolite of P. chlororaphis and chemical analogue to fludioxonil with antifungal activity, in the suppression of the MfCYP51 gene and to investigate if MfCYP51 expression can also be suppressed by Bacillus subtilis. High-performance liquid chromatography (HPLC) detected pyrrolnitrin at 1.75 µg/mg in P. chlororaphis metabolites formulated as Howler EVO (Howler). PRN at 0.1 µg/ml, fludioxonil at 0.1 µg/ml, and Howler applied at a dose that contained 0.1 µg/ml PRN significantly reduced the MfCYP51 gene expression at similar levels in DMI-resistant isolates. Furthermore, MfCYP51 expression in DMI-sensitive and three DMI-resistant isolates treated with Howler (88.1 µg/ml), Theia (209.5 µg/ml), propiconazole (0.3 µg/ml), or the mixture of either Howler or Theia + propiconazole revealed that Howler significantly reduced the MfCYP51 target gene expression in two of three sensitive and all three resistant M. fructicola isolates. On the other hand, Theia showed no suppressive effect and even increased the MfCYP51 gene expression level in two of three resistant isolates. In detached fruit assays on apple with a DMI resistant isolate, only the mixture of Howler + 50 µg/ml propiconazole resulted in synergism. The results indicate that suppression of MfCYP51 target gene is BCA-dependent and can be induced by pyrrolnitrin.

由果核菌(Monilinia fructicola)引起的褐腐病是桃子最重要的经济病害之一。去甲基化抑制剂(DMI)杀真菌剂在美国东南部褐腐病的防治中发挥着重要作用,但在过去 20 年中,有报道称田间分离的过表达 DMI 目标酶编码 MfCYP51 基因的菌株对 DMI 的药效降低。以前,生物控制剂(BCA)绿假单胞菌菌株 AFS009 的代谢物抑制了敏感和抗性果蝇分离株中的 MfCYP51 基因,但不知道是什么分子造成的。本研究的目的是确定在抑制 MfCYP51 基因的过程中是否存在吡咯烷酮(PRN)及其作用,吡咯烷酮是 P. chlororaphis 的一种常见代谢产物,也是具有抗真菌活性的氟虫腈的化学类似物,同时研究枯草芽孢杆菌是否也能抑制 MfCYP51 的表达。高效液相色谱法(HPLC)在配制成 Howler EVO(Howler)的 P. chlororaphis 代谢物中检测到 1.75 µg/mg 的吡咯烷酮。0.1 µg/ml 的 PRN、0.1 µg/ml 的氟虫腈和含有 0.1 µg/ml PRN 的 Howler 都能显著降低抗 DMI 分离物中 MfCYP51 基因的表达量。此外,用 Howler(88.1 微克/毫升)、Theia(209.5 微克/毫升)、丙环唑(0.3 微克/毫升)或 Howler 或 Theia + 丙环唑的混合物处理对 DMI 敏感的分离物和三个对 DMI 抗性的分离物的 MfCYP51 表达情况表明,Howler 能显著降低三个敏感 M. fructicola 分离物中的两个和所有三个抗性 M. fructicola 分离物的 MfCYP51 目标基因表达。另一方面,Theia 没有抑制作用,甚至提高了三个抗性分离物中两个的 MfCYP51 基因表达水平。在对苹果进行的抗 DMI 分离物的分离果实试验中,只有 Howler + 50 µg/ml 丙环唑的混合物产生了增效作用。结果表明,MfCYP51 目标基因的抑制是 BCA 依赖性的,并可由吡咯烷酮诱导。
{"title":"Pyrrolnitrin in <i>Pseudomonas chlororaphis</i> strain AFS009 metabolites reduces constitutive and DMI-induced MfCYP51 gene expression in <i>Monilinia fructicola</i>.","authors":"Johanna Wesche, Peishan Wu, Chaoxi Luo, Guido Schnabel","doi":"10.1094/PDIS-07-24-1470-RE","DOIUrl":"https://doi.org/10.1094/PDIS-07-24-1470-RE","url":null,"abstract":"<p><p>Brown rot, caused by Monilinia fructicola, is one of the most economically important diseases of peach. Demethylation inhibitor (DMI) fungicides play an important part in managing brown rot in the southeastern U.S., but over the last 20 years, reduced efficacy to DMIs has been reported in field isolates overexpressing the DMI target enzyme encoding MfCYP51 gene. Metabolites of the biocontrol agent (BCA) Pseudomonas chlororaphis strain AFS009 suppressed the MfCYP51 gene in sensitive and resistant M. fructicola isolates previousely, but it is not known what molecule was responsible. The goals of this study were to determine the presence and role of pyrrolnitrin (PRN), a common metabolite of P. chlororaphis and chemical analogue to fludioxonil with antifungal activity, in the suppression of the MfCYP51 gene and to investigate if MfCYP51 expression can also be suppressed by Bacillus subtilis. High-performance liquid chromatography (HPLC) detected pyrrolnitrin at 1.75 µg/mg in P. chlororaphis metabolites formulated as Howler EVO (Howler). PRN at 0.1 µg/ml, fludioxonil at 0.1 µg/ml, and Howler applied at a dose that contained 0.1 µg/ml PRN significantly reduced the MfCYP51 gene expression at similar levels in DMI-resistant isolates. Furthermore, MfCYP51 expression in DMI-sensitive and three DMI-resistant isolates treated with Howler (88.1 µg/ml), Theia (209.5 µg/ml), propiconazole (0.3 µg/ml), or the mixture of either Howler or Theia + propiconazole revealed that Howler significantly reduced the MfCYP51 target gene expression in two of three sensitive and all three resistant M. fructicola isolates. On the other hand, Theia showed no suppressive effect and even increased the MfCYP51 gene expression level in two of three resistant isolates. In detached fruit assays on apple with a DMI resistant isolate, only the mixture of Howler + 50 µg/ml propiconazole resulted in synergism. The results indicate that suppression of MfCYP51 target gene is BCA-dependent and can be induced by pyrrolnitrin.</p>","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":null,"pages":null},"PeriodicalIF":4.4,"publicationDate":"2024-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142366221","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 root and stem rot in cowpea (syn. black-eyed pea; Vigna unguiculata) in California. 首次报告镰刀菌(Fusarium falciforme)在加利福尼亚州引起豇豆(同义,黑眼豆;Vigna unguiculata)根腐病和茎腐病。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2024-10-02 DOI: 10.1094/PDIS-08-24-1609-PDN
Kacey Zimmerman, Nicholas E Clark, Thomas A Turini, Andrea Paulk, Johanna Del Castillo Múnera, Ningxiao Li, David Geiser, Frank N Martin, Cassandra L Swett
<p><p>In both April 2018 and September 2019, cowpeas / black-eyed peas (Vigna unguiculata) in one field in Tulare County, California were observed with tap root rot, both underground (foot) and aboveground stem rot, and in some cases canopy decline, compromising bean formation. In both fields, < 5% of plants appeared affected. Foot and stem segments (~1 cm) of 5-10 plants / field were disinfested sequentially with 0.1% Tween 20 (dip), 70% ethanol for 30 s, and 1% sodium hypochlorite for 2 min and placed on 1:10 potato dextrose agar with 0.03% tetracycline and Fusarium selective medium (Leslie and Summerell 2006). Fusarium-like isolates (dominant in isolation plates) were transferred to 0.6% KCl agar, where fusiform, curved macroconidia and varied microconidia in false heads on elongated monophialides were observed, characteristic of the Fusarium solani species complex (FSSC) (Leslie and Summerell 2006). Isolates CS221, CS222, and CS520 (representing different plants and locations) were saved as single hyphal tip cultures. An Illumina-derived genome sequence was assembled (Burkhardt et al. 2019) and partial tef1ɑ and rpb2 sequences (O'Donnell et al. 2022) were extracted from genome sequences in silico. Sequences were 99.9-100% identical to one another and to deposited F. falciforme isolates based on Fusarium ID and Fusarium MLST for tef1ɑ and rpb2, respectively (tef1a accessions: NRRL 28562 and NRRL 32331; rpb2 accession: NRRL 22857), and were deposited in GenBank (accessions in supplementary table). Pathogenicity was evaluated in three-week-old cowpea plants (cv. CB46rk2) in the greenhouse (13.5-33.6℃; 12:12 h L:D). The tap root / stem was wounded (1 mm wide, 2 mm deep) ~ 2 cm below the soil line and drenched with 50 ml of 106 spores / ml 0.1% water agar or with 0.1% water agar (negative control). The trial was arranged in a Randomized Complete Block Design with three blocks and 2-3 plants / isolate / block, and conducted twice. 52 d post-inoculation, below ground tap root / stem rot developed in 83% of F. falciforme-inoculated plants, with lesion lengths ranging from 25.2 ± 4.2 to 29.2 ± 8.0 mm (P = 0.893 for isolate, ANOVA). Canopy decline developed in 33-50% of plants across treatments in trial 1 (P = 0.859 for isolate) but not in trial 2, likely due to cooler conditions in trial 2 (January-March) vs. trial 1 (May-July), which were less stressful. F. falciforme isolates did not affect bean biomass (dry weight) vs. negative controls (12.5-14.8g / plant; P = 0.949 for pathogen treatment). FSSC isolates were recovered from 100% of symptomatic plants in the inoculated treatments but not in negative controls (both trials) and representative isolates from all treatments were confirmed as F. falciforme (tef1a analysis; trial 2 only). This study establishes F. falciforme as a root and stem rot pathogen of cowpea in California-a disease previously attributed to the morphologically and phylogenetically distinct F. phaseoli (syn. F. solani f. sp. phaseol
2018 年 4 月和 2019 年 9 月,加利福尼亚州图莱尔县(Tulare County)一块田里的豇豆/豇豆(Vigna unguiculata)都出现了直根腐烂、地下(足部)和地上茎腐烂,有时冠层衰退,影响豆子的形成。在这两块田里,受影响的植株都小于 5%。用 0.1% 吐温 20(浸泡)、70% 乙醇浸泡 30 秒和 1%次氯酸钠浸泡 2 分钟,依次对每块田 5-10 株植物的根脚和茎段(约 1 厘米)进行消毒,然后将其放在含有 0.03% 四环素的 1:10 马铃薯葡萄糖琼脂和镰刀菌选择性培养基上(Leslie 和 Summerell,2006 年)。将镰刀菌样分离物(在分离板中占优势)转移到 0.6% 氯化钾琼脂中,观察到纺锤形弯曲的大锥体和长单胞上假头的各种微锥体,这是茄科镰刀菌复合菌种(FSSC)的特征(Leslie 和 Summerell,2006 年)。分离物 CS221、CS222 和 CS520(代表不同的植物和地点)被保存为单个菌尖培养物。组装了 Illumina 衍生的基因组序列(Burkhardt 等人,2019 年),并从基因组序列中提取了部分 tef1ɑ 和 rpb2 序列(O'Donnell 等人,2022 年)。根据 Fusarium ID 和 Fusarium MLST,这些序列与 F. falciforme 分离物的 tef1ɑ 和 rpb2(tef1a 编号:NRRL 28562 和 NRRL 32331;rpb2 编号:NRRL 22857)的序列有 99.9%-100%的相同性,并保存在 GenBank 中(编号见补充表格)。在温室(13.5-33.6℃;12:12 h L:D)中对三周大的豇豆植株(cv. CB46rk2)进行了致病性评估。在土壤线下约 2 厘米处的直根/茎受伤(宽 1 毫米,深 2 毫米),并用 50 毫升 106 个孢子/毫升的 0.1% 水琼脂或 0.1% 水琼脂(阴性对照)浸泡。试验采用随机完全区组设计,设三个区组,每个区组 2-3 株/分离株,共进行两次。接种后 52 d,83% 接种 F. falciforme 的植株出现地下直根/茎腐病,病斑长度从 25.2 ± 4.2 mm 到 29.2 ± 8.0 mm 不等(P = 0.893(分离株),方差分析)。在试验 1 中,33%-50% 的植株在不同处理下出现树冠衰退(分离株的 P = 0.859),但在试验 2 中没有出现这种情况,这可能是由于试验 2(1 月至 3 月)与试验 1(5 月至 7 月)相比条件更凉爽,压力更小。与阴性对照相比,F. falciforme 分离物对豆类生物量(干重)没有影响(12.5-14.8 克/株;病原体处理的 P = 0.949)。在接种处理中,100%有症状的植株都能从 FSSC 分离物中回收,而阴性对照中却没有(两个试验),所有处理中的代表性分离物均被确认为镰刀菌(tef1a 分析;仅试验 2)。这项研究确定 F. falciforme 是加利福尼亚州豇豆的根腐病和茎腐病病原菌--这种病以前被认为是形态和系统发育上不同的 F. phaseoli(同义 F. solani f. sp. phaseoli)引起的,但缺乏现代病原学研究(Frate 等人,2018 年;Geiser 等人,2021 年)。这项工作与之前关于 F. falciforme 是豇豆(Ajamu 等人,2023 年)和其他豆类(Sousa 等人,2017 年;Duarte 等人,2019 年)根腐病/茎腐病病原体的报道一致。由于镰刀菌也是加州其他作物(包括甜瓜、番茄和开心果)的病原菌,因此明确病原将有助于准确诊断和有效的轮作管理。
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Plant disease
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