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First report of Diaporthe humulicola causing stem blight in hop plants in Massachusetts.
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2025-03-04 DOI: 10.1094/PDIS-01-25-0146-PDN
Noah Williams, Nolan Stamborski, Chang-Gi Back, Geunhwa Jung
<p><p>Humulus lupulus L., the common hop plant, is a commercially cultivated crop used as a beer flavoring agent that has been grown in Massachusetts since the 1600s (Machado et al., 2019 ; Rumney, 1998). In 2018, Diaporthe leaf spot, caused by the fungal pathogen Diaporthe humulicola, was found at hopyard research plots in Connecticut, followed by reports in multiple New York counties in 2023 (Allan-Perkins et al., 2020; Sharma et al., 2023). Additionally, Diaporthe halo blight, caused by the same fungal pathogen, was reported in Prince Edward Island (PE), Canada (Foster et al., 2024). In August of 2021, severe stem blight symptoms with visible fungal fruiting bodies were observed on >50% of plants at a hopyard in Franklin County, MA. Brown spots were observed on stems. Severe infection presented as widespread dark browning on stems, leaves, and cones. Samples were collected from the hop cultivars Mt Rainier (N=5), Magnum (N=1), and Teamaker (N=6). Under brightfield microscopy (400x) conidia congruent with Diaporthe spp. were observed on stem samples. Diseased stems from all cultivars were peeled and cut into 2 to 4 mm2 pieces, then surface sterilized in 70% ethanol for 60s, and 1% NaOCl for 60s. Samples were rinsed in sterile water, dried for 3 min and placed on potato dextrose agar (PDA). Plates were incubated at 25℃ for 5 days. 11 isolates were obtained from stem tissues of 11 plants. Isolates were cultured on PDA. 10 isolates had morphological traits consistent with the pycnidium of holotype UAMH 12076 (Allans-Perkins et al, 2020). Isolate sequences were identical, using one reinfection tests were performed on detached stem segments obtained from field grown mature plants (≥ 1 year) of cvs. Cascade and Magnum.3 wounding methods and an untreated control were used for pathogenicity testing to simulate field conditions. Each treatment had 6 replicates per cultivar and all treatments had an agar plug of inoculum applied to them, then were incubated at room temperature. Wounding with a knife involved peeling off the outer tissue layer, insect damage was simulated by using a needle to puncture the stem 3 times, and friction damage from hop support ropes was mimicked by rubbing rope against the stem. Plugs were removed 5 days post-application. Pathogenicity was observed as fungal growth and necrosis of plant tissue on all non-control methods in all samples 5 days post plug removal, fulfilling Koch's postulates. The pathogenic organism was reisolated from all inoculated tissues, except the control and identified using morphological characteristics and ITS sequence analysis. DNA amplification of the ITS region was performed according to the methods of White et al. (1990). A maximum likelihood phylogenetic tree based on the region was constructed using the MEGA10 program (Saitou & Nei, 1987). Sequence data was collected from NCBI BlastN, then aligned and trimmed. The ITS sequences (NCBI Acc. Nos. PQ555192, PQ555193, PQ555194) from the 3 isolates (UMAS
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引用次数: 0
First report of Pantoea dispersa causing strawberry root rot in China.
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2025-03-04 DOI: 10.1094/PDIS-11-24-2486-PDN
Peipei Wang, Jiaqi Zhang, Lihong Dong, Yifan Fu, Qinggang Guo, Ping Ma
<p><p>China is the largest strawberry (Fragaria x ananassa) producer worldwide. In 2022 the area devoted to strawberry planting in China was 147.45 thousand hectares. Hebei province is the fourth-largest strawberry planting province in China, where strawberries play a crucial role in the agricultural economy. In November 2023, root rot was observed on strawberry plants (cultivar Benihoppe) in Xushui County (115°35'59″ E, 38°58'39″ N), Hebei province, affecting overall growth and reproduction. Disease incidence in two greenhouses (0.15 ha in size) was up to ~70%, with severe symptoms causing almost complete plant mortality. The initial symptoms included abnormally small new leaves, followed by wilting and collapse of the plants. As the disease progressed, the leaves curled and withered, ultimately leading to plant death. Under humid conditions, a sticky bacterial ooze was observed exuding from the stem base. Dissection of the stem base of diseased plants revealed reddish-brown discolored roots while the roots of healthy plants were white. To isolate the causal agent, 10 root samples from symptomatic plants were randomly selected from two greenhouses and surface sterilized sequentially in absolute ethanol (1 min), 3.125% NaOCl (6 min), and absolute ethanol (30 s), followed by rinsing in sterile water three times (Sahu et al. 2022). Root pieces were then placed on Luria-Bertani (LB) agar and incubated at 25℃ in darkness for 3 days. After isolation of bacteria from the symptomatic tissues, subculturing onto new media was performed to obtain pure cultures. The 10 bacterial isolates selected for further analysis exhibited a similar morphology of circular and smooth colonies that were initially milky white and later were yellow-pigmented. Three representative isolates (CM2402, CM2403, and CM2405) were selected for 16S rDNA sequencing using primers 27F/1492R (27F: 5'-AGAGTTTGATCMTGGCTCAG-3'; 1492R: 5'-GGTTACCTTGTTACGACTT-3') (Lane 1991). BLASTn analysis revealed that the three sequences were identical, with 99.86% (1,420/1,422 bp) identity to <i>Pantoea</i> sp. strain nts-7 (OR392975.1). Biochemical analysis confirmed that the isolates belong to the genus <i>Pantoea</i>, testing positive with the Voges-Proskauer test for production of acetylmethyl carbinol from glucose fermentation, utilization of citrate, D-mannose, inositol, and malonate, positive for β-galactosidase while negative for indole production, phenylalanine deaminase, lysine decarboxylase, urease, H<sub>2</sub>S production, and acid production from raffinose, adonitol, and D-sorbitol (Gavini et al. 1989). Five housekeeping genes (<i>gyrB</i>, <i>fusA</i>, <i>leuS</i>, <i>pyrG</i>, <i>rplB</i>) were amplified and sequenced for further molecular identification (Delétoile et al. 2009). A phylogenetic tree based on the combined sequences of these genes was constructed using the neighbor-joining method, with the isolates clustering with <i>P. dispersa</i>. No sequence variation was detected among
中国是世界上最大的草莓(Fragaria x ananassa)生产国。2022 年,中国草莓种植面积为 147.45 千公顷。河北省是中国第四大草莓种植省份,草莓在该省农业经济中发挥着重要作用。2023 年 11 月,河北省徐水县(东经 115°35'59″,北纬 38°58'39″)的草莓植株(栽培品种 Benihoppe)发生根腐病,影响整体生长和繁殖。两个温室(0.15 公顷)的发病率高达 ~70%,症状严重时植株几乎全部死亡。初期症状包括新叶异常变小,随后植株枯萎和倒伏。随着病情发展,叶片卷曲枯萎,最终导致植株死亡。在潮湿条件下,茎基部会渗出粘稠的细菌性渗液。剖开病株的茎基部,发现根部呈红褐色,而健康植株的根部呈白色。为了分离病原菌,从两个温室中随机选取了 10 个有症状植株的根部样本,依次用绝对乙醇(1 分钟)、3.125% NaOCl(6 分钟)和绝对乙醇(30 秒)进行表面消毒,然后用无菌水冲洗三次(Sahu 等,2022 年)。然后将根块放在 Luria-Bertani (LB) 琼脂上,在 25℃ 黑暗环境中培养 3 天。从有症状的组织中分离出细菌后,在新的培养基上进行亚培养,以获得纯培养物。被选作进一步分析的 10 个细菌分离物表现出相似的形态,即圆形和光滑的菌落,最初为乳白色,后来呈黄色色素沉着。利用引物 27F/1492R(27F:5'-AGAGTTTGATCMTGGCTCAG-3';1492R:5'-GGTTACCTTGTTACGACTT-3')对三个具有代表性的分离物(CM2402、CM2403 和 CM2405)进行了 16S rDNA 测序(Lane,1991 年)。BLASTn 分析表明,这三个序列与泛德菌株 nts-7 (OR392975.1)的序列相同度为 99.86%(1,420/1,422 bp)。生化分析证实,分离物属于泛变形菌属,经 Voges-Proskauer 试验,葡萄糖发酵产生的乙酰甲基甲醇、柠檬酸盐、D-甘露糖、肌醇和丙二酸盐的利用率呈阳性、和丙二酸盐,β-半乳糖苷酶检测呈阳性,而吲哚生产、苯丙氨酸脱氨酶、赖氨酸脱羧酶、脲酶、H2S 生产以及棉子糖、阿糖胞苷和 D-山梨糖醇产酸检测呈阴性(Gavini 等人,1989 年)。1989).为进一步进行分子鉴定,对五个看家基因(gyrB、fusA、leuS、PYRG、rplB)进行了扩增和测序(Delétoile 等人,2009 年)。使用邻接法根据这些基因的组合序列构建了一棵系统发生树,分离物与 P. dispersa 聚类。在 CM2402、CM2403 和 CM2405 分离物之间,未发现这五个基因的序列变异。来自一个代表性分离株(CM2405)的五个看家基因和 16S rDNA 的序列已存入 GenBank,登录号为 PQ100709(gyyr):PQ100709(gyrB)、PQ100708(fusA)、PQ059271(leuS)、PQ100710(PYRG)、PQ100711(rplB)和 PQ095944(16S rDNA)。为了验证科赫假设,对 10 株 1 个月大的草莓植株(栽培品种 Benihoppe)进行了致病性试验,每个处理有三个独立的重复。将土壤从根部抖落,然后将根部浸泡在分离物 CM2402、CM2403 和 CM2405(107 cfu/mL)的细菌悬浮液中 10 分钟。将草莓植株种植到灭菌土壤中,在 25℃、16 小时光照和 8 小时黑暗的光周期下培养。将根部浸泡在消毒水中的草莓植株作为对照。移栽四周后,用细菌悬浮液浸泡过的草莓植株表现出活力差和根部坏死,而对照植株仍无症状。从坏死组织中重新分离出的病原体在形态和分子上与原始菌株完全相同。这些结果证实 P. dispersa 是中国草莓根腐病的病原菌。据报道,P. dispersa 是印度和马来西亚水稻的病原体(Jena 等,2023 年;Toh 等,2019 年),在台湾引起洋葱鳞茎腐烂(Chang 等,2018 年),在中国引起丝核菌褐斑病(Hu 等,2022 年)。据我们所知,这是 P. dispersa 导致草莓根腐病的首次报道。这项研究对于有效管理、促进早期检测、提供有针对性的控制措施以及防止病害进一步蔓延至关重要,最终可减少作物损失并提高可持续性。
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引用次数: 0
First Report of Red Crown Rot of Soybean, caused by Calonectria ilicicola, in Missouri.
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2025-03-04 DOI: 10.1094/PDIS-12-24-2566-PDN
Mandy Bish, Theresa K Herman, Nancy McCoppin, Peng Tian, Steve Clough, Hari Karki
<p><p>In July 2024, soybean (Glycine max) with symptoms of red crown rot, caused by Calonectria ilicicola, were observed in a commercial field with a history of corn-soybean rotation in Marion County, Missouri. Soybean were planted on May 12 and foliar symptoms including interveinal chlorosis and premature senescence, were visible on plants at the R3 growth stage with incidence estimated at 5% of the field. Reddish coloration was observed on the lower stem of symptomatic plants, along with reddish-orange globular perithecia measuring 300-500 µm in height and 250-350 µm in diameter, typical of C. ilicicola infection. Twelve symptomatic plants were collected, and stems were cleaned, followed by immersion in 70% ethanol for 30 seconds, 1.25% NaOCl for 5 minutes, and rinsing three times. Surface disinfected stems were dried at 24°C. After 24 hours, stems were split and placed on water agar (WA) amended with 1% penicillin-streptomycin solution (P4333; Sigma Aldrich). After 4 to 5 days, conidiophores characteristic of C. ilicicola, were observed. Conidia from nine stems were transferred to WA and serial transfers of conidia were made until pure single spore cultures of isolates were obtained and transferred to potato dextrose agar (PDA). On WA, the isolates produced hyaline mycelium that developed conidiophores and conidia as well as reddish brown chlamydospores. Single spore isolates transferred to PDA produced fluffy white mycelium that turned reddish. Three isolates were used for DNA extraction with the Zymo DNA extraction kit (ZD6005). Parts of the internal transcriber region (ITS) (ITS-F2: 5'-TTTACAACTCCCAAACCCCATGTGAAC-3'and ITS-R2: 5'-CTACCTGATTCGAGGTCAA CCAGAA-3') histone 3 (HIS3) (Crous et al. 2004), translation elongation 1α (EF1α) (Carbone and Kohn 1999; O'Donnell et al. 1998) and β-tubulin (TUB2) (Crous et al. 2004; O'Donnell and Cigelnik 1997) genes were amplified and their DNA sequenced. Sequences were processed in Geneious Prime 2024.0 and deposited at NCBI GenBank under the accession numbers PQ390253, PQ390254 and PQ390255 (ITS), PQ519580, PQ519581 and PQ519582 (HIS3), PQ507479, PQ507480 and PQ507481 (EF1α) and PV092563, PV092564 and PV092565 (TUB2). These DNA sequences show 100% identity to C. ilicicola sequences. To fulfill Koch's postulates, eight pots containing three plants (cultivar Williams 82) each were grown for two weeks in a growth chamber at 26°C with a 14-hour photoperiod and watered daily. Six pots were inoculated with 4 mm plugs of mycelium from the distal growing edge of the fungal isolate on PDA plates. The other two pots, treated with 4 mm plugs of PDA-only, served as negative controls. The planting medium at the base of each plant was gently removed, and inoculum was placed on the primary root, approximately 7 mm below the soil level. Five days after inoculation, plants started exhibiting reddish/blackish discoloration at the base of the stems, typical of red crown rot. Then, C. ilicicola was re-isolated from symptoma
2024 年 7 月,在密苏里州马里恩县一块曾进行玉米-大豆轮作的商品田里,大豆(Glycine max)出现了由 Calonectria ilicicola 引起的红冠腐烂病症状。大豆于 5 月 12 日播种,在 R3 生长阶段的植株上可以看到叶片症状,包括叶脉间枯萎和过早衰老,估计发病率为 5%。在出现症状的植株的下部茎干上观察到淡红色,以及橘红色的球状包囊,高度为 300-500 微米,直径为 250-350 微米,这是 C. ilicicola 感染的典型症状。采集 12 株有症状的植物,清洗茎干,然后将其浸泡在 70% 的乙醇中 30 秒,再浸泡在 1.25% 的 NaOCl 中 5 分钟,最后冲洗三次。经过表面消毒的茎秆在 24°C 下烘干。24 小时后,将茎劈开,放在加入 1%青霉素-链霉素溶液(P4333;Sigma Aldrich)的水琼脂(WA)上。4 至 5 天后,可观察到 C. ilicicola 特有的分生孢子器。将 9 根茎上的分生孢子转移到 WA 上,然后连续转移分生孢子,直到获得分离物的纯单孢子培养物并转移到马铃薯葡萄糖琼脂(PDA)上。在 WA 上,分离物产生了透明的菌丝,长出了分生孢子梗和分生孢子以及红褐色衣壳孢子。转移到 PDA 上的单孢子分离物产生的绒毛状白色菌丝变为淡红色。利用 Zymo DNA 提取试剂盒(ZD6005)对三个分离株进行了 DNA 提取。内部转录区(ITS)(ITS-F2:5'-TTTACAACTCCCAAACCCCATGTGAAC-3'和 ITS-R2:5'-CTACCTGATTCGAGGTCAA CCAGAA-3')组蛋白 3(HIS3)(Crous et al.2004)、翻译延长 1α (EF1α)(Carbone 和 Kohn,1999 年;O'Donnell 等人,1998 年)和 β-微管蛋白(TUB2)(Crous 等人,2004 年;O'Donnell 和 Cigelnik,1997 年)基因进行扩增,并对其 DNA 进行测序。序列经 Geneious Prime 2024.0 处理后存入 NCBI GenBank,登录号为 PQ390253、PQ390254 和 PQ390255(ITS)、PQ519580、PQ519581 和 PQ519582(HIS3)、PQ507479、PQ507480 和 PQ507481(EF1α)以及 PV092563、PV092564 和 PV092565(TUB2)。这些 DNA 序列与 C. ilicicola 序列显示出 100%的同一性。为了实现科赫假设,在生长室中种植了八盆植物,每盆包含三株植物(栽培品种为威廉姆斯 82),温度为 26°C,光周期为 14 小时,每天浇水。将真菌分离物远端生长边缘的 4 毫米菌丝塞接种到六个花盆的 PDA 平板上。另外两个花盆只用 4 毫米的 PDA 插条作为阴性对照。轻轻移除每株植物基部的种植介质,将接种体置于主根上,低于土壤水平约 7 毫米。接种五天后,植株的茎基部开始出现红色/黑色变色,这是典型的红冠腐烂病。接种后三周,从有症状的植株中再次分离出 C. ilicicola,并通过分生孢子形态进行了鉴定。这些结果共同证实了密苏里州马里恩县存在伊里奇菌。尽管 1965 年美国首次在花生上证实了 C. ilicicola,但由于它在商业大豆生产中的存在,其症状可能类似于猝死综合症,因此再次引起了人们的关注。该病原体对密苏里州超过 2,000,000 公顷的大豆和大约 8,000 公顷的花生造成了严重威胁。
{"title":"First Report of Red Crown Rot of Soybean, caused by <i>Calonectria ilicicola</i>, in Missouri.","authors":"Mandy Bish, Theresa K Herman, Nancy McCoppin, Peng Tian, Steve Clough, Hari Karki","doi":"10.1094/PDIS-12-24-2566-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-12-24-2566-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;In July 2024, soybean (Glycine max) with symptoms of red crown rot, caused by Calonectria ilicicola, were observed in a commercial field with a history of corn-soybean rotation in Marion County, Missouri. Soybean were planted on May 12 and foliar symptoms including interveinal chlorosis and premature senescence, were visible on plants at the R3 growth stage with incidence estimated at 5% of the field. Reddish coloration was observed on the lower stem of symptomatic plants, along with reddish-orange globular perithecia measuring 300-500 µm in height and 250-350 µm in diameter, typical of C. ilicicola infection. Twelve symptomatic plants were collected, and stems were cleaned, followed by immersion in 70% ethanol for 30 seconds, 1.25% NaOCl for 5 minutes, and rinsing three times. Surface disinfected stems were dried at 24°C. After 24 hours, stems were split and placed on water agar (WA) amended with 1% penicillin-streptomycin solution (P4333; Sigma Aldrich). After 4 to 5 days, conidiophores characteristic of C. ilicicola, were observed. Conidia from nine stems were transferred to WA and serial transfers of conidia were made until pure single spore cultures of isolates were obtained and transferred to potato dextrose agar (PDA). On WA, the isolates produced hyaline mycelium that developed conidiophores and conidia as well as reddish brown chlamydospores. Single spore isolates transferred to PDA produced fluffy white mycelium that turned reddish. Three isolates were used for DNA extraction with the Zymo DNA extraction kit (ZD6005). Parts of the internal transcriber region (ITS) (ITS-F2: 5'-TTTACAACTCCCAAACCCCATGTGAAC-3'and ITS-R2: 5'-CTACCTGATTCGAGGTCAA CCAGAA-3') histone 3 (HIS3) (Crous et al. 2004), translation elongation 1α (EF1α) (Carbone and Kohn 1999; O'Donnell et al. 1998) and β-tubulin (TUB2) (Crous et al. 2004; O'Donnell and Cigelnik 1997) genes were amplified and their DNA sequenced. Sequences were processed in Geneious Prime 2024.0 and deposited at NCBI GenBank under the accession numbers PQ390253, PQ390254 and PQ390255 (ITS), PQ519580, PQ519581 and PQ519582 (HIS3), PQ507479, PQ507480 and PQ507481 (EF1α) and PV092563, PV092564 and PV092565 (TUB2). These DNA sequences show 100% identity to C. ilicicola sequences. To fulfill Koch's postulates, eight pots containing three plants (cultivar Williams 82) each were grown for two weeks in a growth chamber at 26°C with a 14-hour photoperiod and watered daily. Six pots were inoculated with 4 mm plugs of mycelium from the distal growing edge of the fungal isolate on PDA plates. The other two pots, treated with 4 mm plugs of PDA-only, served as negative controls. The planting medium at the base of each plant was gently removed, and inoculum was placed on the primary root, approximately 7 mm below the soil level. Five days after inoculation, plants started exhibiting reddish/blackish discoloration at the base of the stems, typical of red crown rot. Then, C. ilicicola was re-isolated from symptoma","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143543076","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 Rhizoctonia solani AG 2-1 Causing Root and Bulb Rot on Hymenocallis glauca in Mexico.
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2025-03-04 DOI: 10.1094/PDIS-12-24-2775-PDN
José Terrones-Salgado, Cesar Flores-de Los Ángeles, José Francisco Diaz Nájera, Sergio Ayvar Serna, José Luis Arispe Vázquez, Maricela Apáez Barrios, Francisco Javier Sánchez-Ruiz, Carlos David Carretillo Moctezuma, Nicolás Álvarez Acevedo, José Antonio Mendoza Rossano
<p><p>In Mexico, there are 29 native species of the genus <i>Hymenocallis</i>, including <i>H</i>. <i>glauca</i>, which is characterized by a bulb that stores carbohydrates, giving energy for the emergence of foliage and floral scapes; it is the most cultivated species and holds economic value as a potted plant and cut flower (Leszczyñska-Borys and Borys, 2001). In September 2024, a survey was conducted at the Center for Research in Horticulture and Native Plants (18°55'55.6"N 98°24'01.4"W) at UPAEP University, where there was an average temperature of 25 °C and 75% relative humidity (RH) for 10 consecutive days. Approximately 30-day old <i>H</i>. <i>glauca</i> seedlings exhibited symptoms of root and bulb rot in a 0.4 ha area, with a 45% disease incidence. Symptoms included root and bulb rot with constriction at the base of the bulb and the presence of brown mycelia. Symptomatic tissues from 50 seedlings were collected, cut into 5 mm pieces, sterilized with 3% NaClO for a minute, rinsed with sterile distilled water, and placed in Petri dishes with potato dextrose agar (PDA) medium. Samples were incubated in the dark for six days at 28 °C. An isolate was obtained from each diseased seedling using the hyphal tip method. After six days, the colonies consisted of white mycelium that turned brown with age. Right-angle branching hyphae were observed, with slight constriction at the base of the branches. The hyphae were multinucleate, containing four to nine nuclei per cell. After 15 days, some isolates produced dark brown sclerotia. Based on these morphological characteristics, isolates were tentatively identified as <i>Rhizoctonia solani</i> Kühn (Parmeter, 1970). To confirm the anastomosis group (AG), two isolates (RsHg4 and RsHg8) were selected for molecular identification. Genomic DNA was extracted using the CTAB protocol. The ITS region was amplified and sequenced (White et al. 1990) in both isolates, and the sequences were identical. Thus, only the sequence of isolate RsHg8 was deposited in GenBank (PQ524600). BLAST analysis of the partial ITS sequence (639 bp) showed 99.84% similarity with <i>R</i>. <i>solani</i> AG 2-1 isolate (GenBank: JF792354) (Mercado et al. 2012). Phylogenetic analysis of AGs sequences allowed assignment of the isolate RsHg8 to the AG 2-1 clade. Pathogenicity was confirmed by inoculating 50 30-day old <i>H</i>. <i>glauca</i> seedlings, grown in pots with sterile substrate. A 5 mm diameter PDA plug colonized with mycelium from the RsHg8 isolate was placed on each bulb, 10 mm below the soil surface. For control treatment, a PDA plug without fungal growth was placed on the bulb of 25 seedlings. The inoculated seedlings were incubated in a greenhouse at 28 °C and 90% RH. After six days, inoculated seedlings showed root and bulb rot with constriction at the base of the bulb. No symptoms were observed in controls. Fungus was re-isolated from the inoculated seedlings and characterized both morphologically and molecularly, yieldi
{"title":"First Report of <i>Rhizoctonia solani</i> AG 2-1 Causing Root and Bulb Rot on <i>Hymenocallis glauca</i> in Mexico.","authors":"José Terrones-Salgado, Cesar Flores-de Los Ángeles, José Francisco Diaz Nájera, Sergio Ayvar Serna, José Luis Arispe Vázquez, Maricela Apáez Barrios, Francisco Javier Sánchez-Ruiz, Carlos David Carretillo Moctezuma, Nicolás Álvarez Acevedo, José Antonio Mendoza Rossano","doi":"10.1094/PDIS-12-24-2775-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-12-24-2775-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;In Mexico, there are 29 native species of the genus &lt;i&gt;Hymenocallis&lt;/i&gt;, including &lt;i&gt;H&lt;/i&gt;. &lt;i&gt;glauca&lt;/i&gt;, which is characterized by a bulb that stores carbohydrates, giving energy for the emergence of foliage and floral scapes; it is the most cultivated species and holds economic value as a potted plant and cut flower (Leszczyñska-Borys and Borys, 2001). In September 2024, a survey was conducted at the Center for Research in Horticulture and Native Plants (18°55'55.6\"N 98°24'01.4\"W) at UPAEP University, where there was an average temperature of 25 °C and 75% relative humidity (RH) for 10 consecutive days. Approximately 30-day old &lt;i&gt;H&lt;/i&gt;. &lt;i&gt;glauca&lt;/i&gt; seedlings exhibited symptoms of root and bulb rot in a 0.4 ha area, with a 45% disease incidence. Symptoms included root and bulb rot with constriction at the base of the bulb and the presence of brown mycelia. Symptomatic tissues from 50 seedlings were collected, cut into 5 mm pieces, sterilized with 3% NaClO for a minute, rinsed with sterile distilled water, and placed in Petri dishes with potato dextrose agar (PDA) medium. Samples were incubated in the dark for six days at 28 °C. An isolate was obtained from each diseased seedling using the hyphal tip method. After six days, the colonies consisted of white mycelium that turned brown with age. Right-angle branching hyphae were observed, with slight constriction at the base of the branches. The hyphae were multinucleate, containing four to nine nuclei per cell. After 15 days, some isolates produced dark brown sclerotia. Based on these morphological characteristics, isolates were tentatively identified as &lt;i&gt;Rhizoctonia solani&lt;/i&gt; Kühn (Parmeter, 1970). To confirm the anastomosis group (AG), two isolates (RsHg4 and RsHg8) were selected for molecular identification. Genomic DNA was extracted using the CTAB protocol. The ITS region was amplified and sequenced (White et al. 1990) in both isolates, and the sequences were identical. Thus, only the sequence of isolate RsHg8 was deposited in GenBank (PQ524600). BLAST analysis of the partial ITS sequence (639 bp) showed 99.84% similarity with &lt;i&gt;R&lt;/i&gt;. &lt;i&gt;solani&lt;/i&gt; AG 2-1 isolate (GenBank: JF792354) (Mercado et al. 2012). Phylogenetic analysis of AGs sequences allowed assignment of the isolate RsHg8 to the AG 2-1 clade. Pathogenicity was confirmed by inoculating 50 30-day old &lt;i&gt;H&lt;/i&gt;. &lt;i&gt;glauca&lt;/i&gt; seedlings, grown in pots with sterile substrate. A 5 mm diameter PDA plug colonized with mycelium from the RsHg8 isolate was placed on each bulb, 10 mm below the soil surface. For control treatment, a PDA plug without fungal growth was placed on the bulb of 25 seedlings. The inoculated seedlings were incubated in a greenhouse at 28 °C and 90% RH. After six days, inoculated seedlings showed root and bulb rot with constriction at the base of the bulb. No symptoms were observed in controls. Fungus was re-isolated from the inoculated seedlings and characterized both morphologically and molecularly, yieldi","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143543053","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
Flutriafol Drench Provides Season-long Protection of Boxwood Plantings Pre-infected by Calonectria pseudonaviculata in the Mid-Atlantic.
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2025-03-04 DOI: 10.1094/PDIS-12-24-2731-SC
T Michael Likins, Billy Davis, Patrick Anderson, Blake Gillis, Chuanxue Hong

Flutriafol drench application on March 7 without and with additional applications at 8- and 20-week intervals all consistently protected boxwood plantings pre-infected by Calonectria pseudonaviculata in two gardens in Richmond, Virginia through November 21. All drench applications were dosed at 2 ml product with 42% flutriafol and delivered in 250 ml solution per 30.5-cm shrub height in both gardens. While providing season-long protection, this drench dose also resulted in significant phytotoxicity on 30.5-cm tall 'Justin Brouwers' boxwood but not on English and common boxwood, which were 1.4 and 1.1 m tall, respectively. Once the phytotoxicity issue is resolved, only a single drench per year may suffice to protect boxwood plantings in the Mid-Atlantic, which would substantially cut the labor cost and minimize the health impacts commonly associated with current fungicide foliar sprays.

{"title":"Flutriafol Drench Provides Season-long Protection of Boxwood Plantings Pre-infected by <i>Calonectria pseudonaviculata</i> in the Mid-Atlantic.","authors":"T Michael Likins, Billy Davis, Patrick Anderson, Blake Gillis, Chuanxue Hong","doi":"10.1094/PDIS-12-24-2731-SC","DOIUrl":"https://doi.org/10.1094/PDIS-12-24-2731-SC","url":null,"abstract":"<p><p>Flutriafol drench application on March 7 without and with additional applications at 8- and 20-week intervals all consistently protected boxwood plantings pre-infected by Calonectria pseudonaviculata in two gardens in Richmond, Virginia through November 21. All drench applications were dosed at 2 ml product with 42% flutriafol and delivered in 250 ml solution per 30.5-cm shrub height in both gardens. While providing season-long protection, this drench dose also resulted in significant phytotoxicity on 30.5-cm tall 'Justin Brouwers' boxwood but not on English and common boxwood, which were 1.4 and 1.1 m tall, respectively. Once the phytotoxicity issue is resolved, only a single drench per year may suffice to protect boxwood plantings in the Mid-Atlantic, which would substantially cut the labor cost and minimize the health impacts commonly associated with current fungicide foliar sprays.</p>","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143543078","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
Optimal use of insecticide against the small brown planthopper, Laodelphax striatellus, effectively reduces the damage caused by Rice stripe virus. 针对小褐飞虱 Laodelphax striatellus 最佳使用杀虫剂,可有效减少水稻条纹病毒造成的危害。
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2025-03-04 DOI: 10.1094/PDIS-08-24-1707-RE
Takuya Shiba, Masahiro Hirae, Yuriko Hayano-Saito, Yasuo Ohto, Hiroshi Uematsu, Ayano Sugiyama, Mitsuru Okuda

Rice stripe virus disease is one of the most devastating diseases of rice (Oryza sativa) in East Asia. The causal virus, Rice stripe virus (RSV), is persistently transmitted by the small brown planthopper (Laodelphax striatellus, SBPH). Since there is no effective treatment once rice plants are infected with RSV, it is crucial to prevent RSV infection in rice to minimize the damage. This study evaluated the effect of SBPH control with insecticides applied to the nursery box at transplanting and/or sprayed in the field on RSV infection and yield loss. We found that insecticide application either to nursery boxes or in the field reduced the percentage of diseased plants, the magnitude of damage in diseased plants, and yield loss. The combination of nursery box and field insecticide applications was the most effective at controlling the disease as it provided seamless control of SBPH from transplanting to the initial panicle formation stage. The study results offer a detailed description of the process by which chemical control of SBPH reduces damage caused by RSV. We expect these findings to be useful in developing an integrated pest management system for RSV.

{"title":"Optimal use of insecticide against the small brown planthopper, <i>Laodelphax striatellus</i>, effectively reduces the damage caused by Rice stripe virus.","authors":"Takuya Shiba, Masahiro Hirae, Yuriko Hayano-Saito, Yasuo Ohto, Hiroshi Uematsu, Ayano Sugiyama, Mitsuru Okuda","doi":"10.1094/PDIS-08-24-1707-RE","DOIUrl":"https://doi.org/10.1094/PDIS-08-24-1707-RE","url":null,"abstract":"<p><p>Rice stripe virus disease is one of the most devastating diseases of rice (<i>Oryza sativa</i>) in East Asia. The causal virus, Rice stripe virus (RSV), is persistently transmitted by the small brown planthopper (<i>Laodelphax striatellus</i>, SBPH). Since there is no effective treatment once rice plants are infected with RSV, it is crucial to prevent RSV infection in rice to minimize the damage. This study evaluated the effect of SBPH control with insecticides applied to the nursery box at transplanting and/or sprayed in the field on RSV infection and yield loss. We found that insecticide application either to nursery boxes or in the field reduced the percentage of diseased plants, the magnitude of damage in diseased plants, and yield loss. The combination of nursery box and field insecticide applications was the most effective at controlling the disease as it provided seamless control of SBPH from transplanting to the initial panicle formation stage. The study results offer a detailed description of the process by which chemical control of SBPH reduces damage caused by RSV. We expect these findings to be useful in developing an integrated pest management system for RSV.</p>","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143557253","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 Leaf Anthracnose Caused by Colletotrichum fructicola on Tetrastigma hemsleyanum in China.
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2025-03-04 DOI: 10.1094/PDIS-12-24-2728-PDN
Youchao Dang, Meiqin Mao, Jingyi Xu, Fei Xu, Xiaoqing Zhang, Chuan Qi, Bo Zhu, Luping Qin
<p><p>Tetrastigma hemsleyanum in the family Vitaceae, is a rare and endangered medicinal plant endemic in China (Ji et al. 2021). In October 2024, leaf anthracnose was observed in Lishui city (118°96'E, 28°13'N), Zhejiang, affecting T. hemsleyanum plantings over an area of 5.3 × 103 m2. Disease incidence ranged from approximately 30 to 60%. Early symptoms were small circular or irregular brown spots of foliage, gradually expanding in size, then coalescing to form large irregular dark brown spots with grayish white centers causing leaves to senesce and resulting in plant death in severe cases. Leaf pieces (5×5 mm) from nine symptomatic leaves were surface disinfected with 75% ethanol for 30 s, 2.5% NaClO for 1 min, rinsed in sterile water three times, dried, placed on potato dextrose agar medium, and cultured in darkness at 28°C for 5 days. Five isolates (THP10 to THP14) were obtained by the hyphal-tip method from the nine leaves. The colonies were olivaceous to dark gray with white margins and cottony mycelium; reverse sides had black centers. Conidia were single, colorless, cylindrical, 13.37 to 17.89 × 3.91 to 5.73 μm (average 15.42 × 5.11 μm; n=50). The morphological characteristics of the isolates overlapped with those of Colletotrichum species within the C. gloeosporioides complex, including C. fiucticola (Weir et al. 2012). The internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), chitin synthase (CHS),calmodulin (CAL), actin (ACT), and beta-tubulin 2 (TUB2) genes were amplified using ITS1/ITS4, GDF/GDR, CHS-79F/CHS-345R, CL1C/CL2C, ACT-512F/ACT-783R, and T1/Bt2b primer sets, respectively (Weir et al. 2012). Sequences were deposited in GenBank with accession Nos. ITS: PQ571715 - PQ571719; GAPDH: PQ593912 - PQ593916; CHS: PQ593906 - PQ593910; CAL: PQ593900 - PQ593904; ACT: PQ593894 - PQ593898; TUB2: PQ593918 - PQ593922. BLASTn analysis of THP10 sequences had highest matches to the type strain of C. fructicola ICMP 18581 with ITS sequences 100% identical (JX010165; 549/549 bp), GAPDH sequences 99% identical (JX010033; 267/269 bp), CHS sequences 100.00% identical (JX009866; 274/274 bp), CAL sequences 100% identical (JX009676; 731/731 bp), ACT sequences 99% identical (JX009501; 270/272 bp), and TUB2 sequences 100% identical (JX010405; 699/699 bp). A maximum likelihood phylogenetic tree was constructed with the combined sequences data sets using MEGA 11, and the five isolates clustered with C. fructicola (Weir et al. 2012). To test pathogenicity, five isolates of C. fructicola were evaluated, leaves on three healthy 6-month-old potted T. hemsleyanum seedlings were wounded with sterile needles and inoculated with 5 mm diameter mycelial plugs. Sterile PDA plugs served as controls. After inoculation, the plants were incubated at 28°C, 85% relative humidity, with a 12 h photoperiod. The experiment was repeated three times. Symptoms similar to those from the field were observed 21 days after inoculation, whereas control
{"title":"First Report of Leaf Anthracnose Caused by <i>Colletotrichum fructicola</i> on <i>Tetrastigma hemsleyanum</i> in China.","authors":"Youchao Dang, Meiqin Mao, Jingyi Xu, Fei Xu, Xiaoqing Zhang, Chuan Qi, Bo Zhu, Luping Qin","doi":"10.1094/PDIS-12-24-2728-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-12-24-2728-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Tetrastigma hemsleyanum in the family Vitaceae, is a rare and endangered medicinal plant endemic in China (Ji et al. 2021). In October 2024, leaf anthracnose was observed in Lishui city (118°96'E, 28°13'N), Zhejiang, affecting T. hemsleyanum plantings over an area of 5.3 × 103 m2. Disease incidence ranged from approximately 30 to 60%. Early symptoms were small circular or irregular brown spots of foliage, gradually expanding in size, then coalescing to form large irregular dark brown spots with grayish white centers causing leaves to senesce and resulting in plant death in severe cases. Leaf pieces (5×5 mm) from nine symptomatic leaves were surface disinfected with 75% ethanol for 30 s, 2.5% NaClO for 1 min, rinsed in sterile water three times, dried, placed on potato dextrose agar medium, and cultured in darkness at 28°C for 5 days. Five isolates (THP10 to THP14) were obtained by the hyphal-tip method from the nine leaves. The colonies were olivaceous to dark gray with white margins and cottony mycelium; reverse sides had black centers. Conidia were single, colorless, cylindrical, 13.37 to 17.89 × 3.91 to 5.73 μm (average 15.42 × 5.11 μm; n=50). The morphological characteristics of the isolates overlapped with those of Colletotrichum species within the C. gloeosporioides complex, including C. fiucticola (Weir et al. 2012). The internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), chitin synthase (CHS),calmodulin (CAL), actin (ACT), and beta-tubulin 2 (TUB2) genes were amplified using ITS1/ITS4, GDF/GDR, CHS-79F/CHS-345R, CL1C/CL2C, ACT-512F/ACT-783R, and T1/Bt2b primer sets, respectively (Weir et al. 2012). Sequences were deposited in GenBank with accession Nos. ITS: PQ571715 - PQ571719; GAPDH: PQ593912 - PQ593916; CHS: PQ593906 - PQ593910; CAL: PQ593900 - PQ593904; ACT: PQ593894 - PQ593898; TUB2: PQ593918 - PQ593922. BLASTn analysis of THP10 sequences had highest matches to the type strain of C. fructicola ICMP 18581 with ITS sequences 100% identical (JX010165; 549/549 bp), GAPDH sequences 99% identical (JX010033; 267/269 bp), CHS sequences 100.00% identical (JX009866; 274/274 bp), CAL sequences 100% identical (JX009676; 731/731 bp), ACT sequences 99% identical (JX009501; 270/272 bp), and TUB2 sequences 100% identical (JX010405; 699/699 bp). A maximum likelihood phylogenetic tree was constructed with the combined sequences data sets using MEGA 11, and the five isolates clustered with C. fructicola (Weir et al. 2012). To test pathogenicity, five isolates of C. fructicola were evaluated, leaves on three healthy 6-month-old potted T. hemsleyanum seedlings were wounded with sterile needles and inoculated with 5 mm diameter mycelial plugs. Sterile PDA plugs served as controls. After inoculation, the plants were incubated at 28°C, 85% relative humidity, with a 12 h photoperiod. The experiment was repeated three times. Symptoms similar to those from the field were observed 21 days after inoculation, whereas control","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143557245","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
Pyridaben as a novel fungicide against Sphaerotheca fuliginea, the causal agent of cucumber powdery mildew.
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2025-03-04 DOI: 10.1094/PDIS-11-24-2483-RE
Jing Jin, Ge-Ning Song, Shuang-Ying Zhang, Jie Chen

Pyridaben is a novel broad-spectrum insecticide and acaricide that is effective against thrips, mites, aphids and leafhoppers. We accidentally discovered that pyridaben was able to prevent cucumber powdery mildew. In this study, we explored the inhibitory effects and underlying mechanisms of pyridaben against Sphaerotheca fuliginea. Pyridaben effectively inhibited the spore germination and mycelial growth of S. fuliginea with IC50 of 10.83 mg/L and 18.28 mg/L, respectively. The spore germination, mycelial growth, and secondary spore formation of S. fuliginea was severely hindered on cucumber leaves treated with pyridaben at 20 mg/L. SEM revealed that the mycelia growing on pyridaben-treated cucumber leaves were much finer than that growing on untreated leaves. Transcriptomic analysis revealed that pyridaben had a significant impact on the ribosomal function and energy metabolism of the powdery mildew pathogen, restricting its normal physiological activities. In field experiments, the efficacies of pyridaben reached 76.48% in Zhejiang province and 78.40% in Shandong province at the rate of 225 g a.i./ha after two applications, which were comparable to the commercialized fungicides. These results suggest pyridaben is not only an insecticide but also an effective fungicide with the potential for further application in preventing and controlling cucumber powdery mildew.

{"title":"Pyridaben as a novel fungicide against <i>Sphaerotheca fuliginea</i>, the causal agent of cucumber powdery mildew.","authors":"Jing Jin, Ge-Ning Song, Shuang-Ying Zhang, Jie Chen","doi":"10.1094/PDIS-11-24-2483-RE","DOIUrl":"https://doi.org/10.1094/PDIS-11-24-2483-RE","url":null,"abstract":"<p><p>Pyridaben is a novel broad-spectrum insecticide and acaricide that is effective against thrips, mites, aphids and leafhoppers. We accidentally discovered that pyridaben was able to prevent cucumber powdery mildew. In this study, we explored the inhibitory effects and underlying mechanisms of pyridaben against <i>Sphaerotheca fuliginea</i>. Pyridaben effectively inhibited the spore germination and mycelial growth of <i>S. fuliginea</i> with IC<sub>50</sub> of 10.83 mg/L and 18.28 mg/L, respectively. The spore germination, mycelial growth, and secondary spore formation of <i>S. fuliginea</i> was severely hindered on cucumber leaves treated with pyridaben at 20 mg/L. SEM revealed that the mycelia growing on pyridaben-treated cucumber leaves were much finer than that growing on untreated leaves. Transcriptomic analysis revealed that pyridaben had a significant impact on the ribosomal function and energy metabolism of the powdery mildew pathogen, restricting its normal physiological activities. In field experiments, the efficacies of pyridaben reached 76.48% in Zhejiang province and 78.40% in Shandong province at the rate of 225 g a.i./ha after two applications, which were comparable to the commercialized fungicides. These results suggest pyridaben is not only an insecticide but also an effective fungicide with the potential for further application in preventing and controlling cucumber powdery mildew.</p>","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143543080","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 Diaporthe pescicola causing leaf spot on tea (Camellia sinensis) in China.
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2025-03-03 DOI: 10.1094/PDIS-01-25-0119-PDN
Yongtian Zhao, Hong Wang, Taifeng Huang, Ruixing Wang, Li Yang, Yuxin Yang, Lingyu Yang, Fen Wang, Xing'e Wang
<p><p>Duyun Maojian Tea mainly cultivated in Duyun City is one of the ten famous teas in China. A brown leaf spot was discovered in March 2023 in Qiannan Normal University for Nationalities' Maojian Tea experimental field (26.29°N, 107.52°E). The incidence of the disease is about 40% surveyed on about 300 m2 of tea plants. Infected leaves showed light brown, round or irregular lesions. With disease progression, they enlarged and merged, creating dark brown necrotic zones along leaf edges. To identify the responsible pathogen, three symptomatic leaves from the tea nursery base were collected. The samples were cut 3 mm x 3 mm tissues with 75% ethanol for 30 s and 5% sodium hypochlorite for 60 s, rinsing three times with sterile water. Tissues were taken to potato dextrose agar (PDA) medium and incubated dark at 28 °C for 3 days. Three isolates (TF1, TF3 and TF5) with similar morphology were obtained from three samples, respectively. The colony showed white, with short, villous hyphae expanding radially. Notably, mycelium near the disc was erect and sparse 2 cm away. After 30 days, conidia were observed. Alpha conidia (5.9 to 8.6 μm × 2.0 to 2.8 μm (n = 50)) were colorless and transparent, no septum, ellipsoid to spindle-shaped. Beta conidia (11.1 to 25.3 μm × 1.0 to 1.8 μm (n = 50)) were transparent, no septum, filiform, hooklike, tapering towards the ends. The isolates were identified as Diaporthe pescicola based on the morphology (Zhang et al. 2022). Genomic DNA was extracted, the internal transcribed spacer region (ITS), β-tubulin (tub2), and translation elongation factor 1-alpha (TEF) genes were amplified with primer pairs ITS1/ITS4 (White et al. 1990), Bt2a/Bt2b (Glass and Donaldson 1995), and EF1-728F/EF1-986R (Carbone and Kohn 1999), respectively, and sequenced. The ITS (PQ163865, PQ685064 and PQ685065), tub2 (PQ197032, PV055438 and PV055439), and TEF (PQ197029, PV055440 and PV055441) sequences of isolates (TF1, TF3 and TF5) were deposited in GenBank, respectively. BLAST searches showed >97% nucleotide identity to sequences of D. pescicola MFLUCC 16-0105 (ITS, 98.40% to KU557555.1; tub2, 97.40% to KU557579.1; and TEF, 97.63% to KU557623.1). The phylogenetic tree of ITS, tub2, and TEF gene sequences was constructed using MEGA 6 software with the neighbour-joining method (NJ), and the bootstrap method was employed for 1,000 repeated tests. Isolates TF1, TF3, TF5, and D. pescicola DT 1-1 were grouped together in a node with a bootstrap value of 99. Isolates were identified as D. pescicola. Since large quantities of conidia are difficult to produce in cultures, pathogenicity on the wounded leaves of 5-year-old tea plants of Duyun Maojian Tea local population was conducted by inoculating 6 mm mycelium discs of TF1. The control group and the treatment group were inoculated with 3 tea plants, repeated 2 times. All tea plants were incubated at 25 ± 1 °C, 70% humidity, and natural daylight. The onset of symptoms appeared 7 days later. Lesions were br
{"title":"First report of <i>Diaporthe pescicola</i> causing leaf spot on tea (<i>Camellia sinensis</i>) in China.","authors":"Yongtian Zhao, Hong Wang, Taifeng Huang, Ruixing Wang, Li Yang, Yuxin Yang, Lingyu Yang, Fen Wang, Xing'e Wang","doi":"10.1094/PDIS-01-25-0119-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-01-25-0119-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Duyun Maojian Tea mainly cultivated in Duyun City is one of the ten famous teas in China. A brown leaf spot was discovered in March 2023 in Qiannan Normal University for Nationalities' Maojian Tea experimental field (26.29°N, 107.52°E). The incidence of the disease is about 40% surveyed on about 300 m2 of tea plants. Infected leaves showed light brown, round or irregular lesions. With disease progression, they enlarged and merged, creating dark brown necrotic zones along leaf edges. To identify the responsible pathogen, three symptomatic leaves from the tea nursery base were collected. The samples were cut 3 mm x 3 mm tissues with 75% ethanol for 30 s and 5% sodium hypochlorite for 60 s, rinsing three times with sterile water. Tissues were taken to potato dextrose agar (PDA) medium and incubated dark at 28 °C for 3 days. Three isolates (TF1, TF3 and TF5) with similar morphology were obtained from three samples, respectively. The colony showed white, with short, villous hyphae expanding radially. Notably, mycelium near the disc was erect and sparse 2 cm away. After 30 days, conidia were observed. Alpha conidia (5.9 to 8.6 μm × 2.0 to 2.8 μm (n = 50)) were colorless and transparent, no septum, ellipsoid to spindle-shaped. Beta conidia (11.1 to 25.3 μm × 1.0 to 1.8 μm (n = 50)) were transparent, no septum, filiform, hooklike, tapering towards the ends. The isolates were identified as Diaporthe pescicola based on the morphology (Zhang et al. 2022). Genomic DNA was extracted, the internal transcribed spacer region (ITS), β-tubulin (tub2), and translation elongation factor 1-alpha (TEF) genes were amplified with primer pairs ITS1/ITS4 (White et al. 1990), Bt2a/Bt2b (Glass and Donaldson 1995), and EF1-728F/EF1-986R (Carbone and Kohn 1999), respectively, and sequenced. The ITS (PQ163865, PQ685064 and PQ685065), tub2 (PQ197032, PV055438 and PV055439), and TEF (PQ197029, PV055440 and PV055441) sequences of isolates (TF1, TF3 and TF5) were deposited in GenBank, respectively. BLAST searches showed &gt;97% nucleotide identity to sequences of D. pescicola MFLUCC 16-0105 (ITS, 98.40% to KU557555.1; tub2, 97.40% to KU557579.1; and TEF, 97.63% to KU557623.1). The phylogenetic tree of ITS, tub2, and TEF gene sequences was constructed using MEGA 6 software with the neighbour-joining method (NJ), and the bootstrap method was employed for 1,000 repeated tests. Isolates TF1, TF3, TF5, and D. pescicola DT 1-1 were grouped together in a node with a bootstrap value of 99. Isolates were identified as D. pescicola. Since large quantities of conidia are difficult to produce in cultures, pathogenicity on the wounded leaves of 5-year-old tea plants of Duyun Maojian Tea local population was conducted by inoculating 6 mm mycelium discs of TF1. The control group and the treatment group were inoculated with 3 tea plants, repeated 2 times. All tea plants were incubated at 25 ± 1 °C, 70% humidity, and natural daylight. The onset of symptoms appeared 7 days later. Lesions were br","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143543048","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 Pseudomonas syringae pv. atrofaciens Causing Leaf Spot to barley (Hordeum vulgare) in the Republic of Korea.
IF 4.4 2区 农林科学 Q1 PLANT SCIENCES Pub Date : 2025-03-03 DOI: 10.1094/PDIS-09-24-1987-PDN
Sang-Min Kim, Young-Mi Yoon, Yang-Kil Kim, Jong-Ho Park, Su Jwa Seo, Jieun Lee
<p><p>Barley <i>(Hordeum vulgare</i> L.) is one of the staple food in the Republic of Korea, with 68,000 metric tons produced on 24,000 ha in 2022 (Ministry of Agriculture, Food and Rural Affairs 2023). In April 2018, leaf spots with brownish black edges were observed on barley leaves at the early tillering stage in the field of 1,100 m<sup>2</sup> at Iksan-si (35°56'21.4"N 126°55'56.5"E), with an incidence of 5%. Early symptoms on the leaf appeared as small, dark brown to black spots, then enlarged to gray lesions with brownish-black edges. Leaf cuttings (5 x 5 mm) from three symptomatic plants were surface-sterilized with 70% ethanol for 1 min, followed by 1% hypochlorite for 1 min, then macerated in sterile distilled water. The macerates were streaked on Nutrient Agar (Difco, MD, USA) media and incubated at 28 °C in the dark for 2 days. After single colony isolation, three isolates from three different plants were designated as isolates Ik 510-1, Ik511-1, and Ik513-1, then deposited in the Korean Agricultural Culture Collection (https://genebank.rda.go.kr) under the accession numbers KACC 23555, 23556, and 23557, respectively. Bacterial colonies were circular, convex, and cream-colored on NA media and produced fluorescent blue-green pigment on King Agar B media (Millipore, Merck, Darmstadt, Germany). The morphology of the three isolates was identical and showed the same characteristics as <i>Pseudomonas syringae</i> (von Kietzell and Rudolph 1997). Biolog Identification System (Biolog Inc., Calif., USA) and BLAST similarity of the 16S rRNA gene sequence (GenBank accession no. MK140998, MK140999, and MK141000, respectively) with the primer set of F27/R1492 (Heuer et al. 1997) showed that the three isolates belonged to the <i>Pseudomonas syringae</i> species complex, and MLSA (multi locus sequence analysis) with partial sequences of <i>gltA</i>, <i>gapA</i>, <i>gyrB</i>, and <i>rpoD</i> genes (Hwang et al. 2005) showed that the three isolates were on the same clade with <i>Pseudomonas syringae</i> pv. <i>atrofaciens</i> LMG5095<sup>PT</sup>. For confirmation, PCR with the primer set of Psat-F/Psat-R (Animal and Plant Quarantine Agency 2023) amplified the 477 bp amplicons of the target region, thus identifying them as <i>Pseudomonas syringae</i> pv. <i>atrofaciens</i>. To determine the pathogenicity to barley, bacterial suspensions (O.D.<sub>600</sub>=0.1 in PBS, phosphate buffered saline) of the three isolates was infiltrated at approx. 0.05 ml into leaves of three 2-week-old plants 'Keunalbori1ho' grown in pots using a needless syringe, while PBS was infiltrated as negative control, respectively. Infiltrated plants were grown in a growth chamber at 25℃ with a 16-h/8-h light/dark cycle. 5 days after inoculation, the infiltrated areas showed water-soaked symptoms, then dried to gray with characteristic brown-black edges only on leaves infiltrated with bacterial suspension and no symptoms on leaves infiltrated with PBS. The pathogenicity test was
{"title":"First report of <i>Pseudomonas syringae</i> pv. <i>atrofaciens</i> Causing Leaf Spot to barley (<i>Hordeum vulgare</i>) in the Republic of Korea.","authors":"Sang-Min Kim, Young-Mi Yoon, Yang-Kil Kim, Jong-Ho Park, Su Jwa Seo, Jieun Lee","doi":"10.1094/PDIS-09-24-1987-PDN","DOIUrl":"https://doi.org/10.1094/PDIS-09-24-1987-PDN","url":null,"abstract":"&lt;p&gt;&lt;p&gt;Barley &lt;i&gt;(Hordeum vulgare&lt;/i&gt; L.) is one of the staple food in the Republic of Korea, with 68,000 metric tons produced on 24,000 ha in 2022 (Ministry of Agriculture, Food and Rural Affairs 2023). In April 2018, leaf spots with brownish black edges were observed on barley leaves at the early tillering stage in the field of 1,100 m&lt;sup&gt;2&lt;/sup&gt; at Iksan-si (35°56'21.4\"N 126°55'56.5\"E), with an incidence of 5%. Early symptoms on the leaf appeared as small, dark brown to black spots, then enlarged to gray lesions with brownish-black edges. Leaf cuttings (5 x 5 mm) from three symptomatic plants were surface-sterilized with 70% ethanol for 1 min, followed by 1% hypochlorite for 1 min, then macerated in sterile distilled water. The macerates were streaked on Nutrient Agar (Difco, MD, USA) media and incubated at 28 °C in the dark for 2 days. After single colony isolation, three isolates from three different plants were designated as isolates Ik 510-1, Ik511-1, and Ik513-1, then deposited in the Korean Agricultural Culture Collection (https://genebank.rda.go.kr) under the accession numbers KACC 23555, 23556, and 23557, respectively. Bacterial colonies were circular, convex, and cream-colored on NA media and produced fluorescent blue-green pigment on King Agar B media (Millipore, Merck, Darmstadt, Germany). The morphology of the three isolates was identical and showed the same characteristics as &lt;i&gt;Pseudomonas syringae&lt;/i&gt; (von Kietzell and Rudolph 1997). Biolog Identification System (Biolog Inc., Calif., USA) and BLAST similarity of the 16S rRNA gene sequence (GenBank accession no. MK140998, MK140999, and MK141000, respectively) with the primer set of F27/R1492 (Heuer et al. 1997) showed that the three isolates belonged to the &lt;i&gt;Pseudomonas syringae&lt;/i&gt; species complex, and MLSA (multi locus sequence analysis) with partial sequences of &lt;i&gt;gltA&lt;/i&gt;, &lt;i&gt;gapA&lt;/i&gt;, &lt;i&gt;gyrB&lt;/i&gt;, and &lt;i&gt;rpoD&lt;/i&gt; genes (Hwang et al. 2005) showed that the three isolates were on the same clade with &lt;i&gt;Pseudomonas syringae&lt;/i&gt; pv. &lt;i&gt;atrofaciens&lt;/i&gt; LMG5095&lt;sup&gt;PT&lt;/sup&gt;. For confirmation, PCR with the primer set of Psat-F/Psat-R (Animal and Plant Quarantine Agency 2023) amplified the 477 bp amplicons of the target region, thus identifying them as &lt;i&gt;Pseudomonas syringae&lt;/i&gt; pv. &lt;i&gt;atrofaciens&lt;/i&gt;. To determine the pathogenicity to barley, bacterial suspensions (O.D.&lt;sub&gt;600&lt;/sub&gt;=0.1 in PBS, phosphate buffered saline) of the three isolates was infiltrated at approx. 0.05 ml into leaves of three 2-week-old plants 'Keunalbori1ho' grown in pots using a needless syringe, while PBS was infiltrated as negative control, respectively. Infiltrated plants were grown in a growth chamber at 25℃ with a 16-h/8-h light/dark cycle. 5 days after inoculation, the infiltrated areas showed water-soaked symptoms, then dried to gray with characteristic brown-black edges only on leaves infiltrated with bacterial suspension and no symptoms on leaves infiltrated with PBS. The pathogenicity test was ","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143543051","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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