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  • 文章类型: Journal Article
    普通羔羊宿舍(Chenopodium专辑Linn。)是世界上与作物相关的最成问题的杂草之一,由于其生长迅速,高繁殖力,以及对各种条件的广泛容忍。同时,C.专辑也是一种草本植物,这种植物的叶子和嫩芽在人类饮食中被认为是有营养的(Aman等人。2016)。2023年9月,C.专辑植物出现泛黄,发育迟缓,从封丘山药田(北纬34°54\'24\';东经114°34\'57\'')收集到广泛的牙根症状,河南省,中国。在选定的采样地点,我们随机选择了100℃的专辑植物,在0.67公顷的田地上,发病率为73%。发现了属于Meloidogyne属的RKN物种,包括100克10至30厘米土壤层中的平均550个第二阶段少年(J2s)。用Baermann漏斗从新鲜土壤中分离出J2s。用自来水彻底洗涤C.专辑根并解剖。收集不同阶段的线虫并进行形态学鉴定。通过解剖gall获得雌性和卵块。女性是白色的,脖子突出,球形到梨形。女性的会阴模式主要表现出明显的背弓,以正方形或梯形为特征,缺乏明显的侧线。从根gall中分离出的雄性是蠕虫状的,环状的,显示出梯形的唇形区域,包括在侧视图中在顶端的中心凹陷的高头帽。J2s的特点是显眼的,圆形探针旋钮,他们有皱纹的尾巴,透明的区域和钝的尖端。形态学测量在补充材料中描述。所有特征均与南方根结线虫的形态特征一致(Eisenback和Hirschmann1981)。用随后的物种特异性PCR和测序分析完成鉴定。提取了10只雌性个体的基因组DNA,分子鉴定是用未知分枝杆菌特异性引物Mi-F/Mi-R进行的,和Inc-K14-F/Inc-K14-R(Meng等人2004;Randig等人。2002).PCR扩增产生955和399bp片段的分析样品,分别,并且通过序列分析确认扩增子。序列以登录号PP836070和PP836071保存在GenBank中。BLASTn搜索显示与可用的GenBankM.incognita序列具有100%的同一性(登录号。MK410954、OQ427638)。为了验证C.专辑的复制,在温室条件下(明/暗:16h/8h,温度:25-28°C)。五株未接种的植物用作阴性对照。接种后两个月,生长迟缓和根部刮伤症状与野外相似,而对照植物保持无症状。在所有接种的植物中都观察到许多根gall和卵质量。根磨损指数(0至10的标度;Poudyal等人。2005)为~7,线虫繁殖因子(最终种群密度/初始种群密度)为5.3。从根组织中重新分离出的线虫的形态特征与隐身分枝杆菌的描述紧密匹配,履行科赫的假设。致病性测试进行了两次,结果相似。M.incognita是全球许多农作物中具有经济重要性的新兴疾病,并可能造成严重的经济损失(Phani等人。2021)。这种广泛分布的C.album植物可能是病原体的宿主,并充当线虫的替代宿主。这些发现对RKN的综合管理实践具有重要意义,特别是对于受C.专辑感染的作物。据我们所知,这是中国首次报道线虫寄生C.专辑。迫切需要开发有效的短期和长期控制程序来管理隐身病。
    Common lambsquarters (Chenopodium album Linn.) is one of the most problematic weeds associated with crops worldwide due to its fast-growing, high fecundity, and wide tolerance to various conditions. Meanwhile, C. album is also an herbaceous vegetable plant, and the leaves and young shoots of this plant are considered nutritious in the human diet (Aman et al. 2016). In September 2023, C. album plants exhibiting yellowing, stunted growth, and extensive galled root symptoms were collected from a yam field in Fengqiu (34°54\'24\"N; 114°34\'57\"E), Henan Province, China. At the selected sampling site, we randomly selected 100 C. album plants, and the disease incidence was 73% on a 0.67-ha field. A RKN species belonging to the genus Meloidogyne was found, comprising an average of 550 second-stage juveniles (J2s) from 100 g of the 10 to 30 cm soil layer. The J2s were isolated from fresh soil with a Baermann funnel. C. album roots were thoroughly washed with tap water and dissected. Nematodes at different stages were collected and morphologically identified. Females and egg masses were obtained by dissecting galls. Females were white with a protruding neck, globular to pear-shaped. The perineal patterns of females predominantly exhibited a pronounced dorsal arch, characterized by either a square or trapezoid shape, lacking obvious lateral lines. Males isolated from root galls were vermiform, annulated, and showed a trapezoidal labial region, including a high head cap that was concaved at the center of the top end in lateral view. J2s were distinguished by the conspicuous, round stylet knobs, and they had wrinkled tails with a hyaline region and an obtuse tip. Morphological measurements are described in the supplementary material. All features were consistent with the morphological characteristics of Meloidogyne incognita (Eisenback and Hirschmann 1981). Identification was accomplished with subsequent species-specific PCR and sequencing analysis. The genomic DNA of 10 individual females was extracted, and the molecular identification was carried out with M. incognita-specific primers Mi-F/Mi-R, and Inc-K14-F/Inc-K14-R (Meng et al. 2004; Randig et al. 2002). PCR amplification generated 955 and 399 bp fragments for the analyzed samples, respectively, and the amplicons were confirmed by sequence analyses. The sequences were deposited in GenBank under accession number PP836070 and PP836071. BLASTn searches showed 100% identity with available GenBank M. incognita sequences (accession no. MK410954, OQ427638). To verify reproduction on C. album, 10 healthy plants (30 days old) grown in pots with sterilized soil were inoculated with 1,000 M. incognita J2s under greenhouse conditions (light/dark: 16 h/8 h, temperature: 25-28°C). Five uninoculated plants were used as negative control. Two months after inoculation, stunted growth and root-galling symptoms were observed similar to those in field, whereas control plants remained symptomless. Many root galls and egg masses were observed in all inoculated plants. The root galling index (scale of 0 to 10; Poudyal et al. 2005) was ~7 and nematode reproduction factor (final population density/initial population density) was 5.3. The morphological features of the nematodes reisolated from root tissue closely match the description of M. incognita, fulfilling Koch\'s postulates. The pathogenicity test was carried out twice with similar results. M. incognita is an emerging disease of economic importance in many crop plants worldwide, and may cause serious economic losses (Phani et al. 2021). This widely distributed C. album plant is likely a reservoir for the pathogen and serves as an alternate host for nematodes. The findings are significant for the integrated management practices of RKNs, particularly for crops that are infested with C. album. To our knowledge, this is the first report of the nematode parasitizing C. album in China. The development of effective short- and long-term control procedures is urgently needed for managing M. incognita.
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  • 文章类型: Journal Article
    小川贝母是《中国药典》中公认的“川贝母”来源。在中国,它的灯泡已被用作传统的草药咳嗽疗法约2000年。在小金和松潘进行了真菌病调查,四川省,F.unibracteata的初级栽培区,占地150英亩,2022年5月和7月。几乎所有地区都发现了锈病,所有研究地区的发病率从5%到80%不等。患病的叶子在上侧显示黄色斑点,举起buff,金色,或在下侧的粘稠蜡状脓疱。在严重的情况下,感染延伸到茎和叶柄,导致植物枯萎和死亡。精子症,aecia,和telia主要发现在叶子的下面。精孢菌散布在Aecia中,并表现出从蜂蜜黄色到栗色棕色的一系列颜色。它们的横截面直径为94.4至214.3µm,宽度为94.2至197.5µm(n=30)。它们几乎是球形的,嵌入宿主组织中,毛孔有明显的外周。Aecia是半球形的,最初是白色的,外围后来变成黄棕色,并通过中央孔打开。Aeciospores是淡黄色的,细且紧密的疣状,测量20.6至34.1×18.4至30.1µm,细胞壁厚度为1.5至2.4µm(n=51)。在植物枯萎之前,观察到细长的telia,逐渐暴露,然后最终通过表皮的纵向裂缝打开。太阳孢子是单细胞的,深棕色,长方形到椭圆形,在茎上孤独,尺寸为24.7至38.2×19.2至27.8µm(n=130),壁厚为1.6至3.1µm,先端有低透明乳头,中等皱纹,纵向平行脊。这些特征与先前对乌氏霉菌的描述一致(里斯,1917年,庄,2005).引物对LR0R(Moncalvo等人。,1995)/LR5(Vilgalys&Hester,1990)用于扩增和测序来自菌株IS909-3和IS1816的核核糖体RNA基因的大亚基(GenBankPQ008482,PQ008483)。获得的序列与RustHubb中的U.aecidiiformis菌株U1023和UBC19(KR0014142和PUN23000)具有99.9%至100%的相似性(Kaishian等人。,2024).通过形态学检查,主机范围,和序列相似性,我们确定了锈菌的种类是U.A.2023年5月,通过在室内6株健康的4岁F.unbracteata植物上喷洒孢子悬浮液(1×105孢子/mL,在0.05%Tween20溶液中)进行致病性测试。让植物在自然条件下生长,其中昼夜温度范围从9到20℃,平均气温14℃,有利于F.unbracteata的生长。用0.05%Tween20溶液喷洒另外6株幼苗作为对照。三周后,所有受感染的植物都表现出与田间相似的症状,而对照植物保持无症状。显微镜检查和测序证实,病原体形态在田间和接种之间是一致的,符合科赫的假设。尽管以前有报道说,艾氏U形会导致帕氏F.pallidiflora和ussuriensis的锈病(庄,1989年,庄,2005),这是在中国首次报道的U形藻引起的F.unbracteata生锈。这种病菌显著降低了川北木的产量和品质,强调有效识别和控制它的重要性。
    Fritillaria unibracteata Hsiao et K. C. Hsia is a recognized source of \'Chuanbeimu\' in the \'Chinese Pharmacopoeia\'. In China, its bulbs have been used as a traditional herbal cough remedy for about 2,000 years. Surveys for fungal diseases were conducted in Xiaojin and Songpan, Sichuan Province, the primary cultivation region of F. unibracteata, with an area of 150 acres, in May and July 2022. Rust was found in almost all areas and incidence ranged from 5% to 80% in all study areas. Diseased leaves displayed yellow spots on the upper side, and raised buff, golden, or fuscous waxy pustules on the lower side. In severe cases, the infection extended to the stems and petioles, leading to wilting and death of plant. Spermogonia, aecia, and telia were mainly found on the underside of leaves. Spermogonia were scattered among the aecia and exhibited a range of colors from honey-yellow to chestnut-brown. They had a cross-sectional diameter of 94.4 to 214.3 µm height and 94.2 to 197.5 µm in width (n=30). They were nearly spherical, embedded in the host tissue, and had distinct periphysis at the pores. Aecia were hemispherical, initially white, with the peridium later turning yellowish-brown and opening via a central pore. Aeciospores were pale yellow, finely and closely verrucose, measuring 20.6 to 34.1 × 18.4 to 30.1 µm with a cell wall thickness of 1.5 to 2.4 µm (n=51). Prior to plants wilting, elongated telia were observed, gradually exposed, then finally opening through longitudinal cracks in the epidermis. Teliospores were unicellular, dark brown, oblong to oval, and solitary on stems, measuring 24.7 to 38.2 × 19.2 to 27.8 µm (n=130) with a wall thickness of 1.6 to 3.1 µm, with a low hyaline papilla at the apex and were moderately rugose with longitudinal parallel ridges. The characteristics align with previous descriptions of Uromyces aecidiiformi (Rees, 1917, Zhuang, 2005). The primer pair LR0R (Moncalvo et al., 1995)/LR5 (Vilgalys & Hester, 1990) was utilized for amplifying and sequencing the large subunit of the nuclear ribosomal RNA genes from strains IS909-3 and IS1816 (GenBank PQ008482, PQ008483). The obtained sequences showed a high similarity of 99.9% to 100% similarity to strains U1023 and UBC19 of U. aecidiiformis in RustHubb (KR0014142 and PUN23000)( Kaishian et al., 2024). Through examination of morphology, host range, and sequence similarity, we determined the rust species to be U. aecidiiformis. Pathogenicity testing was conducted by spraying a suspension of aeciospores (1×105 spores/mL in 0.05% Tween 20 solution) on six healthy four-year-old F. unibracteata plants indoors in May 2023. The plants were allowed to grow under natural conditions, where the diurnal temperature ranged from 9 to 20℃, with an average temperature of 14℃, which is conducive to the growth of F. unibracteata. Another six seedlings were sprayed with 0.05% Tween 20 solution as controls. After three weeks, all infected plants showed symptoms similar to those seen in the field, while control plants remained symptom-free. Microscopic examination and sequencing confirmed that the pathogen morphology was consistent between the field and the inoculation, meeting Koch\'s postulates. Although U. aecidiiformis has been previously reported to cause rust of F. pallidiflora and F. ussuriensis(Zhuang, 1989, Zhuang, 2005), this is the first report of U. aecidiiformis causing rust on F. unibracteata in China. This pathogen significantly reduces the yield and quality of Chuanbeimu, highlighting the importance of effectively identifying and controlling it.
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  • 文章类型: Journal Article
    艾蒿是一种原产于东亚的多年生草本植物。它是一种重要的传统中药植物,以其浓郁的风味和药用作用而闻名。它含有丰富的活性成分,具有广泛的生物活性,包括消炎药,抗氧化剂,和免疫调节特性。2023年5月至7月,唐河县几个农场(约200亩)的艾吉发生了严重的叶腐病暴发(北纬32°46\'44\”,112°43'13''E),河南省,中国。发病率达到65%(n=200)。淡黄色斑点(直径1-2厘米)首先出现在叶子上,然后扩张形成不规则的黄褐色病变,最终导致整个叶子枯萎。收集患病的叶(30),并在感染和健康组织之间的区域中切成5×5mm2的片。将切下的植物组织在75%乙醇和1%次氯酸钠溶液中灭菌30秒零1分钟,分别。然后用无菌水冲洗组织并置于马铃薯葡萄糖琼脂(PDA)上,然后在25°C下孵育3天。分离的菌株属于镰刀菌属和链格孢属。致病性验证后,获得了25个纯化的镰刀菌菌株。三个代表性菌株(AC-Q,AC-X,来自不同地区的AC-Y)用于进一步研究。每个菌株都形成了丰富的气生菌丝体,最初是白色的,后来发展成紫色的色素。空中分生孢子稀疏分枝,以轮生phialides终止。大分生孢子纤细,直,测量为21.8至47.5×3.1至4.4μm,有两到四个隔片。微分生孢子为棒酸盐,测量值为8.31至11.6×2.1至3.5μm。形态特征与镰刀菌的物种描述一致(Sacc。)尼伦伯格1976年(莱斯利和萨姆梅雷尔,2006).rDNA内部转录间隔区(ITS),β-微管蛋白基因(tub2),翻译延伸因子1-α基因(tef1),calmodulin(cmdlet),扩增RNA聚合酶II最大亚基(rpb1)和RNA聚合酶II第二最大亚基(rpb2)以进行分子鉴定(O'Donnell等人。,2022年)。序列被保存在GenBank中,登录号为OR960548,OR960552,OR960555(ITS),OR972413,OR972414,OR972415(tub2),OR797685,OR797686,OR797687(tef1),OR972410,OR972411,OR972412(cmdlet),PP035106,PP035107,PP035108(rpb1),和PP035109、PP035110、PP035111(rpb2)。AC-Q序列的BLASTn分析显示出99%至100%的相似性与轮虫的序列(菌株CBS576.78)MT010888,rpb1的MT0109566和rpb2的MT010972。用串联序列构建了一个系统发育树(tub2,tef1,cmdlet,rpb1,rpb2),与使用邻居连接方法的类型菌株的序列一起。这三个菌株与轮虫的CBS576.78型菌株形成了进化枝,并与其他镰刀菌分离。这些形态学和分子鉴定表明该病原体为轮虫。通过用分生孢子悬浮液(106分生孢子ml-1)喷洒10个健康的2个月大的盆栽幼苗来测试致病性,和5个幼苗用灭菌水喷雾作为对照。将植物置于28°C和约90%的相对湿度的气候培养箱中。幼苗接种后十天,在处理过的植物上观察到典型的病变,除了对照组。通过形态学和分子鉴定,将重新分离的菌株鉴定为轮虫。履行科赫的假设。已知轮虫会导致玉米镰刀菌穗腐烂,以及中国其他植物的疾病,如芸苔属植物(Akram等人。,2020年)和荆棘(李等人。,2024).这是在世界范围内有关黄曲霉引起叶腐病的第一份报告。病原体的鉴定对于实施减少产量损失的管理方法至关重要。
    Artemisia argyi is a perennial herb native to East Asia. It is an important traditional Chinese medicinal plant known for its strong flavor and medicinal effects. It is rich in active ingredients and has a wide range of biological activities, including anti-inflammatory, antioxidant, and immune regulation properties. From May to July in 2023, a serious leaf rot outbreak occurred on A. argyi in several farms (approximately 200 acres) in Tanghe county (32°46\'44\" N, 112°43\'13\" E), Henan Province, China. The incidence rate reached 65% (n=200). Pale yellow spots (1-2 cm in diameter) first appeared on the leaves, then expanded to form irregular yellowish-brown lesions, eventually causing the entire leaves to wither. Diseased leaves (30) were collected and cut into 5 x 5 mm2 pieces in the areas between infected and healthy tissues. The excised plant tissues were sterilized in 75% ethanol and 1% sodium hypochlorite solution for 30 seconds and one minute, respectively. The tissues were then rinsed with sterile water and placed on potato dextrose agar (PDA) followed by incubating at 25 °C for 3 days. The isolated strains belonged to the genera Fusarium and Alternaria. After pathogenicity verification, 25 purified Fusarium strains were obtained. Three representative strains (AC-Q, AC-X, AC-Y) from different regions were used for further studies. Each strain formed abundant aerial mycelium that was initially white and later developed into purple pigments. Aerial conidiophores were sparsely branched, terminating with verticillate phialides. Macroconidia were slender, straight, and measured 21.8 to 47.5 × 3.1 to 4.4 μm, with two to four septa. Microconidia were clavate and measured 8.31 to 11.6 × 2.1 to 3.5 μm. Morphological characteristics were consistent with the species description of Fusarium verticillioides (Sacc.) Nirenberg 1976 (Leslie and Summerell, 2006). The rDNA internal transcribed spacer (ITS), β-tubulin gene (tub2), translation elongation factor 1-alpha gene (tef1), calmodulin (cmdA), RNA polymerase II largest subunit (rpb1) and RNA polymerase II second largest subunit (rpb2) were amplified for molecular identification (O\'Donnell et al., 2022). The sequences were deposited in GenBank with accession Nos. OR960548, OR960552, OR960555 (ITS), OR972413, OR972414, OR972415 (tub2), OR797685, OR797686, OR797687 (tef1), OR972410, OR972411, OR972412 (cmdA), PP035106, PP035107, PP035108 (rpb1), and PP035109, PP035110, PP035111 (rpb2). BLASTn analysis of AC-Q sequences exhibited 99 to 100% similarity with F. verticillioides sequences (strains CBS 576.78) MT010888 of cmdA, MT0109566 of rpb1, and MT010972 of rpb2. A phylogenetic tree was constructed with concatenated sequences (tub2, tef1, cmdA, rpb1, rpb2), alongside the sequences of the type strains using the neighbor-joining method. The three strains formed a clade with the type strain CBS 576.78 of F. verticillioides, and were separated from other Fusarium spp. These morphological and molecular identifications indicated that the pathogen was F. verticillioides. Pathogenicity was tested on 10 healthy 2-month-old potted seedlings by spraying them with a conidial suspension (106 conidia ml-1), and 5 seedlings were sprayed with sterilized water as a control. The plants were placed in a climate incubator at 28°C and a relative humidity of approximately 90%. Ten days after seedling inoculation, typical lesions were observed on the treated plants, except in the control group. The reisolated strains were identified as F. verticillioides by morphological and molecular characterization, fulfilling Koch\'s postulates. F. verticillioides is known to cause Fusarium ear rot on maize, as well as diseases on other plants in China such as Brassica rapa (Akram et al., 2020) and Schizonepeta tenuifolia (Li et al., 2024). This is the first report of F. verticillioides causing leaf rot on A. argyi worldwide. Identification of the pathogen is crucial for implementing management approaches to reduce yield losses.
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  • 文章类型: Journal Article
    天瓜基里洛威格言。(葫芦科),一种中草药,是安徽省重要的经济作物,中国。近年来,树胶性干枯萎病,一种主要的瓜类疾病,在许多凯里洛威种植园中很普遍。自然感染的茎上的最初症状表现为深棕色的水浸病变,随着疾病的发展,凯里洛威的葡萄藤逐渐枯萎。在树叶上,最初可见棕色水浸的病变,然后病变扩大并合并,导致叶子广泛坏死。关于水果,被白色菌丝体覆盖的病变几乎是圆形的,最初是棕褐色。随后,患病的果实变黑腐烂,俗称果腐病或黑腐病。从有症状的茎中始终分离出Stagonosporopsis样生物,叶子和水果。真菌分离株最初是白色的,后来变成深灰色或黑色,在PDA培养基上具有羊毛感,以絮凝气生菌丝体。选择了来自不同种植园的24个分离株进行进一步的形态学研究。在黄瓜果实上接种3天后,形成了分生孢子和分生孢子。比尼迪亚呈球状至亚球状,棕色,卵形和106.7至213.6μm(平均160.1μm,直径n=50)。分生孢子是透明的,椭圆体,无盐或单隔膜,在隔膜处略微收缩,6.1~13.6×3.5~4.8μm(平均9.9×4.1μm,n=50),并含有两个或更多的油滴。基因组DNA的三个不同位点,包括核核糖体DNA内部转录间隔区(ITS),RNA聚合酶II第二大亚基(RPB2),和β-微管蛋白(TUB2)基因。,使用引物ITS1/ITS4扩增(White等人。1990),RBP2DF/RBP2DR(Lawrenceetal.2013),和T1/β-Sandy-R(O\'Donnell和Cigelnik1997;Stukenbrock等人。2012),分别和测序。在分析ITS的基础上,建立了系统进化树,存放在GenBank中的RPB2和TUB2序列(用于ITS的MW485497-MW485502,MW531661-MW531666用于RPB2,MW531667-MW531672用于TUB2),使用最大似然法。系统发育树表明,分离株与葫芦枯菌一起落入一个分支中。根据形态和分子特征,从赤霉病中获得的分离株被鉴定为南瓜孢霉。对4周龄的木氏T.kirilowii幼苗的茎和叶以及从成年木氏T.kirilowii植物收集的未成熟果实进行致病性测试。表皮,以前用注射器针头受伤,接种直径为5毫米的菌丝体栓,然后将接种区域包裹在浸水的棉花中。用琼脂栓类似地接种对照。在两个垂直方向上测量病变的直径。在PDA培养基上进行从茎和叶病变的再分离。南瓜孢霉,根据其菌落和分生孢子特征重新鉴定,因此,完成了科赫的假设。据报道,在许多宿主中都有由葫芦枯病引起的软糖茎枯病,包括黄瓜,Luffa,南瓜,葫芦,甜瓜,哈密瓜,和西瓜(Jiang等人。2015年;基纳特2011年;赵等人。2019)。据我们所知,这是我国首次报道由葫芦未树引起的赤霉病。该研究为制定和实施有效的管理策略提供了依据。对4周龄的木氏T.kirilowii幼苗的茎和叶以及从成年木氏T.kirilowii植物收集的未成熟果实进行致病性测试。表皮,以前用注射器针头受伤,接种直径为5毫米的菌丝体栓,然后将接种区域包裹在浸水的棉花中。对对照进行类似处理,但是用琼脂栓接种。在两个相互垂直的方向上测量病变的直径。在PDA培养基上重新分离病灶,并根据其菌落和分生孢子特征重新鉴定,以完成科赫的假设。据报道,在许多宿主中都有由葫芦枯病引起的软糖茎枯病,包括黄瓜,Luffa,南瓜,葫芦,甜瓜,哈密瓜,和西瓜(Jiang等人。2015年;基纳特2011年;赵等人。2019)。据我们所知,这是我国首次报道由葫芦未树引起的赤霉病。该研究为制定和实施有效的管理策略提供了依据。
    Trichosanthes kirilowii Maxim. (Cucurbitaceae), one of the Chinese herbal medicines, is an economically important crop in Anhui Province, China. In recent years, gummy stem blight disease, a major disease of cucurbits, was widespread in many T. kirilowii plantations. The initial symptoms on the naturally infected stems appeared as dark brown water-soaked lesions, and as the disease progressed, vines of T. kirilowii gradually withered. On leaves, brown water-soaked lesions were visible initially, and then lesions enlarged and coalesced, resulting in extensive necrosis of leaves. On fruit, lesions covered with the white mycelium were nearly circular and tan to brown initially. Subsequently, the diseased fruit turned black and rotten commonly known as fruit rot or black rot. A Stagonosporopsis-like organism was consistently isolated from symptomatic stems, leaves and fruits. Fungal isolates were initially white and later turned dark grey or black with woolly to floccose aerial mycelium on PDA medium. Twenty-four isolates from different plantations were selected for further morphological studies. Pycnidia and conidia were formed after inoculating on cucumber fruit for 3 days. Pycnidia were globose to sub-globose, brown, ostiolate and 106.7 to 213.6 μm (average 160.1 μm, n = 50) in diameter. Conidia were hyaline, ellipsoidal, aseptate or one-septate, slightly constricted at the septa, 6.1 to 13.6 × 3.5 to 4.8 μm (average 9.9 × 4.1 μm, n = 50), and contained two or more oil drops. Three different loci of the genomic DNA, including the nuclear ribosome DNA internal transcribed spacer (ITS), RNA polymerase II second-largest subunit (RPB2), and β-tubulin (TUB2) genes., were amplified using primers ITS1/ITS4 (White et al. 1990), RBP2DF/RBP2DR (Lawrence et al. 2013), and T1/β-Sandy-R (O\' Donnell and Cigelnik 1997; Stukenbrock et al. 2012), respectively and sequenced. A phylogenetic tree was built based on analysis of ITS, RPB2, and TUB2 sequences that deposited in GenBank (MW485497-MW485502 for ITS, MW531661-MW531666 for RPB2, and MW531667-MW531672 for TUB2), using the maximum likelihood method. The phylogenetic tree showed that the isolates fell into a single clade with S. cucurbitacearum. On the basis of morphological and molecular characteristics, the isolates obtained from T. kirilowii were identified as Stagonosporopsis cucurbitacearum. Pathogenicity tests were carried out on stems and leaves of 4-week-old T. kirilowii seedlings and on immature fruit collected from adult T. kirilowii plants. The epidermis, previously injured with a syringe needle, was inoculated with 5-mm-diameter mycelial plugs, and the inoculated areas were then wrapped in water-soaked cotton. Controls were similarly inoculated with agar plugs. The diameters of lesions were measured in two perpendicular directions. Re-isolations from the stem and leaf lesions were performed on the PDA medium. Stagonosporopsis cucurbitacearum, was re-identified based on its colony and conidial characteristics and, therefore, completed Koch\'s postulates. Gummy stem blight caused by S. cucurbitacearum has been reported in a wide range of hosts, including cucumber, luffa, pumpkin, gourd, muskmelon, cantaloupe, and watermelon (Jiang et al. 2015; Keinath 2011; Zhao et al. 2019). To our knowledge, this is the first report of gummy Stem blight disease on T. kirilowii caused by S. cucurbitacearum in China. The research provides a basis for the development and implementation of effective management strategies. Pathogenicity tests were carried out on stems and leaves of 4-week-old T. kirilowii seedlings and on immature fruits collected from adult T. kirilowii plants. The epidermis, previously injured with a syringe needle, was inoculated with 5-mm-diameter mycelial plugs, and the inoculated areas were then wrapped in water-soaked cotton. Controls were treated similarly but inoculated with agar plugs. Diameters of lesions were measured in two mutually perpendicular directions. Reisolations from the lesions were performed on PDA medium, and was re-identified based on its colony and conidial characteristics to complete Koch\'s postulates. Gummy stem blight caused by S. cucurbitacearum have been reported in a wide range of hosts, including cucumber, luffa, pumpkin, gourd, muskmelon, cantaloupe, and watermelon (Jiang et al. 2015; Keinath 2011; Zhao et al. 2019). To our knowledge, this is the first report of gummy Stem blight disease on T. kirilowii caused by S. cucurbitacearum in China. The research provides a basis for the development and implementation of effective management strategies.
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  • 文章类型: Journal Article
    太白贝母(贝母)是一种贝母,因其镇咳而在中药中常用,祛痰药,和抗高血压特性。2021年和2022年4月,发黄或发紫的发生率为10-30%,枯萎,万源太白北木有死亡症状,四川省。感染的根和鳞茎显示出从棕色到黑色的斑点,还有坏死的腐烂.在严重的情况下,整个灯泡都腐烂了。将15个有症状的灯泡切成0.5×0.5厘米的碎片,在无菌条件下,在75%乙醇中表面灭菌30s,在1%次氯酸钠中灭菌3分钟,用无菌水冲洗3次,空气干燥。将片段置于马铃薯葡萄糖琼脂(PDA)上,并在25°C下在黑暗中孵育7天。获得了六个Clonostachys样单孢子分离株。在25℃黑暗中7天,PDA上的菌落直径达到32至43毫米,毛状至绒毛至颗粒状气生菌丝体,具有白色或浅黄色外观,和反向颜色匹配。在玉米粉-葡萄糖琼脂上,初级分生孢子具有1至3级的黄萎病样结构。柱长36.1至236.3μm。phialides形成2至5,15.3至45.7μm长的螺纹,底座宽1.1~3.4μm,和1.03至2.41μm宽的近开口(n=95)。每个都产生一小滴透明分生孢子。分生孢子为3.7至11.3μm×2.1至4.1μm(n=110)。次级分生孢子显示青霉样结构,柱长23.1至142.3μm。phialides形成于每个metula4至8的压缩螺纹中,长度为7.0~16.0μm,底座宽度为1.3~3.1μm,最宽点为1.8至3.6μm,和0.8至1.8μm附近的开口(n=50)。分生孢子为3.0至6.4μm×1.6至3.4μm(n=65)。形态与先前对Clonostachysrosea的描述一致(Hans-Josef等人。1999).ATP柠檬酸裂解酶(ACL1),β-微管蛋白(TUB2),平移延伸因子1-α(tef1α),并使用引物acl1-230up/acl1-1220low对三个菌株的核核糖体内部转录间隔区(ITS)进行了扩增和测序(Gräfenhan等人。2011),T1/CYLTUB1R(Crous等人2004;O\'Donnell和Cigelnik1997),EF1-728F/EF2(Carbone和Kohn1999;O'Donnell等人。1998),和ITS1/ITS4(White等人1990),分别。Blastn同源性检索显示与玫瑰梭菌的前型菌株(CBS710.86)的相似性>97%。所有序列均已保存在GenBank(PP394342至PP394350和PP396901至PP396903)中。基于组合的ACL1,TUB2,TEF1α的比对,使用贝叶斯分析构建了系统发育树,和通过智商树的ITS序列。该树显示与已知的C.rosea菌株的聚类。通过用菌株WYEB1101的孢子悬浮液(1.0×106孢子mL-1)接种五株健康的5岁的太白北木植物来确认致病性,同时使用无菌水作为对照。接种过程包括将孢子悬浮液倒在受伤的球茎上并用无菌土壤覆盖。随后,所有植物均在适宜太白北木的自然条件下在室内无菌土壤中栽培。致病性测定重复两次。培养20天后,受感染的植物表现出类似于田间观察到的症状,而所有对照植物保持无症状。测序证实了从接种的植物中重新分离出玫瑰梭菌,满足科赫的假设。以前有报道称Clonostachysrosea会导致中草药的根腐病,如黄芪和天麻(Lee等人。2020年;齐等人。2022年)。据我们所知,这是中国首例玫瑰梭菌感染太白北木的报道,突出了这种作物的潜在风险。
    Taibai Beimu (Fritillaria taipaiensis) is a species of Fritillaria commonly used in traditional Chinese medicine for its antitussive, expectorant, and antihypertensive properties. In April of 2021 and 2022, an incidence 10-30% of yellowing or purpling, wilting, and dying symptoms was observed on Taibai Beimu in Wanyuan, Sichuan province. Infected roots and bulbs displayed spots ranging from brown to black, along with necrotic rot. In severe cases, the entire bulbs rotted. Fifteen symptomatic bulbs were cut into 0.5 × 0.5 cm pieces, surface sterilized in 75% ethanol for 30 s and 1% sodium hypochlorite for 3 min under aseptic conditions, rinsed with sterile water 3 times, and air-dried. The segments were placed on potato dextrose agar (PDA) and incubated at 25℃ for 7 days in the dark. Six Clonostachys-like monospore isolates were obtained. Colonies on PDA reached 32 to 43 mm in diameter in 7 days at 25℃ in the dark, felty to tomentose to granulose aerial mycelia with a white or light yellow appearance, and reverse colors matching. On cornmeal-dextrose agar, primary conidiophores had a Verticillium-like structure with 1 to 3 levels. Stipes were 36.1 to 236.3μm long. Phialides formed in whorls of 2 to 5, 15.3 to 45.7μm long, 1.1 to 3.4μm wide at the base, and 1.03 to 2.41μm wide near opening (n=95). Each producing a small hyaline drop of conidia. Conidia were 3.7 to 11.3μm × 2.1 to 4.1μm (n=110). Secondary conidiophores displayed Penicillium-like structures, and stipes were 23.1 to 142.3μm long. Phialides formed in compressed whorls of 4 to 8 per metula, 7.0 to 16.0μm in length, 1.3 to 3.1μm in width at the base, 1.8 to 3.6μm at the widest point, and 0.8 to 1.8μm near opening (n=50). Conidia were 3.0 to 6.4μm ×1.6 to 3.4μm (n=65). The morphology was consistent with the previous description of Clonostachys rosea (Hans-Josef et al. 1999). The ATP citrate lyase (ACL1), β-tubulin (TUB2), translation elongation factor 1-α (tef1α), and the nuclear ribosomal internal transcribed spacer (ITS) of three strains were amplified and sequenced using primers acl1-230up/acl1-1220low (Gräfenhan et al. 2011), T1/CYLTUB1R (Crous et al. 2004; O\'Donnell and Cigelnik 1997), EF1-728F/EF2 (Carbone and Kohn 1999; O\'Donnell et al. 1998), and ITS1/ITS4 (White et al. 1990), respectively. Blastn homology search showed a > 97% similarity to the ex-type strains of C. rosea (CBS710.86). All sequences have been deposited in GenBank (PP394342 to PP394350, and PP396901 to PP396903). A phylogenetic tree was constructed using Bayesian analysis based on the alignment of the combined ACL1, TUB2, tef1α, and ITS sequences through IQ-TREE. The tree displayed clustering with known strains of C. rosea. Pathogenicity was confirmed by inoculating five healthy five-year-old Taibai beimu plants with a spore suspension (1.0 × 106 spores mL-1) of the strain WYEB1101, while sterilized water was used as a control. The inoculation process involved pouring the spore suspension over the wounded bulbs and covering with them sterile soil. Subsequently, all plants were cultivated in sterile soil indoors under natural conditions suitable for Taibai beimu. The pathogenicity assays were repeated twice. After 20 days of cultivation, the infected plants displayed symptoms similar to those observed in the field, while all control plants remained asymptomatic. Sequencing confirmed the re-isolation of C. rosea from the inoculated plants, satisfying Koch\'s hypothesis. Clonostachys rosea has been previously reported to cause root rot of Chinese medicine herb, such as Astragalus membranaceus and Gastrodia elata (Lee et al. 2020; Qi et al. 2022). To our knowledge, this is the first report of C. rosea infecting Taibai Beimu in China, highlighting a potential risk to this crop.
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  • 文章类型: Case Reports
    羟尿症是治疗血液系统疾病的常用药物,但是这种药物引起的下肢溃疡通常很少见,也不会经常被怀疑。我们报道了一名老年患者,患有由羟基脲治疗原发性血小板增多症引起的下肢溃疡。当使用氢氧化物时,密切观察皮肤损伤和迅速处理任何皮肤破裂应防止溃疡。
    Hydroxyuria is a common medication for treating blood system diseases, but ulcers in the lower limbs caused by this medication are often rare and not often suspected. We reported an elderly patient with lower limb ulcers caused by hydroxyurea treatment for primary thrombocytosis. When hydroxide is used, close observation of skin lesions and prompt handling of any skin disruption should prevent ulcers.
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  • 文章类型: Journal Article
    金丝桃在切花和流行花市场上的消费者中越来越受欢迎,因为它是一种观赏木本植物,因为它的花朵浆果和五颜六色的花朵。2019年8月,在昆明的三个商业苗圃(25°05N,102°72'E),云南省,中国。在种植一年后,在大约40%的植物上观察到疾病症状,并且30%的叶被感染。叶子症状开始时很小,水浸在幼叶上的病变后来变得更大,深棕色和坏死。病灶直径0.2~2.8cm。对于病原体分离,从四个不同的苗圃收集了三个有症状的叶子样本。叶子被切成0.5毫米的碎片,表面用70%乙醇灭菌30秒,和3%NaOCl持续5分钟,在无菌蒸馏水中漂洗三次,并铺在马铃薯葡萄糖琼脂(PDA)上(Zhou等人。2023年)。将板在26°C下在黑暗中孵育3天。获得了8个具有可比形态特征的分离株。最初,菌落产生浅灰色至白色的气生菌丝体,5天后变成深灰色。分离物产生透明的,单细胞,直的和圆柱形的分生孢子,平均尺寸为9.7至14.8μm长×3.7至5.6μm宽(n=100)。形态特征与炭疽菌一致。(Bailey和Jeger,1992)。对于分子分析,从三个代表性分离株(XSD1,XSD3和XSD5)中提取基因组DNA,使用引物ITS1/ITS4扩增(Yin等人。2012)和T1/Bt2b(Glass和Donaldson1995),并提交测序(Weir等人。2012).分离物XSD2、XSD3和XSD8的DNA序列是相同的。代表性分离株XSD2的DNA序列保存在GenBank中(登录号。用于ITS的MW202334,和OR347007用于TUB2)。ITS和TUB2序列的MegaBLAST分析显示与C.kahawae菌株ICMP18539具有99.5%和99.3%的相似性(登录号用于ITS的NR_120138.1)和菌株IMI319418(用于TUB2的JX145227.1)。通过在大田健康的黄连成熟叶片上接种病原体进行致病性试验。通过喷洒分离株XSD1,XSD3和XSD5的分生孢子悬浮液(106个孢子/ml)接种十片叶子(两片叶子/植物),并用塑料袋覆盖以保持高湿度48小时,分别。用无菌蒸馏水处理的叶作为对照。所有接种的叶子在接种后10天在23±5°C下表现出与田间观察到的症状相似的症状。在未接种的叶子上没有出现症状。从接种的病叶中重新分离出病原体,并根据形态和分子特征鉴定为C.kahawe。据报道,C.kahawe在意大利的栽培火箭上引起叶斑病(Garibaldi等人。2016),和哥伦比亚树番茄上的炭疽病(Rojas等人。2018),根据我们的知识,这是C.kahawae在世界范围内引起炭疽病的第一份报告。由于黄连具有重要的观赏价值和经济价值,需要调查和监测C.kahawe的分布情况,以便制定有效的疾病管理策略.
    Hypericum chinensis is growing in popularity amongst consumers in cut-flower and pop-flower market as an ornamental woody plant for its florid berry and colorful flower. In August 2019, a new leaf spot disease was observed on H. chinensis in three commercial nurseries in Kunming (25°05\'N, 102°72\'E), Yunnian province, China. Disease symptoms were observed on approximately 40% of the plants one year after planting and 30% of the leaves were infected. Leaf symptoms began as small, water-soaked lesions on young leaves which later became larger, dark brown and necrotic. The lesion size ranged from 0.2 to 2.8 cm in diameter. For pathogen isolation, three samples of symptomatic leaves were collected from four different nurseries. The leaves were cut into 0.5 mm pieces, surface sterilized using 70% ethanol for 30 s, and 3% NaOCl for 5 min, rinsed three times in sterilized distilled water and plated on potato dextrose agar (PDA) (Zhou et al. 2023). The plates were incubated at 26°C in the dark for 3 days. Eight isolates with comparable morphological characteristics were obtained. Initially, colonies produced pale gray to white aerial mycelia, turning dark gray after 5 days. The isolates produced hyaline, single celled, straight and cylindrical conidia, with mean size 9.7 to 14.8 μm long × 3.7 to 5.6 μm wide (n = 100). Morphological characteristics were consistent with Colletotrichum sp. (Bailey and Jeger 1992). For molecular analysis, genomic DNA was extracted from three representative isolates (XSD1, XSD3 and XSD5), amplified using the primers ITS1/ITS4 (Yin et al. 2012) and T1/Bt2b (Glass and Donaldson 1995) and submitted to sequencing (Weir et al. 2012). DNA sequences of the isolates XSD2, XSD3 and XSD8 were identical. DNA sequences of a representative isolate XSD2 were deposited in GenBank (accession no. MW202334 for ITS, and OR347007 for TUB 2). MegaBLAST analysis of the ITS and TUB2 sequences showed 99.5% and 99.3% similarity with C. kahawae strain ICMP 18539 (accession no. NR_120138.1 for ITS) and strain IMI319418 (JX145227.1 for TUB 2). Pathogenicity tests were conducted by inoculating the pathogen on healthy mature leaves of H. chinensis in the field. Ten leaves (two leaves/plant) were inoculated by spraying conidial suspension (106 spores/ml) of isolates XSD1, XSD3 and XSD5, and covered with plastic bags to maintain high humidity for 48 hours, respectively. Leaves treated with sterile distilled water served as a control. All inoculated leaves showed symptoms similar to those observed in the field at 23±5°C 10 days after inoculation. No symptoms developed on non-inoculated leaves. The pathogen was re-isolated from inoculated diseased leaves and identified as C. kahawae based on morphological and molecular characters. C. kahawae has been reported to cause leaf spot on cultivated rocket in Italy (Garibaldi et al. 2016), and anthracnose disease on tree tomato in Colombia (Rojas et al. 2018), to our knowledge, this is the first report of C. kahawae causing anthracnose on H. chinensis worldwide. Due to important ornamental and economic value of H. chinensis, the distribution of C. kahawae needs to be investigated and monitored for effective disease management strategies to be developed.
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  • 文章类型: Journal Article
    Macleayacordata(威尔德。)R.Br。是在中国商业种植的多年生草本植物(罂粟科),已经研究了消肿,排毒,和杀虫效果(Lin等人2018)。2021年8月,本溪县2岁的M.cordata植物中发现了炭疽病,中国东北(北纬41°45'48″,123°69\'15“E)。在每个病叶上观察到数十个不规则的红棕色斑点(3-11毫米)。病灶上覆盖一层灰白色菌丝体。随着疾病的发展,斑点变成坏死和穿孔,或者它们会合并成大的病变,最终导致枯萎的叶子(图。1).2021年,16公顷土地中超过33%的植物被感染。收集患病的叶子,切成3-8毫米的碎片,通过将其浸入1%NaOCl中2分钟进行表面消毒,用无菌蒸馏水冲洗三次。然后用消毒的吸水纸干燥,置于用氯霉素(40mg/L)修正的PDA培养基上,并在25°C的黑暗中以12小时的光周期孵育。获得了20个分离物(BLH1至20),并使用单孢子方法进行了纯化。鉴定分离物BLH12并用于致病性测试。菌落稀疏蓬松,边缘光滑,从最初的白色逐渐变成灰色到浅橙色。殖民地的下侧朝向中心呈浅橙色。分生孢子是单细胞的,圆柱形,透明,两端钝,测量值(15.13±1.14)×(5.80±0.60)μm(n=50)。Pressoria是单细胞的,棕色到深棕色,通常是椭圆形或不规则的,有时是浅叶。未观察到油层。该分离物最初被鉴定为炭疽菌复合物(Prihastuti等人。2009).鉴定如前所述(Weir等人。2012).rDNA内部转录间隔区(OP415560),甘油醛-3-磷酸脱氢酶(OP433642),几丁质合成酶(OP433643),钙调素(OP433644),肌动蛋白(OP433645),谷氨酰胺合成酶(OP433646),β-微管蛋白(OP433647),获得了超氧化物歧化酶(OP433648)基因序列(Carbone&Kohn1999;Weir等人。2012),BLAST搜索显示与类型培养物ICMP18608(JX010244,JX010044,JX009683,JX009443,JX009744,JX010078,JX010389和JX010311)具有99-100%的同源性。结合所有基因座的系统发育分析表明BLH12和C.aenigma的类型菌株聚集在一组中(图2).基于形态特征和系统发育关系,BLH12被鉴定为C.aenigma。对于致病性测试,健康的2岁植物用BLH12孢子悬浮液(1×105/mL)喷雾。对照植物用无菌水喷雾。每个处理有三个重复(每个五株植物)。将所有植物在25°C(12小时光周期和86%相对湿度)下孵育并在7天后检查。该实验重复两次。接种的植物在叶片表面显示出病变,与该领域的类似,而对照植物无症状。按照上述方法成功地重新分离和鉴定了病原体。据报道,这种真菌感染了中国许多木本植物的叶子(Wang等人。2020年;张等人。2021)。这是C.aenigma在M.cordata上引起炭疽病的第一份报告,这将为开发该疾病的有效现场控制实践提供指导。
    Macleaya cordata (Willd.) R. Br. is a perennial herbaceous medicinal plant (Papaveraceae) commercially cultivated in China which has been studied for detumescence, detoxification, and insecticidal effect (Lin et al. 2018). In August 2021, anthracnose was observed in 2-year-old M. cordata plants in Benxi county, northeast China (41°45\'48″N, 123°69\'15″E). Dozens of irregular reddish-brown spots (3-11 mm) were observed on each diseased leaf. The lesions were covered with a layer of gray-white mycelia. As the disease progressed, the spots became necrosis and perforation or they would merged into large lesions, ultimately resulting in wilted leaves (Fig. 1). More than 33% of the plants in a 16-ha field were infected in 2021. The diseased leaves were collected and cut into 3-8 mm pieces, surface-disinfested by immersing them into 1% NaOCl for 2 min, and rinsed three times with sterile distilled water. They were then dried with sterilized absorbent paper, placed on PDA medium amended with chloramphenicol (40 mg/L), and incubated in darkness at 25°C with a 12-h photoperiod. Twenty isolates (BLH1 to 20) were obtained and purified using a single-spore method. Isolate BLH12 was identified and used for the pathogenicity test. Colonies were sparsely fluffy with smooth edges, and gradually became gray to pale orange from the initial white. The underside of the colonies was pale orange towards the center. Conidia were single-celled, cylindrical, and transparent with broadly blunt ends, measuring (15.13 ± 1.14) × (5.80 ± 0.60) μm (n=50). Appressoria were single-celled, brown-to-dark brown, usually elliptical or irregular, and sometimes lobed. Setae were not observed. The isolate was initially identified as Colletotrichum gloeosporioides complex (Prihastuti et al. 2009). The identification was confirmed as described previously (Weir et al. 2012). The rDNA internal transcribed spacer region (OP415560), the glyceraldehyde-3-phosphate dehydrogenase (OP433642), chitin synthase (OP433643), calmodulin (OP433644), actin (OP433645), glutamine synthetase (OP433646), β-tubulin (OP433647), and superoxide dismutase (OP433648) gene sequences were obtained (Carbone & Kohn 1999; Weir et al. 2012), and BLAST searches revealed 99-100% homology with the type culture ICMP 18608 (JX010244, JX010044, JX009683, JX009443, JX009744, JX010078, JX010389, and JX010311). A phylogenetic analysis of combining all loci indicated BLH12 and the type strain of C. aenigma were clustered in one group (Fig. 2). Based on the basis of morphological characteristics and phylogenetic relationships, BLH12 was identified as C. aenigma. For the pathogenicity test, healthy 2-year-old plants were sprayed with a BLH12 spore suspension (1 × 105/mL). Control plants were sprayed with sterile water.There were three replicates (five plants each) per treatment. All plants were incubated at 25°C (12-h photoperiod and 86% relative humidity) and examined after 7 days. The experiment was repeated twice. The inoculated plants showed lesions on the leaf surface, similar to those in the field, whereas the control plants were asymptomatic. The pathogen was successfully reisolated and identified as the methods mentioned above. This fungus reportedly infects the leaves of many woody plants in China (Wang et al. 2020; Zhang et al. 2021). This is the first report of C. aenigma causing anthracnose on M. cordata, which will provide an guideline for developing effective field control practices for the disease.
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  • 文章类型: Journal Article
    Ligulariafischeri(Ledeb。)是菊科的多年生草药植物,在中国作为药用商业种植,装饰性的,和食用植物。在中国东北本溪县的2岁的费氏乳杆菌中观察到叶斑,2021年8月。在感染的叶子上观察到不规则的红棕色斑点,范围从3到11毫米,每片叶子都有几十个斑点(图。1).随着疾病的发展,病斑枯萎,中心倒塌,多个病变合并成较大的病变点,导致树叶枯萎.在生殖阶段,根和茎基部未被感染。2021年,18公顷土地中超过37%的植物被感染。收集10片患病的叶子,切成小片(3-5毫米),通过浸入1%NaOCl2分钟并用无菌蒸馏水冲洗3次进行表面消毒。然后将叶片置于培养皿中的酸化马铃薯右旋糖琼脂(PDA)上,并在黑暗中在25°C下孵育。获得了20个具有相同形态特征的分离株。通过从从水琼脂收集的单个孢子开始每个分离物的新菌落来进一步纯化分离物。随机选择分离株TYTW7进行鉴定和致病性测试。它生长迅速,并产生大量的气生菌丝体,底面为红色红色。分生孢子有许多肥沃的分支,并形成一个细长的柄,尖端有一个sphaeropereduncular囊泡。单间隔分生孢子是圆柱形的,几乎是直的,具有平行的壁和圆形的末端。它们的大小范围为30.35至51.76×2.93至5.01μm(n=100),病原体最初被鉴定为Calonectriasp。(Crous2002;Crous等人。2004;Lombard等人。2015、2016)。根据已发布的方法(Liu和Chen2017;Shao和Li2021)确定了进一步的鉴定确认。包括翻译延伸因子1-alpha的部分基因区域(GenBank登录号。OP290551),组蛋白H3(OP290552),获得钙调蛋白(OP290553)和β-微管蛋白(OP290554),BLAST搜索显示与前型培养物CERC8952(MF527049,MF527065,MF527081和MF527107)具有99-100%的同源性,结合所有基因座的系统发育分析显示,分离株TYTW7和Ca的类型菌株。蒙大拿州聚集在一组中(图。2).根据形态特征和系统发育分析,分离物TYTW7被鉴定为Ca。蒙大拿州将健康的2年生植物用于致病性测试。使用孢子悬浮液(1×105孢子/mL水)接种三株寄主植物;将无菌水喷洒到作为对照的相同数量的植物上。该实验重复三次。将所有植物在27±2°C(12h光周期)下孵育并在7天后进行评估。接种的植物在叶片表面显示出病变,与该领域的类似,而对照组保持无症状.通过测序成功地重新分离和鉴定了病原体,并且没有从无症状对照植物中分离出病原体。据我们所知,这是Ca的第一份报告。蒙大拿州导致费氏落叶斑病。该疾病对生产构成威胁,需要在管理选择上采取更多的控制策略,以最大程度地减少损失。
    Ligularia fischeri (Ledeb.) is a perennial herbal plant of Compositae that is cultivated commercially in China as a medicinal, ornamental, and edible plant. Leaf spots were observed in 2-year-old L. fischeri in Benxi County of northeast China, in August 2021. Irregular reddish brown spots ranging from 3 to 11 mm were observed on infected leaves, and each leaf had dozens of spots (Fig. 1). As the disease progressed, the diseased spots withered and the centers fell out, and multiple lesions merge into large diseased spots, causing leaf wilting. The roots and stem bases were not infected during the reproductive stage. More than 37% of the plants in a 18 ha field were infected in 2021. The ten diseased leaves were collected and cut into small (3-5 mm) pieces, which were surface-disinfested by immersing into 1% NaOCl for 2 min and rinsing with sterile distilled water three times. The leaf pieces were then placed on acidified potato dextrose agar (PDA) in petri plates and incubated in the dark at 25°C. Twenty isolates with the same morphological characteristics were obtained. Isolates were further purified by starting a new colony for each isolate from a single spore collected from water agar. Isolate TYTW7 was randomly selected for identification and pathogenicity testing. It grew rapidly and produced profuse aerial mycelia with a carmine red underside. The conidiophores had many fertile branches and formed an elongated stipe with a sphaeropedunculate vesicle at the tip. The one-septate conidia were cylindrical and almost straight with parallel walls and rounded ends. Their sizes ranged from 30.35 to 51.76 × 2.93 to 5.01 μm (n = 100) and the pathogens were initially identified as Calonectria sp. (Crous 2002; Crous et al. 2004; Lombard et al. 2015, 2016). Further confirmation of the identification was determined according to published method (Liu and Chen 2017; Shao and Li 2021). The partial gene regions including the translation elongation factor 1-alpha (GenBank accession no. OP290551), histone H3 (OP290552), calmodulin (OP290553) and β-tubulin (OP290554) were obtained, and BLAST searches showed 99-100% homology with the ex-type culture CERC 8952 (MF527049, MF527065, MF527081 and MF527107) and phylogenetic analysis combining all loci revealed that the isolate TYTW7 and the type strain of Ca. montana clustered in one group (Fig. 2). Based on morphological characteristics and phylogenetic analysis, isolate TYTW7 was identified as Ca. Montana. Healthy 2-year-old plants were used for the pathogenicity test. A spore suspension (1×105 spores/mL water) was used to inoculate three host plants; sterile water was sprayed on the same number plants serving as a control. The experiment was repeated three times. All plants were incubated at 27±2°C (12h photoperiod) and were evaluated after seven days. The inoculated plants showed lesions on the leaf surface, similar to those in the field, and the control remained symptomless. The pathogens were successfully reisolated and identified by sequencing, and no pathogens were isolated from symptomless control plants. To our knowledge, this is the first report of Ca. montana causing L. fischeri leaf spot. The disease poses a threat to the production and more control strategies are needed on management options to minimize losses.
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  • 文章类型: Case Reports
    Fournier的坏疽是一种罕见的,迅速进步,生殖器的感染性坏死性筋膜炎的暴发性形式,肛周和会阴区。我们介绍了一例阴茎Fournier坏疽,并发急性生殖器溃疡和复发性副异位症的病例,该病例继发于同期的COVID-19和Mpox感染。对于临床医生来说,了解Fournier的坏疽是很重要的,因为早期检测仍然是有效组织和实际上保护生命管理的基石。
    Fournier\'s gangrene is a rare, rapidly progressive, fulminant form of infective necrotising fasciitis of the genital, perianal and perineal regions. We present a case of Fournier\'s gangrene of the penis complicating acute genital ulceration and recurrent paraphimosis that was secondary to contemporaneous COVID-19 and Mpox infection in an otherwise healthy 41-year-old man. It is important for clinicians to be aware of Fournier\'s gangrene, as early detection remains the cornerstone of effective tissue and indeed life conserving management.
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