BACKGROUND & AIMS: :In 2011, common symptoms of grapevine dieback were frequently observed in 2- to 5-year-old table grape (Vitis vinifera L.) cvs. in four vineyards located in northern Tunisia. The symptoms included dead spur and cordons, shoot dieback, and sunken necrotic bark lesions, which progressed into the trunk resulting in the death of large sections of the vine. Longitudinal and transversal sections of cordons and spurs from symptomatic vines revealed brown wedge-shaped cankers of hard consistency. Twelve symptomatic samples from spur and cordons were collected, surface disinfected by dipping into 5% (v/v) sodium hypochlorite for 2 min, and small pieces from the edge of necrotic and healthy tissue were removed and plated onto potato dextrose agar (PDA) at 25°C in the dark. Based on colony and conidia morphological characteristics, isolates were divided in three species, named Diplodia seriata, Botryosphaeria dothidea, and Neofusicoccum luteum. D. seriata colonies were gray-brown with dense aerial mycelium producing brown cylindric to ellipsoid conidia rounded at both ends and averaged 22.4 × 11.7 μm (n = 50). B. dothidea colonies were initially white with abundant aerial mycelium, gradually becoming dark green olivaceous. Conidia were fusiform to fusiform elliptical with a subobtuse apex and averaged 24.8 × 4.7 μm (n = 50). N. luteum colonies were initially pale to colorless, gradually darkening with age and becoming gray to dark gray producing a yellow pigment that diffuses into the agar. Conidia were hyaline, thin-walled, aseptate, fusiform to fusiform elliptical, and averaged 19.8 × 5.5 μm (n = 50). Identity of the different taxa was confirmed by sequence analyses of the internal transcribed spacer (ITS1-5.8S-ITS2) region of the rDNA and part of the elongation factor 1-alpha (EF1-α) gene. BLAST analysis of sequences indicated that six isolates were identified as D. seriata (GenBank: AY259094, AY343353), one isolate as B. dothidea (AY236949, AY786319) and one isolate as N. luteum (AY259091, AY573217). Sequences were deposited in GenBank under accessions from KC178817 to KC178824 and from KF546829 to KF546836 for ITS region and EF1-α gene, respectively. A pathogenicity test was conducted on detached green shoots cv. Italia for the eight Botryosphaeriaceae isolates. Shoots were inoculated by placing a colonized agar plug (5 mm diameter) from the margin of a 7-day-old colony on fresh wound sites made with a sterilized scalpel. Each wound was covered with moisturized cotton and sealed with Parafilm. Control shoots were inoculated using non-colonized PDA plugs. After 6 weeks, discoloration of xylem and phloem and necrosis with average length of 38.8, 17.6, and 11.2 mm were observed from inoculated shoots with D. seriata, N. luteum, and B. dothidea, respectively, and all three fungi were re-isolated from necrotic tissue, satisfying Koch's postulates. Control shoots showed no symptoms of the disease and no fungus was re-isolated. In Tunisia, Botryosphaeria-related dieback was reported only on citrus tree caused by B. ribis (2), on Pinus spp. caused by D. pinea (4), on Quercus spp. caused by D. corticola (3), and on olive tree (Olea europea) caused by D. seriata (1). To our knowledge, this is the first report of D. seriata, B. dothidea, and N. luteum associated with grapevine dieback in Tunisia. References: (1) M. Chattaoui et al. Plant Dis. 96:905, 2012. (2) H. S. Fawcett. Calif. Citrogr. 16:208, 1931. (3) B. T. Linaldeddu et al. J. Plant Pathol. 91:234. 2009. (4) B. T. Linaldeddu et al. Phytopathol. Mediterr. 47:258, 2008.

背景与目标： ：2011年，在2至5岁的食用葡萄（Vitis vinifera L.）CVS中经常观察到葡萄枯死的常见症状。在突尼斯北部的四个葡萄园中。症状包括死刺和警戒线，枝条枯死和凹陷的坏死性树皮病变，这些病变发展到树干中，导致大片葡萄藤死亡。有症状藤蔓的警戒线和刺的纵向和横向截面显示出褐色的楔形溃疡，具有坚硬的一致性。收集十二个有刺和带状疱疹的有症状样品，通过浸入5％（v / v）次氯酸钠中2分钟进行表面消毒，并去除坏死和健康组织边缘的小块并将其铺在马铃薯葡萄糖琼脂（PDA）上在黑暗中25°C下。根据菌落和分生孢子的形态特征，将分离物分为三种，分别为Diplodia seriata，Botryosphaeria dothidea和Neofusicoccum luteum。 D. seriata菌落为灰褐色，具浓密的气生菌丝体，产生棕色圆柱形到椭圆形分生孢子，两端圆形，平均22.4×11.7μm（n = 50）。 B. dothidea菌落最初是白色的，具丰富的气生菌丝体，逐渐变为深绿色的橄榄色。分生孢子为梭形到梭形椭圆形，先端圆形，平均24.8×4.7μm（n = 50）。黄体猪笼草的菌落最初为浅色至无色，随着年龄的增长逐渐变暗，从灰色变为深灰色，产生黄色颜料，其扩散到琼脂中。分生孢子是透明的，卵形的，椭圆形的，梭形到梭形椭圆形的分生孢子，平均为19.8×5.5μm（n = 50）。通过对rDNA的内部转录间隔区（ITS1-5.8S-ITS2）区和部分延伸因子1-alpha（EF1-α）基因进行序列分析，确认了不同分类单元的身份。序列的BLAST分析表明，鉴定出六种分离株为D. seriata（GenBank：AY259094，AY343353），一种分离株鉴定为B. dothidea（AY236949，AY786319），一种分离株鉴定为黄褐猪笼草（AY259091，AY573217）。 ITS区和EF1-α基因的序列分别从KC178817至KC178824和从KF546829至KF546836保藏在GenBank中。对分离的绿芽cv进行了致病性测试。意大利用于八种葡萄球菌的分离株。通过在7天大的菌落边缘上放置一个定植的琼脂塞（直径5毫米），将其接种在用无菌手术刀制成的新鲜伤口部位上，从而对芽进行接种。每个伤口都覆盖有保湿棉，并用Parafilm密封。使用非定殖的PDA塞接种对照芽。 6周后，接种D. seriata，N。luteum和B. dothidea的枝条观察到木质部和韧皮部的变色和坏死的平均长度分别为38.8、17.6和11.2 mm，并且所有三种真菌均重新接种。从坏死组织中分离出来，符合科赫的假设。对照芽未显示出该疾病的症状，并且没有真菌被重新分离。在突尼斯，据报道仅在松树属物种的核糖双歧杆菌（2）引起的柑橘树上发生了与灰霉病有关的死亡。由D. pinea（4）引起，在Quercus spp上。是由D. corticola（3）引起的，以及在橄榄树（Olea europea）上由D. seriata（1）引起的。据我们所知，这是突尼斯的D. seriata，B。dothidea和N. luteum与葡萄死亡相关的首次报道。参考文献：（1）M. Chattaoui等。植物病。 96：905，2012.（2）H. S. Fawcett。加州Citrogr。 （1931）16：208。（3）B. T. Linaldeddu等。 J.植物病理学。 91：234。 2009.（4）B.T. Linaldeddu等。植物病理学。 Mediterr。 47：258，2008年。

BACKGROUND & AIMS: :Apple ring rot caused by the fungus Botryosphaeria dothidea is one of the devastating diseases. Up to date, the responsive mechanism of apple plant to this disease remains unclear. In the present study, an apple CNGC gene (designated as MdCNGC1) was found among highly expressed genes responding to B. dothidea infection. The expression of MdCNGC1 was different among apple cultivars with different resistance to B. dothidea. Intriguingly, MdCNGC1 expression was not induced by other two apple pathogens, Marssonina coronaria and Valsa ceratosperma. Ectopic overexpression of MdCNGC1 in Nicotiana benthamiana conferred elevated susceptibility to bacterial and fungal pathogens. Notably, overexpression of MdCNGC1 reduced salicylic acid (SA) accumulation induced by Alternaria alternata or Pseudomonas syringae. Decreased induction of pathogenesis-related (PR) genes and ROS accumulation were also observed in MdCNGC1-overexpressing plants. Up-regulated scavenging systems as indicated by enhanced expressions of CAT, APX, SOD genes and activities of antioxidative enzymes may in part contribute to reduced ROS accumulation. MdCNGC1 expression in N. benthamiana also decreased flg22 and chitosan-induced callose deposition and lowered the expression of NbPMR4, an ortholog of Arabidopsis callose synthase gene PMR4. These combined results suggested that MdCNGC1 might be a negative factor to plant resistance to bacterial and fungal pathogens.

背景与目标： 真菌Botryosphaeria dothidea引起的苹果环腐病是毁灭性疾病之一。迄今为止，苹果植物对该病的反应机制仍不清楚。在本研究中，发现苹果CNGC基因（命名为MdCNGC1）是在高表达B. dothidea感染的基因中发现的。 MdCNGC1的表达在苹果品种中对B. dothidea的抗性不同。有趣的是，MdCNGC1的表达没有被其他两种苹果病原体，马氏冠状病毒和樱桃小精子所诱导。在本氏烟草中，MdCNGC1的异位过表达赋予了细菌和真菌病原体更高的易感性。值得注意的是，MdCNGC1的过表达减少了交替链霉菌或丁香假单胞菌引起的水杨酸（SA）积累。在过表达MdCNGC1的植物中也观察到致病相关（PR）基因的诱导减少和ROS积累。 CAT，APX，SOD基因表达增强和抗氧化酶活性表明清除系统上调可能部分有助于降低ROS的积累。在本氏烟草中，MdCNGC1的表达也降低了flg22和壳聚糖诱导的call的沉积，并降低了拟南芥call合成酶基因PMR4的直系同源物NbPMR4的表达。这些综合结果表明，MdCNGC1可能是植物对细菌和真菌病原体抗性的负面因素。

BACKGROUND & AIMS: :Pathogenesis-related protein-4 (PR-4) family is a group of proteins with a Barwin domain in C-terminus and generally thought to be involved in plant defense responses. However, their detailed roles are poorly understood in defense of apple plant against pathogenic infection. In the present study, a new PR-4 gene (designated as MdPR-4) was identified from Malus domestica, and its roles in defense responses of apple were investigated. The open reading frame of MdPR-4 gene is of 447 bp encoding a protein of 148 amino acids with a Barwin domain in C-terminus and a signal peptide of 26 amino acids in N-terminus. Sequence and structural analysis indicated that MdPR-4 protein belongs to class II of PR-4 family. The high-level expression of MdPR-4 was observed in flowers and leaves as revealed by quantitative real time PCR. The temporal expression analysis demonstrated that MdPR-4 expression could be up-regulated by Botryosphaeria dothidea infection and salicylic acid (SA) or methyl jasmonate (MeJA) treatment, but suppressed by diethyldithiocarbamic acid (DIECA). In vitro assays, recombinant MdPR-4 protein exhibited ribonuclease activity specific for single strand RNA and significant inhibition to hyphal growth of three apple pathogenic fungi B. dothidea, Valsa ceratosperma and Glomerella cingulata. Moreover, the inhibition was reduced by the presence of 5'-ADP. Taken all together, the results indicate that MdPR-4 protein is involved in the defense responses of apple against pathogenic attack by directly inhibiting hyphal growth, and the inhibition is correlated with its ribonuclease activity, where as MdPR-4 expression is regulated by both SA and JA signaling pathway.

背景与目标： ：发病相关蛋白4（PR-4）家族是一组在C末端带有Barwin域的蛋白，通常被认为与植物防御反应有关。但是，对于苹果植物抵抗病原体感染的作用，人们对其作用的详细了解甚少。在本研究中，从家蝇中鉴定出一个新的PR-4基因（称为MdPR-4），并研究了其在苹果防御反应中的作用。 MdPR-4基因的开放阅读框为447bp，编码148个氨基酸的蛋白质，在C端带有Barwin域，在N端具有26个氨基酸的信号肽。序列和结构分析表明，MdPR-4蛋白属于PR-4家族的II类。如实时定量PCR所揭示，在花和叶中观察到了MdPR-4的高水平表达。时间表达分析表明，MdPR-4表达可被多孢葡萄球菌感染和水杨酸（SA）或茉莉酸甲酯（MeJA）处理上调，但可被二乙基二硫代氨基甲酸（DIECA）抑制。在体外测定中，重组MdPR-4蛋白表现出对单链RNA特异的核糖核酸酶活性，并显着抑制了三种苹果病原真菌B.dothidea，Valsa ceratosperma和Glomerella cingulata的菌丝生长。此外，通过5'-ADP的存在降低了抑制作用。综上所述，结果表明MdPR-4蛋白通过直接抑制菌丝生长而参与苹果对病原体侵袭的防御反应，并且该抑制作用与其核糖核酸酶活性有关，因为MdPR-4的表达受两个SA的调节。和JA信号通路。

BACKGROUND & AIMS: :Botryosphaeria dothidea (Moug.:Fr.) Ces. De Not. causes perennial cankers on apple trees and causes white rot on apple fruit in the field and during storage (1). Prolonged periods of warm wet weather favor rapid disease outbreaks that result in severe losses, which range from 25 to 50% for the southeastern United States (3). A B. dothidea isolate was obtained from decayed 'Fuji' apple fruit exhibiting white rot symptoms from a local farm market in Beltsville, MD, in May 2010. The fruit had characteristic large dark brown lesions with irregular margins and decay expanded unevenly toward the core and the tissue was soft. The pathogen was isolated from symptomatic tissue by spraying the lesion surface with 70% ethanol. The skin with aseptically removed with a scalpel and small pieces of tissue were placed on potato dextrose agar (PDA) and incubated at 20°C. Once fungal growth was evident, the cultures were hyphal-tip transferred to individual PDA plates and incubated at 20°C. The B. dothidea isolate produced black aerial mycelium with a white margin on PDA and had a black reverse. Conidiomata were evident after 10 to 14 days at 20°C only on oatmeal agar. Conidia were hyaline, smooth and straight, fusiform with an subobtuse apex and a truncate base 20 to 26 (24.33) × 4 to 7 (5) μm (n = 50). Genomic DNA was isolated from the fungus and amplified with gene specific primers (ITS 4 and 5) for the ribosomal DNA internal transcribed spacer region ITSI-5.8S-ITS2 as described by White et al. (4). Both forward and reverse strands of the 542-bp amplicon were sequenced and assembled into a contig. The nucleotide sequence (GenBank Accession No. KC473852) indicated 99% identity to B. dothidea isolate CMM3938 (JX513645.1) and to voucher specimens CMW 25686, 25696, and 25222 (FM955381.1, FM955379.1, and FM955377,1). Koch's postulates were conducted using three 'Golden Delicious' apple fruit that were wound-inoculated with 50 μl of a mycelial suspension of the fungus, obtained from aseptically scraping a 7-day-old PDA culture, and was also repeated using 'Fuji' apple fruit. Large, brown, slightly sunken, soft lesions with undefined edges developed 5 days after inoculation at 20°C and water-only inoculated fruit were symptomless. The fungus was reisolated from infected tissue and was morphologically identical to the original isolate from decayed apple fruit. To determine if the B. dothidea isolate was resistant to postharvest fungicides, the minimum inhibitory concentration (MIC) was conducted using the 96 well plate method with a mycelial suspension of the fungus as described by Pianzzola et al. (2). The MIC for the isolate was >1 ppm for Mertect and Scholar and 50 ppm for Penbotec, which are well below the labeled rates for these postharvest fungicides and the experiment was repeated. To our knowledge, this is the first report of B. dothidea causing white rot on apple fruit in Maryland. References: (1) A. R. Biggs and S. S. Miller. HortScience 38:400, 2003. (2) M. J. Pianzzola et al. Plant Dis. 88:23, 2004. (3) T. B. Sutton. White rot and black rot. Pages 16-20 in: Compendium of Apple and Pear Diseases, A. L. Jones and H. S. Aldwinckle, eds. The American Phytopathological Society, St Paul, MN, 1991. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Application. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

背景与目标： ：Botryosphaeria dothidea（Moug.:Fr。）Ces。德不。在田间和贮藏期间会在苹果树上引起多年生溃疡，并在苹果果实上引起白腐（1）。长时间的温暖潮湿天气有利于疾病的快速爆发，从而导致严重的损失，在美国东南部占25％至50％（3）。 2010年5月，从马里兰州贝尔茨维尔的一家当地农产品市场的腐烂的“富士”苹果果实中获得了表现出白色腐烂症状的芽孢杆菌分离株。该果实具有特征性的深黑褐色病灶，边缘不规则，腐烂向核心扩展不均匀并且组织柔软。通过用70％乙醇喷洒病灶表面，从有症状的组织中分离出病原体。用手术刀无菌去除的皮肤和小块组织被放在马铃薯葡萄糖琼脂（PDA）上并在20°C下孵育。一旦明显的真菌生长，将培养物菌丝尖端转移到单独的PDA板中，并在20°C下孵育。 B. dothidea分离株在PDA上产生黑色气生菌丝，白色边缘，黑色反向。在20°C下10-14天后，仅在燕麦琼脂上可见形瘤。分生孢子是透明的，光滑的和笔直的，梭形的，具有近钝的先端和截形的基部20至26（24.33）×4至7（5）μm（n ＝ 50）。从真菌中分离出基因组DNA，并如White等人所述用核糖体DNA内部转录的间隔区ITSI-5.8S-ITS2的基因特异性引物（ITS 4和5）扩增。 （4）。对542bp扩增子的正向和反向链都进行测序并组装成重叠群。核苷酸序列（GenBank登录号KC473852）表明与双歧双歧杆菌分离物CMM3938（JX513645.1）和凭证样本CMW 25686、25696和25222（FM955381.1，FM955379.1和FM955377,1）有99％的同一性。 Koch的假设是使用三种“黄金美味”苹果果实进行的，这些果实用50μl真菌菌丝体悬浮液伤口接种，这种真菌悬浮液是通过无菌刮擦7天大的PDA培养物获得的，并且也使用“富士”苹果进行了重复水果。在20°C接种5天后，出现了大的，褐色的，略微下陷的，柔软的病灶，边缘模糊不清，接种纯水的果实无症状。该真菌是从受感染的组织中重新分离出来的，其形态与从腐烂的苹果果实中分离出来的原始菌株完全相同。为了确定双歧双歧杆菌分离株是否对收获后的杀真菌剂具有抗性，如Pianzzola等人所述，使用96孔板法对真菌的菌丝体悬浮液进行最小抑制浓度（MIC）。 （2）。对于Mertect和Scholar，分离株的MIC大于1 ppm，对于Penbotec，MIC大于50 ppm，远低于这些收获后杀菌剂的标记率，并重复了实验。据我们所知，这是马里兰州B. dothidea在苹果果实上引起白腐的首次报道。参考文献：（1）A. R. Biggs和S. S. Miller。 HortScience 38：400，2003。（2）M.J.Pianzzola等。植物病。 88:23，2004.（3）萨顿（T. B. Sutton）。白腐和黑腐。第16-20页，在《苹果与梨病简编》中，A。L. Jones和H. S. Aldwinckle编辑。美国植物病理学会，明尼苏达州圣保罗，1991年。（4）T. J. White等。第315页中的“ PCR协议：方法和应用指南”。 M.A.Innis等，编辑。学术出版社，加利福尼亚州圣地亚哥，1990年。

BACKGROUND & AIMS: :The family Botryosphaeriaceae comprises a number of species that are associated with the dieback disease of grapevine (Vitis vinifera), referred to as Botryosphaeria canker. To date, there are few effective agents available for the management of this disease. In this study, fungicides were evaluated for controlling the disease using a combination of in vitro tests and field trials. Twenty fungicides registered for use on other diseases in Australian viticulture were tested in vitro for their effect on mycelial growth of four species within the Botryosphaeriaceae. The concentrations of fungicide at which 50% of mycelial growth is inhibited (EC50 values) were significantly affected both by fungicide and isolate (P < 0.001). Differences in sensitivities of the four species to the fungicides were negligible (0.41 to 0.59 mg/liter). The most effective fungicides were fludioxonil, carbendazim, fluazinam, tebuconazole, flusilazole, penconazole, procymidone, iprodione, myclobutanil, and pyraclostrobin, for which EC50 values were <1.0 mg/liter. These fungicides were evaluated under field conditions, in addition to the pruning wound protectants Bacseal Super, Garrison, and ATCS tree wound dressing, as well as the biological control agent Vinevax. In field trials, carbendazim (Bavistin), fluazinam (Shirlan), tebuconazole (Folicur), Garrison, and ATCS tree wound dressing applied to freshly cut pruning wounds were the most effective and reduced infection by Diplodia seriata and D. mutila by 41 to 65%. These results suggest that the occurrence of Botryosphaeria canker on grapevines may be reduced via treatment of pruning wounds with selected fungicides as soon as possible after pruning.

背景与目标： ：Botryosphaeriaceae科包括许多与葡萄（Vitis vinifera）的枯死病有关的物种，被称为Botryosphaeria溃疡病。迄今为止，几乎没有有效的药物可用于治疗这种疾病。在这项研究中，结合体外试验和田间试验评估了杀菌剂对控制疾病的作用。体外测试了20种已注册用于澳大利亚葡萄栽培中其他疾病的杀真菌剂对葡萄孢菌内4种菌丝体生长的影响。抑制菌丝体生长50％的杀菌剂浓度（EC50值）都受到杀菌剂和分离物的显着影响（P <0.001）。四种菌种对杀真菌剂的敏感性差异可忽略不计（0.41至0.59 mg / L）。最有效的杀真菌剂是氟地西尼，多菌灵，氟西那南，戊唑醇，氟西拉唑，戊康唑，普西米酮，异丙隆，Myclobutanil和吡咯菌酯，其EC50值小于1.0毫克/升。除了修剪伤口保护剂Bacseal Super，Garrison和ATCS树伤口敷料以及生物防治剂Vinevax外，还在野外条件下评估了这些杀菌剂。在田间试验中，将多菌灵（Bavistin），氟喹南（Shirlan），戊唑醇（Folicur），加里森和ATCS树形伤口敷料用于新鲜修剪的修枝伤口，是最有效的方法，将Diplodia seriata和mut。D.感染减少了41至65。 ％。这些结果表明，通过在修剪后尽快用精选的杀菌剂处理修剪伤口，可以减少葡萄藤上灰霉病的发生。

BACKGROUND & AIMS: :The influence of carbohydrates: glucose, fructose, galactose, galacturonic acid, xylose, lactose, sucrose, pectin and inulin, were evaluated as sole carbon source for the production of laccases by the ascomycete, Botryosphaeria sp. Veratryl alcohol, a laccase inducer, was added to culture media to study inducible laccase production on the same carbon sources. Inulinase and pectinase were also produced when Botryosphaeria sp. was grown on inulin, and galacturonic acid and pectin, respectively, and their levels were less in the presence of veratryl alcohol. Botryosphaeria sp. produced constitutive laccases on all carbon sources examined, and veratryl alcohol increased the laccase production on most of carbon sources studied except for inulin and galacturonic acid. Evidence is presented that Botryosphaeria sp. is also pectinolytic.

背景与目标： 碳水化合物的影响：葡萄糖，果糖，半乳糖，半乳糖醛酸，木糖，乳糖，蔗糖，果胶和菊粉被评估为唯一的碳源，被子囊菌（Botryosphaeria sp。）生产漆酶。将Veratryl醇（一种漆酶诱导剂）添加到培养基中，以研究在相同碳源上可诱导的漆酶生产。葡萄球菌（Botryosphaeria sp。）也产生菊粉酶和果胶酶。分别在菊粉，半乳糖醛酸和果胶上生长，并且在存在藜芦醇的情况下它们的含量较低。灰霉菌在研究的所有碳源上均产生本构漆酶，而藜芦醇增加了所研究的大多数碳源（菊粉和半乳糖醛酸除外）的漆酶产量。证据表明，Botryosphaeria sp。也是果胶分解的。

BACKGROUND & AIMS: Botryosphaeria dothidea is a latent and important fungal pathogen on a wide range of woody plants. Fruit ring rot caused by B. dothidea is a major disease in China on apple. This study establishes a high quality, nearly complete and well annotated genome sequence of B. dothidea strain sdau11-99. The findings of this research provide a reference genome resource for further research on the apple fruit ring rot pathogen on apple and other hosts.

背景与目标： Botryosphaeria dothidea是许多木本植物上潜在的重要真菌病原体。多角芽孢杆菌引起的果环腐烂是中国苹果的主要病害。这项研究建立了高质量的，几乎完整的和注释良好的双歧杆菌菌株sdau11-99的基因组序列。该研究结果为进一步研究苹果及其他宿主上的苹果果实腐烂病原体提供了参考基因组资源。

BACKGROUND & AIMS: Apple ring rot, caused by Botryosphaeria dothidea, is one of the important diseases in China. This pathogen infects branches and fruit and also results in fruit decay during storage. Biocontrol agents have been proposed to reduce apple decays during storage and are considered as a promising alternative strategy to traditional chemical treatment. In this study, Meyerozyma guilliermondii Y-1, isolated from healthy grape fruit, was firstly evaluated for its biocontrol efficiency against B. dothidea in postharvest apple fruit, and the possible mechanisms were investigated. The results revealed that M. guilliermondii Y-1 treatment effectively reduced apple ring rot caused by B. dothidea in vivo. The disease incidence and lesion diameter were reduced by 32.22% and 57.51% compared with those of control fruit. Furthermore, the use of filtrate and autoclaved culture of M. guilliermondii Y-1 also showed a certain degree of control efficiency against fruit ring rot. M. guilliermondii Y-1 significantly inhibited the mycelial growth and spore generation of B. dothidea in vitro and exhibited an obvious ability to colonize in apple fruit wounds and surface at 25 °C or 4 °C. In addition, M. guilliermondii Y-1 treatment significantly enhanced the activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), phenylalanine ammonialyase (PAL), and polyphenoloxidase (PPO), promoted the total phenolics content, and alleviated lipid peroxidation in apple fruit. As expected, we found that the expression of four pathogenesis-related proteins genes (MdPR1, MdPR5, MdGLU, and MdCHI) was remarkably increased by M. guilliermondii Y-1 treatment. Our data together suggest that M. guilliermondii Y-1 is a potential biocontrol agent against B. dothidea postharvest infection in apple fruit, partially through inhibiting mycelial growth and spore germination of B. dothidea, competing for space and nutrient with pathogen, and inducing resistance in apple fruit by stimulating a series of defense responses.

背景与目标： 苹果多环孢杆菌引起的苹果腐烂病是中国的重要疾病之一。该病原体感染树枝和果实，并在储存过程中导致果实腐烂。已经提出了生物防治剂来减少苹果在贮藏过程中的腐烂，并且被认为是传统化学处理的有希望的替代策略。在这项研究中，首先评估了从健康葡萄果实中分离的Meyerozyma guilliermondii Y-1对苹果果实采后对B. dothidea的生防效果，并研究了可能的机制。结果表明，Guilliermondii Y-1处理可有效减少体内由B. dothidea引起的苹果环腐烂。与对照组相比，病害发生率和病斑直径分别降低了32.22％和57.51％。此外，使用滤液和高压灭菌培养的圭尔蒙德酵母Y-1也显示出对果环腐烂的一定程度的控制效率。 guilliermondii Y-1显着抑制体外双歧杆菌的菌丝体生长和孢子生成，并在25°C或4°C下具有明显的定殖在苹果果实伤口和表面的能力。此外，Guilliermondii Y-1处理显着增强了过氧化氢酶（CAT），超氧化物歧化酶（SOD），过氧化物酶（POD），苯丙氨酸氨解酶（PAL）和多酚氧化酶（PPO）的活性，提高了总酚含量，并且减轻了苹果果实中的脂质过氧化作用。正如预期的那样，我们发现四种致病相关蛋白基因（MdPR1，MdPR5，MdGLU和MdCHI）的表达通过古吉尔蒙德酵母Y-1处理显着增加。我们的数据共同表明，Guilliermondii Y-1是对抗苹果果实中B. dothidea采后感染的潜在生物防治剂，部分是通过抑制B. dothidea的菌丝生长和孢子萌发，与病原体竞争空间和营养物以及诱导抗性通过刺激一系列防御反应来抑制苹果果实中的苦瓜。

BACKGROUND & AIMS: :Gala and Winter Banana apples are important commercial crops in Azurduy and Lima Bamba, which are located in the Department (state) of Chuquisaca, Bolivia. White or bot rot (causal agent Botryosphaeria dothidea (Moug.:Fr.) Ces. De. Not. [anamorph Fusicoccum aesculi Corda]) and black rot (causal agent B. obtusa (Schwein.) Shoemaker [anamorph Sphaeropsis malorum Berk.]) have not been reported previously from Bolivia. Both fungi were isolated from apple fruit and branch cankers in Azurduy, but only B. dothidea was isolated from rotted fruit and limb cankers in Lima Bamba. Both fungi also were isolated from rotted Gala and Winter Banana fruit purchased in the markets in Sucre, Bolivia. Symptoms on fruit consisted of light-to-dark brown lesions that ranged from 3- to 8-cm in diameter. Cankers on limbs were sunken and reddish brown and ranged from 2 to 25+ cm in length and 0.5 to 3 cm in diameter. Neither pathogen produced pycnidia in lesions on rotted fruit, but they often developed in branch cankers. Pseudothecia of B. dothidea and B. obtusa were not observed. Identification of both pathogens was based on descriptions of their anamorphic stages (1). To fulfill Koch's postulates, four healthy Gala apple fruit were inoculated with two isolates of each pathogen by wounding the opposite faces of surface-disinfected fruit with a 5-mm-diameter cork borer and inserting mycelial plugs of the pathogens. Plugs were obtained from the margins of cultures growing on potato dextrose agar (PDA). Wounds were made on the opposite sides of each fruit, a mycelial plug of one of the pathogens was inserted in one wound, and on the opposite side, a plug of sterile PDA was inserted as a control. Each plug containing fungal mycelium or sterile PDA was covered with a plug of trimmed apple tissue, and the apple fruit were incubated in a moist chamber at 17 to 20°C for 10 days. Six branches on two young apple trees growing outdoors in a nursery were inoculated in a similar manner with one isolate of each pathogen: bark was wounded with a 5-mm-diameter cork borer, and the wounded area was inoculated with a plug of PDA containing the pathogen or a plug of sterile PDA for the control. The inoculated sites were wrapped with masking tape to prevent dehydration. Within 10 days, all fruit wounds inoculated with isolates of each pathogen developed brown lesions up to 5 cm in diameter. Each pathogen was reisolated from tissues in which it had been inoculated, but not from any of the noninoculated control sites. Within 6 to 8 weeks, all but one wound on branches inoculated with each pathogen developed depressed canker lesions up to 2 cm in length. Each pathogen was reisolated from the canker produced by inoculation with that pathogen, but not from any of the control sites. Reference: (1) T. B. Sutton. White rot and black rot. Pages 16-20 in: Compendium of Apple and Pear Diseases, A. L. Jones and H. S. Aldwinckle, eds. The American Phytopathological Society, St. Paul, MN, 1991.

背景与目标： ：Gala和Winter Banana苹果是Azurduy和Lima Bamba的重要商业作物，它们位于玻利维亚Chuquisaca的省（州）。白腐病或腐烂病（病原体Botryosphaeria dothidea（Moug.:Fr。）Ces。De。Not [anamorph Fusicoccum aesculi Corda]）和黑腐病（病原体B. obtusa（Schwein。））鞋匠[Anamorph Sphaeropsis malorum Berk。] ）以前尚未从玻利维亚报道。两种真菌均从苹果果实和Azurduy的树枝状溃疡中分离出来，但仅B. dothidea从腐烂的水果和肢体鳞茎中在利马班巴中分离出来。两种真菌也均从玻利维亚苏克雷市场上购买的腐烂的联欢晚会和冬季香蕉果实中分离出来。水果的症状包括直径从3到8厘米不等的浅棕色至深褐色的病变。肢体上的溃疡呈凹陷和红褐色，长度在2至25厘米之间，直径在0.5至3厘米之间。两种病原体均未在腐烂果实的病斑中产生粘虫病，但它们通常在分支溃疡中生长。没有观察到假双歧杆菌和钝双歧杆菌的假性皮病。两种病原体的鉴定均基于其变形阶段的描述（1）。为了满足科赫的假设，通过用5毫米直径的软木塞打伤表面消毒过的水果的相对表面，并插入病原体的菌丝体塞，在四种健康的嘎拉苹果果实中接种两种分离的每种病原体。从在马铃薯葡萄糖琼脂（PDA）上生长的培养物的边缘获得塞子。在每个水果的相对侧制作伤口，将一种病原体的菌丝体塞插入一个伤口中，并在相对侧插入无菌PDA塞作为对照。每个装有真菌菌丝体或无菌PDA的塞子都用修剪过的苹果组织塞子覆盖，并将苹果果实在潮湿的室内于17至20°C孵育10天。用每种病原体的一种分离物，以相似的方式接种在苗圃中生长在室外的两棵年轻苹果树上的六个树枝上：用直径5毫米的软木塞打伤树皮，并用装有PDA的插头接种伤处用于控制的病原体或无菌PDA的塞子。接种部位用胶带包裹，以防脱水。在10天之内，接种了每种病原体的所有水果伤口都出现了直径达5厘米的棕色损伤。每种病原体都是从已经接种过的组织中分离出来的，而不是从任何未接种的对照位点中分离出来的。在6至8周内，除接种了每种病原体的树枝上的一个伤口外，所有伤口均出现了长达2 cm的凹陷性溃疡病。从通过接种该病原体产生的溃疡病中分离出每种病原体，但没有从任何对照位点中分离出病原体。参考：（1）T. B. Sutton。白腐和黑腐。第16-20页，在《苹果与梨病简编》中，A。L. Jones和H. S. Aldwinckle编辑。美国植物病理学会，明尼苏达州圣保罗，1991年。

BACKGROUND & AIMS: :Panicle and shoot blight of pistachio caused by species of Botryosphaeriaceae fungi remains one of the major diseases of pistachio in California, with the highest potential for crop destruction because buds and fruit clusters (panicles) are killed. Pistachio fruit can also be damaged by various insect pests. For instance, hemipteran insects are considered major pests of pistachio and can attack maturing fruit, causing necrotic areas on the fruit epicarp (epicarp lesion). Surveys in pistachio orchards showed a putative association of punctures of pistachio fruit with infection by Botryosphaeriaceae fungi, resulting in panicle and shoot blight. This study was undertaken to determine any association of Botryosphaeriaceae fungi with fruit bearing epicarp lesions and whether hemipteran insects can actively carry propagules of the pathogens to healthy fruit and cause disease. Three large insects of the order Hemiptera (Thyanta pallidovirens, Acrosternum hilare, and Leptoglossus clypealis) were used in pathogen-spread and infection experiments. The most frequently isolated species of the family Botryosphaeriaceae in pistachio, Neofusicoccum mediterraneum, was used in these transmission experiments. Significantly higher incidence of fruit infected by N. mediterraneum was recorded when pistachio clusters were sprayed with a spore suspension of N. mediterraneum and caged with the above insects than the level of infected fruit sprayed similarly with the pathogen but caged without any insects. Furthermore, increasing the number of insects caged with pistachio clusters resulted in an increase of insect feeding punctures and, subsequently, disease incidence. Similarly, increasing the number of wounds per fruit made with a needle, simulating injuries caused by the insect's stylet, resulted in significantly higher incidence of infected fruit than for wounded fruit with fewer injured sites. In addition, bird-injured fruit were also associated with higher incidence of panicle blight than fruit not injured by birds, especially in orchards with high disease incidence. In laboratory experiments, wounding or applying fruit sap to nonwounded fruit resulted in significantly higher incidence of infected fruit than in nonwounded fruit without sap. In field experiments, wounding of fruit before or after inoculation with N. mediterraneum resulted in higher disease incidence than nonwounded and inoculated fruit. Additionally, fruit sap enhanced initial mycelial growth of N. mediterraneum. Protecting fruit clusters from injuries by hemipteran insects or birds using mesh cages significantly reduced the incidence of panicle and shoot blight. The association of wounding by hemipteran insects and birds with high incidence of infected pistachio fruit may explain partially the broad distribution of Botryosphaeria panicle and shoot blight among pistachio orchards of the central San Joaquin Valley, even in areas where inoculum sources are not nearby.

背景与目标： ：由葡萄球菌属真菌引起的开心果的穗状茎和茎叶枯萎病仍是加利福尼亚州开心果的主要疾病之一，由于杀死芽和果实簇（穗），其对农作物的破坏潜力最高。阿月浑子果实也可能被各种害虫破坏。例如，半翅目昆虫被认为是开心果的主要害虫，可侵袭成熟的果实，在果实果皮（表皮病）上造成坏死区域。阿月浑子果园的调查显示，开心果刺穿与Botryosphaeriaceae真菌感染有关，导致穗和枯萎病。这项研究的目的是确定葡萄球菌真菌与带有果皮果皮损伤的果实之间是否存在关联，以及半翅目昆虫是否可以将病原体的繁殖体主动携带到健康的果实上并引起疾病。在病原体传播和感染实验中使用了半翅目的三种大型昆虫（Thyanta pallidovirens，Acrosternum hilare和Leptoglossus clypealis）。在这些传播实验中，使用了开心果中最频繁分离的葡萄球菌科物种，Neofusicoccum mediterraneum。当用开心果簇用中生猪笼草的孢子悬浮液喷洒并用上述昆虫进行笼养时，记录到被中生猪笼草感染的水果的发生率要比类似地喷洒病原体但笼中没有任何昆虫的被感染果实的水平高。此外，被开心果簇包裹的昆虫数量的增加导致昆虫摄食穿刺的增加，进而导致疾病的发生。同样，模拟用昆虫的探针引起的伤害，增加每个针头刺伤每个水果所产生的伤口数量，会导致感染水果的发生率明显高于受伤部位较少的受伤水果。此外，与没有受到鸟类伤害的果实相比，鸟类受伤的果实与穗枯萎病的发病率也更高，尤其是在发病率高的果园中。在实验室实验中，对未受伤的水果进行创伤或应用水果树汁会导致感染水果的发生率显着高于没有树液的未受伤水果。在田间试验中，接种中性猪笼草前后对水果的伤害要比未受伤和接种的水果高。此外，水果树液增强了中性猪笼草的初始菌丝生长。使用网状笼子保护果实免受半球虫或鸟类伤害，显着降低了穗和枯萎病的发生率。半昆虫昆虫和鸟类造成的伤害与开心果受感染的发生率高有关，这可能部分解释了葡萄孢的穗状花序广泛分布和圣华金山谷中部开心果果园的枯萎病，即使在附近没有接种源的地区也是如此。

BACKGROUND & AIMS: :Botryosphaeria lutea (anamorph Fusicoccum luteum) most easily is distinguished from other Botryosphaeria spp. by a yellow pigment that is formed in young cultures. This fungus has been reported from a number of cultivated hosts in New Zealand and Portugal. During a survey of Botryosphaeria fungi that occur on native Acacia species in Australia, a yellow pigment was observed in some cultures. These isolates were morphologically similar to B. lutea, but the pigment differed slightly from the one formed by authentic B. lutea isolates. Preliminary data also revealed small differences in ITS rDNA sequence data. The aim of this study was to determine whether these small differences were indicative of separate species or merely variations within B. lutea. Anamorph, teleomorph and culture morphology were compared between B. lutea and Acacia isolates from Australia. Sequence data of two other genome regions, namely the β-tubulin and EF1-α gene and intron regions, were combined with ITS rDNA sequence data to determine the phylogenetic relationship between these isolates. Isolates of B. lutea and those from Australian Acacia species were not significantly different in spore morphology. The yellow pigment, however, was much more distinct in cultures of B. lutea than in cultures from Acacia. There were only a few base pair variations in each of the analyzed gene regions, but these variations were fixed in the two groups in all regions. By combining these data it was clear that B. lutea and the isolates from Acacia were distinct species, albeit very closely related. We, therefore, propose the new epithet B. australis for the fungus from Australia. Botryosphaeria australis also was isolated in this study from exotic Sequoiadendron trees in Australia. Re-analyses of GenBank data in this study showed that B. australis also occurs on other native Australian hosts, namely a Banksia sp. and a Eucalyptus sp., as well as a native Protea sp. in South Africa and on Pistachio in Italy. These records from GenBank have been identified previously as B. lutea. The common occurrence of B. australis on a variety of native hosts across Australia suggests that this fungus is native to this area.

背景与目标： ：Botryosphaeria lutea（变形金缕梅）最容易区别于其他Botryosphaeria spp。由年轻文化中形成的黄色颜料形成。已经从新西兰和葡萄牙的许多栽培寄主中报告了这种真菌。在对澳大利亚本地相思树种上发生的灰霉病真菌进行的一项调查中，在某些文化中观察到黄色素。这些分离物在形态上与黄褐芽孢杆菌相似，但是色素与真正的黄褐芽孢杆菌分离株形成的色素略有不同。初步数据还显示了ITS rDNA序列数据的微小差异。这项研究的目的是确定这些小的差异是指示单独的物种还是仅代表黄褐变内的变异。比较了来自澳大利亚的黄杆菌和金合欢分离株的无性型，远形型和培养物形态。将其他两个基因组区域（即β-微管蛋白和EF1-α基因和内含子区域）的序列数据与ITS rDNA序列数据相结合，以确定这些分离株之间的系统发育关系。黄褐芽孢杆菌和澳大利亚金合欢种的分离物在孢子形态上没有显着差异。然而，黄色的色素在黄褐芽孢杆菌培养物中比在阿拉伯树胶培养物中明显不同。在每个分析的基因区域中仅有几个碱基对变异，但是这些变异在所有区域的两组中固定。通过综合这些数据，可以清楚地看出，黄褐芽孢杆菌和金合欢的分离物是不同的物种，尽管密切相关。因此，我们为来自澳大利亚的真菌提出了新的上皮B. australis。这项研究还从澳大利亚的异国红杉树中分离出了澳大利亚灰霉病菌。对GenBank数据进行的重新分析显示，澳大利亚双歧杆菌也存在于其他澳大利亚本土寄主，即Banksia sp.。和一个桉树种，以及一个本地的普罗梯亚树种。在南非和意大利的开心果上。这些来自GenBank的记录以前被鉴定为B. lutea。在澳大利亚的各种本地寄主中普遍存在澳大利亚芽孢杆菌，这表明该真菌是该地区的原生真菌。

BACKGROUND & AIMS: :Fungal effectors play important roles in host-pathogen interactions. Botryosphaeria dothidea is an ascomycetous fungus that is responsible for the diseases of hundreds of woody plant species, including apple ring rot, which seriously affects apples worldwide. However, little is known about the effectors of B. dothidea. In this study, we analyzed the B. dothidea genome and predicted 320 candidate effector genes, 124 of which were successfully amplified and cloned. We investigated the effects of these genes on plant cell death in Nicotiana benthamiana while using a transient expression system. Twenty-four hours after initial inoculation with Agrobacterium tumefaciens cells carrying candidate effectors, the infiltrated leaves were challenged with A. tumefaciens cells carrying the BAX gene. In total, 116 candidate effectors completely inhibited, while one partially inhibited, the programmed cell death (PCD) of N. benthamiana induced by BAX, whereas seven candidate effectors had no effect. We then further tested seven candidate effectors able to suppress BAX-triggered PCD (BT-PCD) and found that they all completely inhibited PCD triggered by the elicitors INF1, MKK1, and NPK1. This result suggests that these effectors were activated in order to suppress pathogen-associated molecular pattern-triggered immunity. The signal peptides of these candidate effectors exhibited secretory activity in yeast (pSUC2 vector). Moreover, the respective deletion of Bdo_11198 and Bdo_12090 significantly reduced the virulence of B. dothidea. These results suggest that these effectors play important roles in the interaction of B. dothidea with its hosts.

背景与目标： ：真菌效应子在宿主-病原体相互作用中起重要作用。 Botryosphaeria dothidea是一种子囊菌，负责数百种木本植物的病害，包括严重影响全球苹果的苹果环腐病。然而，关于双歧杆菌的效应子知之甚少。在这项研究中，我们分析了B. dothidea基因组并预测了320个候选效应基因，其中124个被成功扩增和克隆。我们使用瞬时表达系统调查了这些基因对本生烟草中植物细胞死亡的影响。在初始接种携带候选效应子的根癌农杆菌细胞后二十四小时，用携带BAX基因的根癌农杆菌细胞攻击浸润的叶片。总共有116种候选效应物完全抑制了BAX诱导的本氏烟草的程序性细胞死亡（PCD），而部分抑制了BAX诱导的本氏烟草的程序性细胞死亡（PCD），而有7种候选效应物没有任何作用。然后，我们进一步测试了7种能够抑制BAX触发的PCD（BT-PCD）的候选效应子，发现它们都完全抑制了由激发子INF1，MKK1和NPK1触发的PCD。该结果表明，激活这些效应子是为了抑制病原体相关的分子模式触发的免疫力。这些候选效应子的信号肽在酵母（pSUC2载体）中表现出分泌活性。此外，Bdo_11198和Bdo_12090的各自缺失显着降低了十二指肠杆菌的毒力。这些结果表明，这些效应子在B. dothidea与其宿主的相互作用中起重要作用。

BACKGROUND & AIMS: :Blueberry (Vaccinium spp.) production in southwest and northeast China has grown to over 100,000 ha in the last 20 years thanks to the fruit's high nutritional and economic value. As blueberry acreage increases, the diversity of diseases and challenges for control are gaining more attention. In August 2010, stem and branch blight occurred on Highbush Blueberries (Vaccinium corymbosum L.) at commercial farms in Lijiang and Zongdian, Yunnan Province (southwestern China), with crop damage ranging from 10 to 15%. Typical symptoms of the disease were blight and dieback on the stems with lesions extending along entire branches. Diseased samples (phloem and xylem sectors in the wood) were washed with running tap water, disinfected with 2% sodium hypochlorite, then 70% alcohol, rinsed in sterile distilled water, plated on potato dextrose agar (PDA), and incubated at 28°C. Fungal isolates developed copious, white aerial mycelium that became dark gray after 4 to 5 days and formed black pycnidia after 18 days. Conidia were hyaline, aseptate, thin walled, fusiform, and measured 21 to 27 × 4 to 6 μm. Identity was confirmed by analysis of the ribosomal DNA internal transcribed spacer region ITSI-5.8S -ITS2 with primers ITS1 and ITS4. BLAST searches showed 99% identity with Botryosphaeria dothidea isolates from GenBank (Accession Nos. AB693904 and JF800139). Representative sequences of B. dothidea from Highbush Blueberries from China were deposited into GenBank (Accession No. JX096631). On the basis of morphological and molecular results, the fungus isolated from diseased Highbush Blueberries stem was confirmed to be B. dothidea. Pathogenicity tests were conducted on 2-year-old blueberry seedlings (Highbush Blueberries). Mycelial plugs (2 to 3 mm in diameter) of B. dothidea from actively growing colonies (PDA) were applied to same-size bark wounds in the center of the stems. Inoculation wounds were wrapped with Parafilm. Control seedlings received sterile PDA plugs. Inoculated and control seedlings (five each) were kept in a greenhouse and watered as needed. After 12 days, all of the inoculated but none of the control blueberry seedlings showed dark vascular stem tissue. B. dothidea was reisolated from symptomatic tissues, thus fulfilling Koch's postulates. No symptoms were visible in the control seedlings. B. dothidea has been reported as a pathogen of sycamore (3), olives (1), and peach (2). However, no research has been conducted on stem blight of blueberry caused by B. dothidea in southwest or mainland China. To our knowledge, this is the first report of B. dothidea on blueberry in southwest China. References: (1) M. Chattaoui, et al. Plant Dis. 95:770, 2011. (2) Y. Ko et al. Plant Pathol. Bull. 1:70, 1992. (3) E. Turco, et al. Plant Dis. 90:1106, 2006.

背景与目标： ：由于该水果的高营养价值和经济价值，在过去的20年中，西南和东北地区的蓝莓（Vaccinium spp。）产量已增长到100,000多公顷。随着蓝莓种植面积的增加，疾病的多样性和控制挑战日益受到关注。 2010年8月，云南丽江和宗甸（中国西南部）的商业农场的高灌木蓝莓（越桔）发生了茎叶和枝叶枯萎病，对作物的危害范围为10％至15％。该病的典型症状是茎枯萎和枯萎，病斑沿整个分支延伸。患病样品（木材中韧皮部和木质部部分）用流动的自来水洗涤，先用2％次氯酸钠消毒，再用70％酒精消毒，再用无菌蒸馏水冲洗，然后涂在马铃薯葡萄糖琼脂（PDA）上，并在28°C下孵育C。真菌分离物形成大量的白色气生菌丝体，在4至5天后变成深灰色，在18天后形成黑色的粘虫。分生孢子是透明的，腹水的，薄壁的，梭形的，大小为21至27×4至6μm。通过用引物ITS1和ITS4分析核糖体DNA内部转录的间隔区ITSI-5.8S-ITS2，证实了同一性。 BLAST搜索显示与GenBank中的Botryosphaeria dothidea分离株具有99％的同一性（登录号AB693904和JF800139）。来自中国的Highbush蓝莓的B. dothidea的代表性序列被保藏到GenBank（登录号JX096631）。根据形态学和分子结果，从患病的Highbush蓝莓茎中分离出的真菌被确认为B. dothidea。对2岁的蓝莓幼苗（Highbush蓝莓）进行了致病性测试。将来自活跃生长菌落（PDA）的B. dothidea菌丝体塞（直径2到3 mm）应用于茎中央相同大小的树皮伤口。接种伤口用Parafilm包裹。对照幼苗接受无菌的PDA塞。接种和对照的幼苗（每棵5株）都放​​在温室中，并根据需要浇水。 12天后，所有已接种但没有对照的蓝莓幼苗均显示出深色的维管干组织。从症状组织中分离出双歧杆菌，从而满足了科赫的假设。在对照幼苗中没有可见的症状。据报道，B。dothidea是美国梧桐（3），橄榄（1）和桃子（2）的病原体。然而，在西南或中国大陆尚未对由多虫双歧杆菌引起的蓝莓枯萎病进行过研究。据我们所知，这是B. dothidea在中国西南蓝莓上的首次报道。参考文献：（1）M. Chattaoui等。植物病。 95：770，2011.（2）Y.Ko等。植物病理学。公牛。 1:70，1992.（3）E.Turco等人。植物病。 90：1106，2006。

BACKGROUND & AIMS: :Botryosphaeria dothidea (Moug.: Fr.) Ces. & De Not has a worldwide distribution infecting species from over 80 genera of plants (1). Apart from being an important pathogen of apple trees in many countries, B. dothidea can cause pre- and postharvest decay on apple fruit (2). It has been known to cause canker and dieback of forest trees in Serbia (3), but has not been recorded either on apple trees or apple fruit. In December 2010, apple fruit cv. Idared (Malus × domestica Borkh.) with symptoms of white rot were collected from one storage in the area of Svilajnac in Serbia. The incidence of the disease was low but the symptoms were severe. Affected fruit were brown, soft, and almost completely decayed, while the internal decayed tissue appeared watery and brown. A fungus was isolated from symptomatic tissue of one fruit after surface sterilization with 70% ethanol (without rinsing) and aseptic removal of the skin. Small fragments of decayed tissue were placed on potato dextrose agar (PDA) and incubated in a chamber at 22°C under alternating light and dark conditions (12/12 h). Fungal colonies were initially whitish, but started turning dark gray to black after 5 to 6 days. Pycnidia were produced after 20 to 25 days of incubation at 22°C and contained one-celled, elliptical, hyaline conidia. Conidia were 17.19 to 23.74 μm (mean 18.93) × 3.72 to 4.93 μm (mean 4.45) (n = 50). These morphological characteristics are in accordance with those described for the fungus B. dothidea (4). Genomic DNA was isolated from the fungus and internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The nucleotide sequence has been assigned to GenBank Accession No. KC994640. BLAST analysis of the 528-bp segment showed a 100% similarity with several sequences of B. dothidea deposited in NCBI GenBank, which confirmed morphological identification. Pathogenicity was tested by wound inoculation of five surface-sterilized, mature apple fruit cv. Idared with mycelium plugs (5 mm in diameter) of the isolate grown on PDA. Five control fruit were inoculated with sterile PDA plugs. After 5 days of incubation in plastic containers, under high humidity (RH 90 to 95%) at 22°C, typical symptoms of white rot developed on inoculated fruit, while wounded, uninoculated, control fruit remained symptomless. The isolate recovered from symptomatic fruit showed the same morphological features as original isolate. To the best of our knowledge, this is the first report of B. dothidea on apple fruit in Serbia. Apple is widely grown in Serbia and it is important to further investigate the presence of this pathogen in apple storage, as well as in orchards since B. dothidea may cause rapid disease outbreaks that result in severe losses. References: (1) G. H. Hapting Agriculture Handbook 386, USDA, Forest Service, 1971. (2) A. L. Jones and H. S. Aldwinckle Compendium of Apple and Pear Diseases. APS Press, St. Paul, MN, 1990. (3) D. Karadžic et al. Glasnik Šumarskog Fakulteta 83:87, 2000. (4) B. Slippers et al. Mycologia 96:83, 2004.

背景与目标： ：Botryosphaeria dothidea（Moug .: Fr.）Ces。 ＆De Not在全球范围内感染80多种植物的物种（1）。除了在许多国家/地区是苹果树的重要病原菌外，多虫双歧杆菌还可以导致苹果果实收获前和收获后的腐烂（2）。在塞尔维亚（3）已知会引起森林树木的萎缩和枯萎（3），但在苹果树或苹果果实上均未见记录。 2010年12月，苹果果实简历。从塞尔维亚的Svilajnac地区的一个仓库中收集了具有白腐病症状的Idared（Malus×domestica Borkh。）。该病的发生率低，但症状很严重。受影响的果实为棕色，柔软，几乎完全腐烂，而内部腐烂的组织则呈水状和褐色。在用70％乙醇（无需冲洗）进行表面灭菌并无菌去除皮肤后，从一种水果的症状组织中分离出一种真菌。将腐烂组织的小片段放在马铃薯葡萄糖琼脂（PDA）上，并在22°C的暗室中于明暗条件下交替孵育（12/12 h）。真菌菌落最初呈白色，但在5至6天后开始从深灰色变成黑色。在22°C下孵育20至25天后，产生了碧萝cn，并含有单细胞，椭圆形，透明分生孢子。分生孢子为17.19至23.74μm（平均18.93）×3.72至4.93μm（平均4.45）（n = 50）。这些形态特征与描述真菌B. dothidea（4）的特征一致。从真菌中分离出基因组DNA，并用引物ITS1 / ITS4扩增rDNA的内部转录间隔区（ITS）。该核苷酸序列已分配给GenBank登录号KC994640。 528 bp片段的BLAST分析显示，其与NCBI GenBank中存放的多个B. dothidea序列具有100％的相似性，这证实了形态学鉴定。通过伤口接种五种经表面灭菌的成熟苹果果实简历来测试其致病性。厌倦了在PDA上生长的菌株的菌丝体塞（直径5毫米）。用无菌PDA塞接种了五个对照水果。在塑料容器中孵育5天后，在22°C的高湿度（RH 90至95％）下，接种的果实会出现典型的白腐症状，而受伤，未接种的对照果实仍无症状。从有症状果实中分离出的分离株表现出与原始分离株相同的形态特征。据我们所知，这是B. dothidea对塞尔维亚苹果果实的首次报道。苹果在塞尔维亚广泛种植，重要的是要进一步调查这种病原体在苹果储藏室以及果园中的存在，因为多头芽孢杆菌可能引起快速的疾病暴发，并造成严重的损失。参考文献：（1）G. H. Hapting农业手册386，美国农业部，林业局，1971年。（2）A. L. Jones和H. S. Aldwinckle《苹果和梨病简编》。 APS出版社，明尼苏达州圣保罗，1990年。（3）D.Karadžic等。 GlasnikŠumarskogFakulteta 83:87，2000。（4）B. Slippers等。 Mycologia 96：83，2004年。

BACKGROUND & AIMS: :ABSTRACT Laboratory and field experiments were conducted to study the dispersal of Botryosphaeria dothidea conidia using single-drop impactions and natural and simulated precipitations. For laboratory studies, 200 single drops were released from a height of 1 m on infected pistachio nuts. On pieces of photographic film, 50% of the droplets were collected within 20 mm (average droplet travel distance) of the target area, and the droplets ranged from 0.041 to 3.19 mm in diameter, with an average of 0.3 mm. Each droplet carried an average of 23 B. dothidea conidia. In 3 years of field experiments, rainwater was collected in funnels connected to bottles positioned at different heights inside the tree canopy and at different distances away from the edge of tree canopy in three commercial pistachio orchards in San Joaquin, Yolo, and Glenn counties in California. Numbers of conidia in rainwater varied among and within sampling seasons by sampling dates and orchards. Up to 67,000 conidia/ml were obtained in rainwater samples collected from an orchard in Yolo County. Rainwater from orchards in Yolo and Glenn counties contained a consistently higher number of conidia than rainwater collected from the orchard in San Joaquin County. Variation in numbers of conidia also existed among heights where bottles were located. There were significantly more conidia in rainwater collected inside than outside tree canopies. Inside tree canopies, bottles located at 100 and 150 cm above ground collected more B. dothidea conidia than those placed at 50 and 200 cm. Conidia were collected as far as 1 m from the tree canopy edge. Based on data from the Glenn County orchard, a linear relationship between number of conidia (Y) and rainfall amount (X) in millimeters was determined as Y = 240X - 3,867, with r(2) = 0.91, which meant that a minimum of 16.1 mm of rain was needed to disperse conidia of B. dothidea. The power law model best described the dispersal gradients of B. dothidea propagules in the 1999-2000 and 2001-02 sampling seasons, with r(2) values of >/=0.73, whereas the exponential law model fit best for the 2000-01 data, with r(2) values of >/=0.81. In a rain simulation experiment, the intensity of the rain generated by a nozzle at 138 kPa of pressure inside the tree canopy was approximately five times higher than rain recorded outside the tree canopy. Rain removed up to 65% of conidia from infected fruit. These results confirmed that B. dothidea is a splash-dispersed pathogen with relatively short distances of spore dispersal within pistachio orchards. Only pycnidia are present in pistachio orchards; therefore, the results also indicate that inoculum of B. dothidea should be entirely splashed dispersed.

背景与目标： ：摘要进行了实验室和现场实验，使用单滴撞击以及自然和模拟的降水来研究灰霉菌分生孢子的扩散。为了进行实验室研究，在受感染的开心果上从1 m的高度释放了200个单滴。在摄影胶片上，在目标区域的20毫米（平均液滴行进距离）内收集了50％的液滴，液滴的直径范围为0.041至3.19 mm，平均为0.3 mm。每个液滴平均携带23个B. dothidea分生孢子。在3年的野外试验中，在加利福尼亚州圣华金，约洛和格伦县的三个开心果果园中，雨水收集在与树冠内不同高度且距树冠边缘不同距离的瓶子相连的漏斗中。雨水中分生孢子的数量在采样季节之间和采样季节之内随采样日期和果园的不同而变化。从Yolo县的一个果园收集的雨水样品中可获取高达67,000分生孢子/ ml。与从圣华金县果园收集的雨水相比，来自尤洛和格伦县果园的雨水所含的分生孢子数量一直较高。分装瓶高度之间的分生孢子数量也存在差异。内部收集的雨水中的分生孢子明显比外部树冠更多。在树冠内部，位于地面100和150厘米处的瓶子比放置在50和200厘米处的瓶子收集到的双歧杆菌分生孢子更多。从树冠边缘到1 m处收集分生孢子。根据格伦县果园的数据，分生孢子数（Y）和降雨量（X）的线性关系以毫米为单位确定为Y = 240X-3,867，r（2）= 0.91，这意味着最小需要16.1毫米的雨水来分散B. dothidea的分生孢子。幂律模型最好地描述了1999-2000年和2001-02采样季节中B. dothidea繁殖体的扩散梯度，r（2）值> / = 0.73，而指数规律模型最适合2000-01 r（2）值> / = 0.81。在降雨模拟实验中，在树冠内部压力为138 kPa时，喷嘴产生的降雨强度大约是树冠外部记录的降雨强度的五倍。雨水从受感染的水果中清除了最多65％的分生孢子。这些结果证实了双歧杆菌是一种散布在病菌中的散布病原体，其开心果园内的孢子散布距离相对较短。开心果园中仅存在吡虫啉；因此，该结果还表明，B。dothidea的接种物应完全飞溅分散。