• 【曝气和藜芦醇对子囊灰霉菌产生两种漆酶的影响。】 复制标题 收藏 收藏
    DOI:10.1016/s0141-0229(00)00274-x 复制DOI
    作者列表:Dekker RF,Barbosa AM
    BACKGROUND & AIMS: :The ascomycete, Botryosphaeria sp, produced two extracellular constitutive laccases (PPO-I and PPO-II) active toward the substrates: 2, 2(1)-azino-bis(3-ethyl-benzthiazoline-6-sulfonic acid) [ABTS], and 2,6-dimethoxyphenol (DMP), respectively. The production of both laccases increased when the fungal isolate was grown in the presence of veratryl alcohol, and resulted in optimal laccase production (100- and 25- fold, respectively) at 40 mM. The effect of aeration on growth and laccase production was studied in baffled flasks, and showed that aeration of the cultures increased the production of both enzymes 4-5 fold in the presence of veratryl alcohol. Both laccases were susceptible to inhibition by azide, acetate and chloride anions. Veratryl alcohol inhibited the laccase-catalyzed polymerization of DMP. Growing cultures of Botryosphaeria sp. produced an exopolysaccharide of the beta-glucan type whose synthesis was depressed when grown in the presence of veratryl alcohol.
    背景与目标: :子囊孢子虫(Botryosphaeria sp)产生了两种对底物有活性的细胞外组成型漆酶(PPO-I和PPO-II):2,2(1)-叠氮基双(3-乙基-苯并噻唑啉-6-磺酸)[ABTS ]和2,6-二甲氧基苯酚(DMP)。当真菌分离物在藜芦醇存在下生长时,两种漆酶的产量均增加,并在40 mM时产生最佳漆酶产量(分别为100倍和25倍)。在带挡板的烧瓶中研究了通气对生长和漆酶产生的影响,结果表明,在存在藜芦醇的情况下,对培养物通气可使两种酶的产量增加4-5倍。两种漆酶均易于被叠氮化物,乙酸根和氯离子抑制。藜芦醇抑制漆酶催化的DMP聚合。 Botryosphaeria sp。的日益增长的文化。产生β-葡聚糖类型的胞外多糖,当在藜芦醇存在下生长时其合成受到抑制。
  • 【系统发育和形态重新评估引起芒果病的葡萄球菌物种。】 复制标题 收藏 收藏
    DOI:10.3852/mycologia.97.1.99 复制DOI
    作者列表:Slippers B,Johnson GI,Crous PW,Coutinho TA,Wingfield BD,Wingfield MJ
    BACKGROUND & AIMS: :Species of Botryosphaeria are among the most serious pathogens that affect mango trees and fruit. Several species occur on mangoes, and these are identified mainly on the morphology of the anamorphs. Common taxa include Dothiorella dominicana, D. mangiferae (= Natrassia mangiferae), D. aromatica and an unidentified species, Dothiorella 'long'. The genus name Dothiorella, however, is acknowledged as a synonym of Diplodia. This study aimed to characterize and name the Botryosphaeria spp. associated with disease symptoms on mangoes. To achieve this isolates representing all four Dothiorella spp. mentioned above were compared with the anamorphs of known Botryosphaeria spp., based on conidial morphology and DNA sequence data. Two genomic regions were analyzed, namely the ITS rDNA and beta-tubulin regions. The morphological and molecular results confirmed that the fungi previously identified from mango as species of Dothiorella belong to Fusicoccum. Dothiorella dominicana isolates were identical to isolates of F. parvum (teleomorph = B. parva). A new epithet, namely F. mangiferum, is proposed for isolates previously treated as D. mangiferae or N. mangiferae. Isolates of D. aromatica were identified as F. aesculi (teleomorph = B. dothidea). A fourth Fusicoccum sp. also was identified as those isolates previously known as Dothiorella 'long'. A key is provided to distinguish these species based on anamorph morphology in culture. This study provides a basis for the identification of Botryosphaeria species from mango, which is important for disease control and to uphold quarantine regulations.
    背景与目标: :Botryosphaeria的物种是影响芒果树和水果的最严重的病原体之一。芒果上有几种物种,这些物种主要是根据无性型的形态鉴定的。常见的分类群包括Dothiorella dominicana,D。mangiferae(= Natrassia mangiferae),D.aromana和一种不明物种,Dothiorella'long'。然而,Dothiorella属被认为是Diplodia的同义词。这项研究旨在表征和命名Botryosphaeria spp。与芒果的疾病症状有关。为实现此目的,分离株代表了所有四个Dothiorella spp。根据分生孢子的形态和DNA序列数据,将上述的与已知的Botryosphaeria spp。的变形体进行了比较。分析了两个基因组区域,即ITS rDNA和β-微管蛋白区域。形态学和分子结果证实了先前从芒果中鉴定为多硫杆菌属的真菌属于镰刀菌。 Dothiorella dominicana分离株与细小镰刀菌的分离株相同(teleomorph = B. parva)。对于先前被视为芒果D. mangiferae或芒果N. mangiferae的分离株,提出了一种新的称谓,即芒果F. mangiferum。 D.aromatica的分离物被鉴定为F. aesculi(teleomorph = B. dothidea)。第四个Fusicoccum sp。也被鉴定为以前称为Dothiorella'long'的分离株。提供了根据培养中的变形形态来区分这些物种的关键。这项研究为鉴定芒果中的葡萄球菌提供了基础,这对于控制疾病和维护检疫法规很重要。
  • 【葡萄孢杆菌十一月在葡萄树上:带有Dothiorella anamorph的新物种。】 复制标题 收藏 收藏
    DOI:10.3852/mycologia.97.5.1111 复制DOI
    作者列表:Luque J,Martos S,Phillips AJ
    BACKGROUND & AIMS: :Botryosphaeria viticola sp. nov., isolated from pruned canes of Vitis vinifera in NE Spain, is described and illustrated. Phylogenetic analysis based on ITS and EF1-alpha sequences and morphological characters of both anamorph and teleomorph confirmed this taxon to be included within the group of Botryosphaeria species with Dothiorella anamorphs. It is related most closely to B. sarmentorum and B. iberica from which it differs in morphological characters of the teleomorph and DNA sequences.
    背景与目标: :葡萄孢杆菌(Botryosphaeria viticola sp。) nov。,从西班牙东北部的葡萄树的修剪的藤茎中分离出来,进行了描述和说明。根据ITS和EF1-alpha序列以及无性型和远形型的形态学特征进行的系统发育分析证实,该分类单元被包括在具有Dothiorella anamorphes的葡萄球菌物种组中。它与S. sarmentorum和B. iberica关系最密切,它们在远形和DNA序列的形态特征上有所不同。
  • 【全基因组识别八种酒渣鼻科中包含蛋白家族基因的溶素基序,并分析中国白梨对病原性真菌灰霉病的反应。】 复制标题 收藏 收藏
    DOI:10.1186/s12864-020-07032-9 复制DOI
    作者列表:Chen Q,Li Q,Qiao X,Yin H,Zhang S
    BACKGROUND & AIMS: BACKGROUND:Lysin motif-containing proteins (LYP), which act as pattern-recognition receptors, play central roles in growth, node formation, and responses to biotic stresses. The sequence of Chinese white pear genome (cv. 'Dangshansuli') along with the seven other species of Rosaceae has already been reported. Although, in these fruit crops, there is still a lack of clarity regarding the LYP family genes and their evolutionary history. RESULTS:In the existing study, eight Rosaceae species i.e., Pyrus communis, Prunus persica, Fragaria vesca, Pyrus bretschneideri, Prunus avium, Prunus mume, Rubus occidentalis, and Malus × domestica were evaluated. Here, we determined a total of 124 LYP genes from the underlined Rosaceae species. While eighteen of the genes were from Chinese white pear, named as PbrLYPs. According to the LYPs structural characteristics and their phylogenetic analysis, those genes were classified into eight groups (group LYK1, LYK2, LYK3, LYK4/5, LYM1/3, LYM2, NFP, and WAKL). Dispersed duplication and whole-genome duplication (WGD) were found to be the most contributing factors of LYP family expansion in the Rosaceae species. More than half of the duplicated PbrLYP gene pairs were dated back to the ancient WGD (~ 140 million years ago (MYA)), and PbrLYP genes have experienced long-term purifying selection. The transcriptomic results indicated that the PbrLYP genes expression was tissue-specific. Most PbrLYP genes showed differential expression in leaves under fungal pathogen infection with two of them located in the plasmalemma. CONCLUSION:A comprehensive analysis identified 124 LYP genes in eight Rosaceae species. Our findings have provided insights into the functions and characteristics of the Rosaceae LYP genes and a guide for the identification of other candidate LYPs for further genetic improvements for pathogen-resistance in higher plants.
    背景与目标: 背景:含赖氨酸基序的蛋白质(LYP),作为模式识别受体,在生长,结节形成和对生物胁迫的响应中起着核心作用。中国白梨基因组(c山苏里)与其他七种蔷薇科的序列已被报道。尽管在这些水果作物中,关于LYP家族基因及其进化史仍然缺乏明确性。
    结果:在现有研究中,评估了八种蔷薇科物种,即Pyrus communis,Prunus persica,Fragaria vesca,Pyrus bretschneideri,Prunus avium,Prunus mume,Rubus occidentalis和Malus×domestica。在这里,我们从下划线的蔷薇科物种中确定了总共124个LYP基因。其中有18个基因来自中国白梨,名为PbrLYPs。根据LYPs的结构特征和系统发育分析,将这些基因分为八类(LYK1,LYK2,LYK3,LYK4 / 5,LYM1 / 3,LYM2,NFP和WAKL)。分散复制和全基因组复制(WGD)被发现是蔷薇科物种LYP家族扩张的最主要因素。重复的PbrLYP基因对中有一半以上可追溯到古代WGD(约1.4亿年前(MYA)),并且PbrLYP基因经历了长期的纯化选择。转录组结果表明PbrLYP基因表达是组织特异性的。大多数PbrLYP基因在真菌病原体感染下在叶片中显示差异表达,其中两个位于浆膜中。
    结论:综合分析鉴定了八种蔷薇科的124个LYP基因。我们的发现为酒渣鼻LYP基因的功能和特性提供了见识,并为其他候选LYP的鉴定提供了指导,以进一步遗传改良高等植物中的病原体抗性。
  • 【植物致病性真菌灰葡萄孢的新型线粒病毒的分子特征。】 复制标题 收藏 收藏
    DOI:10.1007/s00705-020-04886-y 复制DOI
    作者列表:Liu H,Liu M,Zhu H,Zhong J,Liao X,Zhou Q
    BACKGROUND & AIMS: :Here, a novel mycovirus, Botryosphaeria dothidea mitovirus 1 (BdMV1), was isolated from a phytopathogenic fungus, Botryosphaeria dothidea, and its molecular characteristics were determined. BdMV1 has a genome of 2,667 nt that contains a single large open reading frame (ORF) using the fungal mitochondrial genetic code. The ORF encodes an RNA-dependent RNA polymerase (RdRp) of 727 amino acids with a molecular mass of 81.64 kDa. BLASTp analysis revealed that the RdRp domain of BdMV1 has 39.59% and 39.18% sequence identity to Plasmopara viticola associated mitovirus 43 and Setosphaeria turcica mitovirus 1, respectively. Phylogenetic analysis further suggested that BdMV1 is a new member of the genus Mitovirus within the family Mitoviridae. To the best of our knowledge, this is the first report of a mitovirus in B. dothidea.
    背景与目标: :在这里,从一种植物病原性真菌Botryosphaeria dothidea中分离出了一种新型的分枝杆菌Bothosphaeria dothidea线粒体病毒1(BdMV1),并确定了其分子特性。 BdMV1具有2,667 nt的基因组,其中包含使用真菌线粒体遗传密码的单个大开放阅读框(ORF)。 ORF编码一个727个氨基酸的RNA依赖性RNA聚合酶(RdRp),分子量为81.64 kDa。 BLASTp分析显示,BdMV1的RdRp结构域分别与葡萄膜疟原虫相关的线粒体病毒43和粘粒线粒体线粒体病毒1具有39.59%和39.18%的序列同一性。系统发育分析进一步表明,BdMV1是线粒体病毒科中线粒体病毒属的新成员。据我们所知,这是首次在轮状芽胞杆菌中出现线粒体病毒的报道。
  • 【苹果圆环病的病原体Botryosphaeria dothidea和B. kuwatsukai的比较基因组学揭示了致病性相关基因的物种扩展和物种形成过程中毒力基因含量的变化。】 复制标题 收藏 收藏
    DOI:10.5598/imafungus.2018.09.02.02 复制DOI
    作者列表:Wang B,Liang X,Gleason ML,Zhang R,Sun G
    BACKGROUND & AIMS: :Ring rot, one of the most destructive diseases of apple worldwide, is caused primarily by Botryosphaeria dothidea and B. kuwatsukai. Here, we sequenced the genomes of B. dothidea strain PG45 (44.3 Mb with 5.12 % repeat rate) and B. kuwatsukai epitype strain PG2 (48.0 Mb with 13.02 % repeat rate), and conducted a comparative analysis of these two genomes, as well as other sequenced fungal genomes, in order to understand speciation and distinctive patterns of evolution of pathogenicity-related genes. Pair-wise genome alignments revealed that the two species are highly syntenic (96.74 % average sequence identity). Both species encode a significant number of pathogenicity-related genes, e.g. carbohydrate active enzymes (CAZYs), plant cell wall degrading enzymes (PCWDEs), secondary metabolites (SMs) biosynthetic enzymes, cytochrome P450 enzymes (CYPs), and secreted peptidases, in comparison to all additional sequenced fungal species involved in various life-styles. The number of pathogenicity-related genes in B. dothidea and B. kuwatsukai is higher than other genomes of Botryosphaeriaceae pathogens (Macrophomina phaseolina and Neofusicoccum parvum), suggesting a secondary round of Botryosphaeria-lineage expansion in the family. There were, however, also significant differences in the genomes of the two Botryosphaeria species. Botryosphaeria kuwatsukai, which infects only apple and pear, apparently lost a set of SMs genes, CAZYs and PCWDEs, possibly as a result of host specialization. Botryosphaeria kuwatsukai contained significantly more transposable elements and higher value of repeat induced point (RIP) index than B. dothidea. Our results will be instrumental in understanding how both phytopathogens interact with their plant hosts and in designing efficient strategies for disease control and molecular breeding to help ensure global apple production and food security.
    背景与目标: :腐烂病是全球范围内最具破坏力的苹果病之一,主要是由Botryosphaeria dothidea和B. kuwatsukai引起的。在这里,我们对B. dothidea菌株PG45(44.3 Mb,重复率5.12%)和B. kuwatsukai表型菌株PG2(48.0 Mb,13.02%重复率)的基因组进行了测序,并对这两个基因组进行了比较分析。与其他测序真菌基因组一样,以便了解致病性相关基因的物种形成和独特的进化模式。成对基因组比对显示这两个物种具有高度同义性(平均序列同一性为96.74%)。两种物种都编码大量的致病性相关基因,例如。与涉及各种生活方式的所有其他测序真菌物种相比,碳水化合物活性酶(CAZYs),植物细胞壁降解酶(PCWDEs),次生代谢产物(SMs)生物合成酶,细胞色素P450酶(CYPs)和分泌的肽酶。 B. dothidea和B. kuwatsukai中与致病性相关的基因的数量高于其他Botryosphaeriaceae病原体的基因组(Macrophomina phaseolina和Neofusicoccum parvum),表明该家族中出现了第二轮Botryosphaeria世系的扩展。但是,两种葡萄球菌物种的基因组也存在显着差异。只感染苹果和梨的Botryosphaeria kuwatsukai显然丢失了一组SMs基因,CAZYs和PCWDEs,这可能是宿主特化的结果。黑僵菌(Botryosphaeria kuwatsukai)比双歧杆菌(B. dothidea)含有更多的转座因子和更高的重复诱导点(RIP)指数值。我们的结果将有助于理解两种植物病原体如何与其植物宿主相互作用,以及设计有效的疾病控制和分子育种策略,以帮助确保全球苹果生产和食品安全。
  • 【毛白杨中的水杨酸是由多孢灰霉菌感染诱导的远程信号分子。】 复制标题 收藏 收藏
    DOI:10.1038/s41598-018-32204-9 复制DOI
    作者列表:Li YX,Zhang W,Dong HX,Liu ZY,Ma J,Zhang XY
    BACKGROUND & AIMS: :The salicylic acid (SA) plays a critical role during the establishment of systemic acquired resistance (SAR) in uninfected plant tissues after localised exposure to a pathogen. Here, we studied SA in Populus tomentosa infected by the plant pathogen Botryosphaeria dothidea. The accumulation of SA and methyl salicylate (MeSA) occurred in chronological order in P. tomentosa. The SA and MeSA contents were greater at infected than uninfected sites. Additionally, a gene expression analysis indicated that SA might be accumulated by phenylalanine ammonialyase (PAL) and converted to MeSA by SA carboxyl methyltransferase (SAMT), while MeSA might convert to SA by SA-binding protein 2 (SABP2). The expressions of SAMT at infected sites and SABP2 at uninfected sites, respectively, were significantly up-regulated. Thus, SA might be converted to MeSA at infected sites and transported as a signalling molecule to uninfected sites, where it is converted to SA for SAR. Moreover, the expressions of pathogenesis-related genes PR-1, PR-2 and PR-5 in P. tomentosa were up-regulated by the B. dothidea infection. Our study determined that variations in SA and MeSA contents occur at infected and uninfected sites in poplar after pathogen infection and contributed to the remote signals for poplar SAR.
    背景与目标: 水杨酸(SA)在局部暴露于病原体后在未感染的植物组织中建立系统性获得性耐药(SAR)的过程中起着至关重要的作用。在这里,我们研究了被植物病原体Botryosphaeria dothidea感染的毛白杨中的SA。 SA和水杨酸甲酯(MeSA)的积累按时间顺序发生在毛白杨中。被感染部位的SA和MeSA含量高于未感染部位。此外,基因表达分析表明,SA可能通过苯丙氨酸解氨酶(PAL)积累,并通过SA羧甲基转移酶(SAMT)转化为MeSA,而MeSA可能通过SA结合蛋白2(SABP2)转化为SA。分别在感染位点的SAMT和未感染位点的SABP2表达明显上调。因此,SA可能会在感染部位转化为MeSA,并作为信号分子转运到未感染部位,然后在SA转化为SA。此外,致病性相关基因PR-1,PR-2和PR-5在毛白僵菌中的表达受B. dothidea感染的影响而上调。我们的研究确定,病原体感染后,SA和MeSA含量的变化发生在杨树的受感染和未感染部位,并有助于杨树SAR的远程信号。
  • 【突尼斯的葡萄藤的溃疡病和溃疡病与Botryosphaeria dothidea,Diplodia seriata和Neufusicoccum luteum有关。】 复制标题 收藏 收藏
    DOI:10.1094/PDIS-05-13-0530-PDN 复制DOI
    作者列表:Chebil S,Fersi R,Yakoub A,Chenenaoui S,Chattaoui M,Melki I,Zemni H,Rhouma A,Durante G,Zacchi E,Mliki A
    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。
  • 【苹果环核苷酸门控离子通道基因对灰霉病菌感染具有高度响应性,提高了本氏烟草对细菌和真菌病原体的敏感性。】 复制标题 收藏 收藏
    DOI:10.1016/j.plantsci.2018.01.009 复制DOI
    作者列表:Zhang W,Dong C,Zhang Y,Zhu J,Dai H,Bai S
    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。
  • 【从家蝇(Malus domestica)鉴定出的PR-4基因参与了对灰霉病的防御反应。】 复制标题 收藏 收藏
    DOI:10.1016/j.plaphy.2012.10.016 复制DOI
    作者列表:Bai S,Dong C,Li B,Dai H
    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。
  • 【马里兰州苹果果实上出现白色腐烂的Botryosphaeria dothidea的首次报道。】 复制标题 收藏 收藏
    DOI:10.1094/PDIS-01-13-0053-PDN 复制DOI
    作者列表:Jurick WM 2nd,Vico I,Gaskins VL,Janisiewicz WJ,Peter KA
    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。
  • 【评估葡萄藤中灰霉病的杀菌剂管理。】 复制标题 收藏 收藏
    DOI:10.1094/PDIS-11-11-0998-RE 复制DOI
    作者列表:Pitt WM,Sosnowski MR,Huang R,Qiu Y,Steel CC,Savocchia S
    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。
  • 【碳水化合物碳源对灰霉菌产生组成型和诱导型漆酶的影响。】 复制标题 收藏 收藏
    DOI:10.1002/jobm.200310250 复制DOI
    作者列表:Alves da Cunha MA,Barbosa AM,Giese EC,Dekker RF
    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。
  • 【苹果果实环腐病的潜在病原体Botryosphaeria dothidea sdau11-99的基因组装配和注释。】 复制标题 收藏 收藏
    DOI:10.1094/PDIS-06-20-1182-A 复制DOI
    作者列表:Yu C,Diao Y,Lu Q,Zhao J,Cui S,Peng C,He B,Liu HX
    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。

  • 【Meyerozyma guilliermondii Y-1的生物防治效率对由Botryosphaeria dothidea引起的苹果采后腐烂的作用及其可能的作用机理。】 复制标题 收藏 收藏
    DOI:10.1016/j.ijfoodmicro.2020.108957 复制DOI
    作者列表:Huang Y,Sun C,Guan X,Lian S,Li B,Wang C
    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。

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