蓝莓枝枯病病原菌鉴定

 蓝莓 (Vaccinium spp.),是一类属于杜鹃花科 (Ericaceae) 越橘属 (Vaccinium) 的多年生小浆果类果树,广泛地应用于医药、保健、化妆品和环境保护等各方面,联合国粮农组织将其果实列为人类世界五大健康食品之一,也是我国南方酸性土丘陵地区值得发展的经济作物。全国已超过 10 个省份开始大面积的蓝莓商业化栽培,预计未来 10 年将超过 10 万 hm^{2 [1]}。但是,随着生产面积的不断扩大,蓝莓病害也日趋严重,严重影响和制约蓝莓产业的发展。2010年至今,在云南省昆明市周边及丽江市蓝莓种植区发生一种病害,主要为害嫩枝、枝条和主干,发病初期,感病枝条上产生红褐色圆或椭圆形坏死斑,随后逐渐扩大,造成枝条干枯死亡,并可扩展造成主干枯萎。调查发现,该病发病率通常在 10％～20％,重者可达 25％ 以上,可引起蓝莓整株枯萎死亡。本文通过病原菌分离、致病性测定及传统、分子鉴定,明确了该病的病原,以期为病害防治提供理论依据。     

蓝莓间座壳芽枯病病原菌鉴定及其生物学特性

<正>蓝莓(Vaccinium corymbosum L.),属于杜鹃花科(Ericaceae)越橘属(Vaccinium)多年生小浆果类果树,果实蓝色,营养丰富,具有良好的保健作用。随着蓝莓的大面积商业化种植,其病害的发生也愈加频繁。2013年7月对沈阳市蓝莓种植园病害调查时,发现一种新病害。该病主要危害基生枝中上部芽眼,受害芽坏死,芽眼周围出现褐色或红褐色椭圆形病斑,凹陷,后期中部呈灰白色,病斑     

韩国首次报道蓝莓红环斑病毒侵染蓝莓

蓝莓红环斑病毒(Blueberry red ringspot virus,BRRSV)属Caulimovididae科大豆萎黄(退绿)斑驳样病毒属(Soymovirus),引起蓝莓(Vaccinium corymbosum)叶、茎和果的红环斑病。美国、日本、捷克、斯洛文尼亚、     

蓝莓枝干溃疡病病原鉴定

 越橘为杜鹃花科 (Ericaceae) 越橘属 (Vaccinium) 小浆果类果树,其中的蓝果类型俗称“蓝莓”。自21 世纪以来,蓝莓这一新兴果树在中国开始了大面积的产业化生产栽培,据统计,到2008年,全国超过10个省份开始了商业化栽培,总栽培面积由2000年的24 hm2快速发展到2 000 hm2,预计未来10年内将超过10万 hm2。随着生产面积的不断扩大,各种病虫害的危害以及如何进行有效防治的问题显得越来越突出。然而,我国对蓝莓病虫害的研究很少,病虫害对于蓝莓产业发展的潜在威胁随着栽培面积的迅速增加而增大［1］。蓝莓枝干溃疡病是近年来在我国辽宁地区发生的一种为害严重的新病害。国外许多学者曾对蓝莓枝干溃疡病进行过研究报道,明确Botryosphaeria属真菌为该病病原,并对其病原学、致病性及发生流行规律进行了大量研究［2,3］,而我国尚未见相关研究报道。据此,作者采集病害枝干样品,分离获得病原菌,通过致病性测定、病原菌培养特性观察并结合分子生物学研究,旨在明确该病害的症状、病原菌种类,为其防治提供理论依据。     

云南蓝莓病虫害调查及防治方法

为了解云南蓝莓栽培中的病虫害危害情况,于2008-2012年对云南省5个蓝莓主栽区的蓝莓病虫害进行了系统调查。记录危害蓝莓的病虫害23种,其中真菌病害7种,病毒病害3种,生理性病害3种,虫害10种。在系统调查病虫害危害程度的基础上,提出了蓝莓的缺铁症和斑喙丽金龟为当前防治的主要对象,并对其发生、危害情况及其相应的防治技术措施作了概述。     

不同栽培模式下蓝莓园果蝇数量的动态规律

蓝莓作为新兴的经济作物,因其极高的营养价值和经济价值,种植面积逐年增加。但近几年,果蝇成为蓝莓上的主要害虫,严重影响了我国蓝莓产业的发展。为了明确青岛地区为害蓝莓的果蝇种类及发生动态规律,2016年5-12月,在青岛沃林蓝莓采摘园,选择暖棚、冷棚和大田3种蓝莓栽培环境,在相同高度悬挂糖醋液诱集瓶,监测果蝇发生的种类及数量动态规律。共进行了33次调查,诱集果蝇11 709头。结果表明,青岛地区为害蓝莓的果蝇主要有黑腹果蝇Drosophila melanogaster Meigen、铃木氏果蝇Drosophila suzukii Matsumura、海德氏果蝇Drosophila hydei Sturtevant和伊米果蝇Drosophila immigrans Sturtevant 4种。在3种不同的种植环境中,为害蓝莓的优势种为黑腹果蝇,共计6 878头,占总数量的59%,平均23.7头/5瓶,其次是铃木氏果蝇,共计4 372头,占总数量的37%,平均14.7头/5瓶,海德氏果蝇和伊米果蝇发生数量较少,共369头和90头,分别占总数量的3%和1%,平均1.2头/5瓶和0.4头/5瓶。黑腹果蝇在6-10月发生数量一直处于较高水平,而铃木氏果蝇在6-7月和10-11月出现2个明显的发生高峰,8-9月是发生的低谷期。冷棚诱集到果蝇数量为5 736头,占总数量的49%,大田为3 746头,占总数量的32%,暖棚为2 227头,占总数量的19%,暖棚能更好地减少果蝇的发生。本文讨论了不同栽培措施对蓝莓园果蝇种类和数量动态的影响,以期为蓝莓果蝇的监测和综合防治提供依据。     

麻江县蓝莓枝枯病和果腐病病原鉴定及害虫调查

2015年5月,贵州省黔东南州麻江县蓝莓(Vaccnium uliginosum)种植地发生了2种重要真菌病害和1种鳞翅目幼虫导致的虫害。采用ITS保守序列测序和形态学鉴定的方法对致病的病原微生物进行了分类鉴定。结果表明,其病原菌为乌饭树拟茎点霉(Phomopsis vaccinii)和葡萄座腔菌(Botryosphaeria dothidea),导致的真菌病害分别为为蓝莓枝枯病和果腐病;害虫为绿尾大蚕蛾(Actias selene ningpoana Felder)的幼虫。     

蓝莓蜜蜂授粉与病虫害绿色防控技术集成示范效益分析

利用意大利蜜蜂在怀宁县蓝莓主栽品种奥尼尔、巴尔德温、杰兔上进行蜜蜂授粉与病虫害绿色防控集成技术融合大面积示范。结果显示,示范区蓝莓的坐果率、平均单果重、单株平均产量等比仅靠风媒自花授粉都有不同程度提高,单位面积蓝莓鲜果产量比自然授粉(不放蜂)、病虫害常规防治对照区增加65.8%,示范区绿色防控技术能有效地防治蓝莓病虫害并保护天敌,病虫害总体防效87.5%,鲜果损失率3.4%;绿色防控技术集成示范区比常规防治区减少农药制剂使用量41.3%,其效益比自然授粉(不放蜂)、病虫害常规防治对照区增加约1倍、投入产出比高出50.9%。     

基于胶体金免疫层析法快速检测蓝莓中的百菌清残留

为建立蓝莓样品中百菌清残留快速筛查方法，以农药百菌清为目标分析物，系统研究了胶体金标记参数及样品前处理方法对胶体金免疫层析方法 (colloidal gold immuno-chromatographic assay, GICA)的影响。结果表明：以25 nm的胶体金颗粒标记百菌清单克隆抗体作为检测探针，分别将包被原百菌清-BSA (1 mg/mL)和羊抗鼠IgG抗体(0.1 mg/mL)包被于硝酸纤维膜(NC膜)，形成检测线(T线)和质控线(C线)，组装成百菌清胶体金免疫层析检测试纸条。蓝莓样品经酸化乙腈提取，双蒸水(dd H2O)稀释后，应用该纸条对蓝莓中百菌清残留肉眼观察检出限(LOD)为0.1 mg/kg (T线完全消线)，可实现15 min内蓝莓中百菌清的定性与半定量分析，同时，试纸条对样品中4-羟基百菌清、五氯硝基苯、多菌灵和腐霉利的检测不存在交叉反应。蓝莓中百菌清添加回收试验的胶体金免疫层析检测试纸条测试结果与超高效液相色谱-三重四级杆串联质谱仪(UPLC-MS/MS)方法的检测结果一致。这两种方法都可以成功地应用于蓝莓中百菌清的检测，胶体金免疫层析检测试纸条有助于现场检...     

解淀粉芽胞杆菌JK10防治蓝莓根癌病的机理研究

本研究旨在揭示解淀粉芽胞杆菌JK10防治蓝莓根癌病的生防机制。利用改良发酵培养基对解淀粉芽胞杆菌JK10菌株进行发酵,用盐酸沉淀、硫酸铵沉淀、乙酸乙酯萃取、大孔树脂吸附四种方法对JK10发酵液中的活性物质进行分离,利用MALDI-TOF-MS对活性最高的粗提物进行分析明确其种类;对JK10进行绿色荧光蛋白标记,明确其在蓝莓根际土及组织内的定殖能力;使用高通量测序技术对蓝莓健康根组织、根癌病根组织及JK10处理的根癌病根组织内生细菌群落结构进行分析。结果表明,四种方法得到的粗提物对根癌土壤杆菌均有良好的拮抗效果,其中大孔树脂吸附法得到的粗提物抑菌活性与产率均最高。JK10发酵液中能检测出bacillomycin D和fengycin两类脂肽类化合物、大环内酯类抗生素Divergolide D和聚酮类化合物Bacillaene;菌株JK10能够稳定定殖于蓝莓根际土、蓝莓根组织细胞间隙和蓝莓叶中,定殖数量在接种35 d后趋于稳定,在蓝莓根际土和蓝莓叶中分别为1×10⁶、4.0×10⁵cfu/g;蓝莓根癌病根组织在施入生防菌JK10菌株后,相比自然条件下蓝莓根癌病根组织内生细菌菌落结构发生改变,物种多样性增加,结构更加趋向于健康蓝莓根组织内生细菌。综上所述,解淀粉芽胞杆菌JK10通过产生对蓝莓根癌病菌有抑制活性的物质,稳定定殖在蓝莓根际土及蓝莓根、叶组织中,改变蓝莓根内生细菌的群落结构而达到防治蓝莓根癌病的目的。     

Multiple Alternaria species groups are associated with leaf blotch and fruit spot diseases of apple in Australia

Leaf blotch and fruit spot of apple caused by Alternaria species occur in apple orchards in Australia. However, there is no information on the identity of the pathogens and whether one or more Alternaria species cause both diseases in Australia. Using DNA sequencing and morphological and cultural characteristics of 51 isolates obtained from apple leaves and fruit with symptoms in Australia, Alternaria species groups associated with leaf blotch and fruit spot of apples were identified. Sequences of Alternaria allergen a1 and endopolygalacturonase gene regions revealed that multiple Alternaria species groups are associated with both diseases. Phylogenetic analysis of concatenated sequences of the two genes resulted in four clades representing A. arborescens and A. arborescens‐like isolates in clade 1, A. tenuissima/A. mali isolates in clade 2, A. alternata/A. tenuissima intermediate isolates in clade 3 and A. longipes and A. longipes‐like isolates in clade 4. The clades formed using sequence information were supported by colony characteristics and sporulation patterns. The source of the isolates in each clade included both the leaf blotch variant and the fruit spot variant of the disease. Alternaria arborescens‐like isolates were the most prevalent (47%) and occurred in all six states of Australia, while A. alternata/A. tenuissima intermediate isolates (14%) and A. tenuissima/A. mali isolates (6%) occurred mostly in Queensland and New South Wales, respectively. Implications of multiple Alternaria species groups on apples in Australia are discussed.     

山茱萸褐斑病病原鉴定

 山茱萸(Cornus officinalis)为山茱萸科(Cornaceae)山茱萸属植物, 其成熟干燥果肉(俗称枣皮)为我国常用名贵中药材。河南省南阳市(西峡、内乡和南召等县)为我国山茱萸主产区之一, 仅西峡县截止1990年已发展到500万株, 年产枣皮315万kg, 产值达4 200万元, 居全国首位。近年, 作者对西峡县山茱萸产区进行病害调查时发现, 由链格孢菌(Alternaria sp.)引起的山茱萸褐斑病发生普遍, 该病严重发生时可致植株95%以上叶片枯死, 落果率高达87%, 造成山茱萸大幅减产, 给当地果农带来严重经济损失。迄今, 未见链格孢菌引起的山茱萸病害报道。本研究对该病的病原菌进行分离鉴定, 以期为病害的防控提供依据。     

水稻新病害叶鞘黑斑病的病原鉴定

 2011年7月,在湖南省花垣县水稻上发现一种新的病害,症状表现为叶鞘上出现长椭圆形黑斑,病健交界模糊。分离到的4隔孢弯孢霉(HNHY001)经离体和活体人工接种均可产生黑斑;另一种黑孢霉菌株(HNHY002)不引致叶鞘黑斑。从湖南省栽培水稻150份种子样本中共分离到弯孢霉菌株27个,其中4个菌株具有4隔孢子。这4个菌株中1个菌株不产生子座,其余3个均产生分枝子座。经人工接种除不产生子座的外,其余3个菌株均能产生典型的叶鞘黑斑。HNHY001的rDNA-ITS序列(GenBank登录号JQ360963)经BLAST搜索,与之最接近的几个序列为膝曲旋孢腔菌(Cochliobolus geniculatus)及其有丝分裂产孢种—4隔孢弯孢霉。鉴于单独以及与其它4隔孢弯孢霉菌株进行对峙培养均未产生有性阶段,建议根据此菌无性阶段特征即分生孢子较直、4横隔、子座分枝等特征,鉴定为膝曲旋孢腔菌有丝分裂产孢种之一的假弯孢(C.fallax)。水稻叶鞘黑斑病系国内外首次报道。     

Pathogenic variation of Alternaria species associated with leaf blotch and fruit spot of apple in Australia

Four Alternaria species groups (A. longipes, A. arborescens, A. alternata/A. tenuissima and A. tenuissima/A. mali) are associated with leaf blotch and fruit spot of apple in Australia. There is no information on the variability of pathogenicity among the species and isolates within each species causing leaf blotch or fruit spot. We used a detached leaf assay and an in planta fruit inoculation assay to determine the pathogenicity and virulence of the four Alternaria species. Our results showed that isolates within the same species were not specific to either leaf or fruit tissue and showed great variability in pathogenicity and virulence, indicating cross-pathogenicity, which may be isolate dependent rather than species dependent. Generally, virulence of A. tenuissima and A. alternata isolates on leaf and fruit was higher than other species. Isolates of all species groups were pathogenic on leaves of different cultivars, but pathogenicity on fruit of different cultivars varied among isolates and species. Implications of our findings on prevalence of the diseases in different apple-producing regions in Australia and the development of targeted disease management of the diseases are discussed.     

Alternaria Leaf Spot on Three Species of Pelargonium Caused by Alternaria alternata in Japan

A new disease of pelargonium (Pelargonium domesticum Bailey), ivy geranium (P. peltatum (L.) L'Hér. ex Ait.) and scented geranium (P. graveolens L'Hér.), primarily causing brown spots on leaves, was found in Kawasaki-shi in Kanagawa Prefecture and Tachikawa-shi in Tokyo. An Alternaria sp. was consistently isolated from these diseased leaves, and the isolates were pathogenic to their host leaves. Based on morphological characteristics, the causal fungus in all three cases was identified as Alternaria alternata (Fr.) Keissler. Because Alternaria leaf spot of geranium by A. alternata has already been reported, the pathogenicity of isolates from four groups of genus Pelargonium was investigated. The isolates from scented geranium were pathogenic only to their original host, but the isolates from pelargonium, ivy geranium and geranium were pathogenic to all groups of pelargonium. This is the first report of this disease on pelargonium, ivy geranium and scented geranium caused by A. alternata in Japan. We propose the names for these diseases as Alternaria leaf spot of pelargonium (kappan-byo), Alternaria leaf spot of ivy geranium (kappan-byo) and Alternaria leaf spot of scented geranium (kappan-byo). Received 11 December 2000/ Accepted in revised form 19 July 2001     

Application timing of herbicides,glyphosate and atrazine,sway respective epidemics of foliar pathogens in herbicide-tolerant rapeseed

Rapeseed (Brassica napus) production in Australia relies heavily on triazine-or glyphosate-tolerant cultivars. For 14 triazine-tolerant cultivars, disease development of Neopseudocercosporella capsellae (white leaf spot), Alternaria brassicae and A. japonica (Alternaria leaf spot), and Hyaloperonospora brassicae (downy mildew) were all dependent upon herbicide application timing (p < 0.001), with significant differences between cultivars (p < 0.001) and a significant interaction (p < 0.001) between herbicide application timing and cultivars. Atrazine applied preinfection by N. capsellae, A. brassicae, or A. japonica enhanced disease incidence, severity, and leaf collapse, while atrazine application postinfection for these same pathogens reduced all three disease parameters. However, for H. brassicae, application of atrazine after, and especially prior to, infection resulted in lower disease incidence, severity, and leaf collapse. Application of glyphosate on five glyphosate-tolerant cultivars for N. capsellae resulted in significant differences (p < 0.05) between glyphosate application treatments, and between host cultivars in terms of incidence and consequent leaf collapse. For A. brassicae, A. japonica, and H. brassicae, glyphosate resulted in significant differences (p < 0.001) across application timings between cultivars, and a significant interaction (p < 0.001) between herbicide application timings and cultivars. Glyphosate applied on glyphosate-resistant rapeseed after, and especially prior to, attack by H. brassicae, reduced downy mildew. These are the first studies to highlight how the timing of application of triazine or glyphosate in relation to pathogen infection is critical to the susceptibility of rapeseed to white leaf spot, Alternaria leaf spot, and downy mildew. This new understanding offers fresh possibilities for improved management of these diseases in herbicide-tolerant rapeseed crops.     

苍术黑斑病病原鉴定

 The pathogen causing dark leaf spot on Atractylodes lancea was isolated from diseased leaf. Based on morphological characteristics, pathogenicity test, the sequence of ribosomal DNA-ITS, the pathogen was identified as Alternaria tenuissima (Kunze) Wiltshire. This is the first report that this fungus infects A. lancea.     

苎麻叶斑病病原菌鉴定

 苎麻\[Boehmeria nivea（L.）Gaud\]属荨麻科（Urticaceae）苎麻属（Boehmeria）,多年生宿根性草本植物。我国苎麻栽培历史悠久,病害及病原种类多。2010年,在重庆璧山丁家镇高古村苎麻种植区发现一种新病害,该病菌主要为害苎麻叶片,病害蔓延迅速且危害严重,严重时病叶率达50%以上,给当地苎麻产业造成严重损失。为了明确该病害的病原菌种类,本文通过形态学鉴定和致病性测定,结合分子生物学技术对病原菌进行系统鉴定,确定该叶斑病的病原菌为多主棒孢（Corynespora cassiicola）。     

Identification of putative defense-related genes in Japanese pear against <Emphasis Type="Italic">Alternaria alternata</Emphasis> infection using suppression subtractive hybridization and expression analysis

The Japanese pear pathotype of Alternaria alternata, a toxin-dependent necrotrophic pathogen, causes black spot of Japanese pear by producing the host-specific AK-toxin. Pre-inoculation with nonpathogenic A. alternata or pretreatment with an elicitor prepared from A. alternata reduced disease symptoms caused by the pathogen. Salicylic acid- and jasmonic acid-dependent signaling pathways are not involved in the induced resistance to infection by the pathogen. The expression of multiple defense-related genes in Japanese pear leaves inoculated with nonpathogenic A. alternata was examined using suppression subtractive hybridization. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank database as accessions DC993229–DC993535.