拮抗植物病原真菌链霉菌菌株ZH-356的鉴定及其生防评价

[目的]对实验室分离到的菌株ZH-356进行鉴定并评价其对植物病原真菌的生物防治效果,为研发针对植物真菌病害的生防菌剂提供理论指导。[方法]通过平板对峙法确定菌株ZH-356抗菌谱,并通过16S rRNA基因序列分析确定其种属,利用离体枝条的苹果树腐烂病菌感染预防试验和患腐烂病苹果树的防治试验评价其生防效果。[结果]菌株ZH-356鉴定为链霉菌属,与直丝紫链霉菌(Streptomyces rectiviolaceus)相似性最高,为99.71%。抗菌谱试验表明,菌株ZH-356对苹果树腐烂病菌、小麦赤霉病菌、小麦根腐病菌和番茄早疫病菌等多种植物病原真菌均具有较强的抑制作用,这种抑制作用可导致苹果树腐烂病菌菌丝变粗、交叉扭曲、分支变少且容易断裂。此外,ZH-356产生的抑菌活性物质对温度和酸碱度具有高度稳定性,并且该活性物质只存在于其胞内,只有当ZH-356遇到植物病原真菌时才会被分泌出来以抑制它们的生长。在离体枝条的苹果树腐烂病菌感染预防试验中,ZH-356对苹果树腐烂病防效可达94%以上,而在患腐烂病苹果树的防治试验中,ZH-356菌制剂对苹果树腐烂病的防效高达100%。[结论]链霉菌ZH-356抑菌谱广,对多种植物病原真菌均具有良好的拮抗活性,可作为防治植物真菌病害的生防菌株,为基于ZH-356菌株的生防菌剂的开发和防治苹果树腐烂病等植物真菌病害奠定了基础。     

三种生物源农药对桃树蚜虫的防治效果研究

为筛选生产上防治桃树蚜虫的生物源农药,开展了1.5%除虫菊素水剂、0.3%苦参碱水剂和5%桉油精可溶液剂对桃树蚜虫的田间防效试验。结果表明,3种生物源农药中以0.3%苦参碱水剂防治效果最好,药后1 d防效为70.12%,药后7 d防效为91.96%,与对照药剂70% 吡虫啉水分散粒剂防效相当,具有较好的速效性和持效性。苦参碱对桃树安全,是防治桃蚜的理想药剂,生产上推荐使用浓度为1 000倍液。     

农用硫酸链霉素防治芦笋茎枯病试验初报

通过室内孢子萌发抑制作用的测定和田间防治的试验,测定农用硫酸链霉素对芦笋茎枯病的防治作用。结果表明,农用硫酸链霉素200 ppm对芦笋茎枯病菌的孢子萌发和病斑形成与扩展均具有明显的抑制作用,田间防效达64%,略优于70%甲基托布津可湿性粉剂800倍液的防效,可作为一种防治芦笋茎枯病的生物制剂。     

贝莱斯芽孢杆菌TMQ-KSL-1分离鉴定及其发酵液对番茄根结线虫的生防作用

镰刀菌(Fusarium spp.)和根结线虫(Meloidogyne spp.)都是植物的重要病原物,这两种病原物在寄主植物中存在着非常复杂的互作关系,可导致严重的植物土传病害。为探寻对番茄根结线虫病害具有高效防治作用的优良菌株,本研究以禾谷镰刀菌(Fusarium graminearum)为靶标病菌,采用平板稀释涂布法从多年种植番茄的设施大棚土壤中分离和筛选到一株抑菌效果较好的生防细菌菌株TMQ-KSL-1,根据形态特征、生理生化特性和16S rRNA基因测序对该菌株进行鉴定;测定不同浓度的发酵液及发酵上清液对根结线虫卵孵化率以及根结线虫二龄幼虫死亡率的影响,通过盆栽实验分析其发酵液对根结线虫病害的防治效果。结果表明,菌株TMQ-KSL-1具有较强的杀线虫活性,其发酵液和发酵上清液处理48 h线虫卵孵化抑制率分别为94.76%和90.72%;处理24 h番茄根结线虫二龄幼虫的校正死亡率分别为100%和97.37%;菌株TMQ-KSL-1发酵液100倍稀释液、200倍稀释液对番茄根结线虫病害防治效果分别为59.54%和12.14%,且100倍液处理防效与阿维菌素500倍液处理防效(6...     

瓜类细菌性果斑病菌拮抗细菌的筛选及其抑菌作用

从土壤中筛选出抑制哈密瓜细菌性果斑病的拮抗菌株,明确其分类地位,测定其生长条件及防病效果。采用梯度稀释法及平板对峙法进行菌株的分离及筛选,综合形态学观察、生理生化测定以及分子鉴定等多方面对菌株进行鉴定,确定生防菌种属,并测定其最适生长条件,利用平板对峙法测定其抑菌作用,并分别采用针刺接种法、叶面喷施法对生防菌的离体叶片和盆栽防病试验进行测定。本研究从土样中共分离出196株细菌,经对峙筛选发现有20株细菌具有抑菌效果,其中编号为131、791的两株细菌对哈密瓜细菌性果斑病病原菌具有较好的平板拮抗效果,抑菌圈直径分别为（19.03±0.13）mm,（17.55±0.29）mm。经鉴定菌株131为沙福芽孢杆菌（Bacillus safensis）,菌株791为贝莱斯芽孢杆菌（B. velezensis）。菌株131的最适生长条件为NB培养基,培养18 h,温度37℃,pH 7,接种量为2%。菌株791的最适生长条件为KB培养基,培养18 h,温度37℃,pH 6,接种量为1%。菌株131和791对哈密瓜细菌性果斑病的离体叶片防效分别为83.33%和87.53%,室内盆栽防效分别为69.86%和77.99%,同时也对多种病原真菌具有抑制作用。菌株131和791均为对哈密瓜细菌性果斑病具有较好防效的芽孢杆菌（Bacillus）,对瓜类作物细菌性果斑病的防治提供了理论补充,奠定了果斑病害生物防治的基础。     

氯虫苯甲酰胺对苹果树桃小食心虫及金纹细蛾的控制作用

2006～2007年,对35%氯虫苯甲酰胺水分散粒剂进行田间药效试验。结果表明,35%氯虫苯甲酰胺水分散粒剂是防治苹果树桃小食心虫Carposina niponensisi Walsingham和金纹细蛾Lithocolletis ringoniella Mats的有效药剂。其有效成分为35～50mg/kg处理防治桃小食心虫,药后5d的防效在83%～100%之间,药后35d防效在77%以上,整体防效不及对照药剂4.5%高效氯氰菊酯乳油30mg/kg处理;10～20mg/kg处理防治金纹细蛾,药后5～15d防效在58%～100%之间,各期防效优于生产常用药剂25%灭幼脲悬浮剂125mg/kg处理。     

应用11371复合剂防治人参锈腐病的研究

人参锈腐病是栽培参的主要病害,发病率一般在60—80％,对栽培参的损失很大。由于该病是土传病害,而且致病菌复杂,多年来尚无较好的防治方法。本文介绍了使用抗生素11371为主的复合药剂防治人参锈腐病的情况;几年来田间防效稳定在45％以上,最高防效可达70％,并且用药当年可增产15％左右。     

银杏叶枯病的田间药剂防治试验

用 4种药剂和 2种药剂组合防治银杏叶枯病的田间试验。示范结果表明 :扑海因 +甲基托布津 ,敌力脱防效最好 ;与其它的参试药剂相比 ,其防效有显著差异。从始病期 (4月中下旬 )开始 ,每隔 1 5 d左右 ,用 2 5 %敌力脱 2 0 0 0倍液 ,5 0 %扑海因 1 0 0 0倍液 + 70 %甲基托布津 1 0 0 0倍液 (按 1∶ 1的体积比混合 )交替喷药 1次 ,特别是在 6月 1 0日和 7月 30日左右的发病高峰前必须重点施药 ,防治效果就可达到 85 %以上 ,有效控制叶枯病的发生。研究结果还表明 ,喷药的时间不同 ,防治效果不同 ;用药时能加强病害综合防治的其它措施 ,防治效果也会有显著的提高。     

苹果树腐烂病菌拮抗放线菌的筛选、鉴定及防效

以苹果树腐烂病菌为靶标菌,通过对峙法和生长速率法对分离自苹果树根际土壤的放线菌进行筛选,对筛选出的拮抗菌株通过形态学和分子生物学特征进行鉴定,并测定了拮抗菌ZZ-9发酵滤液对苹果树腐烂病菌孢子萌发和菌丝生长的影响及离体枝条防效.结果表明: 经对峙初筛,15株放线菌对苹果树腐烂病菌具有抑菌作用,占所分离株数的18.8%,其中抑制率>50%的有8株.复筛结果表明,ZZ-9对腐烂病菌抑制率最高,达96.4%,显著高于其他菌株;通过培养特征、生理生化特性及16S rDNA序列分析将菌株ZZ-9初步鉴定为娄彻氏链霉菌,其在GenBank上的序列登录号为KT986228;不同稀释倍数的ZZ-9发酵滤液对腐烂病孢子萌发及菌丝生长均有明显的抑制作用,其中50倍发酵滤液对孢子萌发抑制率和菌丝生长抑制率均达80%以上,且受抑制菌丝颜色加深,分支增多,末端膨大、畸形,出现原生质浓缩与释放现象;离体枝条防效试验表明,菌株ZZ-9发酵原液对苹果树腐烂病防效可达75%以上,表明该菌株可作为防治苹果树腐烂病的生防菌株.     

农用硫酸链霉素防治芦笋茎枯病试验初报

通过室内孢子萌发抑制作用的测定和田间防治的试验,测定农用硫酸链霉素对芦笋茎枯病的防治作用。结果表明,农用硫酸链霉素２００ｐｐｍ对芦笋茎枯病菌的孢子萌发和病斑形成与扩展均具有明显的抑制作用,田间防效达６４％,略优于７０％甲基托布津可湿性粉剂８００倍液的防效,可作为一种防治芦笋茎枯病的生物制剂。     

上海市进口火龙果软腐病病害分析

应用病原形态学和真菌rDNA-ITS分子标记及Fusarium oxysporum种特异性序列分析, 对引起上海市水果市场销售的进口火龙果软腐的市场病害进行了分离与鉴定, 明确了两种主要的致病真菌为尖孢镰刀菌(Fusarium oxysporum)和单隔镰刀菌(Fusarium dimerum)。其中, F. oxysporum为引起进口火龙果采后软腐的最主要病原真菌, 该菌最适生长温度和致病温度均为25 °C, 光照条件下的致病性强, 在15 °C和35 °C条件下致病性明显降低, 5 °C和45 °C条件下该菌无法正常生长和致病, 并且该菌还能够引起香蕉、番茄、葡萄等多种水果腐烂。系统研究引起进口火龙果采后软腐病病原真菌的生物学特性, 为进口火龙果采后病原真菌有效控制方法的制定提供了有益的参考。     

First Report of Dragon Fruit (Hylocereus undatus) Anthracnose Caused by Colletotrichum truncatum in China

Anthracnose disease was detected from dragon fruit (Hylocereus undatus) at a market of Yuanjiang County, Yunnan Province, China. The results of pathogenicity test, morphology studies and sequence analyses based on ITS and β‐tubulin loci indicated that the disease was caused by Colletotrichum truncatum. The pathogen produced elliptic, yellow spots with chlorotic halos on the surface of the fruit, and the lesion become depressed gradually. Grey to black acervuli appeared on the lesion surface in concentric circles later. This is the first report of dragon fruit anthracnose caused by this pathogen in China.     

短波紫外线照射对‘金都’火龙果采后保鲜的影响

以‘金都’火龙果(Hylocereus polyrhizus ‘Jindu’)果实为试材,采用波长254 nm紫外杀菌灯为辐射源,给予不同剂量短波紫外线(Ultraviolet-C,UV-C)照射处理,探讨低剂量UV-C对火龙果采后保鲜的影响及作用机理。结果表明,不同剂量UV-C照射处理能有效抑制‘金都’火龙果果实腐烂和电导率上升,降低果实TSS含量,其中1.0 kJ·m–2紫外线辐照效果最好。1.0 kJ·m–2 UV-C处理能极显著提高贮藏期火龙果的SOD和CAT活性,显著提高贮藏早中期的几丁质酶活性和PPO活性,β-1,3葡聚糖酶活性在贮藏后期也显著高于对照,但降低了火龙果贮藏中期(第6天)的POD活性。此外,1.0 kJ·m–2 UV-C处理显著提高火龙果贮藏期H2O2含量(除第6天外),对果实的失水率无影响。采后火龙果应用适当剂量UV-C照射可提高抗病性,延长贮藏保鲜期。     

Fusarium fujikuroi associated with stem rot of red‐fleshed dragon fruit (Hylocereus polyrhizus) in Malaysia

Stem rot was recorded as one of serious diseases of red‐fleshed dragon fruit, (Hylocereus polyrhizus), in Malaysia. Fusarium fujikuroi was recovered from stem rot lesion of H. polyrhizus and the species was identified using TEF1‐α sequence and mating study. From maximum likelihood phylogenetic tree using combined TEF1‐α and β‐tubulin sequences, the F. fujikuroi isolates from stem rot were grouped according to three geographical locations, namely Peninsular Malaysia, Sabah and Sarawak. Phylogenetic analysis indicated that F. fujikuroi isolates from stem rot of H. polyrhizus were clustered separately from F. fujikuroi isolates from rice because of intraspecific variation. From amplification of MAT allele‐specific primers, 20% of the isolates carried MAT‐1 allele while 80% carried MAT‐2 allele. From isolates that carried MAT‐1 allele, 65% crossed‐fertile with MP‐C (mating population of F. fujikuroi) tester strain while for MAT‐2 allele, 56% crossed‐fertile with MP‐C. None of the isolates were identified as MP‐D (mating population of F. proliferatum). Pathogenicity test conducted on 40 representative isolates showed that the stem rot symptoms were similar with the symptoms observed in the field, and can be categorized as low, moderate and high aggressiveness, which indicated variation in pathogenicity and virulence among the isolates. This study provides novel findings regarding Fusarium species associated with stem rot of H. polyrhizus and indicated that F. fujikuroi as a new causal pathogen of the disease.     

Fusarium incarnatum is associated with postharvest fruit rot of muskmelon (Cucumis melo)

Postharvest fruit rot was observed on muskmelon (Cucumis melo) on market shelves at a melon farm in the city of Hatyai, Songkhla, Thailand. Diseased muskmelon fruit displayed cotton-like mycelia on wounds and had brown to dark brown internal tissue. Based on morphological characteristics and molecular analysis of internal transcribed spacer (ITS) and translation elongation factor 1-α (TEF1-α) DNA sequences, the fungal pathogen was identified as Fusarium incarnatum. This species belongs to the Fusarium incarnatum-equiseti species complex (FIESC). A pathogenicity test was conducted to verify Koch's postulates, and F. incarnatum was observed to cause fruit rot on muskmelon; symptoms of the disease were similar to those seen in the field. However, only artificially wounded melons became infected, suggesting that F. incarnatum is an obligate wound-infecting pathogen. To our knowledge, this is the first report of F. incarnatum causing fruit rot of muskmelon in Thailand.     

凤梨小果芯腐病病原菌凤梨镰孢菌在中国的首次发现

近年,在广东徐闻凤梨种植基地生产的凤梨果实在采后贮藏过程中发生一种果肉腐烂病害,发病率达10%左右,并有逐年加重危害的趋势。为了明确该病的病原菌,本文通过果实组织分离和接种试验获得1个致病菌株,通过形态学观察和TEF-1α序列分析,确定该病的病原菌为凤梨镰孢菌Fusarium ananatum,这是该菌所致凤梨小果芯腐病在国内的首次报道。     

First morphogenetic identification of <i>Fusarium solani</i> isolated from orange fruit in Egypt

Losses due to postharvest decay may occur at any time during postharvest handling, from harvest to consumption affecting the produce quality and quantity. Accurate identification of the pathogen causing postharvest disease is essential to the selection of an appropriate disease control approach. Nine isolates of Fusarium recovered from orange fruit were identified as Fusarium solani. The fungus is involved with fruit decay. The obtained cultures were purified and grown on potato-dextrose agar (PDA), malt yeast agar (MYA), and Czapek's nutrient media (CNM) under light for identification. A pathogenicity test was carried out to fulfil Koch's postulates. The pathogen could only enter ripe orange fruit through wounds and cracks causing the rot disease. The identification of the fungal isolates was confirmed to be F. solani by DNA sequencing, which was 99 to 100% homologous to those deposited in the Gen- Bank. The identity of nine fungal isolates was confirmed to be F. solani by DNA sequencing of the internal transcribed spacer (ITS) rDNA region (GenBank Accession Nos. DQ486874 to DQ486881 and KC758879). To our knowledge, this is the first morphogenetic identification of F. solani isolated from orange fruit in Egypt.     

An integrated approach with Trichoderma harzianum DGA01 and hot water treatment on control of crown rot disease and retention of overall quality in banana

A biological control of crown rot disease of banana fruit was analysed using an integrated approach combining hot water treatment and Trichoderma harzianum strain DGA01. Treated fruits were stored at 22–25 °C and 90–95% relative humidity for 2 weeks. The bioefficacy of fungal antagonist in vitro towards crown rot-causing pathogens, namely Lasiodiplodia theobromae, Thielaviopsis paradoxa, Colletotrichum musae and Fusarium verticillioides, was enhanced by 11.41% following hot water treatment (50 °C, 20 min). DGA01 germinated on the fruit 48 h after inoculation and parasitised the pathogen. Postharvest application showed that hot water treatment and conidial suspension of DGA01 (10⁶ ml^?1) applied singly performed significantly better than the untreated control in reducing the incidence of crown rot, but were not as effective as the fungicide. The combination of hot water treatment and DGA01 gave 93% control of fruit decay which was comparable with fungicide treatment of 95%. The quality of fruit was markedly improved in hot water treatment + DGA01 as compared to those dipped in fungicide solution. The inconsistencies of single treatments, by DGA01 or hot water dips, in controlling crown rot such as variation in severity of disease among treatments and within a treatment, were lessened by dipping the fruit in DGA01 conidial suspension following hot water treatment.     

苹果轮纹病病害名称之规范

苹果轮纹病(apple ring rot)为苹果重要病害,严重危害果实和枝干,甚至造成幼树枯死等。苹果轮纹病可引起果实腐烂、枝干疣突、溃疡、粗皮等症状,导致苹果轮纹病病害汉语名称使用混乱,如苹果轮纹病、苹果干腐病、苹果果实轮纹病、苹果枝干轮纹病、轮纹烂果病等。病原拉丁学名的使用亦相当混乱,葡萄座腔菌Botryosphaeria dothidea、贝林格葡萄座腔菌B. berengeriana、贝林格葡萄座腔菌梨生专化型B. berengeriana f. sp. pyricola、七叶树壳梭孢Fusicoccum aesculi、锹冢大茎点霉Macrophoma kuwatsukai、梨生囊孢Physalospora pyricola和梨生球座菌Guignardia pyricola等学名都在被使用。本文根据近些年来国内外的研究新进展,认为苹果轮纹病是一类复合病害,与欧美国家发生的白腐病为同一种病害,其病原包括葡萄座腔菌B. dothidea和锹冢葡萄座腔菌B. kuwatsukai。建议将该复合病害汉语统称为苹果轮纹病,英文采用apple ring rot。鉴于两种病原在我国苹果产区的普发性,建议在病害发生规律及抗病育种等研究中,对于葡萄座腔菌和锹冢葡萄座腔菌均需充分重视。     

Identification and mapping of fruit rot resistance QTL in American cranberry using GBS

Sustainability of the cranberry industry is threatened by widespread and increasing losses due to fruit rot in the field as well as increasing restrictions on fungicide inputs. Breeding for resistance offers a partial solution but is challenging because fruit rot is caused by a complex of pathogenic fungi that can vary by location and from year to year. We identified four genetically diverse germplasm accessions that exhibit broad-spectrum fruit rot resistance under field conditions. Three of these accessions were used in biparental crosses to develop four populations segregating for resistance. Genotyping by sequencing was used to generate single-nucleotide polymorphism (SNP) markers for development of high-density genetic maps and quantitative trait locus (QTL) analyses. Nineteen QTL associated with fruit rot resistance, distributed on nine linkage groups, were discovered in our populations. Three of these QTL matched previously reported fruit rot resistance QTL. Four newly reported QTL found on linkage group 8 (Vm8), which explain between 21 and 33% of the phenotypic variance for fruit rot, are of particular interest to our breeding program. The populations described herein were also phenotyped for other horticulturally important traits, and QTL associated with yield and berry weight were identified. These QTL provide markers for candidate gene discovery and for future breeding efforts to enhance and pyramid disease resistance and other traits into elite horticultural backgrounds.