杧果与Fusarium mangiferae在转录水平上的互作机制

基于Illumina HiSeq~(TM)2000平台,对健康与感病杧果顶芽的转录组进行测序,采用BLAST软件将获得的Unigene与NCBI-nr、Swiss-Prot、KEGG和COG数据库进行比对,根据基因功能注释后分析杧果病健组织的差异表达基因(DEGs),并对DEGs进行GO和Pathway富集分析。结果表明:2个样品共获得119 815条Unigene,N50为1 546 bp,平均片段长度为880 bp;鉴定了29 878个DEGs,对随机挑选的11个差异表达基因进行了qRT-PCR验证,结果与转录组一致。以corrected P-value≤0.05为阈值的代谢途径有22条,其中大多数代谢途径与植物的抗逆响应密切相关;153个DEGs参与了淀粉和蔗糖代谢途径,DEGs主要是编码糖类异构酶、水解酶和转移酶等,参与葡萄糖水解、细胞碳水化合物、丙酮酸盐和核苷酸代谢等生物进程;在抗氧化生物过程中,编码活性氧代谢相关酶且log_2Ratio10或-10以上的DEGs有20个,14个属上调表达,表明活性氧代谢在杧果与病原互作过程中起到重要的调解作用;F.mangiferae侵染杧果后,以log_2Ratio10或-10为筛选条件,共获得酚类代谢相关差异表达基因53个,其中40个DEGs上调表达,推测杧果可能是通过合成加固细胞壁的木质素或生成抑菌作用的酚类化合物来提高对病原菌的抵抗能力;杧果与F.mangiferae互作过程中,钙信号传导、SA信号途径和丝裂原活化蛋白信号传导途径相关基因表达下调,造成了下游植物抗病基因RPM1的表达受到抑制,这可能是杧果畸形病的主要成因之一。     

芦笋茎枯病病原菌及其生物学研究

芦笋茎枯病是一种分布广、为害重的毁灭性病害。研究证实,临沂市芦笋茎枯病病原为Phoma asparagi Sacc.。该菌分生孢子发芽和菌丝生长适温为23—26℃,致死温度分别为50℃10分钟和60℃10分钟,发芽和生长最适pH为7。分生孢子生长在分生孢子器内,单个分生孢子常温下可活10天,菌块内可活25天,孢子器内则长达240天。分生孢子在蒸馏水中不发芽,加糖液发芽率略有提高,在文竹和芦笋茎煮液中发芽率最高;条件适宜时2小时产生芽突,5小时发芽率达98.21%。     

法国向日葵种子中向日葵黑茎病菌的首次截获与检测

从法国进境向日葵种子中分离到3株疑似向日葵黑茎病的菌株,对所有菌株进行形态学、致病性测定和分子序列比对分析。分离菌菌落乳白色或象牙色至灰白色,有大量黑褐色分生孢子器产生,分生孢子器球形至扁球形,内含无色单胞、卵圆形分生孢子,有明显或不明显油球;针刺接种4片真叶向日葵幼苗的下胚轴,7～9d后茎部产生典型黑茎病黑色椭圆形病斑,病斑上着生黑色分生孢子器;菌丝DNA用ActF1/R1和ITS1/ITS4扩增,序列与NCBI基因库中P.macdonaldii序列相似性为98%～100%。形态学、分子生物学及致病性检测结果显示,截获的3株菌均为向日葵黑茎病菌。     

一种新的香蕉黑斑病病原菌鉴定及生物学特性研究

 在广西香蕉种植区发现一种引起香蕉黑斑病的新病原。通过对病原菌分离培养、致病性测定、形态学观察、分子生物学鉴定及对该病原菌生物学特性进行初步研究,表明引起该病害的病原菌为芒果球座菌(Guignardia mangiferae A.J. Roy)。该病原菌菌落为深橄榄色,产生棒状子囊、梭形子囊孢子及倒梨形分生孢子;其最适生长温度为28℃,最适pH 值为6;病原菌生长较好的碳源为果糖和阿拉伯糖,氮源为蛋白胨和酵母膏;在全光条件下,病原菌菌丝扩展速度最快;菌丝的致死温度为54℃。由G. mangiferae引起的香蕉黑斑病,在国内外为首次报道。     

茎基腐亚洲镰孢菌侵染抗感小麦品种的组织病理学观察

小麦茎基腐是由多种镰孢菌侵染的世界性土传病害,亚洲镰孢菌(Fusarium asiaticum)是我国冬小麦主产区茎基腐镰孢菌的优势种群,对小麦生产造成巨大损失。本研究利用绿色荧光蛋白报告基因标记亚洲镰孢菌,研究其侵染抗感小麦的病理组织学过程,建立了茎基腐病菌与寄主互作的直观性的研究体系,对病害防治及抗病育种具有重要意义。基于PEG-CaCl_2介导原生质体转化法将gfp导入亚洲镰孢菌株CF0915,对转化子进行荧光表达、PCR验证、遗传稳定性、生长特性及致病力分析,选取与野生型表现相近的转化子进行侵染分析。结果表明,绿色荧光蛋白基因(gfp)与潮霉素基因(hyg)PCR扩增表明gfp已整合入真菌基因组中,转化子菌丝与分生孢子表现强烈绿色荧光信号,gfp能够在转化子中稳定遗传,菌落形态、生长速度及致病力与野生型菌株无显著差异;将gfp标记病菌分生孢子接种感病品种1 d后,大量孢子附着于根毛及根表皮细胞开始萌发,接种2 d后观察到抗性品种分生孢子萌发;感病品种接种3 d后,菌丝直接侵入表皮细胞或沿表皮细胞间层定殖生长,扩展至皮层组织,8 d后菌丝从根部迅速扩展至茎基部,至第10 d大量菌丝充塞根皮层细胞,叶鞘维管束也被菌丝侵染,并产生大量大型分生孢子,植株表现褐色病斑,14 d后根部及茎维管束被大量菌丝体填充,而后产生大量厚垣孢子,至25 d大部分感病品种幼苗萎蔫死亡;与感病品种相比,抗性品种在整个侵染过程中表现时间滞后。本研究对引起茎基腐病的亚洲镰孢菌侵染小麦的组织学过程观察,为病菌致病机理的阐释及抗病资源的利用提供了重要理论依据。     

杧果炭疽菌拮抗细菌的筛选鉴定及其防病效果

为寻找对杧果炭疽菌Colletotrichum gloeosporioides有效的生防菌株,从瓜叶白粉病病斑上分离得到104株细菌,通过平板对峙培养筛选出16株拮抗细菌,分别测定其在8种不同培养基中的抑菌活性,筛选出有较强抑菌作用的9A、26A和4N菌株进行室内、果园防病试验,并对具有较好防治作用的9A、26A菌株进行了鉴定。结果显示,拮抗菌9A、26A和4N分别在南瓜汁、红薯汁和马铃薯汁培养基上的抑菌活性最高,对杧果炭疽菌菌丝生长抑制率分别为88.83%、87.83%和85.87%;室内9A、26A和4N菌株菌液处理离体果实,24 h后接种炭疽菌分生孢子液,对杧果炭疽病的防治效果均达100%,与咪鲜胺对照组相同;在果园9A和26A的防病效果分别为98.4%和90.5%,显著高于对照组的防效78.8%,而4N的防效只有58.2%。根据形态、生理生化特征及16S rDNA 序列分析结果,将菌株9A和26A鉴定为枯草芽胞杆菌Bacillus subtilis。研究表明,菌株9A和26A具有较好的生防利用价值。     

新疆榆树黄萎病病原菌鉴定

2013年以来,在石河子大学农学院试验站榆树苗上,出现了一种病害,可导致叶片褪绿,变黄,萎蔫,甚至枯死,剖茎检查可见维管束变成褐色。采用常规组织分离法对病茎组织进行分离、纯化获得单孢纯培养菌株;通过常规纸钵撕底蘸根法对其进行致病性测定;用形态学和rDNA-ITS序列分析方法对病原菌进行鉴定。结果表明,分离菌株的菌落形态、分生孢子梗和分生孢子的形态都与大丽轮枝菌(Verticillium dahliae)一致;经分子生物学鉴定,该菌的rDNA-ITS序列与中国棉花黄萎病菌(V.dahliae)的ITS序列(登录号EU 835817.1)相似性达99.0%,故将引起新疆榆树黄萎病的病原菌鉴定为大丽轮枝菌(V.dahliae)。     

杧果炭疽病菌Colletotrichum asianum和C.fructicola的危害特征及对杀菌剂的敏感性初探

通过离体针刺接种法初步探讨了广西杧果炭疽病的主要致病菌Colletotrichum asianum和C.fructicola在不同品种杧果上的危害症状及致病力,并用生长速率法测定这2种病菌代表菌株对咪鲜胺和吡唑醚菌酯的敏感性。结果显示:C.asianum和C.fructicola侵染同一品种杧果的症状仅有细微的差别;每种病菌对不同品种杧果的致病力有较明显的差异,其中C.asianum对‘桂七杧’的致病力最强,对‘台农一号杧’的致病力最弱,C.fructicola对‘金煌杧’的致病力最强,对‘四季蜜杧’的致病力最弱。咪鲜胺对C.asianum和C.fructicola的代表菌株的EC 50分别为0.0395~0.1322μg/mL、0.0571~0.1574μg/mL,吡唑醚菌酯对C.asianum和C.fructicola的代表菌株的EC 50分别为0.0109~0.5616μg/mL、0.1511~0.4474μg/mL,其中C.asianum对吡唑醚菌酯的最大EC 50是最小EC 50的51.5倍,存在抗药性风险。研究结果为杧果炭疽病的正确诊断、防治等提供理论依据。     

芦笋茎枯病防治对策

芦笋茎枯病是一种毁灭性的病害 ,严重影响芦笋高产、稳产。生产上 ,如何防治芦笋茎枯病 ,对提高产量、增加笋农经济收入 ,具有重大的现实意义。本文就芦笋茎枯病提出综合防治对策。1　病因芦笋茎枯病由天冬茎点霉菌寄生引起 ,以分生孢子器或菌丝在病残体上越冬 ,翌年温度达到 5℃时 ,病菌开始活动 ,15℃时散出孢子 ,侵染刚萌发的芦笋。其发病适温为 2 0～ 30℃ ,最适 2 4℃。分生孢子器随雨水滴溅或空气传播进行再侵染。其发病原因主要与种子品种、气候、土壤、施肥以及栽培方式等因素有关。2　防治对策在防治对策上应采取以农业防治为基础 …     

贝莱斯芽胞杆菌HN-2的鉴定及对杧果炭疽菌的抑菌活性研究

菌株HN-2为本实验室分离得到的一株生防细菌，通过采用形态学观察结合现代分子生物学的手段鉴定生防菌HN-2为贝莱斯芽胞杆菌Bacillus velezensis，以杧果炭疽菌Colletotrichum gloeosporioides Penz作为靶标，其发酵上清液的正丁醇萃取粗提物的活性最好，抑菌圈大小为20.93 mm，半最大效应浓度（EC₅₀）为70.62 μg/mL。通过飞行时间质谱（MALDI-TOF-MS）数据结合基因检测的结果分析发现，正丁醇提取物中主要活性成分为脂肽类物质，其中含有表面活性素（surfactin）、伊枯草菌素（iturins）和泛革素（fengycin）等。通过显微观察发现正丁醇粗提物可以造成杧果炭疽病菌菌丝扭曲、膨大、畸形，从而抑制杧果炭疽病菌的生长，菌株HN-2正丁醇提取物处理后的杧果15 d内未出现杧果炭疽病病状，对杧果果实具有较好的保护作用。贝莱斯芽胞杆菌HN-2的主要活性物质为脂肽类物质，其对植物病原真菌有较好的防治效果，具有进一步深入研究和开发应用的潜力。     

芒果疮痂病菌生物学特性研究

 本文主要研究了温度、pH、光照、维生素对芒果疮痂病菌(Sphaceloma mangiferae Bit.&Jenk.)孢子萌发、营养生长及孢子形成的影响,在14种半固体、固体培养基上对产孢量作了比较,还对湿度和营养对孢子萌发的影响做了试验。湿病原菌分生孢子萌发的温度范围为12℃~37℃,最适28℃;pH值范围为3~9,最佳pH 5;以自由水或饱和湿度条件下萌发率最高;连续黑光灯光照促进萌发;1%的葡萄糖液在28℃或33℃下明显增加了萌发率。该菌营养生长的温度范围为4℃~37℃,22℃~33℃生长良好,最适为28℃,与Bitancourt的报道基本相似;pH值范围为2~11,最佳为pH 5;黑光灯连续光照有利生长,VB₂、VB₆及V A对生长均有促进作用,在试验的培养基中PDA是生长最好的。该菌孢子形成的温度范围为12℃~33℃,最适温度为28℃;pH值范围为5~9,最佳为pH 7;光暗12 h交替处理比连续光照更利于产孢;VB₁及VA可促进产孢,但只有VA (50μg/L)既促进生长,又促进产孢;所试验的培养基中以PDA₊牛肉浸膏及PDA的产孢量最多,胡萝卜块是固体培养基中较理想的。     

Integrated Control of Powdery Mildew on Field-grown Mango Trees by Foliar Sprays of Mono-potassium Phosphate Fertilizer, Sterol Inhibitor Fungicides and the Strobilurin Kresoxym-methyl

Foliar sprays of solutions of di-potassium hydrogen orthophosphate, K2HPO4 (DKP) and potassium di-hydrogen orthophosphate, KH2PO4 (MKP), commercial systemic fungicides, and an alternating treatment with phosphate fertilizer and systemic fungicides inhibited development of the powdery mildew fungus, Oidium mangiferae, on flowers and bloom clusters of field-grown mango trees. The effectiveness of the alternation treatments with an appropriate systemic fungicide and 1% solution of mono-potassium phosphate (MKP) in controlling powdery mildew on bloom clusters was similar to that of the commercial treatment with the systemic fungicides. However, application of the systemic fungicides alone, on the same dates on which they were applied in the alternation treatment, without application of the phosphate treatment was significantly less effective in controlling the disease than either the phosphate or the alternation treatment. This indicates that the use of phosphate fertilizer has a significant role in disease control and that it can reduce the number of fungicide treatments necessary against powdery mildew by up to 50%. These results were confirmed in large-scale demonstration trials conducted in commercial orchards in 1994 and 1995. Our 1997 findings also revealed that tank-mix treatments of 1% MKP solution with half the recommended quantity of sterol inhibitor fungicide applied at 14-day intervals provided a protection against powdery mildew comparable with or superior to that given by the standard fungicides-based treatment applied at 7-day intervals. Tank-mix treatments of MKP (1%) with sterol inhibitor at the recommended rate or with the new strobilurin Kresoxym-methyl (BAS 490F, strobi), or the BAS 490F alone, were the most effective, and provided >95% protection against O. mangiferae, compared with the control. Phosphate solutions were found not to be phytotoxic to plant tissue. These treatments affected the yield of mango trees: a significantly lower yield was observed on control untreated trees, probably because of powdery mildew infection on flowers. The inhibitory effectiveness of phosphate salts makes them a potential major component of an integrated pest management program.     

Epidemiological aspects of mango malformation disease caused by Fusarium mangiferae and source of infection in seedlings cultivated in orchards in Egypt

Mango malformation, caused by the fungus Fusarium mangiferae , is one of the major diseases of this crop occurring worldwide. This study was conducted to investigate aspects of the epidemiology, survival and spread of the pathogen in general and specifically in seedlings, the majority of which are cultivated in infected orchards in Egypt. Survival of conidia of a representative isolate (506/2) declined very rapidly in soil under summer conditions (1·6 weeks for 50% population decline), but significantly less in controlled and winter conditions (17·9 and 15·0 weeks, respectively, for 50% population decline). Likewise, inoculum survival in naturally infected panicles on the soil surface declined faster than in those buried at 30-cm depths. Natural infections were evaluated on fruits and seeds in a heavily infected and a healthy orchard. In infected trees, the skins of all sampled fruits within a 2-m radius of infected panicles were infected, but the pathogen was not detected in the seeds, seed coats or flesh. The pathogen was not detected in any parts of fruits from a healthy orchard. Vegetatively malformed mango seedlings, growing under infected trees bearing infected panicles, were sampled in two locations in Egypt to determine whether infection in seedlings was systemic (evenly distributed within plant tissue) or whether the pathogen originated from malformed panicles. According to PCR-specific primer amplification, the pathogen was detected in 97% of seedling apical meristems, declining gradually to 5% colonization in roots. It was concluded that inoculum of the pathogen originates from infected panicles and affects seedlings from the meristem, with infections descending to lower stem sections and roots. Minor infections of roots may occur from inoculum originating from infected panicles, but the pathogen is not seedborne.     

A review of the pest management situation in mango agroecosystems

Integrated pest management programs for mango must be based on sampling and on economic thresholds, and must take into account the effects of cultural practices, horticultural sprays and disease control on pest and natural enemy interactions. An analysis of the mass of information available on the different mango pests,viz., fruit flies(Bactrocera sp.,Ceratitis sp.,Anastrepha sp.), mango seed weevil(Sternochetus mangiferae), thrips(Frankliniella spp.), gall midges(Procontarinia sp.), scales, mites and mealybugs is given, as well as different examples for future entomological research.     

福建果树根部的芒果半轮线虫记述

 芒果半轮线虫(Hemicriconemoides mangiferae Siddiqi,1961)在福建省一些果园发生。寄主有龙眼(Euphoria longana)、荔枝(Litchi chinensis)、芒果(Mangifera indica)、橄榄(Canarium album)、枇杷(Eriobotrya japonica)和葡萄(Vitis vinifera)等果树。雌虫唇部正面、环纹和阴门,雄虫侧带以及幼虫角质膜刺突等细微结构用扫描电子显微镜观察。     

基于Histone H3、EF-1α和β-tubulin基因部分序列的芒果畸形病菌鉴定与系统发育研究

<正>芒果畸形病是芒果生产中重要的病害之一,主要表现为花畸形和芽畸形,受害果园常因畸形花絮无法挂果而造成减产,我国攀西地区的部分果园株发病率高达100%,给当地的芒果产业带来严重经济损失。该病自1891年首次在印度发现后,陆续在世界芒果产区发生危害~([1])。引起芒果畸形病的病原菌有Fusarium mangiferae、F.mexicanum、     

Natural occurrence of Beauveria bassiana on adults of the invasive mango seed weevil Sternochetus mangiferae (Coleoptera: Curculionidae) in Brazil

Journal of Plant Diseases and Protection - The mango seed weevil Sternochetus mangiferae (Fabricius) (Coleoptera: Curculionidae) was detected in Brazil in 2014, becoming a potential threat to mango...     

A new Fusarium lineage within the Gibberella fujikuroi species complex is the main causal agent of mango malformation disease in Brazil

Mango malformation is a serious disease in tropical and subtropical areas of the world and has been attributed to various Fusarium spp., including F. mangiferae , F. proliferatum , F. sacchari , F. sterilihyphosum and F. subglutinans . Isolates of Fusarium associated with mango malformation from Brazil, Egypt, India, South Africa and the United States were evaluated through amplified fragment length polymorphisms (AFLPs) and partial DNA sequences of the genes encoding β-tubulin ( tub2 ) and translation elongation factor 1-α ( tef1 ). These techniques were used to delimit species and to estimate the genetic and phylogenetic relatedness of the isolates. In the AFLP analysis, most of the Brazilian isolates formed a unique cluster. Additionally, one small cluster was formed by isolates of F. sterilihyphosum from Brazil and South Africa, and another by isolates of F. mangiferae from Egypt, India, South Africa and the United States. In the phylogenetic analysis, most of the Brazilian isolates represented a new phylogenetic lineage in the Gibberella fujikuroi species complex, where they formed a sister clade to F. sterilihyphosum. Representatives of both clades were pathogenic to mango (cv. Tommy Atkins) and Koch's postulates were completed for isolates belonging to the new lineage and to F. sterilihyphosum . Thus, most of the mango malformation disease in Brazil is due to a distinct phylogenetic lineage of Fusarium , and to a lesser extent by F. sterilihyphosum. The new phylogenetic lineage identified in this study, together with F. mangiferae and F. sterilihyphosum , are the only known taxa of Fusarium proven to be capable of causing mango malformation.     

Antifungal activity of Datura stramonium, Calotropis gigantea and Azadirachta indica against Fusarium mangiferae and floral malformation in mango

Floral malformation caused by Fusarium mangiferae is a serious threat to mango cultivation in various countries. Different long-term measures suggested to control it were found to be unsuccessful. Present studies clearly showed strong antifungal activity of a concoction brewed from Datura stramonium, Calotropis gigantea, Azadirachta indica (neem) and cow manure (T₁) followed by methanol-water (70/30 v/v) extracts of Datura stramonium, Calotropis gigantea and Azadirachta indica (T₂) against Fusarium mangiferae. Optimal control of floral malformation was found in trees sprayed with T₁ followed by T₂ at bud break stage and again at fruit set stage when compared with the control. All the malformed buds or panicles completely dried two days after foliar spray with T₁ or T₂. In the trees treated with T₁ at fruit set stage, flower abscission was observed from the fourth day after spraying and all flowers dropped by the ninth day without requiring any manual de-blossoming, whereas in the control, the malformed panicles remained green and competed with the growing fruits for plant nutrients. In vitro culture of fresh malformed tissues in MS media along with T₁ or T₂ showed no growth of any fungus in the media. However, in vitro culture of the completely dry malformed tissues in MS media after foliar treatment with T₁ or T₂ revealed growth of F. mangiferae on the twenty fifth day indicating that the concoction-brewed compost (T₁) or methanol-water (70/30 v/v) extracts (T₂) could not completely eliminate the pathogen but helped in controlling malformation by suppressing the activity of F. mangiferae. Mango trees sprayed with T₁ and T₂ revealed significant differences in percent fruit set and retention when compared with the control. This could be due to observed higher levels of nitrogen, phosphorus, potassium, calcium, magnesium, copper, zinc, iron and manganese in T_1, followed by T₂ when compared with T₃ (control). Among the different fruit quality parameters analysed, the total flavonoids were found to be significantly higher in T₁ and T₂ when compared with T_3. The study proved that the concoction-brewed compost (T₁) is effective, inexpensive, easy to prepare and constitutes a sustainable and eco-friendly approach to control floral malformation in mango when it is sprayed at bud break stage and again at fruit set stage. In this present study, exogenous treatment of emerging buds with (T_c) further proved that with increase in the number of malformed panicles/tree the number of buds developing into healthy panicles/tree decrease.