星油藤青枯病病原菌鉴定

 星油藤(Plukenetia volubilis L.) 是一种重要的藤本油料植物,在我国华南地区广泛种植。青枯病是近两年在海南星油藤种植区发生的新病害,为探究星油藤青枯病菌的基本特性及种下分化情况,本研究对分离的6株代表菌株进行了相关分析。细菌学鉴定及致病性测定结果表明,该病害是由类茄科雷尔氏菌(Ralstonia pseudosolanacearum)侵染引起。同时,从传统分类及分子生物学不同层面分析了星油藤青枯病菌的遗传分化情况。生理小种及生化变种的测试结果表明,星油藤青枯病菌属于1号生理小种和生化变种Ⅲ;16S rDNA和egl基因部分序列聚类分析显示,星油藤青枯病菌属类茄科雷尔氏菌演化型Ⅰ即亚洲分支菌株,序列变种34。     

Direct quantitative real‐time PCR assay for detection of the emerging pathogen Neonectria neomacrospora

The epidemic outbreak in northern Europe of Neonectria neomacrospora, the causal agent of dieback in Abies spp., led the European and Mediterranean Plant Protection Organization (EPPO) to include the pathogen on its alert list in 2017. Effective monitoring of this pathogen calls for a rapid and sensitive method of identification and quantification. A probe‐based real‐time PCR (qPCR) assay based on the β‐tubulin gene was developed for the detection and quantification of N. neomacrospora in infected wood samples, and directly for ascospores. This study presents the first published species–specific molecular detection assay for N. neomacrospora. The analytical specificity was validated on taxonomically closely related fungal species as well as on 18 fungal species associated with the host (Abies sp.). The analytical sensitivity was tested on naturally infected wood, on purified pathogen DNA in a matrix of host DNA and on N. neomacrospora ascospores for detection of airborne inoculum. The latter was tested both with a DNA extraction step prior to qPCR and without DNA extraction by direct qPCR on collected ascospores. The assay was specific to N. neomacrospora, with a sensitivity of 130 fg purified DNA, or 10 ascospores by direct qPCR. Omitting DNA extraction and amplifying directly on unpurified ascospores improved assay sensitivity significantly.     

Characterization and pathogenicity of six Cytospora strains causing stem canker of wild apple in the Tianshan Forest,China

Cytospora species are capable of causing destructive cankers of stems belonging to a wide range of woody plant species. In severe cases, cankers may lead to dieback of twigs and branches. Little is known about the Cytospora species causing canker disease of wild apple (Malus sieversii) trees in the Wild Fruit Forest Reserve in Tianshan Forest, Xinjiang Uygur Autonomous Region, China. In this study, six Cytospora isolates belonging to two species were isolated from cankerous lesions of wild apple twigs. Based on multi‐locus phylogenetic analysis using three DNA markers (ITS, tef1‐α and tub2) and morphological characterization, these isolates were identified as Cytospora mali and Cytospora parasitica. Temperature trials (15, 20, 25 and 30°C) showed that the optimal growth temperature for six isolates was 25°C. At a variety of temperatures, C. mali isolates tended to grow faster than isolates of C. parasitica, with the C. mali isolate, EGI1 performing better than others with regard to growth rate. Morphological observations showed that these species exhibited a single locule without conceptacles, and the conidia length was 3–5 μm. In vitro inoculation trials of twigs and leaves of M. sieversii seedlings revealed that the C. mali isolates were highly virulent phytopathogenic fungi, whereas the C. parasitica isolates were less virulent. The isolate EGI1 was the most virulent isolate among the six isolates. This paper presents the first report of pathogenic Cytospora spp. of the M. sieversii Tianshan Wild Fruit Forest Reserve of Yili, Xinjiang in China. It will aid in the understanding of how apple tree cankers are induced and provide disease management guidelines for M. sieversii forest conservation.     

新疆部分地区苹果、核桃和杨树腐烂病的病原鉴定

 树木腐烂病在新疆主林果种植区均有不同程度的发生,近年呈加重趋势。本研究在调查了新疆阿克苏、和田、伊犁、喀什和库尔勒等地区苹果树、核桃树和杨树3种林果腐烂病田间发病情况的基础上,重点对14个样本点的样品进行病原物的分离与鉴定。结果表明,各样本点3种林果均有不同程度的腐烂病发生,分离鉴定结果为引起苹果树腐烂病病原菌主要为Valsa mali var. mali(分离频率为64.5%),其次是V. mali var. pyri(25.8%)、V. malicola(3.2%)和V. nivea(6.5%);引起核桃树和杨树腐烂病病原菌为V. sordida(分离频率为83.3%和96%)和V. nivea(分离频率为16.7%和4%)。离体枝条及苹果果实上的致病性测定结果表明,3种林果上分离得到的病原菌均能感染原寄主。引起新疆部分地区苹果树腐烂病的主要病原菌是Valsa mali var. mali,核桃树和杨树腐烂病的主要病原菌是V. sordida。同时,基于实际生产中存在感病枯死杨树枝干作为苹果园、核桃园树体支撑木的现象,推测3种林果腐烂病菌间可能存在交叉感染的潜在风险。     

番茄棒孢叶斑病病原鉴定及生物学特性研究

本研究采用PSA培养基对上海地区的番茄棒孢叶斑病的病原菌进行分离、培养及致病性测定,通过形态学和分子生物学方法鉴定病原菌,并对该病菌的生物学特性进行研究。结果表明:番茄棒孢叶斑病病原菌菌落呈灰褐色,菌丝质地致密、绒毛状;分生孢子单生或串生,圆柱形或倒棍棒形、顶端钝圆,基部平截,呈半透明至浅褐色,假隔膜4～10个,直或稍弯曲,基脐加厚,深褐色。结合病原菌rDNA ITS序列测定、比对后,确定该病原为多主棒孢Corynespora cassiicola。该病菌的菌丝生长最适温度为25～30℃,最适pH范围为4～8,产孢的最适温度约为25℃,最适pH范围为5～9,麦芽糖和乳糖为碳源的培养基适宜菌丝生长,可溶性淀粉和乳糖溶液中孢子萌发率最高,光暗交替条件适宜菌丝生长,孢子在水滴中极易萌发。这是该病在上海地区的首次报道。     

Fungal colonization patterns in necrotic rootstocks and stem bases of dieback‐affected Fraxinus excelsior L.

European ash (Fraxinus excelsior) trees currently face the major threat of ash dieback caused by an invasive fungus, Hymenoscyphus fraxineus. Collar rots in F. excelsior have been increasingly associated with infections by this pathogen. However, the aetiology of the collar rots is still unclear and remains heavily debated. In contrast to most studies of this kind, entire rootstocks of four diseased ash trees were dug out to examine necrotic tissues in these rootstocks and stem bases in detail and to sample necrotic wood for fungal isolation. With the aid of morphological and molecular identification techniques, five to twelve fungal taxa were detected per tree. Members of the Nectriaceae family and Botryosphaeria stevensii, the causal agent of stem and branch cankers on many tree species, were frequently isolated from outer xylem. In contrast, H. fraxineus was the dominating species in interior wood layers. Microsatellite genotyping of 77 H. fraxineus isolates helped to identify up to six different genotypes per tree. The role of H. fraxineus and other isolated fungi in the aetiology of ash collar rots are discussed.     

粤北地区杉木真菌性病害种类

对粤北地区 ( 小坑林场、龙山林场、大坑山林场 ) 杉木（Cunninghamia lanceolate）真菌性病害进行全面调查，共采集 86 份标本，分离病组织获得菌株 162 份，主要归属于炭疽菌属（Colletotrichum）、拟盘多毛孢属（Pestalotiopsis）、腐皮壳属（Diaporthe）、镰刀菌属（Fusarium）、黑孢菌属（Nigrospora）、拟茎点霉属（Phomopsis）、叶点霉属（Phyllosticta）以及散斑壳属（Lophodermium）等。根据病害症状粤北地区主要杉木病害包括：杉木立枯病、杉木炭疽病、杉木溃疡病、杉木顶枯病、杉木梢枯病、杉木赤枯病、杉木球果坏死。各林场杉木林中，杉木炭疽病普遍发生、分布广；种子园中杉木球果病害危害严重，直接影响球果的产量和质量；在大坑山林场 4-5 年生的林分中，杉木顶枯病大面积发生，但在其它地区少有发生；此外，杉木溃疡病、杉木赤枯病和杉木立枯病在个别无性系或者苗圃中零星发生，但危害较轻。     

Multilocus PCR assay reveals high diversity of vegetative compatibility types in populations of Cryphonectria parasitica in Croatia

The ascomycete fungus Cryphonectria parasitica, causal agent of chestnut blight, is probably one of the best known invasive fungal pathogens in forests of Europe and North America. Mycovirus that reduces virulence of C. parasitica can be used as a biocontrol agent of the chestnut blight. However, anastomosis‐mediated virus transmission is limited by a vegetative (in)compatibility (vc) system involving at least six known diallelic vic genetic loci. This study looked at vegetative compatibility (vc) diversity in two populations of C. parasitica in Croatia. For that purpose, a PCR assay was validated and implemented using already known/published and newly designed primers for amplification of six known vic loci. The vc genotypes determined by PCR for 158 C. parasitica isolates investigated in this study were in complete agreement with the vc genotypes determined by pairwise co‐culturing of the same isolates, revealing the specificity and accuracy of the PCR‐based molecular vic genotyping assay. Twenty‐six unique vc genotypes were found among 158 isolates, and 19 vc types per population, which makes Croatian C. parasitica populations among the most diverse in Europe regarding the number of vc types and genetic diversity. Low values of multilocus linkage disequilibrium suggest sexual reproduction as a major contributor to high C. parasitica genetic diversity in studied populations.     

高产奶牛孕检后同期发情程序选择的研究

为了研究适用于高产奶牛的同期发情程序方案,将218头膘情良好、子宫和卵巢发育正常的不同胎次荷斯坦牛随机分成两组,试验1组为106头,试验2组为112头,分别采用两种同期程序(0.5.6.8和0.7.8.9)进行处理,测定情期受胎率,用于评价两种同期程序对高产奶牛的适用性。结果表明,试验2组情期受胎率平均为41.96%,显著高于试验1组的平均情期受胎率30.19%(P<0.01);将参与试验的218头牛根据胎次分成1胎组、2胎组和3胎及以上组,分析不同胎次对输精效果的影响,2胎情期受胎率最高为40.68%,与1胎情期受胎率存在显著差异(P<0.05),3胎及以上的情期受胎率最低为28.79%,与2胎、1胎存在极差异显著(P<0.01)。     

Intermediary metabolic response and gene transcription modulation on the Sub-Antarctic notothenioid Eleginops maclovinus (Valenciennes, 1930) injected with two strains of Piscirickettsia salmonis

Pathogen interactions with cultured fish populations are well studied, but their effects on native fishes have not been characterized. In Chile, the disease caused by bacterial species Piscirickettsia salmonis represents one of the main issues and is considered to be one of the important pathogens in the field of aquaculture. They have been found to infect native fish. Therefore, it is necessary to understand the impact of P. salmonis on native species of local commercial value, as well as the potential impact associated with the emergence of antibiotic-resistant strains of P. salmonis. Due to this purpose, the native fish Eleginops maclovinus was used in our study. Fish were randomly distributed in tanks and intraperitoneally inoculated with two strains of P. salmonis. No mortality was recorded during the experiment. Cortisol, glucose and total α-amino acid levels increased in fish injected with AUSTRAL-005 strain compared to sham-injected and LF-89-inoculated fish. Moreover, results showed an increase in the activity of carbohydrates and lipids metabolism in liver; and an increase in the carbohydrates, lipids and total α-amino acid metabolism in muscle after injection with AUSTRAL-005. Our results suggest that P. salmonis modulates the physiology of E. maclovinus and the physiological impact increase in the presence of the antibiotic-resistant strain AUSTRAL-005.