拟康宁木霉Tk1的分离鉴定、拮抗作用及其生物学特性

为了分离利用高效植物病害生物防治菌株，从广东药用植物金丝皇菊根部分离获得一株木霉菌株Tk1。经过形态观察、rDNA-ITS和EF-1α序列分析明确其为拟康宁木霉Trichoderma koningiopsis。用平板对峙培养法测定，该菌株对金丝皇菊枯萎病菌（尖孢镰刀菌Fusarium oxysporum）和柑橘炭疽病菌（胶胞炭疽菌Colletotrichum gloeosporioides）的抑制率分别为68.5%和77.1%，且在后期能完全覆盖或部分覆盖病原真菌。其无菌发酵液对金丝皇菊枯萎病菌的抑制率为45.55%。该菌株营养生长和产孢的最适碳源分别为乳糖和葡萄糖，最适氮源是酵母膏；在pH 4~11的培养基上均可生长、产孢，最适pH为7；12 h交替光照条件下菌丝的生长速度最快，全光照条件下产孢数量最大；该菌株的致死温度为73℃处理10 min。     

油茶内生拮抗细菌的筛选、鉴定及防效

为筛选出对油茶炭疽菌有拮抗效果的生防菌株，以油茶健康叶为对象，平板对峙法筛选，利用形态和生理生化特征以及16S rDNA基因和gyrA基因序列对菌株进行鉴定，测定其抑菌活性，并将该菌株与枯草芽胞杆菌Y13复配进行林间防效试验。结果表明，对油茶炭疽病菌拮抗作用效果较强的内生菌株为HBMC—B05；菌落为乳白色，不透明，边缘不整齐，中间有凸起；分子鉴定结果显示菌株HBMC—B05为贝莱斯芽胞杆菌Bacillus velezensis，其对果生炭疽菌Collectotrichum fructicola，抑菌率可达到81.31%，对另外3种油茶炭疽病病原菌抑菌率可达到70%以上，对其他4种致病菌也有抑制作用；复合菌剂对油茶病害的田间防治效果达到59%以上。该菌株的抑菌具有广谱性，可为油茶病害的生物防治扩充菌种资源，具有广泛的应用前景。     

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。     

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。     

牦牛源正呼肠孤病毒2型的检测和分离鉴定

旨在调查川西北牦牛哺乳动物正呼肠孤病毒（MRV）的感染情况并分离病毒。采用RT-PCR方法，对采自川西北15个牧场的72份牦牛腹泻粪便样本和其中5个牧场的15份腹泻牦牛血清样本进行MRV检测，阳性样本进一步用分型PCR确定其血清型。结果显示，粪便样本中MRV检出率为20.83%（15/72），血清2型的比例为60%（9/15）；血清样本中MRV检出率为40%（6/15），血清2型的比例为83.33%（5/6）；未检测到其他血清型。成功地从腹泻粪便中分离到1株MRV血清2型毒株（TCID₅₀为4×10^-8.56·mL^-1），并获得长度为23 587 bp的分离株全基因组，该分离株与中国猪源毒株的遗传关系最近；与GenBank中所有的MRV S1基因相比，该分离株有4个独特的氨基酸突变。本研究从牦牛中检测到MRV，并分离到1株牛源MRV血清2型毒株，为进一步研究MRV血清2型生物学特性奠定了基础。     

鸡粪除臭菌的分离筛选及除臭效果分析

以鸡粪堆肥为材料,筛选高效除臭菌株并构建复合菌系,为鸡粪的无害化处理提供具有除臭功能的菌种资源。首先,采用驯化富集法、平板稀释法、产气试验、除氨试验、除硫化氢试验以及纤维素降解试验筛选出鸡粪堆肥中的除臭菌;其次,通过16S rRNA基因测序鉴定菌株;最后,通过拮抗试验构建除臭菌系。研究分离得到5株具有除臭功能的菌株,分属于芽孢杆菌属(MS03,Bacillus sp.)、贝莱斯芽孢杆菌(MS07,Bacillus velezensis)、耐寒短杆菌(MS11,Brevibacterium frigoritolerans)、木糖葡萄球菌(MS42,Staphylococcus xylosus)和变异棒杆菌(MS82,Corynebacterium variabile)。复合菌系C2(MS03+MS07)和C4(MS03+MS82)的综合除臭率均高于60%,且明显高于单株菌株及其他复合菌系。研究表明,这两个由不同除臭菌组合而成的复合菌系在无害化处理鸡粪方面具有较大的潜力和应用前景。     

云南和广东省流行性出血病病毒血清6型毒株(EHDV-6)的分离与遗传特性

【目的】分离流行于我国南方地区的流行性出血病病毒血清6型毒株(EHDV-6),掌握分离毒株的遗传特征。【方法】对设立于我国云南和广东省的哨兵动物定期采血,进行EHDV感染情况监测与病毒分离培养。通过血清中和试验确定分离EHDV的血清型,采用一步法RT-PCR对分离的EHDV-6毒株基因节段2、3、6(Seg-2、-3、-6)进行扩增与序列分析。【结果】2012—2016年,从云南省和广东省的哨兵动物中分离出25株EHDV毒株,其中,11株为EHDV-6型。中国EHDV-6型毒株的Seg-2、-3与-6均属Eastern型,核酸序列相似性分别为99.1%、98.9%和98.8%,在系统发生树上分别聚为1个独立的中国支系。在日本引起疫病暴发的EHDV-6型毒株与中国EHDV-6型毒株具有最近的亲缘关系。日本EHDV-6型毒株在系统发生树上处于中国支系内部,Seg-2、Seg-3与中国毒株的核酸序列相似性分别为98.5%和93.9%。【结论】云南和广东省流行的EHDV-6型毒株核酸与氨基酸序列高度相似;日本和中国的EHDV-6型毒株具有最近的亲缘关系,提示中日之间可能存在EHDV的流动。研究结果为进一步开展我国EHDV-6型毒株流行病学分析、致病性、病原学诊断与疫苗制备等研究提供了基础。     

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.