黑胫病菌（Leptosphaeria biglobosa）在油菜叶片和茎中的侵染过程观察

为明确黑胫病菌（Leptosphaeria biglobosa）在甘蓝型油菜叶片和茎中的侵染及扩展过程，利用绿色荧光蛋 白（GFP）标记的黑胫病菌株接菌油菜叶片，利用激光共聚焦显微镜观察菌株在油菜叶片和茎中的侵染过程。结果 表明，接种油菜叶片7 h后，分生孢子萌发并长出芽管；17 h后，芽管侵入气孔；24 h后，分生孢子全部萌发；36 h后萌 发的芽管形成菌丝；120 h后，菌丝在叶片表皮细胞间隙蔓延，并侵入叶肉细胞。13 d后，菌丝侵入茎部皮层组织； 15 d后，菌丝在皮层细胞间隙蔓延，并侵染至茎表皮；21 d后，菌丝侵染至维管组织；23 d后，菌丝侵染至茎韧皮部； 25 d后，茎导管被侵染，并向木质部扩展。本研究发现的L. biglobosa 在油菜叶片和茎中的侵染过程，可为油菜与黑 胫病菌互作的研究、黑胫病致病机理及防治提供参考。     

谷氨酰胺对斜带石斑鱼GF-1细胞中C-Myc蛋白的表达与神经坏死病毒复制的影响

为探讨C-Myc表达、谷氨酰胺代谢和神经坏死病毒复制三者之间的关系,本研究首先克隆了斜带石斑鱼鳍条细胞(GF-1)中的C-Myc基因(GF-1-C-Myc),结果显示GF-1-CMyc基因cDNA全长814 bp,开放阅读框(ORF)为285 bp,编码95个氨基酸(aa),有亮氨酸拉链结构域与螺旋-环-螺旋(HLH)结构域。实验表达和纯化了GF-1-C-Myc蛋白,并制备其多克隆抗体。采用实时定量PCR技术(qRT-PCR)与免疫印迹法(WB)检测了GF-1-C-Myc基因的表达和神经坏死病毒的复制。结果显示,缺乏谷氨酰胺会同时抑制GF-1-C-Myc基因的表达和神经坏死病毒(NNV)的复制,添加谷氨酰胺可同时促进GF-1-C-Myc的表达和NNV的复制;此外,NNV感染可上调GF-1-C-Myc基因的表达,并显著消耗GF-1细胞培养液中的谷氨酰胺。研究表明,GF-1-C-Myc基因可调控宿主谷氨酰胺代谢,从而有利于神经坏死病毒的复制。本结果为防控NNV的感染提供了参考。     

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。     

稻曲病菌致病机制研究进展

 稻曲病是一种水稻穗部病害,在世界各水稻主产区均有发生,已成为世界性病害,在我国也属于水稻主要病害之一。稻曲病的发生严重影响水稻产量和稻米品质。研究其病原菌致病机制,对找寻防治药物的特异性靶标具有重要作用,可为抗病分子育种提供新的思路,为稻曲病菌防治药剂研发提供新的靶标。本文综述了稻曲病菌侵染过程、致病相关基因功能研究、水稻响应稻曲病菌侵染等方面的研究进展,提出进一步研究的策略。现有研究表明,稻曲病菌能成功侵染水稻孕穗期雄蕊的花丝引起发病。病原菌可通过抑制多个水稻免疫途径实现成功侵染;还可模仿胚珠受精激活水稻灌浆、糖代谢等途径为稻曲球的形成提供营养物质;全基因组测序完成和基因编辑技术在基因敲除中的成功应用,为稻曲病菌功能基因研究提供了很好的基础。稻曲病菌独特的侵染过程和营养利用机制是未来研究的重要方向。     

多孢木霉HZ-31菌株侵染对野燕麦生理机制的影响

选用青海农田常见杂草野燕麦作为靶标杂草,盆栽接种后测定了多孢木霉HZ-31菌株对野燕麦体内各种生理指标的影响,分析了寄主与病原菌相互作用的生理变化机制。结果表明,多孢木霉HZ-31菌株侵染野燕麦后,对植株生理机制防御酶的影响是多方面的,PAL和POD活性增加,以抵御菌株侵染,其余生理防御指标包括MDA含量、CAT、PPO、SOD活性、可溶性蛋白含量和叶绿素含量则受到菌株侵染的破坏,含量减少、活性降低。被抑制的防御酶作用效应大于被促进的防御酶。HZ-31菌株的侵染钝化或抑制了野燕麦体内主要酶类,中断相应的酶促反应,引起植物广泛的代谢变化,包括细胞膜透性改变、干扰光合作用、蛋白质合成、酚类物质代谢等生理方面的变化。     

PVX病毒载体介导2种马铃薯Y病毒科病毒VPg蛋白表达诱导烟草叶片系统坏死的研究

芜菁花叶病毒（Turnip mosaic virus, TuMV）和槟榔坏死环斑病毒（Areca plam necrotic ringspot virus, ANRSV）是马铃薯Y病毒科（Potyviridae）中分别可诱导十字花科作物和槟榔产生坏死病症的2种不同属病毒。VPg（Viral protein genome linked）作为与马铃薯Y病毒科病毒基因组5′端的结合蛋白,在病毒复制、翻译和运动等侵染过程中发挥重要作用。本研究利用In-Fusion克隆策略分别构建了可表达3′端融合Strep蛋白标签序列的ANRSV和TuMV VPg全长编码基因的马铃薯X病毒（Potato virus X, PVX）载体pPVX-VPg-AR和pPVX-VPg-Tu。通过农杆菌渗透注射接种本生烟（Nicotiana benthamiana）,利用RT-PCR、Strep蛋白标签亲和层析和Western blot等方法证明了PVX病毒载体能够介导VPg-AR和VPg-Tu蛋白在本生烟中系统表达,且发现这2种VPg蛋白的过量表达诱导本生烟叶片产生了不同程度的系统性坏死病症。进一步采用DAB和NBT染色,在产生坏死症状叶片中检测到大量活性氧（reactive oxygen species, ROS）的积累。因此,推测VPg可能是TuMV和ANRSV诱发植物病症产生的一个决定因子,为后续研究这2种病毒诱导症状形成机制奠定了基础。     

Role of basidiomycete fungi in the grapevine trunk disease esca

Phaeomoniella chlamydospora and wood-rotting basidiomycete fungi, namely Fomitiporia spp., are known esca pathogens. However, the effect of their mixed infections and the sequence of infection on disease development is unclear. To determine the effects of single and co-inoculations on symptoms, potted Vitis vinifera ‘Crimson Seedless’ was inoculated with P. chlamydospora either alone or in combination with one of four basidiomycetes: Coprinellus radians, Fomitiporia langloisii, F. polymorpha and the novel species Tropicoporus texanus. Basidiomycetes were isolated from vines with foliar symptoms of esca in California and Texas. In sequential co-inoculations, the effects of different sequences of infection (P. chlamydospora first, basidiomycete 6 months later; and vice versa) were tested, compared to simultaneous co-inoculations. Plants inoculated with P. chlamydospora either alone or in combination with a basidiomycete (in any sequence) did not differ significantly in the length of black-streaking lesions. In plants inoculated only with a basidiomycete, the appearance of large brown lesions, coupled with the absence of this wood symptom from control plants, suggests that C. radians, F. langloisii and T. texanus are pathogenic. Foliar symptoms resembling those of esca in the field (marginal and/or interveinal scorching, combined with red and/or yellow discoloration) were statistically more frequent among plants inoculated with F. polymorpha or T. texanus, either simultaneously or following P. chlamydospora, compared to single inoculations. Sequential co-inoculations of a basidiomycete before or after P. chlamydospora were associated with similar lesion lengths, suggesting that basidiomycetes may not require infection by P. chlamydospora in order to extensively colonize the wood.     

Three‐dimensional visualization of green fluorescence protein‐labelled Edwardsiella tarda in whole Medaka larvae

The invasive fish pathogen Edwardsiella tarda is common in aquatic environments and causes the environmentally and economically destructive emphysematous putrefactive disease called edwardsiellosis. In order to understand the organism's infection pathway, medaka larvae (Oryzias latipes) were immersion‐infected with E. tarda labelled with green fluorescence protein (GFP) and then visualized in three dimensions under confocal laser microscopy and light‐sheet fluorescence microscopy. Confocal microscopy revealed GFP‐labelled E. tarda in the mouth, head, gill bridges, gill cover, skin, membrane fin, gastrointestinal tract and air bladder, and in the caudal vein, somite veins, caudal artery and caudal capillaries. Light‐sheet microscopy additionally showed GFP‐labelled E. tarda in the pharyngeal cavity, muscle of the pectoral fin and cardiac atrium and ventricle. These findings suggest that during its infection of fish, E. tarda initially adheres to, and invades, the epithelial cells of the skin, gills and gastrointestinal tract (through the pharyngeal cavity); E. tarda then enters the blood vessels to access organs, including the air bladder and heart.     

First report of Fusarium oxysporum species complex infection in zebrafish culturing system

Fusarium oxysporum species complex (FOSC) is a highly diverse fungus. Recently, F. oxysporum infection was identified from zebrafish (Danio rerio) culturing system in Korea. Initially, a rapid whitish smudge was appeared in the water with the fungal blooming on walls of fish tanks. Microscopic studies were conducted on fungal hyphae, colony pigmentation and chlamydospore formation and the presence of macro‐ and microspores confirmed that the isolated fungus as F. oxysporum. Furthermore, isolated F. oxysporum was confirmed by internal transcribed spacer sequencing which matched (100%) to nine F. oxysporum sequences available in GenBank. Experimental hypodermic injection of F. oxysporum into adult zebrafish showed the development of fungal mycelium and pathogenicity similar to signs observed. Histopathologic results revealed a presence of F. oxysporum hyphae in zebrafish muscle. Fusarium oxysporum growth was increased with sea salt in a concentration‐dependent manner. Antifungal susceptibility results revealed that F. oxysporum is resistant to copper sulphate (up to 200 μg mL^?1) and sensitive to nystatin (up to 40 μg mL^?1). This is the first report of FOSC from zebrafish culture system, suggesting it appears as an emerging pathogen, thus posing a significant risk on zebrafish facilities in the world.