Fusarium oxysporum对苗期大豆根部的致病性和侵染规律

Fusarium oxysporum对苗期大豆具有较强的致病作用。病原菌自伤口或直接侵入大豆幼苗根部的皮层组织,随着病原菌的侵入和扩展,细胞出现坏死,组织发生崩解,根部组织表现红褐色病斑。环境因素对病原菌的侵染时期和在大豆体内潜育时间的长短有着明显的影响。侵染时期主要受病原菌侵入前自身所处状态和寄主的影响,侵入后潜育时间的长短则受植株状态的影响。土壤环境条件直接影响着病原菌的侵入和寄主的抗病性,间接影响着病原菌在大豆植株体内的扩展和病症的表现。     

Phytotoxicity on cotton ex-plants of an 18.5 kDa protein from culture filtrates of Verticillium dahliae

A phytotoxic protein that evokes the typical symptoms of Verticillium wilt disease in seedlings of Gossypium hirsutum L. (Upland cotton) was isolated from culture filtrates of Verticillium dahliae. The protein was purified by ammonium sulfate precipitation, Sephadex-G100 fractionation, and native PAGE. The 18.5 kDa protein, designated VD18.5, appears to be a single subunit protein with an isoelectric point between 3 and 5. VD18.5 induces symptoms of leaf dehydration, chlorosis, necrosis and stem discoloration in seedlings of the disease susceptible cotton cultivar Siokra 1–4. The LD₅₀ of VD18.5 on protoplasts of Siokra 1–4 was 18 μg mL⁻¹. VD18.5 had no noticeable effect on Pima S-7, which is a disease resistant cultivar. Phytotoxic activity was partially destroyed at high temperature and was abolished by digestion with proteinase K. Mass spectrometry fingerprinting and protein sequence data from VD18.5 yielded no significant matches when submitted to the Mascot search engine and NCBI non-redundant protein databases, respectively. These results suggest that VD18.5 is a novel protein that may be involved in the development of some of the symptoms associated with Verticillium wilt disease in the cotton plant.     

Homolog of FlhDC, a master regulator for flagellum synthesis: required for pathogenicity in Erwinia carotovora subsp. carotovora

 Erwinia carotovora subsp. carotovora (Ecc) is a causal agent of soft-rot diseases in a wide variety of plants. Here, we have isolated nonmotile mutants in Ecc by in vivo insertional mutagenesis using a transposon Tn5. The sequence disrupted by the Tn5 insertion in YMU1 and YMU5 mutants was highly homologous to that of flhC and flhD genes, respectively. They are involved in the initiation of the expression of flagellum-related genes in many gram-negative bacteria such as Escherichia coli and Salmonella. With electron microscopy, the flhC and the flhD homolog mutants were shown to be aflagellate. Furthermore, the virulence of these mutants was greatly reduced in Chinese cabbage and potato compared to that of the parental strain. These results suggest that flagellar formation is required for the pathogenicity of Ecc. Received: November 5, 2002 / Accepted: December 2, 2002 Acknowledgments This research was supported in part by Grant-in-Aid (12052210) and by a grant from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (13073).     

大豆种子寄藏真菌致病性测定方法比较

寄藏真菌可侵染大豆种子,降低种子的萌芽及活力,影响种子的品质和商品等级。为探讨不同接种方法对寄藏真菌致病性的影响,在实验室条件下比较了种子接种法和植株接种法(切顶端接种法、伤口接种法、切茎接种法和下胚轴接种法)对寄藏真菌致病性的影响。结果表明:各方法中寄藏真菌均对大豆产生致病性,并能显著区分寄藏真菌间致病性差异。其中切顶端接种法相比其他3种植物接种方法更简单有效而准确,而种子接种法更适合在室内进行,且周期短,相比植株接种法方便快捷。     

Effects of different NS genes of avian influenza viruses and amino acid changes on pathogenicity of recombinant A/Puerto Rico/8/34 viruses

To examine the effects of the NS1 and NEP genes of avian influenza viruses (AIVs) on pathogenicity in mice, we generated recombinant PR8 viruses containing 3 different NS genes of AIVs. In contrast to the reverse genetics-generated PR8 (rPR8) strain and other recombinant viruses, the recombinant virus rPR8-NS(0028), which contained the NS gene of A/chicken/KBNP-0028/2000 (H9N2) (0028), was non-pathogenic to mice. The novel single mutations of 0028 NS1 to corresponding amino acid of PR8 NS1, G139D and S151T increased the pathogenicity of rPR8-NS(0028). The replacement of the PL motifs (EPEV or RSEV) of pathogenic recombinant viruses with that of 0028 (GSEV) did not reduce the pathogenicity of the viruses. However, a recombinant virus with an EPEV-grafted 0028 NS gene was more pathogenic than rPR8-NS(0028) but less than rPR8. The lower pathogenicity of rPR8-NS(0028) might be associated with the lower virus titer and IFN-β level in the lungs of infected mice, and be attributed to G139, S151 and GSEV-PL motif of NS1 gene of 0028. In conclusion we defined new amino acid residues of NS1 related to mice pathogenicity and the presence of pathogenic NS genes among low pathogenic AIVs may encourage continuous monitoring of their mammalian pathogenicity.     

青贮玉米蠕形菌的分离鉴定及其致病性分析

本研究为初步明确引起青贮玉米(Zea Mays L.)叶斑病的蠕形菌种类,采集疑似病斑叶片样本,对病原菌进行分离纯化,形态特征观察,ITS-rDNA序列和GADPH基因序列系统发育分析及致病性测定。形态学和分子生物学鉴定结果表明分离得到的142株病原菌为3属6种真菌,其中麦根腐平脐蠕孢(Bipolaris sorokiniana)和玉米生平脐蠕孢(B.zeicola)为优势病原菌,分别为77株和48株;其次为大斑病菌(Exserohilum turcicum)9株,疣状弯孢(Curvularia verruculosa)2株,新月弯孢(C.lunata)3株,穗状弯孢(C.spicifera)3株。致病性测定发现麦根腐平脐蠕孢和玉米生平脐蠕孢对青海省8个主栽青贮玉米品种均有致病性,其余4种病原菌的致病性各有差异。本研究初步明确引起青贮玉米叶斑病的蠕形菌种类及其致病性,为后续开展病害诊断及其综合防控奠定了基础。     

河北省玉米小斑病菌优势生理小种鉴定及ITS序列分析

2007~2016年在河北各地采集玉米小斑病标样,对分离出的356株玉米小斑病菌菌株进行生理小种鉴定。结果表明,在鉴定的年度范围内,玉米小斑病菌T小种、C小种、S生理型、O小种的分离频率在年度间存有差异,O小种的平均分离频率94.94%,是河北省玉米小斑病菌的优势小种;O小种对主栽品种郑单958和自交系C103的致病力呈下降趋势,在C103上致病力下降幅度小于郑单958。对河北省石家庄、保定、衡水、沧州邯郸、邢台采集的45株菌株进行ITS序列分析构建UPGMA进化树,分析表明,河北省内玉米小斑病菌株的遗传进化与地域有一定关系,差异不明显。     

豫皖小麦田麦根腐平脐蠕孢交配型基因检测

麦根腐平脐蠕孢是一种可以引起多种植物病害的病原真菌,广泛分布于世界各地,对全世界的禾谷类作物生产带来了不可估量的损失。子囊菌交配型基因具有一定的广泛性,其编码的产物是调控子囊菌有性生殖的重要转录因子。为了对河南、安徽小麦田分离获得的麦根腐平脐蠕孢群体的MAT基因型进行探索,对在河南、安徽两省不同地区分离获得33株麦根腐平脐蠕孢菌株,利用麦根腐平脐蠕孢菌的MAT特异性引物进行基因型检测,并对不同交配型代表菌株进行生长形态、孢子形态观察和致病力测定,对麦根腐平脐蠕孢菌 MAT-2基因进行了系统发育分析。结果表明,河南、安徽两省存在两种MAT基因型的麦根腐平脐蠕孢群体,含有 MAT-2基因的菌株21株,含 MAT-1基因的菌株12株,比例为7∶4（ MAT-2∶ MAT-1）。两种交配型菌株的生长形态、孢子形态和致病力无显著差异,整体分析结果与物种的分子系统学分析表现出来的亲缘关系基本一致。     

玉蜀黍尾孢菌CzVelB基因功能分析

以获得的野生型和CzVelB敲除突变体菌株为供试菌株,分别对其进行生物学及致病力测定,在此基础上分析毒素和细胞壁降解酶的活性。结果表明,与野生型相比,敲除CzVelB后菌丝生长速率及颜色无明显差异,但产孢量下降56.6%,且孢子萌发率下降,萌发过程滞后1 h。致病性分析发现,敲除CzVelB后菌株致病性明显下降,病情指数、病斑大小,突变体均低于野生型。分析毒素对叶片的致病性发现,突变体所产生的毒素低于野生型产生的毒素。敲除突变体菌株ΔCzVelB及野生型菌株在活体内外均能产生5种细胞壁降解酶,其中敲除CzVelB后,5种离体病原菌细胞壁降解酶活性明显下降。分析侵染过程中5种细胞壁降解酶发现,CzVelB主要通过影响CX和PMG在接种前期12 h内起重要作用;在接种后期(72～96 h),CzVelB主要通过影响PGTE的活性调控玉蜀黍尾孢菌致病力。     

黄瓜根腐病致病病原的鉴定

在天津市及周边地区采集病株进行组织分离，同时分离到两类病原菌，经初步的生物学特性鉴定表明，引起黄瓜根腐病的主要病原菌为甜瓜疫霉、尖孢镰刀菌和茄病镰刀菌。经接种试验，认为甜瓜疫霉为主要致病菌。