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。     

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。     

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。     

Influence of the major nitrite transporter NirC on the virulence of a Swollen Head Syndrome Avian Pathogenic E. coli (APEC) strain

Avian Pathogenic Escherichia coli (APEC) strains are extra-intestinal E. coli that infect poultry and cause diseases. Nitrite is a central branch-point in bacterial nitrogen metabolism and is used as a cytotoxin by macrophages. Unlike nitric oxide (NO), nitrite cannot diffuse across bacterial membrane cells. The NirC protein acts as a specific channel to facilitate the transport of nitrite into Salmonella and E. coli cells for nitrogen metabolism and cytoplasmic detoxification. NirC is also required for the pathogenicity of Salmonella by downregulating the production of NO by the host macrophages. Based on an in vitro microarray that revealed the overexpression of the nirC gene in APEC strain SCI-07, we constructed a nirC-deficient SCI-07 strain (ΔnirC) and evaluated its virulence potential using in vivo and in vitro assays. The final cumulative mortalities caused by mutant and wild-type (WT) were similar; while the ΔnirC caused a gradual increase in the mortality rate during the seven days recorded, the WT caused mortality up to 24 h post-infection (hpi). Counts of the ΔnirC cells in the spleen, lung and liver were higher than those of the WT after 48 hpi but similar at 24 hpi. Although similar number of ΔnirC and WT cells was observed in macrophages at 3 hpi, there was higher number of ΔnirC cells at 16 hpi. The cell adhesion ability of the ΔnirC strain was about half the WT level in the presence and absence of alpha-d-mannopyranoside. These results indicate that the nirC gene influences the pathogenicity of SCI-07 strain.     

Hemagglutinin glycosylation modulates the pathogenicity and antigenicity of the H5N1 avian influenza virus

The location and number of glycosylation in HA proteins exhibit large variations among H5 subtype avian influenza viruses (AIVs). To investigate the effect of glycosylation in the globular head of HA on the pathogenicity and antigenicity of H5N1 AIVs, seven rescued AIVs differing in their glycosylation patterns (144N, 158N and 169N) within the HA globular head of A/Mallard/Huadong/S/2005 were generated using site directed mutagenesis. Results showed that loss of glycosylation 158N was the prerequisite for H5 AIV binding to the α2,6-linked receptor. Only in conjunction with the removal of the 158N glycosylation, the H5 AIVs harboring both 144N and 169N glycosylations obtained an optimal binding preference to the α2,6-linked receptor. Compared with the wild-type virus, growth of viruses lacking glycosylation at either 158N or 169N was significantly reduced both in MDCK and A549 cells, while replication of viruses with additional glycosylation 144N was significantly promoted. Mutant viruses with loss of 158N or 169N glycosylation sites showed increased pathogenicity, systemic spread and pulmonary inflammation in mice compared to the wild-type H5N1 virus. In addition, chicken studies demonstrated that inactivated de-glycosylation 169N mutant induced cross-reaction HI and neutralization antibody against various clades of H5N1 AIVs. Moreover, this type of glycan pattern vaccine virus provided better cross-protection in chickens compared to wild-type vaccine virus. Thus, the glycosylation alteration of HA should be considered in the global surveillance and vaccine design of H5 subtype AIVs.     

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.     

东北地区小麦赤霉病镰孢菌种群及其致病性测定

为了明确东北地区小麦赤霉病镰孢菌的种群组成及分布,分别从辽宁省沈阳市、黑龙江省哈尔滨市和密山市、内蒙古自治区扎兰屯市、呼伦贝尔市和通辽市等地区采集小麦赤霉病病样,经单孢分离纯化共得到118株镰孢菌,传统形态学鉴定的基础上,采用基因组DNA的EF-1α序列分析技术进行了镰孢菌种类的辅助鉴定,确定属于7个种:禾谷镰孢菌为优势种,分离频率为64.41%,藤仓镰孢菌为次优势种,分离频率为18.64%;燕麦镰孢菌、尖孢镰孢菌、木贼镰孢菌、锐顶镰孢菌和轮枝镰孢菌的分离频率分别为5.08%,3.39%,3.39%,3.39%,1.70%。同时,在玉米成株期进行了致病性测定,结果表明,小麦赤霉病镰孢菌可以侵染玉米,较玉米茎腐镰孢菌的致病力低。     

用不同类型的水稻鉴别品种鉴定江苏稻瘟病菌的致病性

根据DNA指纹图谱和系谱, 从1999年采集的96个江苏稻瘟菌株中选择44个菌株进行致病性测定. 它们被鉴定为7个生理小种. 用日本清泽系列单基因鉴别品种和LTH近等基因系(NILs)鉴别品种分析, 分别产生27和10个致病类型, CO39 NILs对江苏菌株几乎没有鉴别作用. 分析结果可见供试菌株之间致病性差异极大. 研究表明, 发展包含较多抗病     

松球壳孢菌的致病性研究

实验证明 ,国内供试松球壳孢菌 (Sphaeropsissapinea)的致病力明显高于美国的B型菌株 ,与A型相似 ,致病性较强。B型也能无伤侵入 ,只是发病率明显低于A型。伤口并不是病菌侵入的唯一途径 ,自新梢伸长至针叶发育完全期间 ,病菌可直接侵入树木当年嫩梢并使之枯萎 ;此后 ,随着梢部组织的不断成熟与老化 ,伤口对病菌侵入与危害的作用才得以显现出来 ,无伤接种时不发病或只产生部分枯叶。气孔也是病菌侵入的途径之一。病菌不但可侵入当年新稍 ,而且还可于枝梢的 2a和 3a生处有伤侵入 ,使之染病甚至死亡。油松(Pinustabulaeformis)、赤松 (P .densiflora)、彰武松 (P .densifloravar .zhangwuensis)较樟子松 (Pinussylvestrisvar.mongolica)抗病 ,且彰武松主要表现为“耐病”。幼龄的樟子松较大龄的抗病。树势削弱的枝梢抗病性能降低 ,发病快而重。潜育期和繁殖期的长短与寄主、树势、组织的成熟程度和气温等有关