Selection forMeloidogyne incognita virulence against resistance genes from tomato and pepper and specificity of the virulence/resistance determinants

Experiments were designed to analyze the relationships between the root-knot nematodeMeloidogyne incognita and resistant tomato and pepper genotypes. From a natural avirulent isolate, near-isogenic nematode lineages were selected with virulence either against the tomatoMi resistance gene or the pepperMe3 resistance gene. Despite the drastic selection pressure used, nematodes appeared unable to overcome the pepperMe1 gene, therefore suggesting some differences in the resistance conferred byMe1 andMe3 in this species. Nematodes virulent onMi-resistant tomatoes were not able to reproduce onMe1-resistant nor onMe3-resistant peppers, and nematodes virulent onMe3-resistant peppers were not able to reproduce onMi-resistant tomatoes nor onMe1-resistant peppers. These results clearly demonstrate the specificity ofM. incognita virulence against resistance genes from both tomato and pepper, and indirectly suggest that gene-for-gene relationships could occur between these two solanaceous crops and the nematode.     

Application of polyclonal and monoclonal antibodies for the detection of Xanthomonas campestris pv. campestris in crucifer seeds using immunofluorescence microscopy

Polyclonal and monoclonal antibodies (PCAs and MCAs) were tested for the detection ofXanthomonas campestris pv.campestris (Xcc) in cabbage seeds using immunofluorescence microscopy (IF). It was concluded that PCA 94, MCAs 20H6, 2F4, 18G12 and a mixture of MCAs 20H6, 18G12, 2F4 and 16B5 could be used to detect Xcc in seed extracts when 5 min and 2.5 h shaking of seeds are used as extraction methods. The reliability of confirming suspect colonies with MCAs and PCA 94 in IF depended in part on the seed lot tested and the antibody used. Some virulent Xcc strains derived from seed lots, did not react with MCAs 10C5, 2F4, 18G12, 17C12 and 16B5. On the other hand, saprophytic isolates obtained from one seed lot cross-reacted with MCA 17C12 and to a lesser extent with MCAs 2F4, 18G12 and PCA 94. No relationship was found between IF-reactions of Xcc strains using MCAs and reactions of Xcc strains in pathogenicity testing. Xcc andX. c. pv.amoraciae (Xca) could in general not be distinguished on the basis of reactions with MCAs and PCAs. Also in pathogenicity tests Xcc and Xca were hard to distinguish.     

1993年全国小麦秆锈菌种群动态分析

１９９３年我国小麦秆锈病发生较轻，从８个省１８个区县中的４７个品种上采集到的９５个标样中分离到菌株３３６个，鉴定出２１Ｃ３ＣＫＲ、２１Ｃ３ＣＫＨ、２１Ｃ３ＣＴＲ、２１Ｃ３ＣＴＨ、２１Ｃ３ＣＦＲ、２１Ｃ３ＣＦＨ、３４Ｃ１ＭＫＲ、３４Ｃ１ＭＫＨ和３４Ｃ２ＭＫＲ等９个致病类型；２１Ｃ３ＣＫＲ的出现频率为６５．２％，居于首位，２１Ｃ３ＣＤＨ为１４．５％，２１Ｃ３ＣＴＲ为１０．７％，２１Ｃ３ＣＦＲ为４．２％     

生物间遗传学在防治河北省小麦叶锈病研究中的应用

作者应用生物间遗传学的观点和方法,对河北省的小麦品种与叶锈菌的相互作用进行了分析。供试的叶锈菌是来自河北省的177个标样,供试的28个小麦品种包括河北省的主要生产品种、区试品种(系)抗源和新鉴别寄主。用毒力频率法分析品种与叶锈菌群体的关系,有6个品种即鉴61、保麦2号、翼植88-5163、石86-2848、唐86-4043和百农3217表现较抗病。根据这6个品种对叶锈菌群体的反应、列出33种不同的毒性公式,在此基础上,根据若干品种对叶锈菌群体同时表现抵抗或感染计算抗性组合,为品种合理布局选择最佳组合,在小种鉴定方面,用新鉴别寄主对177个菌株的反应鉴定出频率在3.4%以上的小种8个,其中以V1,9-16和V1,9-12,14-16的频率较高,分别为13.6%和10.7%,也是近两年来北方冬麦区常出现的小种类型。在小种鉴定的基础上,根据28个品种与6个优势小种互作所出现的浸染型来推导品种和叶锈菌的基因型,共推导出7个抗性基因分散在不同的品种中。     

The Control of Rinderpest in Tanzania between 1997 and 1998

In January 1997, Tanzania requested international assistance against rinderpest on the grounds that the virus had probably entered the country from southern Kenya. Over the next few months, a variety of attempts were made to determine the extent of the incursion by searching for serological and clinical evidence of the whereabouts of the virus. At the clinical level, these attempts were hampered by the low virulence of the strain, and at the serological level by the lack of a baseline against which contemporary interpretations could be made. Once it became apparent that neither surveillance tool was likely to produce a rapid result, an infected area was declared on common-sense grounds and emergency vaccination was initiated. The vaccination programme had two objectives, firstly to prevent any further entry across the international border, and secondly to contain and if possible eliminate rinderpest from those districts into which it had already entered. On the few occasions that clinical rinderpest was subsequently found, it was always within this provisional infected area. Emergency vaccination campaigns within the infected area ran from January to the end of March 1997 but were halted by the onset of the long rains. At this time, seromonitoring in two districts showed that viral persistence was still theoretically possible and therefore a second round of emergency vaccination was immediately organized. Further seromonitoring then indicated a large number of villages with population antibody prevalences of over 85%. These populations were considered to have been `immunosterilized'. Although no clinical disease had been observed in them, it was decided to undertake additional vaccination in a group of districts to the south of the infected area. Serosurveillance indicated that rinderpest could have been present in a number of these districts prior to vaccination. Serosurveillance in 1998 suggested that numerous vaccinated animals had probably moved into districts outside the infected and additional vaccination areas, but did not rule out the continued presence of field infection.     

Genomic and Pathogenic Studies on a Glycoprotein E Variant Field Isolate of Bovine Herpesvirus 1

Glycoprotein E-negative (gE–) laboratory strains of bovine herpesvirus 1 (BHV-1) were recently introduced as novel marker vaccines, allowing serological discrimination between vaccinated and naturally infected animals on the basis of lack or presence of antibodies against gE epitopes. The applicability of this approach is based on the genetic stability of the gE. However, mutant field variants of BHV-1 with a variable response in anti-gE ELISA have been isolated. The molecular characterization of a gE variant field isolate (Salwa strain) is presented here. By comparing the gE nucleotide and amino acid sequences of the Salwa strain with those of the wild strain Jura, ten mutated bases were found in the gE strain of Salwa, six of which alter the amino acid sequence, leading to changes in five amino acids. Both strains caused respiratory disease in experimentally infected calves, but Salwa generated slightly milder signs. Both viruses were excreted in nasal and ocular discharges, and were reactivated by dexamethasone treatment. In conclusion, the rather close similarities observed in the gE gene structure and pathogenicity features of the gE mutant and of the wild strain of BHV-1 confirm the genetic stability of gE. The findings indicate that the Salwa isolate is virulent, but less virulent than wild strains. Our data support the use of gE-negative marker vaccines in eradication programmes.     

表达猪瘟兔化疫苗株E0蛋白的PK-15细胞系构建

将克隆到pGEM T-easy载体中的E0基因双酶切回收后，连接到增强型绿色荧光蛋白（EGFP）中，利用Lipofectamine^TM2000将重组子转染PK-15、BHK-21、VERO 3种细胞，直接在荧光显微镜下用蓝光激发进行观察，在3种细胞中都可以观察到绿色荧光，而且在转染后的24、48、72 h 3个时间点上，观察到的荧光细胞数量逐渐增多，在72 h时荧光细胞的数量在3种细胞中都达到最多，总的来看，在PK 15中荧光细胞的数量最多。通过对细胞荧光的定位发现，重组质粒的荧光主要分布在胞浆内，而且细胞核周围的荧光较强，胞核与胞浆的界限明显，尤其是在PK-15细胞和VERO细胞中这种现象尤为明显。未携带E0目的DNA的pEGFP-N1 Vector转染3种细胞后，都可以观察到绿色荧光，但是整个细胞中是均匀出现荧光的。经酶切、PCR、单抗间接免疫荧光检测，PK-E0细胞中有大量的HCLV的E0表达，单纯的PK-15细胞中没有E0的表达。HCLV株在PK-E0细胞中从48h开始到72h的滴度比在PK-15细胞中的滴度要高。证明PK-E0细胞中E0蛋白的大量表达增加了HCLV在细胞中的复制。     

Construction of Brucella ClpS Gene Mutant Strain and Its Effect on Virulence

The present study aimed to determine the role of ClpS gene,and to analyse the impact of ClpS mutation on the virulence of Brucella.A ClpS gene mutant strain,named ΔClpS was constructed by homologous recombination technology.The bacterial growth kinetics,the LPS synthesis ability and the survival ability of bacterial within macrophages as well as the virulence in mouse model were measured.In addition,the difference between parent strain 2308 and the mutant strain ΔClpS were compared.The results showed that under the same culture conditions,no difference in bacterial concentration was observed between 2308 and ΔClpS strains.The silver staining examination showed that the expression level of LPS extracted from two strains were similar,indicating ClpS gene mutation did not alter the growth rate and LPS synthesis ability of Brucella. In the cell infection assay,the survival ability of ΔClpS strain in cells was extremely significantly lower than that of 2308 strain at 72 h after infection (P<0.01).The results of mouse infection experiment showed that in the first week after infection,no significant difference in spleen weight and bacterial concentration between 2308 and ΔClpS strains infected mice was observed.However,at 4 weeks after infection,the bacterial concentration in spleen of ΔClpS infected mice was 10^3.93 CFU/g spleen,which was significantly lower than that of 2308 strain (10^6.68 CFU/g spleen,P<0.01).The spleen weight of ΔClpS infected mice was also remarkably lower than that of 2308 strain (P<0.01).In summary,the results suggested that the ClpS gene of Brucella did not play a role in Brucella growth rate and ability of LPS synthesis,whereas ClpS gene mutation decreased the ability of Brucella colonization in mouse spleen.     

口蹄疫重组鸡痘病毒活载体疫苗在豚鼠、猪体内的毒性、分布及抗体消长规律

利用临床观察、病理解剖、PCR、RT-PCR、ELISA、中和抗体检测等方法,对口蹄疫(FMD)重组鸡痘病毒(FPV)在豚鼠、仔猪体内的毒性、分布以及抗体消长规律进行研究。结果表明,FMD重组鸡痘病毒免疫的动物在整个试验期间,未表现出明显的临床症状和不良反应;病理组织切片检测无明显的组织学变化;PCR、RT-PCR检测证明,豚鼠、猪免疫FMD重组鸡痘病毒后,在心脏、肝脏、脾脏、肺脏、肾脏、肌肉、脑、肠系膜淋巴结内检测到FPVDNA和FMDV DNA,且在大部分组织能存在3 d左右;FMD重组鸡痘病毒均可诱导免疫动物产生较高水平的抗FMDV特异性抗体和中和抗体,验证了所构建重组FPV的生物安全性及良好的免疫原性,为其他哺乳动物实验提供了必要的基础数据。     

4株鸭源肠球菌的鉴定和致病性

对临床分离的4株鸭源肠球菌郑1株、郑2株、郑3株、北京株和1株粪肠球菌参考菌株进行了系统鉴定,并用SDS-PAGE和Western-blot技术对各菌株细胞壁蛋白图谱进行比较分析。结果5个菌株的形态、染色、生理生化特性均与粪肠球菌特性一致;它们均对青霉素、万古霉素和庆大霉素敏感而对四环素耐药;5个菌株人工感染雏鸭及小白鼠均有致病性,但各菌株间致病力存在差异,北京株最强,参考株最弱,其余3株介于北京株和参考株之间;各菌株的细胞壁蛋白经SDS-PAGE在相对分子质量33 370~131 690之间均显示数十条蛋白带,其中郑2株和北京株在相对分子质量66 840处均有1条染色较深的蛋白带,而用Western-blot分析显示抗北京株胞壁蛋白抗体只能检测到北京株相对分子质量为66 840的抗原蛋白。以上结果表明,这5个被检菌株为致病性粪肠球菌,且致病性以北京株最强。