Accessions of Australian Nicotiana species suitable as indicator hosts in the diagnosis of plant virus diseases

When screening the genusNicotiana for sensitive and differential hosts for a group of mechanically transmissible plant viruses with narrow host ranges, development of systemic symptoms was alsmost exclusively observed in species of three closely related sections of the subgenusPetunioides. These species wereN. miersii (sectionAcuminatae),N. bigelovii andN. clevelandii (Bigelovianae) andN. benthamiana, N. cavicola, N. ingulba, N. occidentalis, N. rosulata andN. rotundifolia (Suaveolentes). Except forN. benthamiana andN. clevelandii, which are already known for their large virus ranges, they are new experimental hosts that appeared very useful for detection of viruses and for differentiation of viruses that closely resemble each other in host range. Accessions of the same species often varied largely in local and systemic viral response. EspeciallyN. benthamiana-9,N. miersii-33 andN. occidentalis-37B (code numbers given by Tobacco Research Laboratory, Oxford, N.C., USA) are recommended for routine inoculation tests. The sensitiveSuaveolentes species mentioned are native to the arid parts of Australia. Collections of these species deserve attention in studies on virus diseases of unknown etiology where experimental hosts are lacking.Samenvatting In het genusNicotiana werden vatbare, gevoelige en differentiële toetsplanten opgespoord door toetsing van vooral de collectie van het Tobacco Research Laboratory (Oxford, N.C., VS) met een aantal virussen waarvoor weinig of geen bruikbare toetsplanten bekend waren. Systemische reacties werden bijna uitsluitend in soorten van drie nauw verwante secties van het subgenusPetunioides waargenomen. Deze soorten warenN. miersii (sectieAcuminatae),N. bigelovii enN. clevelandii (Bigelovianae) enN. benthamiana, N. cavicola, N. ingulba, N. occidentalis, N. rosulata enN. rotundifolia (Suaveolentes). Behalve voor virusvermeerdering bleken deze soorten ook zeer geschikt te zijn voor differentiatie van virussen of virusstammen met een bijna gelijke waardplantenreeks.Collectienummers van één en dezelfde soort konden sterk in lokale en systemische reactie verschillen. VooralN. benthamiana-9,N. miersii-33 enN. occidentalis-37B kunnen voor routine-inoculaties worden aanbevolen. Met uitzondering vanN. benthamiana enN. clevelandii zijn de genoemde soorten of genotypen nieuwe experimentele waardplanten voor de virologie. De soorten die van de sectieSuaveolentes worden genoemd, zijn inheems in de woestijnachtige gebieden van Australië. Verzamelingen daarvan verdienen nadere aandacht bij de diagnostiek van virusziekten waarvoor nog geen experimentele waarplanten voorhanden zijn.     

模拟降水量对毛乌素沙柳幼苗蒸发蒸腾的潜在影响

选择毛乌素沙地优势灌木沙柳为对象,研究157.5、315、472.5和630 mm 4种人控年降水量,对沙柳幼苗蒸发蒸腾作用的影响.结果表明,沙柳蒸发蒸腾量随着降水量的增加而逐渐增大,从157.5 mm增至630 mm,日蒸散量分别为1.63、3.11、4.37和5.66 mm,其中沙地蒸发量占主要部分.少量降水时,幼苗蒸腾速率明显呈现"午睡”现象,而充足降水则能消除"午睡”现象.相关分析结果表明,环境因子,如沙地含水率、太阳有效辐射、大气温度和饱和水汽压差,以及植物因素,如叶面积指数、叶片温度和气孔导度等,均能显著影响沙柳的蒸腾作用.     

福建马铃薯S病毒的分子鉴定及发生情况

为明确福建省马铃薯S病毒(PVS)的发生与分布情况,对福建省马铃薯主要种植区的PVS进行了鉴定和普查.在利用电镜技术和传统生物学方法鉴定的基础上,克隆了PVS外壳蛋白(cp)基因,依据PVS外壳蛋白氨基酸序列建立了PVS不同分离物的系统进化树.研究表明,利用PVS 外壳蛋白氨基酸序列分析可准确鉴定PVS,同时可分析不同分离物间的分子差异.利用病毒特异性引物和DIG标记的PVS cp基因为探针,分别利用RT-PCR技术和核酸斑点杂交技术(NASH)对PVS进行了检测,并对检测技术进行了改进.调查结果表明,PVS在福建省广泛分布,发病率最高可达80%以上,当地农家自留种可能是田间PVS的主要来源.     

桃红叶植原体检测及鉴定

对表现红叶的桃植株进行植原体16SrRNA基因PCR扩增,得到1.2kb的特异片段.将此片段与pGEM T Easy载体连接并转化到大肠杆茵DH5α感受态细胞中.通过酶切、PCR鉴定,对筛选得到的重组阳性克隆进行序列测定及同源性比较分析,确定该株系属于翠菊黄化植原体组(Aster yellows group,16SrI).在国内首次报道了翠菊黄化组中的植原体侵染桃树.     

中国禽(番鸭)呼肠孤病毒分离株S1基因全序列分析

采用RT-PCR技术,分别以DRV-YH、DRV-YJL两株中国禽(番鸭)呼肠孤病毒RNA为模板,扩增了S1全基因的cDNA片段.将S1 cDNA克隆到T载体后进行序列测定,测序结果表明所扩增的cDNA片段长1643个核苷酸,包含了完整的S1基因的三个开放阅读框架(ORF1,ORF2和ORF3)和基因两端的非编码区.核苷酸序列比较分析结果表明:DRV-YH与ARV-S1133,176分别有6个,10个核苷酸的差异,DRV-YJL与ARV-S1133,176分别有8个,12个核苷酸的差异,DRV-YH与DRV-YJL有4个核苷酸的差异.     

中国禽(番鸭)呼肠孤病毒YB株S4基因序列分析

应用非免疫番鸭胚增殖中国禽(番鸭)呼肠孤病毒YB分离株,用LSTRIAZOL提取病毒RNA,反转录-聚合酶链反应(RT-PCR)扩增中国禽(番鸭)呼肠孤病毒YB分离株S4基因节段cDNA.将S4cDNA克隆到PMDl8-T载体上,并进行了鉴定和核苷酸序列测定.序列分析结果,克隆的S4cDNA共1 124bp,包括非编码区和完整的阅读框架.分子进化系统分析表明该毒株与禽呼肠孤病毒(鸡呼肠孤病毒)的亲缘关系较远,DRV-YB与DRV-89330同缘率为93.3%.     

Association of aster yellows subgroup 16SrI-B phytoplasmas with a disease of Rehmannia glutinosa var. purpurea

A new phytoplasma disease of Rehmannia glutinosa var. purpurea was observed in the Czech Republic in 1998. Infected plants showing severely proliferating shoots, leaves reduced in size with vein clearing and chlorosis, shortened internodes and virescent petals died in advanced stages of the disease. Electron microscopy examination of the ultra-thin sections revealed the presence of numerous polymorphic bodies in phloem tissue of leaf midribs and petioles. The disease was successfully transmitted from infected plant via a dodder bridge into periwinkle ( Catharanthus roseus ). The phytoplasma aetiology of this disease was further confirmed by polymerase chain reaction (PCR) using universal primers R16F2/R16R2. Restriction fragment length polymorphism (RFLP) analysis of amplification products indicated the presence of aster yellows related phytoplasmas (16SrI-B) in naturally infected samples of R. glutinosa var . purpurea and in symptomatic periwinkle after dodder transmission of the agent. A comparison of the amplified sequence with 17 sequences available in the GenBank confirmed the classification of the phytoplasma in the subgroup 16SrI-B. This is the first report of natural occurrence of phytoplasma-associated disease in R. glutinosa var. purpurea.     

葡萄保鲜中的硫伤害研究

离体葡萄果实接触二氧化硫气体,在发生伤害的情况下,可引起细胞汁液pH的降低和组织质膜差别透性的增加,这种变化在一定范围内与硫伤害程度相关。电导率在伤害出现前有一峰值,而pH值呈较缓的降低趋势。以电导率变化做为细胞功能恢复的参照指标,结果表明“无核白”葡萄比“沙力皮”葡萄有较强的伤害恢复能力。     

Emended descriptions of indole negative and indole positive isolates of Brachyspira (Serpulina) hyodysenteriae

Two type/reference strains of Brachyspira (B.) hyodysenteriae, 14 Belgian and German indole negative, and 14 Belgian, German and Swedish indole positive field isolates of strongly β-haemolytic intestinal spirochaetes were compared by pulsed-field gel electrophoresis (PFGE) patterns, biochemical reaction patterns, 16S rDNA sequences and MIC determinations of six antibacterial substances. Three tests for indole production, including a spot indole test, were compared with congruent results. All field isolates were classified as B. hyodysenteriae due to a high genetic and phenotypic similarity with the type strains. The Belgian and German indole negative isolates had identical and unique PFGE patterns for the tested restriction enzymes MluI and SalI, as well as identical 16S rDNA sequences, and they could not be differentiated by any of the methods used. Seven unique PFGE patterns were achieved from the 14 indole positive field isolates. The patterns were identical and unique for epidemiologically related isolates. Type/reference strains and isolates without known relation to other tested isolates showed unique banding patterns. The MICs of tylosin, tiamulin, erythromycin, clindamycin, carbadox and virginiamycin were determined in broth for all isolates. In contrast to Belgian and German isolates, the majority of the Swedish field isolates were susceptible to tylosin, erythromycin and clindamycin. Probable pathways of infection for some of the Swedish isolates were determined. The PFGE patterns of epidemic clones of B. hyodysenteriae remained stable for a period of up to 8 years. In vivo development of resistance to macrolide and lincosamide antibiotics due to use of tylosin was clearly indicated for two epidemic clones.     

Molecular and biological characterisation of Cryptosporidium in pigs

OBJECTIVE: Genetic and biological characterisation of 12 isolates of Cryptosporidium from pigs and comparing them with Cryptosporidium isolates from humans and cattle. DESIGN: Cryptosporidium isolates from pigs were compared with those obtained from human and cattle using rDNA sequence analysis. The infectivity of two of the porcine isolates was determined in neonatal mice and the clinical history of the infected pigs recorded. RESULTS: Pig-derived isolates of Cryptosporidium exhibited two distinct genotypes; a porcine genotype and a bovine genotype, which is common to cattle and other livestock. The porcine genotype did not produce any infection in neonatal mice whereas the bovine genotype did. CONCLUSION: Two distinct genetically and biologically differing strains of Cryptosporidium appeared to be associated with acute diarrhoea in pigs. Whether Cryptosporidium was a primary or secondary pathogen is unclear but warrants further investigation. As the bovine genotype is known to infect humans, the results suggest that pigs can act as reservoirs of cryptosporidial infections for humans and other live-stock. The zoonotic potential of the pig-adapted genotype is uncertain and requires further study.