Some coat protein constituents from strawberry latent ringspot virus expressed in transgenic tobacco protect plants against systematic invasion following root inoculation by nematode vectors

The coding sequences in RNA2 for the coat proteins (CP) of strawberry latent ringspot virus (SLRSV) were modified and amplified using polymerase chain amplification reactions (PCR) to facilitate their expression inAgrobacterium tumefaciens-transformedNicotiana tabacum Xanthi-nc. The coding sequences for the smaller capsid protein (S, 29kDa) and that for the theoretical precursor of L and S (P, 73kDa) had ATG initiation codon sequences added at the 5-proximal Ser/Gly (S/G) cleavage site in the unmodified sequence. The sequence coding for the larger of the two proteins of mature SLRSV capsids (L, 44kDa) had an ATG codon added at its 5 S/G site and a TAG stop codon sequence added at the 3-proximal S/G site. The P, L and S proteins were expressedin planta to a maximum concentration of 0.01 % of total extractable proteins but did not assemble into virus-like particles. When challenged by mechanical inoculation with virus particles or viral RNA, and compared with control plants, tobacco plants (primary transgenic clones or S1 and S2, kanamycin-resistant seedlings) expressing the virus capsid subunits separately, or their precursor, decreased the accumulation of SLRSV particles in inoculated leaves and fewer plants became invaded systemically. In experiments in which the roots of seedlings were exposed to SLRSV-carrying vector nematodes (Xiphinema diversicaudatum), SLRSV was detected in the roots of non-transformed control tobacco plants (6/20) and in transgenic tobacco expressing the L protein (7/40), but not in any of 25 tobacco plants expressing the S protein or in 35 expressing the P protein. This is the second example of CP-mediated resistance to virus inoculation by nematode vectors.     

Comparison of the effects of salicylic acid and ethephon with virus-induced hypersensitivity and acquired resistance in tobacco

The induction of a hypersensitive reaction in Samsun NN tobacco by tobacco mosaic virus (TMV) at 20°C leads to the development of both localized and systemic acquired resistance, and is associated with the appearance of pathogenesis-related proteins (PR's) and large increases in peroxidase activity and ethylene production. Salicylic acid (SA) induced a similar resistance in treated plant parts and occasionally also in untreated upper leaves of plants of which three lower leaves had been injected. SA also induced the same four PR's, but these were confined to the treated leaves. Thus, the connection between the presence of PR's and the reduction of TMV multiplication and spread may not be direct.In contrast to TMV, SA did not stimulate ethylene production and hardly increased peroxidase activity. Induction of acquired resistance and PR's by SA developed equally well at 20°C and at 32°C. However, pricking leaves with needles moistened with the ethylene-releasing compound ethephon mimicked TMV infection in inducing acquired resistance and PR's in both treated and untreated leaves at 20°C, but not at 32°C. Ethephon increased peroxidase activity at both temperatures, but only at 20°C dit it induce changes in both the anodic and the cathodic isoenzymes that were similar to those induced as a result of TMV infection. SA induced PR's and reduced TMV multiplication in Samsun tobacco, and inhibited virus spread in Samsun NN at 32°C.These observation indicate that neither the induction of PR's, nor the development of acquired resistance is temperature-sensitive. On the other hand, the effects of ethephon are temperature-sensitive in the same way as the hypersensitive response to TMV. It can thus be hypothesized that ethylene, produced naturally during the hypersensitive reaction of tobacco to TMV, leads to the temperature-sensitive synthesis or release of a presumably benzoic acid-type compound that functions as the natural inducer of PR's and acquired resistance. Although vanillic acid has been shown to accumulate in hypersensitively reacting tobacco leaves, it produced none of the effects of SA, and thus cannot be the natural inducer.Samenvatting Inductie van een overgevoeligheidsreactie in Samsun NN-tabak door tabaksmozaïekvirus (TMV) bij 20°C leidt tot de ontwikkeling van een verworven resistentie die zowel lokaal als systemisch werkzaam is, en gaat samen met het verschijnen van pathogenesis-related proteins (PR's) en sterke toename in de activitieit van peroxidase en de produktie van ethyleen. Salicylzuur (SA) induceerde een vergelijkbare resistentie in behandelde plantedelen en af en toe ook in niet behandelde bovenbladeren van planten waarvan drie onderbladeren waren ingespoten. SA induceerde ook dezelfde vier PR's, maar deze waren beperkt tot de behandelde bladeren. Er bestaat dus geen directe samenhang tussen de aanwezigheid van PR's en de remming van de vermeerdering en uitbreiding van TMV in de plant.In tegenstelling tot TMV stimuleerde SA de ethyleenproduktie niet en verhoogde het de peroxidaseactiviteit nauwelijks. Inductie van verworven resistentie en PR's door SA trad even goed op bij 32°C als bij 20°C. Net als infectie met TMV leidde aanprikken van bladeren met naalden die gedoopt waren in een oplossing van ethefon — waaruit in het blad ethyleen vrijkomt — echter tot inductie van verworven resistentie en PR's in zowel behandelde als onbehandelde bladeren bij 20°C, maar niet bij 32°C. Ethefon verhoogde de peroxidaseactiviteit bij beide temperaturen, maar alleen bij 20°C induceerde het veranderingen in zowel de anodische als de kathodische isoënzymen die vergelijkbaar waren met die welke geïnduceerd werden als gevolg van infectie met TMV. SA induceerde PR's en verminderde de vermenigvuldiging van TMV in Samsun tabak, en remde de uitbreiding van het virus in Samsun NN bij 32°C.Deze waarnemingen tonen dat noch de inductie van PR's, noch de ontwikkeling van verworven resistentie een temperatuurgevoelig proces is. Daarentegen zijn de effecten van ethefon op dezelfde wijze temperatuurgevoelig als de overgevoeligheidsreactie op TMV. Men kan daarom veronderstellen dat ethyleen, dat op natuurlijke wijze geproduceerd wordt tijdens de overgevoeligheidsreactie van tabak op TMV, aanleiding geeft tot een temperatuurgevoelig proces, namelijk de synthese of het vrijkomen van een verbinding, vermoedelijk een benzoëzuurderivaat, dat fungeert als de natuurlijke inductor van PR's en verworven resistentie. Hoewel is aangetoond dat vanillinezuur zich ophoopt in overgevoelig reagerende tabaksbladeren, veroorzaakte deze verbinding geen enkel van de effecten van SA. Vanillinezuur kan dus niet de natuurlijke inductor zijn.     

Pelargonium flower-break and pelargonium line pattern viruses in the Netherlands; purification,antiserum preparation,serological identification,and detection in pelargonium by ELISA

Two viruses, detected frequently in the Netherlands in pelargonium, were identified by serology and test plant reactions. Antisera were prepared and an ELISA procedure was developed to detect the viruses in pelargonium.One of the viruses, PFBV-N, proved to be pelargonium flower-break virus. With the antiserum to PFBV-N, it could be detected reliably throughout the year inPelargonium zonale Springtime Irene.The other virus, PLPV-N, was serologically closely related to pelargonium line pattern virus (PLPV) and to pelargonium ring pattern virus (PRPV), as were an old virus isolate from Saturnus, collected in the Netherlands in 1971 (L128), and PLPV isolates from Yugoslavia (PLPV-Y) and Denmark (PLPV-D). There were only minor differences in host-plant reactions between the virus isolates. Based on these tests, PLPV and PRPV are considered as isolates of the same virus, for which, for practical reasons, the name pelargonium line pattern virus is proposed.PLPV could be reliably detected by ELISA inP. zonale Springtime Irene and Amanda throughout the year with only a few exceptions. InPelargonium peltatum Tavira, however, reslts were erratic due to uneven distribution of virus in the plant. Best results were obtained when petioles of fully expanded leaves were tested.     

高寒地区不同建植期人工草地群落垂直结构和生产力变化的研究

对高寒地区不同建植期人工草地群落垂直结构和生产力的变化研究结果表明：（1）随着人工草地建植时间的延长,人工草地群落垂穗披碱草高度异质性逐渐减小.（2） 不同建植期（1999、1998、1997年建植）人工草地垂穗披碱草和群落平均地上生物量均主要分布在0 ～20cm冠层中,约占平均地上生物量的61.03%、90.64%、84.55%和67.47%、92.54%、86.08%.（3） 不同建植期 （1999、1998、1997年建植）人工草地群落平均地下生物量均主要分布在0～10cm土层中,约占地下总生物量的85.97%、81.73%和78.47%.（4） 建植的人工草地如果其组分单一,即物种的丰富度很低时,均匀度增加,物种对环境资源的竞争力和利用率提高,导致土壤资源库中某些营养成分缺乏,草地群落初级生产力因此而降低。     

陇南山区生态旅游资源评价及开发策略

陇南山区生态环境脆弱,社会经济落后,同时又是生态旅游资源富集区,正处在西安旅游区、成都旅游区和兰州旅游区的包围之中,区位条件和客源市场较佳,实施生态旅游开发将对发展地方经济和提高游客生态环境意识意义重大。在对其旅游资源、区位条件、客源市场进行定性与定量评价的基础上,提出开发森林生态游、休闲度假游、避暑消夏游、三国怀古游、长征体验游等特色旅游项目,设计出不同尺度、不同主题的若干旅游线路,并针对发展生态旅游所面临的问题提出了相关对策。     

樟子松在西北干旱沙区的光合日变化特征

在甘肃省民勤治沙站 ,用 Li- 62 0 0便携式光合仪分别测定人工栽植的 4龄、9龄和 2 0龄樟子松的净光合速率日进程 ,其结果表明 :在干旱沙区 ,9龄以上樟子松的光合速率在午前随光强和气温而上升 ,午后下降。其 4龄、9龄和 2 0龄樟子松的最大净光合速率分别为 8.7μmol· m- 2 · s- 1、62 .3μmol· m- 2 · s- 1、1 9.7μmol· m- 2 · s- 1。     

关中西部四灌区旱情规律分析

为给关中西部灌区水资源合理调控提供依据，在简述干旱及其分类的基础上，指出了关中西部各类干旱多源于气象干旱。对该区7个气象站的年降水进行分析表明，各气象站年降水量的变差系数均为0．25，偏态系数均为0．5。以兴平气象站作为代表站对不同历时的旱情进行了分析，结果显示11月到来年2月发生重旱和特旱的概率较大，在30％左右，特别是12月份，概率达54％，且以特旱为主；7、8月份以中旱和重旱为主；夏玉米播前发生特旱的概率较高，达27％，拔节孕穗和抽穗开花期以特旱为主，概率达33％；冬小麦的越冬和麦黄期以特旱为主，发生概率在30％左右。     

西北干旱半干旱地区发展苜蓿草业的优势条件及前景分析

分析了西北干旱半干旱地区发展苜蓿产业和气候资源优势、土地资源优势、苜蓿的生长特性及其与这一地区地理条件的匹配优势，同时论述了种植苜蓿对这一地区的生态效益及经济意义，全面分析了这一地区发展苜蓿草产业的可行性和广阔的市场前景。     

Engineering of a Single Chain Variable Fragment Antibody Specific for the Citrus tristeza virus and its Expression in Escherichia coli and Nicotiana tabacum

Citrus tristeza virus (CTV) is one of the most destructive citrus virus diseases in the world. The construction of an engineered antibody, EMBL accession number AJ278109, able to specifically recognize its antigen, i.e. the coat protein of CTV, directly on infected plant material without any purification or manipulation of the entire woody plant. The potential uses of this engineered antibody are discussed.     

胶体金免疫层析法快速检测烟草环斑病毒

采用柠檬酸三钠还原法制备胶体金颗粒 ,标记烟草环斑病毒的抗体 ,制成免疫层析检测试纸条。检测粗提纯病毒的灵敏度为 1 0 0 0ng/ml,病汁液稀释 1 0 0 0倍后仍可快速检出。对大豆病种子、烟草冻干病叶等不同材料进行检测也有良好的效果 ,1～ 2min即可出现结果。用 9种不同的病毒进行测试 ,未出现非特异性反应。