Biochemical properties of the pathogenesis-related proteins from tobacco

This review describes the discovery and identification of the pathogenesis-related proteins (PRs) from tobacco. In crude leaf extracts the PRs are distinguished from the proteins in uninfected plants by their solubility at pH 3, resistance to a range of proteases, and mobility in polyacrylamide gels upon electrophoresis (PAGE) in non-denaturing conditions. PAGE has been used as a qualitative and semi-quantitative assay for PRs, and their migration in gels made from different acrylamide concentrations has been used to identify charge and size isomers and electrophoretically identical PRs in different tobacco cultivars. The subunit composition and molecular weight (mol. wt) of the four PRs identified first in Xanthi-nc were determined by SDS-PAGE; staining the gels has shown that these same four proteins in Samsun NN did not contain carbohydrate, lipid or nucleic acid, nor were they isozymic forms of twenty five enzymes known to increase in activity following infection with TMV. Evidence suggests that most of the PRs in Xanthi-nc and Samsun NN are extracellular.The purification of several PRs from Xanthi-nc, Samsun NN and other tobaccos is described, as well as their mol. wt, subunit and amino acid composition. PRs 1a, b and c consist of a single polypeptide and have similar mol. wt and amino acid compositions. Antisera prepared against purified Xanthi-nc b₁ protein have been used to determine serological relationships between PRs and form the basis of a very sensitive quantitative assay using ELISA. The regulation of synthesis of some PRs has been shown to involve translational control.     

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.     

Seasonal pattern of shoot emergence and its endogenous control in horsenettle (Solanum carolinense L.)

Controlling established horsenettle plants is achieved by suppressing shoot emergence from root systems. The seasonal pattern of shoot emergence and its possible endogenous control in horsenettle ( Solanum carolinense L.) were investigated. The shoot emergence period in an undisturbed population was limited to a seven-week period from mid-April, and a little longer in tilled conditions. Detached roots showed very high shoot-sprouting ability under 15–30°C throughout the year. In shoot clipping experiments, new shoots sprouted only from the stem and not from the root when attached to shoots, whether above-ground or underground. On the contrary, new shoots sprouted from the roots when all parts of the shoots were clipped off. From these results, the limited shoot emergence period in horsenettle is thought to be initiated by temperatures necessary for sprouting and is ended by a growth correlation effect between early emerged and matured shoots.     

The rates of maize growth and lignin biosynthesis change after root-applied chalcone

The effects of root-applied chalcone at 0.15 mmol L⁻¹ on the growth and lignin biosynthesis in maize were investigated. The contents of 4-coumarate: CoA ligase (4CL, EC 6.2.1.12) substrates in maize shoots were increased more rapidly in the samples with chalcone application than in the control and the increase occurred at ≤ 3 h after the application (HAA). The lignin content was reduced by chalcone at ≤ 6 HAA. The shoot growth was suppressed by chalcone at ≤ 9 HAA. Consequently, the results suggest that chalcone suppressed maize growth by inhibiting monolignol biosynthesis.     

Growth loss of Lebanon cedar (Cedrus libani) stands as related to periodic outbreaks of the cedar shoot moth (Dichelia cedricola)

An outbreak ofDichelia cedricola (Diakonoff) (Lep.: Tortricidae), the cedar shoot moth (CSM), began in spring 1998 and lasted 3 years. This was the first monitored outbreak of the CSM in Isparta, Turkey. Tree crowns recovered to near normal condition by the middle of each growing season (in early June) during the outbreak. Tree volume and volume element increments were examined throughout the outbreak cycle from 1954 to 2001. In the past, CSM activity in stands of Lebanon cedar (Cedrus libani A. Rich.) was assessed through radial increment analyses. Cedar tree ring chronologies were analyzed for evidence of the CSM. Tree-ring chronologies from nonhost cedar (nondefoliated sample trees) were used to estimate potential growth in the host cedar (defoliated sample trees) during current and past outbreaks; all trees selected were the same subspecies and varieties. Regional outbreaks of the CSM were identified by synchronous and sustained growth periods of the trees. In 2001, increment cores were collected from 17 host and 16 nonhost dominant or codominant trees and annual radial growth indices from 1954–2001 were calculated for each of two host and two nonhost sample plots. Growth functions were defined as the cumulative sum of radial, height, and volume increment, and were graphically compared between CSM host cedar and nonhost cedar trees. Tree ring evidence suggests that a large-scale outbreak occurred in 1955 (from 1955 to 1966) and a small outbreak occurred in 1985 (1985–1990) and in 1998 (1998-continued) in the study area. The average diameter growth reductions around 1955, 1985 and 1998 were 40%, 46% and 7% of potential, respectively. It was concluded that a narrow latewood band is significant indicator of defoliation by the CSM and the outbreaks appear to be associated with dry winter and spring weather prior to the autumn and winter in which wood feeding occurred. http://www.phytoparasitica.org posting Nov. 24, 2004.     

Relevance of the variability of kidding day in Creole goats in Guadeloupe

The results of 5732 records of kids born between 1985 and 1996 at Gardel Agricultural Experiment Station (INRA) in Guadeloupe, were used in order to estimate the effect of kidding day (KD) on individual preweaning growth performances, total productivity of Creole goats and litter size. The flock was subjected to a restricted mating season for a long time, by using male effect. The results of the fixed linear model showed a highly significant (P<0.001) effect of KD on growth rate and total productivity of does. Live weights of kids born around the 21st day of the kidding period (KP) was 4% to 7% higher than those of kids born the first day of the KP. For total productivity of does, this ratio did not reached more than 4%. The optimum at 70 days of age occurred around 14th day of the KD with 3% of improvement of total productivity. No effect was observed upon litter size. The genetic (co)variance components were estimated by six different Individual Animal Models. The heritability (h²) estimated from the best model, was h_D²=0.25±0.05 for genetic direct effect; h_M²=0.09±0.04 for genetic maternal effect and the genetic correlation between direct and maternal effects was −0.86±0.12. The use of KD could be highly recommended in a breeding program in this population of Creole meat goats, since it is quite easy to record under commercial conditions as a character related to reproductive performance.     

荔枝生产相关的水分生理指标远程监测研究

利用植物远程监测系统对荔枝园中的大气温湿度、土壤湿度、光照强度、大气蒸汽压差(VPD)等环境因子和茎干直径微变化、果实生长、叶片温度等树体生理指标进行了监测,发现该系统能准确及时并无伤害地记录他们的实时和周期性变化。据获得的数据初步表明,当果实处于快速膨大期,主茎的加粗生长明显缓慢,一旦果实采收后,茎干加粗生长则有一个迅速上升的变化;当土壤湿度高于33%(v/v)时,主茎的生长受到明显的抑制;VPD白天上升,夜间下降,当夜间VPD始终高于0,就形成所谓的“夜间干燥”,VPD的这种变化对荔枝主茎和果实的生长以及叶气温差(LATD)都有较大的影响,如主茎和果实的日间收缩量随VPD的增大而增大,夜间空气干燥不但抑制了主茎生长,也使日最大LATD下降。据此认为,果实和茎干的生长之间存在竞争关系,土壤湿度过高或者夜间空气干燥对荔枝的生长不利。