Bacterial diseases of potatoes: from classical phytobacteriology to molecular pathogenicity

Various bacterial pathogens attack potato plants and tubers. These pathogens includeErwinia, Corynebacterium (Clavibacter), Psuedomonas andStreptomyces. Over the past few years there have been significant advances in the molecular biological analysis of several of these pathogens and this is now helping us to understand the major aspects of virulence mechanisms. However, to date, such information is not sufficiently useful to allow us to intervene rationally in these potato diseases, except by the standard practises of good husbandry.     

Genetic mapping and utilization of quantitative trichome-mediated insect resistance in potato

Introgression of trichome-mediated insect resistance from the wild speciesSolanum berthaultii has become a major focus of the potato improvement program at Cornell University during the past twelve years. Several quantitative characters are involved in this resistance which is effective against a wide range of pest types. Correlative biochemical assays have been developed to assay specific components of the resistance, and the effects of the resistance on the target pests have been studied. Quantitative laboratory assays and specific measurements of insect behavior and biology have increased the precision of selection and enable the investigation of the genetic control of the resistance.We are currently using restriction fragment length polymorphisms (RFLPs) for genetic mapping of factors controlling the trichome traits fromS. berthaultii. Backcrosses to both the wild and the cultivated species parents have been evaluated for phenotypes contributing to the resistance mechanism, including trichome density, sucrose ester and polyphenol oxidase production by the trichomes, and the enzymatic browning reaction responsible for insect entrapment. Genetic maps are being developed for these progenies, using RFLP markers previously mapped in potato. Field and greenhouse trials under insect infestations are also being conducted with the mapping progeny. Our goal is to locate genes responsible for quantitative insect resistance by correlating RFLP variation at mapped loci with the trichome phenotypes and insect resistance. Genetic markers for these traits will be useful in transfer of the effective wild chromosomal segments into and among tetraploid potatoes, and for a better understanding of the resistance mechanism.     

Characterisation of energy-dependent efflux of imazalil and fenarimol in isolates of Penicillium italicum with a low,medium and high degree of resistance to DMI-fungicides

Differential accumulation of [¹⁴C]imazalil and [¹⁴C]fenarimol by germlings of wild-type and DMI-resistant isolates ofPenicillium italicum was studied at various pH values. At pH 7 and 8 the low-resistant isolate E_300–3 accumulated 22% and 35%, respectively, less imazalil than the wild-type isolate W₅. Imazalil accumulation at pH 5 and 6 was similar. Isolate E_300–3 also accumulated less fenarimol as compared with the wild-type isolate. This difference was much more obvious than for imazalil and was observed at all pH values tested. Differences in accumulation of both imazalil and fenarimol between low (E_300–3), medium (H₁₇) and high resistant (I₃₃) isolates were not observed. These results suggest that decreased accumulation of DMIs is responsible for a low level of resistance only and that additional mechanisms of resistance might operate in isolates with a medium and high degree of resistance. With all isolates fenarimol accumulation was energy-dependent. This was not obvious for imazalil.The wild-type and DMI-resistant isolates had a similar plasma membrane potential as determined with the probe [¹⁴C]tetraphenylphosphonium bromide ([¹⁴C]TPP⁺). Various test compounds, among which ATPase inhibitors, ionophoric antibiotics and calmodulin antagonists, affected the accumulation of [¹⁴C]TPP⁺, [¹⁴C]imazalil and [¹⁴C]fenarimol. No obvious correlation between the effects of the test compounds on accumulation levels of the fungicides and [¹⁴C]TPP⁺ could be observed. These results indicate that the plasma membrane potential does not mediate the efflux of DMI fungicides byP. italicum.     

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.     

Prospects for improving potato crop protection against aphid vectors

Conventional ways to control aphid vectors on potato crops rely essentially on cultural and chemical methods. The extensive use of insecticides, too often applied regardless of real needs and consequences, has resulted in more and more failures in both controlling aphids and virus spread. This paper stresses the need to introduce more rationale in the operation of existing methods by improving the efficiency of some of them like forecasting systems, to assess the potential of others like fertilization and semiochemicals and to put most efforts on resistant cultivars so that integrated pest management can be operated in the near future.     

New genes for disease resistances through somatic hybridization

Somatic hybridization, a process of combining protoplasts from different plants, can provide new sources of disease resistances for plants. In the case of wild and cultivatedSolanum species, the hybrids express resistances from each partner in the fusion and can often be crossed with cultivars to improve agronomic characteristics of the tubers. Restriction fragment length polymorphism (RFLP) analyses can provide a means for determining that the plants being investigated are actually hybrids as well as a means for following the introgression of DNA into progeny lines. These points are addressed in this paper with specific reference to somatic hybrids betweenSolanum brevidens and potato.     

Effective irrigation uniformity as related to root zone depth

Summary In models used for relating the yield to irrigation uniformity it has been assumed that the spatial distribution of irrigation water, as measured at the soil surface, is indeed the water distribution at any depth throughout the root zone. In the present paper the distribution of infiltrated water within the soil bulk, as determined by an analytic solution of the two-dimensional unsaturated flow equation, did not conform to this assumption. A new alternative definition of irrigation uniformity is proposed under the assumption that water uptake by roots does not affect the flux distribution within the soil profile. In this analysis the spatial distribution of irrigation water flux at the soil surface, which is the upper boundary condition of the flow equation, is assumed to be a sine function. The solution to this problem indicates that there is a damping effect, which increases with soil depth, on the surface flux fluctuations. Furthermore, the actual irrigation uniformity at a given depth below the soil surface depends upon the initial uniformity at the surface and the distance between adjacent water sources. The closer the water sources are to each other, the shallower is the depth needed to damp the oscillations down to a certain level. This may explain why the actual uniformity of drip irrigation is high while the detailed distribution is very nonuniform and on the other hand, why the actual uniformity of sprinkler guns is low while the detailed actual distribution is close to uniform. Two uniformity coefficients are derived in this study: 1. A depth dependent coefficient which is made up of the damping factor that multiplies the flux fluctuations at the soil surface; 2. An effective uniformity coefficient, which is an average of the depth dependent coefficient over a part or the entire root zone. Different degrees of uniformity are expected when water is applied by different irrigation systems having similar uniformity coefficients at the soil surface, but dissimilar distances between the emitters. Assuming that crop yield depends to some extent on the uniformity of water depth actually available to the roots, the yields associated with such irrigation systems will probably also vary.     

Nutritional stress in ficea sitchensis plantations in coastal British columbia: The effects of gaultheria shallon and declining site fertility

Examples of nutritional stress in conifer seedlings caused by competing ericaceous species (e.g. Calluna and Kalmia), have been reported in several parts of the world. Nutritional stress (primarily N deficiency) has been reported in Sitka spruce (Picea sitchensis) plantations growing in association with an ericaceous species, salal (Gaultheria shallon), in coastal British Columbia. Nutritional interference by salal was investigated on a chronosequence of sites up to 10 yr after clearcutting and slashburning. No direct evidence for an allelopathic contribution to the N stress was obtained. However, the rapid accumulation of salal fine roots and rhizomes, and the nutrients contained therein, provides a partial explanation for the observed stress symptoms. Soil analyses and seedling bioassays demonstrated a reduction in fertility in the period 8 to 10 yr after clearcutting and slashburning in comparison to the period 2 to 4 yr, which is believed to impose further nutritional stress on Sitka spruce. It is concluded that the nutritional stress in these Sitka spruce plantations is caused by a combination of (1) salal competition for nutrients and their subsequent immobilization in salal biomass, and (2) declining site fertility caused by the termination of the flush of nutrients (the assert period) that occurs in the immediate post-clearcutting and slashburning period. Sustaining good growth of plantations under such circumstances will require site nutrient management as well as vegetation management.     

Changes in the plasma concentrations of prolactin,luteinising hormone,progesterone and d‐(β)‐hydroxybutyrate in turkey hens (meleagris gallopavo), during treatment of broodiness under commercial conditions

1. An experiment was done under commercial conditions to investigate the physiological effects of isolating broody turkey hens, for 72 h, in sand and wire floored pens on the third, 10th and 16th weeks of production.

2. Hens identified as broody and removed from the flock had higher plasma prolactin concentrations than the laying hens at each of the three experimental stages.

3. Confinement in sand and wire floored pens, induced a decline in plasma prolactin concentrations. This decline probably impeded immediate development of broody behaviour. Alternately, levels of prolactin higher than those of laying hens were again measured 7 and 14 days after treatment during third week but not after the 10 th and 16th week of production.

4. Confinement did not induce consistent changes in luteinising hormone (LH) and progesterone concentrations from one period to an other.

5. An increase in the plasma concentration of D‐(/?)‐hydroxybutyrate was observed in the hens which had an egg present in the oviduct on day 2, 3 and/or 4 of the treatment. Subsequently, a decrease in ovulation rate was observed in the hens with higher concentrations of D‐(β)‐hydroxybutyrate while under treatment, during the 10th week of production.

6. These data confirm that the effectiveness of the traditional methods for broodiness prevention under commercial conditions is related to the induction of a decrease of prolactin.     

Detection of squash mosaic virus in seeds of melon (Cucumis melo) by enzyme-linked immunosorbent assay (ELISA)

Squash mosaic virus (SqMV, comovirus) is seed-transmitted in severalCucurbitaceae. Therefore, the use of virus-free seed is important to prevent establishment of this virus in the Netherlands and to avoid spread to other countries.This study was undertaken to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of SqMV in melon seeds. An antiserum was produced to a serotype 1 isolate from melon. Two ELISA variants were investigated viz. an ELISA variant with simultaneous incubation of sample and enzyme conjugate (ELISA 1) and an ELISA variant with successive incubation of sample and enzyme conjugate (ELISA 2). The sensitivity of ELISA was tested by mixing fluor of ground infected and non-infected seeds in different proportions. SqMV was detected by both ELISA variants at dilutions of 1 160 (1 part of infected flour mixed with 159 parts of non-infected flour) or higher after a substrate incubation period of 4 h. However, ELISA 1 gave relatively higher absorbance values than ELISA 2 for nearly all dilutions. Since ELISA 1 is also faster than ELISA 2, ELISA 1 is advised for routine testing. In these test, using subsamples of 100 melon seeds SqMV is detected reliably. ELISA 1 is now used in the Netherlands for routine-indexing of melon seed lots for SqMV.Samenvatting Het pompoenemozaïekvirus gaat over met het zaad van verscheideneCucurbitaceae. Het gebruik van virusvrij zaad is belangrijk om te voorkomen dat het virus zijn intrede doet in Nederland en zich naar andere landen verspreidt.Een antiserum werd geproduceerd tegen een serotype 1 isolaat van meloen. Met behulp van dit antiserum werd een ELISA ontwikkeld om pompoenemozaïekvirus in zaden van meloen aan te tonen. Twee varianten van ELISA werden vergeleken, namelijk een variant waarbij monster en enzymconjugaat gelijktijdig geïncubeerd werden (ELISA 1) en een variant waarbij monster en enzymconjugaat na elkaar geïncubeerd werden (ELISA 2). De gevoeligheid van de ELISA varianten werd uitgetoetst door meel van zieke zaden in verschillende verhoudingen te mengen met meel van gezonde zaden. Het pompoenemozaïekvirus werd met beide ELISA varianten aangetoond in verdunningen van 1 160 (1 deel meel van zieke zaden gemengd met 159 delen meel van gezonde zaden) of hoger na 4 uur incubatie met substraat. ELISA 1 gaf doorgaans hogere extinctiewaarden dan ELISA 2 voor bijna alle verdunningen. Omdat ELISA 1 ook nog sneller is dan ELISA 2, wordt ELISA 1 aanbevolen voor routinematig gebruik. Wanneer voor routinematig gebruik 100 meloenezaden per submonster getoetst worden, kan het pompoenemozaïek virus betrouwbaar worden aangetoond. In Nederland worden momenteel per zaadpartij 20 submonsters van 100 zaden getoetst.