Physiological effects of arginine vasotocin and mesotocin in cockerels

1. The effects of continuous infusion of 0.1, 1.0 and 10.0 mU/min/kg body weight of arginine vasotocin (AVT) or mesotocin (MT) on cardiovascular and thermoregulatory responses, on plasma osmolality and ionic composition and on plasma concentrations of AVT, MT, prolactin and aldosterone, were investigated in conscious White Leghorn cockerels.

2. Neither of the peptides, at any dose, affected cardiovascular functions, plasma ions and osmolality. Infusion of MT at the rate of 10 mU/min/kg body weight increased respiratory rate. Both peptides at doses of 1 and 10 mU/min/kg reduced the temperatures of the comb and shank but had no effect on the skin and cloaca.

3. Doses of 0.1 and 1.0 mU MT/mu/kg reduced plasma aldosterone and at 10 mU/min/kg increased plasma AVT. At any given dose MT had no effect on plasma prolactin. AVT at 0.1 and 1.0 mU/min/kg of AVT reduced plasma MT. AVT at 1.0 mU/min/kg increased plasma prolactin and at 10 mU/min/kg reduced plasma aldosterone.

4. During saline infusion, plasma MT was positively correlated with plasma AVT and negatively correlated with respiratory rate and cloacal temperature. Plasma AVT showed a positive correlation with plasma MT and aldosterone and a negative correlation with respiratory rate and skin temperature.

5. During saline infusion, there was no significant correlation between cardiovascular functions, or plasma osmolality and ionic composition and plasma MT or AVT.

6. The present study suggests that interrelationships between circulating concentrations of AVT and MT do exist and that AVT affects     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

Effects of Latent Infection of Stock Plants and Abundance of Thrips on the Occurrence of <Emphasis Type="Italic">Tomato spotted wilt virus </Emphasis>in Chrysanthemum Fields

DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001     

Suppression of Verticillium Wilt in Eggplant by Some Fungal Root Endophytes

One hundred and twenty-three fungal isolates were obtained from 225 root segments of eggplants, melon, tomato, strawberry and Chinese cabbage, grown as bait plants in a mixed soil made up of samples from different fields in Shizuoka, Japan. Isolates belonging to Mycelium radicis atrovirens (MRA), including Phialocephala fortinii, were the most prevalent in all the five bait plants. Eleven of the 123 isolates, after being inoculated onto axenically reared eggplant seedlings, almost completely suppressed the pathogenic effects of a post-inoculated, virulent strain of Verticillium dahliae. Seven of these 11 isolates had come from the roots of eggplant and included Heteroconium chaetospira, P. fortinii, and unidentified species of Fusarium, Penicillium, Trichoderma and MRA. P. fortinii, H. chaetospira, a non-sporulating isolate with white mycelium (SWM) and MRA were easily reisolated from root segments. Hyphae of H. chaetospira, P. fortinii and SWM colonized the root tissues of eggplant without causing apparent pathogenic symptoms. The mechanisms by which these endophytes confer resistance to infection by V. dahliae are unknown but the effectiveness of these fungi in a laboratory setting indicates that they have potential as biocontrol agents and merit further investigation.     

Movement of Xanthomonas campestris pv. vitians in the stems of lettuce and seed contamination

Xanthomonas campestris pv. vitians , the causal agent of bacterial leaf spot of lettuce (BLS), can be seedborne, but the mechanism by which the bacteria contaminates and/or infects lettuce seed is not known. In this study, the capacity of X. campestris pv. vitians to enter and translocate within the vascular system of lettuce plants was examined. The stems of 8- to 11-week-old lettuce plants were stab-inoculated, and movement of X. campestris pv. vitians was monitored at various intervals. At 4, 8, 12 and 16 h post-inoculation (hpi), X. campestris pv. vitians was recovered from 2 to 10 cm above (depending on stem length) and 2 cm below the inoculation site. Xanthomonas campestris pv. vitians was also recovered from surface-disinfested stem sections of spray-inoculated plants. Together, these results are consistent with X. campestris pv. vitians invading and moving systemically within the vascular system of lettuce plants. To investigate the mechanism of seed contamination, lettuce plants at the vegetative stage of growth were spray-inoculated with X. campestris pv. vitians and allowed to develop BLS. Seed collected from these plants had a 2% incidence of X. campestris pv. vitians external colonization, but no bacteria were recovered from within the seed.     

Indole-3-acetic acid biosynthesis in <Emphasis Type="Italic">Aciculosporium take</Emphasis>, a causal agent of witches' broom of bamboo

 Aciculosporium take (Ascomycota; Clavicipitaceae) is a causal agent of witches' broom of bamboo plants. The symptoms of this disease are believed to be induced by plant hormones, particularly auxins. Indole-3-acetic acid (IAA) was identified in cultures of this fungus in an l-tryptophan-supplemented liquid medium. IAA production was confirmed on 30 isolates of A. take from various hosts and locations at levels up to 1 mg/l. The biosynthetic pathway of IAA in A. take culture was examined by analyzing intermediate products and by feeding experiments. The results showed that the indole-3-pyruvic acid pathway (l-tryptophan → indole-3-pyruvic acid → indole acetaldehyde → IAA) was the dominant pathway in A. take. Received: June 3, 2002 / Accepted: July 25, 2002     

Inconspicuous restraint of rice seedling growth by root-infecting fungi in soil of a rice paddy field

 Rice seedling growth, estimated by plant height and root development and discoloration, was better in pasteurized soil than in unpasteurized soil obtained from a flooded rice field. Rice seedlings also grew better in sterilized soil modified by adding roots harvested from the pasteurized soil than in soil modified by adding roots harvested from the unpasteurized soil. The results demonstrate that seedling growth in the rice field soil was inhibited by soil microorganisms, even though no typical symptoms such as seedling blight or damping-off appeared. Pythium aristosporum is suggested to be involved in the inhibition. Thus, it appears that inconspicuous restraint of rice seedling growth could occur in soils of rice paddy fields. Received: May 20, 2002 / Accepted: October 16, 2002 Acknowledgments The authors thank Dr. T. Ichitani, former professor at Osaka Prefectural University, for providing an isolate of Pythium aristosporum for comparison, and Mr. Mitsuaki Sato of Akita Prefectural College of Agriculture for technical assistance.