University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln Papers in Plant Pathology Plant Pathology Department 1949 Mechanical Transmission of a Plant Tumor Virus to an Insect Vector Karl Maramorosch Myron K. Brakke University of Nebraska-Lincoln L. M. Black Follow this and additional works at: https://digitalcommons.unl.edu/plantpathpapers Part of the Plant Pathology Commons Maramorosch, Karl; Brakke, Myron K.; and Black, L. M., "Mechanical Transmission of a Plant Tumor Virus to an Insect Vector" (1949). Papers in Plant Pathology. 181. https://digitalcommons.unl.edu/plantpathpapers/181 This Article is brought to you for free and open access by the Plant Pathology Department at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Papers in Plant Pathology by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. Published in SCIENCE, New Series, Vol. 110, No. 2850 (Aug. 12, 1949), pp. 162-163 162 162 SCIENCE SCIENCE August 12, 1949, Vol. 110 ions on orientation of the water molecule by hydration the food intake of the animals and a loss in weight. with their influence on the amount of coagulation during The data show that the antivitamin effect of neopyri- freezing. As predicted, coagulation varies with salt con- thiamine is more pronounced than that of oxythiamine. centration and goes through a minimum at low concen- trations. Under favorable conditions coagulation can be TABLE 1 entirely inhibited. These effects would appear to have COMPARATIVE ANTITHIAMINE EFFECT OF OXYTHIAMINE various biological applications. AND NEOPYRITHIAMINE W\Vtof Wt. of. No. of References mice at start Inci- NO daysNo. ofbe- u 1. DJATSCHKOWSKY,S. J. Kolloid Z., 1932, 59, 76. Substance of of injections dence of urvival 2. DORSEY,N. E. Properties of ordinary water-substance. tested e of anti- polyneu- fore poly- time i New York: Reinhold, 1940. P. 397. vitamin ritis dee ~g % 3. Ibid., p. 503. g % ~developed 4. HAZEL, F. J. Phys. colloid Chem., 1947, 51, 415. 5. WALKER, H. W. J. Phys. colloid Chem., 1947, 51, 451. Oxythiamine 9 10.6-13.6 0 ... 13-21 Neopyri- thiamine 9 10.2-13.8 100 5-7 7-8 Antithiamine Effect of Oxythiamine and This manifested itself in the of A development polyneuritic Neopyrithiamine. Comparative Study' symptoms in the animals treated with neopyrithiamine and in a shorter survival period. The polyneuritic syn- Albert J. Eusebi and Leopold R. Cerecedo drome was similar to that described by Morris (1). In Department of Biochemistry, Fordham University, addition, we observed that the mice were apt to hold New York their heads on one side, and that they developed com- plete paralysis of the hind legs. The controls main- According to Wilson and Harris (3), pyrithiamine does tained their weight, and they survived until they were not have the structure assigned to it. These workers have sacrificed two weeks after the last animal in the corre- synthesized a new compound which they call "neopyri- sponding experimental group had died. thiamine," and which shows chemical and physical prop- erties that would be expected from the pyridine analogue References of thiamine. When this compound became available,2 an 1. MORRIS,H. P. Vitamins and Hormones, 1947, 5, 175. 2. SOODAK,M. and L. R. J. Amer. experiment was set up with a view of comparing its CERECEDO, chem. Soc., 1944, 66, 1988; Fed. 293. antithiamine effect with that of which was Proc., 1947, 6, oxythiamine, 3. WILSON, A. N. and HARRIS, S. A. Amer. Chem. Soc. shown in this laboratory to act as an antagonist for Abstr., Washington meeting, Sept. 1948. thiamine (2). Mice of the Swiss and C-57 Black strains from our stock colony were used. The animals were placed at Mechanical Transmission of a Plant Tumor on a thiamine-deficient diet which consisted of weaning Virus to an Insect Vector' casein (Labco) 25%, sucrose 53%, hydrogenated vege- table oil 10%, lard 5%, Osborne and Mendel salt mix- Karl Maramorosch,2 M. K. Brakke, and L. M. Black ture3 5%, and Ruffex 2%, and contained the following Brooklyn Botanic Garden, New York supplements per kg of diet: riboflavin 10 mg, pyridoxin 10 calcium 100 40 mg, pantothenate mg, ca-tocopherol mg, Most plant viruses transmitted by leafhoppers have not 3-carotene4 20 mg, vitamin D (Drisdol) 5000 units, and been demonstrated to be infective in extracts. The virus choline chloride 1.5 When the animals had become g. dealt with in this paper causes tumors in certain sus- in were maintained on 1 of thi- steady weight, they gsg ceptible plants, is carried by leafhoppers, and like others amine in solution subcutane- per day, injected aqueous in that group has proven difficult to detect in extracts. ously for one week. Daily injections of oxythiamine or It is our purpose to recount briefly the variety of meth- neopyrithiamine were then started together with 1 [(g ods that failed to transmit the virus and to report on the of thiamine in a ratio (thiamine: antivitamin) of 1: 50. success of insect-to-insect transmission by injection. Control animals, three for each group, received thiamine The virus, known as wound-tumor Aureogenus alone. The results are shown in Table 1. virus, inagnivena Black (2), is transmitted from plant to plant In the case of both compounds the effect on the food by the leafhoppers Agalliopsis novella (Say), Agallia intake was apparent overnight. There was a drop in constricta Van Duzee, and Agallia quadripunctata (Pro- 1 This investigation was aided by grants from the National vancher). Attempts by the authors to transmit the virus Institutes of Health and the John and Mary R. Markle Foun- dation. 1 This work was supported in part by a grant from the aWe are indebted to Dr. K. Folkers of Merck & Co., Inc. American Cancer Society in accordance with a recommenda- for the neopyrithiamine. tioa by the Committee on Growth, National Research Council. 8The quantity of manganese was doubled. 2 Fellow in Cancer Research, American Cancer Society. 4A mixture of 90% beta and 10% alpha carotene (GBI) Present address: The Rockefeller Institute for Medical Re- was used. search, New York City. August 12, 1949, Vol. 110 SCIENCE 163 from to have been mechanically plant plant made, using kept as controls. All insects were tested individually on various methods of both with untreated inoculation, juice two sets of crimson clover plants over a period of two from infected and also after the addition of ma- plants months, and the plants were then observed in the green- terials to the juice to alter the osmotic pressure, pH, and house for an additional period of two months. None of oxidation-reduction potential with the aim of stabilizing the control insects became infective, but some insects in- the virus. Juice has been rubbed onto the leaves and in- oculated with each dilution of viruliferous insect extract into the stems and crowns of jected plants by different up to and including 10-3 did. There was a long incuba- techniques. Several species of susceptible plants have tion period before inoculated insects were able to infect been used in these experiments, but an infection has healthy plants with wound-tumor virus. never been obtained by any of these methods. The in- These experiments show that wound-tumor virus can oculation of with from infective insects also plants juices be detected in extracts from insects, and the experiments failed to produce infection, even though such juice was also make possible the determination of some of the proved to have infective virus when tested by the suc- physical and chemical properties of the virus. The fact cessful described technique here. Leafhoppers fed on that three leafhopper-transmitted plant viruses have juices from diseased plants, from viruliferous insects, and proved transmissible by this method of insect inoculation on concentrates of these juices prepared by high speed suggests that the method may succeed in certain instances centrifugation did not become infective. It is possible, where other techniques have failed. of course, that more extensive trials with these procedures or variations of them might yield positive results. References Storey (5) was the first to transmit a plant virus from 1. BLACK,L. M. Phytopathology, 1941, 31, 120. 2. Amer. J. 408. insect to insect by inoculating virus-free vector leafhop- Bot., 1945, 32, 3. HOLMES, F. O. The filterable viruses. 2. Ber- Cicadujina mbila with from Suppl. pers, Naude, juice insects gey's Manual of determinative bacteriology (6th Ed.). carrying maize-streak virus, Fractilinea maidis (Holmes) 1948. Baltimore: Williams and Wilkins. Pp. 1127- McKinney (3). Using fine glass capillaries, Black (1) 1286. 4. MARAMIOROSCH,K. 14. the aster-yellows virus (Chlorogenus calliste- Phytopathology, 1949, 39, transmittee 5. STOREY,H. H. Proc. roy. Soc. Lond., Ser. B., 1933, 113, phi H. var. vulgaris H.) by injecting juice from virulifer- 463. ous into nonviruliferous leafhoppers (Macrosteles divisies Uhler). In view of these earlier successes, it seemed The Estimation of to Chromatographic Lysine logical attempt transmission of the virus by injecting and Some healthy vector leafhoppers with extracts from insects car- Applications of the Method1 rying wound-tumor virus. Anthony A. Albanese2 and Marilyn Lein Three hundred and fifty nonviruliferous leafhoppers (Agallia constricta Van Duzee) were caged on crimson Department of Pediatrics, New York clover plants (Trifolium incarnatum L.) showing pro- University College of Medicine nounced symptoms of wound-tumor disease and were kept In there for four weeks at about 25? C. Maramorosch had exploring the various published chromatographic it occurred to previously (4) obtained good transmission of the virus techniques, us that paper chromatography of the salts of at 25? C with this vector, after a minimum incubation copper amino acids might provide a simple means of some period in the insect of two weeks.