Blackeye Cowpea Mosaic Virus in Florida: Host Range and Incidence in Certified Cowpea Seed
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ZETTLER, EVANS: COWPEA MOSAIC VIRUS 99 BLACKEYE COWPEA MOSAIC VIRUS IN FLORIDA: HOST RANGE AND INCIDENCE IN CERTIFIED COWPEA SEED F. W. Zettler and I. R. Evans separate and distinct members of the PVY-group of plant viruses. Plant Pathology Department This study was designed to assess Florida cer Gainesville tified cowpea seed for the presence of CV and to compare the host range of CV with that of ascer Abstract Sixteen lots of certified cowpea tained BYMV isolates. (Vigna unguiculata L. Walp.) seed were assayed for the presence of blackeye cowpea mosaic virus Materials and Methods (CV). Although all three samples from Georgia Samples of certified cowpea seed were obtained had a high incidence (>13%) of this virus, no during the autumn of 1969 from M. V. Meadows, CV was found in seed from California, Oklahoma Florida Department of Agriculture, Tallahassee, or Texas. In a host range study, an isolate of CV and tested for the presence of seed-borne CV. Each was compared with three isolates of bean yellow of the cultivars tested proved susceptible to CV in mosaic virus (BYMV). CV proved distinct from preliminary trials. The single noncertified seed BYMV in that only the former infected cowpeas sample tested was from a field of 'Knuckle Purple systemically whereas only the latter systemically Hull* cowpeas in Bradford County, Florida. The infected plants of pea (Pisum sativum L.), bean plants in this field were inspected by the senior (Phaseolus vulgaris L.), white sweet clover (Meli- author during the summer of 1969 and determined lotus alba Desr.), crimson clover (Trifolium to be uniformly infected with CV. incarnatum L.), and broadbean (Vicia faba L.). All seeds were planted, four seeds per 4-inch pot, in a greenhouse free from plants infected Virus diseases are major limiting factors in with CV. Seedlings were inspected for seed-borne the production of cowpeas in the southeastern virus when the first trifoliolates had fully ex United States, and certain horticulturally desir panded. Seed-borne infection by CV was deter able cultivars such as 'Knuckle Purple Hull' are mined on the basis of symptoms expressed on the subjected to repeated outbreaks of virus contained unifoliates and first trifoliolate leaves (Fig. 1). in planting stock. Kuhn et ah (15) lists six cowpea That the observed symptoms were CV-induced was viruses of the Southeast, at least three of which indicated by electron microscopic examination of can be introduced into fields through the planting leaf extracts stained in phosphotungstate for the of infected seed. A flexous-rod virus transmitted presence of flexuous-rod virus particles. Such by aphids appears to be the most important of particles were seen whenever extracts from leaves these seed-borne viruses from the standpoints of with symptoms were examined but never from being prevalent and affecting cowpea yields (10, leaves without such symptoms. 11, 12). This virus, herein called blackeye cowpea The CV isolate used in the host range study mosaic (CV), was first described from Florida by was originally obtained from field-infected 'Knuckle Anderson (1, 2). Later workers in the Southeast Purple Hull' cowpeas in Florida. Some of the (10, 11, 12, 15) have referred to this virus as a properties of this isolate have been described strain of bean yellow mosaic virus (BYMV) on previously (6, 16). In addition, this virus is seed- the basis of a report (3) that it was serologically borne in 'Ramshorn Blackeye' cowpeas (28 of 130 related to BYMV. However, Anderson (1) was seed harvested from plants inoculated with CV in unable to infect peas and beans with his virus of a greenhouse). Three isolates of BYMV were cowpeas, whereas other BYMV isolates, including used in this study. One isolate was obtained from that described by Corbett (4) from lupines, nor field-grown Crotalaria mucronata Desv. near mally infect these plants. Later, Edwardson et al. Gainesville, Florida. An isolate from Wisconsin (6) reported marked differences between CV- and was obtained from D. J. Hagedorn of the Uni BYMV- induced cylindrical inclusions and sug versity of Wisconsin, Madison, and an isolate gested that these two viruses are cytologically from Kentucky was obtained from S. Diachun of the University of Kentucky, Lexington. These isolates have been described previously (7, 8). Florida Agricultural Experiment Stations Journal Series The following species were tested for suscepti- No. 4610. 100 FLORIDA STATE HORTICULTURAL SOCIETY, 1972 bility to CV and to at least one of the above mentioned BYMV isolates: alfalfa (Medicago sa- tiva L.); bean ('Red Kidney', 'French Horticul ture/ 'Perry Marrow/ Michelite 62'); blue lupine (Lupinus angustifolius L.); broadbean ('Long- pod') ; cowpea ('Ramshorn Blackeye/ 'Black Local'); crimson clover; cucumber (Cucumis sati- vus L. 'A & C); globe amaranth (Gomphrena globosa); pea ('Alaska/ 'Midfreezer/ 'Ranger/ 'Thomas Laxton'); rattlebox (Crotalaria specta- bilis Roth); squash (Cucurbita pepo (L.) Alef. 'Early Prolific Straightneck'); tobacco (Nicotiana tabaeum L. 'Havana 425'); white clover (Tri- folium repens L.) and white sweet clover. All mechanical inoculations were made from expressed juice diluted with tap-water. Six hun dred-mesh carborundum dusted upon the leaves of test plants was used as an abrasive. All host range studies were checked by back inoculations to 'Red Kidney' bean and 'Ramshorn Blackeye' cowpea seedlings grown from virus-free certified stock from California. Results A high proportion (>13%) of the plants from Florida and Georgia seed were virus-infected and developed prominent symptoms similar to those described by Kuhn et al. (15) for CV (Fig. 1). The presence of symptoms on the primary Fig. 1. "Knuckle Purple Hull' cowpea seedling from CV- leaves of all infected plants indicated that these infected seed. Note symptoms on unifoliates and trifoliolates. plants resulted from infected seed. None of the plants from California, Oklahoma or Texas-grown peas but the three BYMV isolates did not. Ander seed developed such symptoms. son (1, 2) reported that while susceptible to CV, Of 236 seedlings of 'Knuckle Purple Hull' cow- broadbean is difficult to infect. CV did not infect peas from Florida, 30.9% were CV-infected. Sim any of the bean cultivars systemically in this com ilarly, 17.1% of 369, 18.0% of 355 and 13.6% of parative study, although this virus did induce 315 seedlings of 'Knuckle Purple Hull' cowpeas localized areas of veinal chlorosis on inoculated from three lots of Georgia seed were CV-infected. unifoliates. The three BYMV isolates, however, Conversely, none of two lots of 'Knuckle Purple readily moved systemically in all bean cultivars Hull' seed from Oklahoma or one from Texas during these trials. Later studies with CV re were infected. No infected seedlings were observed vealed that this virus was able to induce systemic in single lots of 'Lady' cowpeas from Oklahoma, mosaic symptoms in 'Red Kidney/ and Terry or in 'Texas Cream 40' or 'Floricream' cowpeas Marrow' beans, although systemic symptoms oc from California. Also, single seed lots of 'Lady/ curred relatively infrequently among inoculated 'Floricream/ 'Champion Cream' and three lots of plants. 'Texas Cream 40' from Texas produced no CV- All three BYMV isolates and CV infected blue infected plants. All seed lots except those from lupines systemically. This plant proved hyper Florida were certified according to the provisions sensitive to BYMV, as reported by Corbett (4), of the Florida Certification Seed Law (9). and died shortly after inoculation. The Kentucky Substantial differences in the host ranges of and Wisconsin BYMV isolates and CV infected C. CV and BYMV were noted. The BYMV isolates spectabilis systemically but symptoms differed in infected broadbean, white sweet clover, crimson that the CV did not induce foliar distortion as clover and four pea cultivars, whereas CV infected did the BYMV isolates. none of these plants. Conversely, CV infected cow- Neither CV nor the BYMV isolates infected ZETTLER, EVANS: COWPEA MOSAIC VIRUS 101 cucumber, squash, globe amaranth, alfalfa, tobacco, success that such programs have had in con or white clover. trolling seed-borne viruses elsewhere. Bean grow In recovery trials, 'Ramshorn Blackeye' cow- ers, for example, are protected from purchasing peas did not become systemically infected when planting stock infected with bean common mosaic inoculated from BYMV-inoculated plants nor did virus by a 1% limit of infected plants in fields 'Red Kidney' bean plants when inoculated from to be harvested for certified seed (14). According CV-inoculated plants. to these standards, none of the certified seed lots from Georgia in this study could have been sold to Florida growers for seed. Discussion This study shows that CV and BYMV have Literature Cited dissimilar host ranges and thus are distinct as 1. Anderson, C. W. 1955. Vigna and crotalaria viruses in suggested by Edwardson et al. (6). The assump Florida. II. Notions concerning cowpea mosaic virus (Marmor vignae). Plant Dis. Reptr. 39:349-353. tion that CV is a strain of BYMV is based upon 2. , 1955. Vigna and crotalaria viruses in Florida. III. Notations concerning identification difficul a reported serological relationship between these ties, indicator plants, possible vector relationships, and two viruses (3). However, both viruses appear to virus maintenance. Plant Dis. Reptr. 39:354-357. 3. Corbett, M. K. 1957. Serological and morphological be members of the PVY-group of plant viruses relationships of plant viruses. Florida Univ. Agr. Exp. Sta. Annu. Rep. 1956:117-118. and thus might be expected to have common anti 4. , 1958. A virus disease of lupines caused gens despite having different host ranges (13). by bean yellow mosaic virus. Phytopathology 48:86-91. 5. , and J. R. Edwardson. 1957. Virus The apparent inability of BYMV to infect cowpeas diseases of yellow lupines: Preliminary investigation on control by the use of a protecting border. Soil and Crop Sci.