. AAA- A sawax‘ FOR msmm mus mus: VECTORS: occugamc ON THE cumvmap BLUEBERRY VACCEWM coamaowm_ LENNAEUS ‘fi-n- *A... ~ _ u -. Thesh ’50:. ‘Ehe Degree 0? Me 3. WICEIIEAN STATE UNWEBEETY Thomas Lee Burger 1966 LIBRARY Ill!lllllllfljfllflllllwlfllfllmflWWII WWW University ABSTRACT A SURVEY FOR POSSIBLE VIRUS DISEASE VECTORS OCCURRING ON THE CULTIVATED BLUEBERRY Vaccinium corymbosum Linnaeus by Thomas Lee Burger During the spring and summer of 1964 a survey of possible virus disease vectors occurring on cultivated blueberry, Vaccinium corymbosum Linnaeus, was conducted in southwestern Michigan. Objectives of the survey were to correlate differences occurring between counties and between farms in the counties as to species of Arthropods present and virus disease incidence. Observations on other Arthropods were also made. Survey sites were chosen on the bases of acreage and disease incidence appearing in areas under intensive cultivation of the high- bush blueberry. At the conclusion of the study the results indicate that: 1. Scaphytopius magdalensis (Prov.), a leafhopper known to be a vector of stunt disease in New Jersey, is present in Michigan. 2. Separation of Scaphytopius magdalensis (Prov.) from other species of the same genus occurring in Michigan, has been accomplished on the bases of anatomical characteristics and color variation. 3. There are a number of insect species present in Michigans' cultivated blueberry fields, capable of transmitting virus diseases. 4. Virus disease incidence is increasing annually in Michigans' cultivated blueberry fields. Thomas Lee Burger Shoestring, a virus disease of highbush blueberries, has remained at a low incidence level in Berrien County, Michigan which may be due to the absence of Myzus scammelli, an aphid. A SURVEY FOR POSSIBLE VIRUS DISEASE VECTORS OCCURRING ON THE CULTIVATED BLUEBERRY Vaccinium corymbosum Linnaeus By Thomas Lee Burger A THESIS Submitted to Michigan State University in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Entomology 1966 ACKNOWLEDGEMENTS The author is grateful to Dr. Gordon E. Guyer, Chairman of the Entomology Department, for providing financial assistance for this study and for serving on the author's guidance committee. my sincere appreciation goes to my major professor, Dr. A. J. Howitt, under whose constant advice and suggestions this investigation was made. I am greatly appreciative of Drs. Paul H. Wooley and James Bath, for their technical assistance and valuable suggestions, and to people of the Entomology and Natural Science Departments for their technical assistance. Special thanks are extended to John W. Nelson, Research Director of the Michigan Blueberry Growers' Association and the many members of the association, for their interest, assistance, and added financial aid given the author. Finally, my personal thanks are due to my parents, and to Mr. and Mrs. Ralph Gies, for their constant encouragement and assistance through my years in college. ii TABLE OF CONTENTS Page INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . 1 Background History . . . . . . . . . . . . . . . . . . . 1 Distribution and Economic Importance . . . . . . . . . . . 3 Nature of the Problem . . . . . . . . . . . . . . . . . . 5 REVIEW OF LITERATURE . . . . . . . . . . . . . . . . . . . . . 7 Blueberry Viruses and Vectors . . . . . . . . . . . . . . 7 Insect Survey Equipment . . . . . . . . . . . . . . . . . 11 MATERIALS AND METHODS . . . . . . . . . . . . . . . . . . . . 12 RESULTS AND DISCUSSION . . . . . . . . . . . . . . . . . . . . 18 General Arthropod Survey . . . . . . . . . . . . . . . 18 Possible Virus Disease Vectors - Aphids . . . . . . . . . 18 Possible Virus Disease Vectors - Leafhoppers . . . . . . . 25 Rearing Experiments with Leafhoppers . . . . . . . . . . . 28 SUMMARY AND CONCLUSIONS . . . . . . . . . . . . . . . . . . . 37 LITERATURE CITED . . . . . . . . . . . . . . . . . . . . . . . 39 APPENDIX I . . . . . . . . . . . . . . . . . . . . . . . . . . 41 APPENDIX II . . . . . . . . . . . . . . . . . . . . . . . . . 43 iii LIST OF TABLES Table Page 1. Comparison of Counties and Farms Showing Differences in Numbers of Masonaphis pepperi (MacGillivray) Collected . 20 2. Comparison of Counties and Farms Showing Differences in Numbers of Myzus scammelli (Mason) Collected . . . 21 3. The Comparison of Virus Disease Incidence in Five Counties (1959-1965). Data was Obtained from Division of Plant Industry Records . . . . . . . . . 22 4. Leafhopper Species, with Total Number of Each Species, Found in the Five-County Survey . . . . . . . . . . . 26 iv Plate LIST OF PLATES Map of Michigan Showing Blueberry Areas D-Vac Model 12 Sampler Used in Survey for Collecting Taxon Found in Cultivated Blueberry Fields Collecting Sample From Cultivated Blueberry Bush Showing Ease by Which Center of the Bush can be Sampled . . . . . . . . . Copulation Between Male and Female of Scaphytopius magdalensis (Prov.) . Nymphs of Scaphytopius magdalensis (Prov.) Showing Second Instar Nymph with Black Markings and Third Instar Nymph with Red . . . . . . . . . Nymphs of Scaphytopius magdalensis (Prov.) Showing Black-Banded First Instar Nymph, Red-Banded Second and Third Instar Nymphs and Sand Stone Colored Fifth Instar Nymph . . Female of Scaphytopiys magdalensis (Prov.) Showing Feeding and Ovipositing on Cultivated Blueberry, Vaccinium corymbosum Linnaeus . Male of Graphocephala coccinea (Forst.) Showing Coloration and Feeding on Cultivated Blueberry, Vaccinium corymbosum Linnaeus Nymph of Graphocephala coccinea (Forst.) Showing Coloration Occurring in Fifth Instar . . Page 13 15 31 32 33 34 35 36 INTRODUCTION Background History Vaccinium corymbosum Linnaeus, the cultivated highbush blue- berry, has been an economic asset to the agriculture of Michigan, con- tributing over $5,000,000 annually to the State's economy. The acreage of cultivated blueberries has been restricted to certain areas of southwestern Michigan where climatic and soil conditions favorable for growing highbush blueberries are present. In general, blueberries re- quire a temperate climate, a soil with low pH, and a high water table. These agronomic characters are present only in the southwestern section of the State near Lake Michigan. While great advances have been made in agriculture pertaining to blueberry production, there are some factors which cause a reduction in potential production that need investigation. One such area of lower production includes fungus, bacterial and virus diseases of blueberries. In the last 10 to 15 years, there has been an increase of not only numbers of diseased bushes but also an increase of new blueberry diseases. The diseases of the highbush blueberry are not restricted to Michigan alone, but may be found in other blueberry-growing areas of the United States including New Jersey, which competes with Michigan as a major producer of cultivated blueberries. Research entomologists and plant pathologists in New Jersey have 1 investigated the spread of diseases, particularly virus diseases. One virus disease, commonly known as blueberry stunt, which has seriously threatened the blueberry industry in New Jersey, has been the object of research efforts the past 15 to 20 years. The disease, first ob- served in 1928, was described and named by Wilcox (1942) when its virus nature was demonstrated by budding techniques. Further investigations have been conducted since 1943 by research personnel employed by the New Jersey Agricultural Experiment Station. They have determined that an insect vector known as Scaphytopius magdalensis (Prov.) is respon- sible for the transmission of blueberry stunt disease; this vector be- longs to the family Cicadellidae in the class Insecta. Determination of this species of leafhopper as the insect vector was not accomplished without extensive investigation and elimination of many of the other possible disease vectors found in the insect class. Insects are not the only biological agents suspected of the transmission of the different virus diseases. Investigations involving nematode, mechanical and root grafting transmission are currently being conducted. In 1963, the Michigan Blueberry Growers' Association proposed that a research project should be initiated to determine how virus diseases are being spread in commercial plantations in Michigan. Be- fore detailed investigations could be initiated, it was necessary to determine the possible virus disease vectors present in commercial plantings. The preliminary survey program was divided into two parts: the first to determine by survey techniques the 'nematode species present in the fields, and the second to survey and determine possible insect vectors present in the blueberry fields. During the 1964 growing season, a survey of the insect fauna was taken. The objectives were to obtain a cross-section of the insect fauna present in the fields throughout the growing season, which is the most probable time for disease transmissions. Another objective of the survey was to eliminate and reduce the number of species of insects that would be required for further tests on transmission of blueberry virus diseases, such as stunt, red ringspot, shoestring, mosaic, and necrotic ringSpot. Distribution and Economic Importance The highbush blueberry is native from northern Florida to southern Maine and westward to southern Michigan. Michigan and New Jersey lead in the production of cultivated blueberries, with other areas such as Indiana, North Carolina and Washington having sizeable acreage under cultivation. Smaller areas under cultivation can be found in New York, Pennsylvania, Ohio, several New England states and Oregon as reviewed by Dorr (1965). In Michigan, there is a distinct demarcation between areas where cultivated highbush blueberries and native lowbush blueberries are found (Plate 1). The line runs from Bay City southwest across the state to a point just north of Grand Rapids, then northwest along the isothermal line to the shore of Lake Michigan. The northerly projec- tion on the west side of the state is due to the moderating influence of Lake Michigan. The areas under the highest cultivation of highbush blueberries are confined to five counties: Berrien, Van Buren, Allegan, Ottawa and Muskegon (Plate 1). The 1964 census of the Michigan Blueberry - Boundary of high cultivation °° Divisan of highbush from lowbush ' Survey areas Plate 1.--Map of Michigan showing blueberry areas. Growers' Association shows that 9,000 acres of land in these five counties are under cultivation of the highbush blueberry. Total acreage of highbush blueberries is increasing annually in Michigan, but due to the requirements needed for growing, the increase is small. In 1965, the cultivated blueberry industry was valued at over five million dollars (Hobein, 1965, personal interview). Nature of the Problem Members of the Michigan Blueberry Growers' Association and the Departments of Botany and Plant Pathology and Entomology hypothesized that production of the highbush blueberry crop could be increased by improved disease control, particularly in the area of insect-transmitted viruses. In order to provide information on this problem, a program was initiated to survey the insect fauna present in cultivated blue- berry fields, and more specifically, to determine the presence of pos- sible virus disease vectors. Since 1942, when Wilcox (1942) first discovered and isolated the stunt disease of highbush blueberries in New Jersey, several re- search workers have carried on a continuous search for insect vectors of blueberry virus diseases. Tomlinson, Marucci and Doelhert (1950) reported transmission of blueberry stunt by a complex of Scaphytopius magdalensis (Prov.) and S. verecundus (Van Duz.). One of the principal reasons the survey was undertaken was to determine whether these insect vectors were present in Michigan. The last insect survey of Michigan's highbush blueberries was taken by Tuttle (1947), the main collecting method being a sweep net. Since then, many new and more advanced collecting techniques have been introduced. The research project was designed to determine: (1) possible new pests that have entered into the blueberry insect problems; (2) possible disease vectors present; and (3) distribution of possible vectors of blueberry diseases. This survey was conducted on two farms in each of the five counties that contain the highest acreage of cul- tivated blueberries in Michigan (Plate 1). LITERATURE REVIEW Blueberry Viruses and Vectors Virus diseases have become a serious problem to the cultivated highbush blueberry industry, not only in Michigan, but also in New Jersey, where most of the diseases were first noticed. One such disease, called stunt, was first observed in 1928 and was later described and named by Wilcox (1942) when its virus nature was demonstrated by transmission through budding. Wilcox also explained that the general effect of the stunt diseases is a reduction in length and vigor of new growth, a moderate stimulation of branching, and the production of small, unmarketable fruit. At that time, Wilcox had no proof as to the means of dissemination of the disease, but suspected that some insect was the vector. L. O. Kunkel (Tomlinson 35 31 1951) of the Rockefeller Institute confirmed, by means of graft and dodder transmissions, the conclusion reached by Wilcox that stunt is caused by a virus. Johnston 25 £1 (1945) stated that, while stunt disease is present in a number of Michigan blueberry fields, it has spread very slowly. Johnston further stated that a quarantine had been enacted, which pro- hibited the shipment of nursery stock into the state from any outside point, or from point to point within the state, except under, or in compliance with, the quarantine regulations. From this quarantine stemmed the annual inspection of fields in Michigan by inspectors from 7 the State Department of Agriculture as insurance against the rapid Spread of stunt disease in Michigan. A search for insect vectors was initiated by the New Jersey Agricultural Experiment Station in 1943. Tomlinson (1947) initiated a general insect control experiment in relation to stunt disease in hopes of limiting the spread of stunt in fields by controlling the insect vector or vectors, but his efforts were of little avail. Marucci (1947) published results of a Cicadellidae survey in blueberry fields and Tuttle (1947) of a survey of insects in Michigan blueberry fields. The two surveys differed greatly as to the species and number of Species of Cicadellidae found in blueberry fields. In 1944, C. A. Doehlert, in a preliminary experiment with leaf- hoppers, successfully transmitted stunt from a diseased blueberry bush to healthy plants, as reported by Marucci E£.§L (1947). This led to subsequent studies by Marucci SE EL until Tomlinson SE 3; (1951) re- ported that a complex of Scaphytgpius magdalensis (Prov.) and S. verecundus (Van Duz.) was responsible for the transmission of blueberry stunt disease in New Jersey. Later research revealed that it was S. magdalensis that was a vector of stunt and not S. verecundus (Hutchinson 1955 and Maramorosch 1955). Goheen (1953) reported that stunt, a virus disease of the yellows type, is prevalent in New Jersey and North Carolina and that entire fields may become so badly diseased that little or no crop is produced. Goheen also stated that the disease occurs in Massachusetts, New York, Michigan, Maryland, and eastern Canada and could be spread from bush to bush by dodder, a parasitic seed plant found in some blueberry fields. Hutchinson 95 31 (1954) was apparently the first to investigate a disease locally known as "ringspot." They report that symptoms are suggestive of other described ringspot diseases caused by viruses. They are most apparent in the fall when infected leaves may Show red rings or jagged patterns of the oak-leaf type associated with the veins or both. In the spring, small red circles and dots may be seen on the leaves. In grafting experiments, small chlorotic dots were the first symptoms of ringspot. These later changed to red dots, after which red rings and the oak-leaf patterns appeared. Hutchinson further states that ringspot was transmitted to 10 of 13 two-year-old Cabot plants whip-grafted in April, 1952. Clear virus symptoms were evident within five months. The presence of red ringspot in wild blueberry in New Jersey may mean that it has spread from the wild to commercial plantings (Stretch, 1965). Red ringspot has not been reported in the other blueberry-growing areas, except Michigan, where red ringspot was found in plants whose origin could be traced to New Jersey. No reports of red ringspot in wild or in most commercial plantings in Michigan, North Carolina, Massachusetts, Washington and Oregon may mean that the vector is not present in those areas. Varney (1957) was the first to report the occurrence of mosaic and shoestring, two virus diseases of cultivated blueberry in New Jersey. Shoestring had been assumed to be a virus and mosaic a genetic disorder, but Varney presented evidence that both are of virus origin. Shoestring virus symptoms appear on both twigs and leaves. Red streaks appear first on new twigs in the spring but may become masked later in the summer. Severely infected leaves are narrow and pointed or strap- like; moderately affected leaves are wavy and distorted. Varney inoculated two Jersey and two Burlington varieties of blueberry bushes 10 in the spring of 1953 by chipbuds from Cabot plants showing typical shoestring symptoms. No symptoms appeared the first year, but the following Spring red streaks, red veinbanding and leaf distortion appeared on the inoculated bushes. Mosaic disease symptoms on blueberry, as reported by Varney (1957), are shown by the leaves of affected bushes which are strikingly mottled with yellow and yellow-green areas. The degree of mottling varies with leaf position and blueberry variety. Varney inoculated healthy bushes at budbreak in the greenhouse by means of chipbuds from a Stanley bush showing symptoms typical of mosaic. Twelve out of 13 inoculated bushes showed symptoms in two to three weeks after inocula- tion. Necrotic ringspot, a virus disease of cultivated blueberries, reported occurring in New Jersey by Varney SE S; (1960) and in Michigan, causes eventual death to the bush. Symptoms of the disease are ex- pressed by reduction in Size of leaves which have chlorotic areas appearing as rings or lines. The chlorotic areas may drop-out of the leaves, leaving a tattered, shot hole, appearance. In transmission tests conducted by Griffin SE a; (1963), a nematode, Xiphinema americanum (Cobb), transmitted the disease from infected blueberry bushes to cucumber plants in greenhouse experiments. Transmission of the disease from one bush to another by S, americanum has not been demonstrated. Le Clug (1964) estimated the average annual loss from potential blueberry production, due to fungus, bacteria and virus diseases, at nearly two million dollars. ll Insect Survey Equipment The use of the vacuum-sweeping technique for surveying for pos- sible disease vectors was demonstrated by Maki (1965). Vacuum-sweeping is a superior method, as compared to the sweep net, for collecting Cicadellidae, Aphididae, Cercopidae and many other families of insects that could be possible vectors of blueberry Virus diseases. MATERIALS AND METHODS The survey program for possible virus vectors was carried out in the five counties with highest acreage of cultivated highbush blue- berries in Michigan: Berrien, Van Buren, Allegan, Ottawa and Muskegon (Plate 1). Two farms in each of the counties were selected as sampling sites. They were selected on the basis of disease incidence in the field, one field of low disease incidence and the other of high, to determine the difference in species and population size of insects be- tween the two fields. Ten bushes were sampled in each field and were picked on the basis of different types of conditions that existed in that particular field, such as low, wet areas; high, dryer areas; and areas of vigorous growth, to obtain a representative cross-section of the insect fauna. Jersey, the major variety of highbush blueberries, in Michigan, made up the bulk of the bushes sampled. The D-Vac Model 12 sampler, a vacuum sweeper-like machine, was employed for sampling the bushes for Arthropods. The D-Vac (Plate 2) consists of a power source, a blower, and a fiberglass housing for holding the sample bags. A 1.26 cubic inch, three-quarter horsepower, two-cycle gasoline engine supplies the power. A fine mesh nylon net, capable of holding mites, was fastened to the fiberglass housing, the open end of which measures 0.929 square feet. The entire device weighs only 15-1/2 pounds. Samples were taken by directing the open end of the device 12 13 , \ ’ ‘ "x, .3." 3’31". s. :- .a 'pa . ’ f ' V I ‘ .1 A I ' Plate 2.--D-Vac Model 12 Sampler used in survey for collecting taxon found in cultivated blueberry fields. 14 toward the bush and sweeping from bottom to top and repeating until the entire bush was covered (Plate 3). This sampling method not only enabled the outside of the bush to be sampled easily, but also the center, where the younger and more succulent canes are found. Each bush took approximately one minute to sample, varying a little as to size of bush. After 10 bushes in the field had been sampled, the machine was kept running and the larger debris was checked and thrown out. The remaining material in the net was then emptied into pint jars partially filled with K-A-A-D, a fixative consisting of 1 part Kerosene, 7 parts 95% ethyl alcohol, 2 parts glacial acetic acid and 1 part diopane, and then labeled as to date, locality and farm. K-A-A-D was used to keep the soft-bodied insects from becoming deformed so that subsequent identification could be handled more easily. After remaining in K-A-A-D from six to eight hours, the material was washed and stored in 80% ethyl alcohol. The two farms in each county were sampled on the same day; all ten farms were sampled within three consecutive days. The sampling periods, which varied between 15 and 20 days, ran from May 7 through September 12, 1964. This gave assurance of not missing the appearance and disappearance of some particular species of Arthropod in the field. The first problem encountered was the separation of the insects from the debris, which consisted of leaves, twigs, and blossoms, on which and in which the smaller insects would be caught. It was also found that the samples were too large, which meant more time and work would be required for separation. To solve these problems, a sub- sampling technique was employed. 15 0 ~ ' I! \M' ‘~ 2 ¥ L ,1 1'. - ' 1 I ‘ ‘ $7 i" o '[ :2 9 I I Plate 3.--Collecting sample from cultivated blueberry bush showing ease by which center of the bush can be sampled. 16 To determine the size of the subsample needed for accurate determination of the Arthropods present, each of the seventy samples was graded with respect to size. It was determined that the largest sample would fit inside a petri dish which had a radius of 2.75 in. and a total area of 23.77 in. The petri dish was divided into eight equal parts and the number of each species was counted in each section. Through a comparison of the individual section counts with total counts, it was demonstrated that 95% accuracy of total insect species present in the sample could be obtained by counting only one section of the plate. Each subsample was removed from the entire sample by placing a wedge-Shaped cup over the subsample that was randomly chosen. The rest of the sample was then washed back into the pint jar and the subsample into a smaller petri dish for easier sorting. The figures resulting from the tally were then multiplied by 8 to give an estimate of the total number in the entire sample. All 70 samples were subsampled in the above manner. Each subsample was sorted and separated into the different classes and orders of Arthropods and stored in vials of 80% ethyl alcohol for further taxonomic classification. Subsamples were examined under a binocular scope to assure complete removal of such specimens as mites, thrips, aphid nymphs, etc., from the petri dish. Identification of Arthropods in the samples was done by the author with the exception of the Aphididae, Coleoptera, Lepidoptera, Hymenoptera and Acarina. The other Arthropods presented in this thesis were submitted to persons in the Entomology Department at Michigan State University for final confirmation. The insect survey data were analyzed for information that might l7 answer the following questions: (1) What possible virus disease vectors are present in the fields, and (2) What difference is there, if any, between high and low virus disease incidence fields as far as Species of insects are concerned? A discussion of the results of these analyses are presented in the following sections of the thesis. RESULTS AND DISCUSSION General Arthropod Survey The author has found, as did Maki (1965), that the vacuum-sweep method of surveying is ideal for collecting such taxa as Cercopidae, Cicadellidae, Aphididae, acalypterate Diptera and Acarina as indicated by the taxon collected (Appendix II). The fluctuation in population of each taxon represented could be due to such factors as insecticide application, weather, and predators or parasites. The application of insecticides is probably the most critical factor, since greater numbers of individual taxon were collected in the spring before dusting was initiated, and in the fall after dusting had been terminated. Tables in Appendix II list the taxa present in the samples. Totals are given for each farm sampled, with each table comparing the two farms sampled in the county. Possible Virus Disease Vectors - Aphids In the survey for possible insect virus disease vectors, emphasis was placed on collecting two major families of insects: Cicadellidae and Aphididae. The latter are discussed in this section. After the 1964 survey, it was apparent that only 2 species, Masonaphis pepperi (MacGillivray) and Myzus scammelli (Mason), colonized on high- bush blueberry in Michigan. The other aphids, which were collected during their transient activity on blueberry, can not be disregarded 18 19 as possible virus vectors. This study was restricted to the 2 Species closely associated with cultivated blueberries as a point for research embarkation. The transient aphid collections have not been identified but have been retained for future use should efforts with M, scammelli and M. pepperi as vectors of blueberry viruses prove futile. Masonaphus pepperi was the predominate species found colonizing on highbush blueberry. Each county and farm surveyed contained M. pepperi (Table l). The largest number of specimens was collected in the two counties with the highest virus disease incidence as shown in Table 3. Myzus scammelli, on the other hand, was not found in every county or on every farm surveyed, as Shown in Table 2, and was not found as numerous as M. pepperi. To verify the findings that M. scammelli was not present in Berrien County, a follow-up survey was conducted on 10 additional farms in Berrien County during the summer of 1965. The results of that survey were also negative as to the presence of M, scammelli. Along with this follow-up survey a few farms in La Porte County, Indiana, which adjoins Berrien County at the Indiana-Michigan border, were surveyed and found to be free of this species. A comparison of Tables 1 and 2 with Table 3 indicates a marked difference between the presence of the two species of aphids and the disease incidence of all viruses in each of the five counties. There is good correlation between the absence of M. scammelli and the low incidence of shoestring virus in Berrien County. Survey results from La Porte County compare with Berrien County in this respect. Comparisons of Tables 2 and 3 Show that the presence of M, scammelli and the spread of shoestring to be quite closely correlated by the fact that, where M. scammelli is found, shoestring incidence is also 20 TABLE l.—-Comparison of counties and farms Showing differences in numbers of Masonaphis pepperi (MacGillivray) collected Masonaphis pepperi Farm Sampling Dates Berrien County, Mich., 1964 5/8 5/27 6/15 7/6 7/27 8/18 9/10 Chikaming - 24 - - 16 - - Hutchinson - - 72 104 40 24 - Van Buren County, Mich., 1964 5/7 5/25 6/16 7/8 7/28 8/19 9/11 Hartman - - - 24 16 - - Wakeman - — 40 - 72 110 24 Allegan County, Mich., 1964 5/10 5/25 6/16 7/8 7/29 8/19 9/11 Wadsworth 8 32 96 384 216 848 192 Double A - - - 144 40 16 8 Ottawa County, Mich., 1964 5/11 5/26 6/17 7/9 . 7/29 8/20 9/12 De Pree - - 16 384 496 352 - Boo Hoot - - 112 16 - 48 16 Muskegon County, Mich., 1964 5/11 5/26 6/18 7/9 7/29 8/20 9/12 Derkse - 8 - _ - - - Pauls 8 - - 16 16 16 8 21 TABLE 2.--Comparison of counties and farms showing differences in numbers of Myzus scammelli (Mason) collected Myzus scammelli Farm Sampling Dates Berrien County, Mich., 1964 5/8 5/27 6/15 7/6 7/25 8/18 9/10 Chikaming - - - - - - - Hutchinson - - - - - - - Van Buren County, Mich., 1964 5/7 5/25 6/16 7/8 7/28 8/19 9/11 Wakeman - - - 16 - 8 8 Hartman - - - _ - - - Allegan County, Mich., 1964 5/10 5/25 6/16 7/8 7/29 8/19 9/11 Wadsworth - 8 16 - - - - Double A - - - 8 40 - - Ottawa County, Mich., 1964 5/11 5/26 6/17 7/9 7/29 8/20 9/12 De Pree - - 24 - - _ 3 B00 Hoot 24 16 16 - - - - Muskegon County, Mich., 1964 5/11 5/26 6/18 7/9 7/29 8/20 9/12 Derkse - - 16 16 - - _ Pauls - 8 16 24 - - - 22 woomo>pow mommomwn mouw> no~.o qu nso.o mNH nom.o Ham nmm.o mama omHN Hmuoe m~.o ma ao.o mm Ho.o a am.~ mma HNm cowmxmsz m~.o Haw mo.o mm on.o Hos mH.H cam cam msmuuo qN.o mm mo.o om mo.o om mm.o mm mam sameH< HH.o em mo.o as ma.o em mH.o mm com amusm am> oo.o - No.0 m oo.o - Hm.a am we ewauamm Hoaa nom.o amm ano.o Ema amq.o NooH amm.o amm «com Hmuoa oq.o mma oo.o - oo.o 0H no.0 Hwa wwm aowmxmsz mN.o Hmm mo.o Hm oo.H sow Hm.o mmm sow «gauge SH.o mm Na.o mm «0.0 w ma.o Ne omm gammaaa om.o moH HH.o an mm.o mod om.o 50H mNm sauna cm> eo.o q mN.o mm no.0 m «H.o SH NHH cmauamm coma BAH.o Ham amo.o aha pam.o HHS Boa.o amm moom Hades NH.o mo Ho.o N No.0 o mm.o HNN Sam cowmxmsz mm.o mam ea.o aw Na.o mom «a.o mms was mzmuuo So.o mm ao.o Nm so.o SH om.o on com :mwmaaa mo.o as oo.o om no.0 om m~.o oma mam cmusm cm> mo.o a H~.o mN oo.o - oH.o ma NmH amauumm amma <\ma ma «\mo mm «\mn ma <\mn ma emsm>a=m Rococo mmuo< owmmoz .>.m.m.z wawuumoonm uaoum mouooom mnumsvaH ucmflm mo coamw>wn Eopm voofimuno mm3 moan Amomfiummmfiv mmfiucooo o>Hw CH oocoofioaw ammomwo mspfl> mo GOmfiumano oSH .m mqmuam mommomwa msuw> 000.0 000 000.0 05 000.0 000 05.0 0000 5000 00000 00.0 00 00.0 - 00.0 0 00.0 000 005 00000002 00.0 050 00.0 00 00.0 500 00.0 500 000 030000 05.0 50 50.0 00 00.0 00 00.0 500 000 0000000 00.0 00 50.0 00 00.0 5 00.0 000 050 00000 00> 00.0 0 00.0 a 00.0 a 00.0 00 00 0000000 0000 000.0 005 000.0 00 005.0 000 000.0 000 5000 00000 00.0 00 00.0 H 00.0 H 00.0 000 005 00000002 00.0 050 00.0 00 00.0 000 00.0 000 000 030000 00.0 0 00.0 0 00.0 00 00.0 00 050 0000000 00.0 0 00.0 0 00.0 00 00.0 000 000 00000 00> 00.0 0 05.0 0 50.0 N 05.0 05 00 0000000 0000 050.0 500 000.0 00 000.0 055 000.0 0000 0000 00000 00.0 50 00.0 H 00.0 - 05.0 000 050 00000002 00.0 000 00.0 00 00.0 050 00.0 000 005 030000 00.0 50 00.0 0 50.0 00 05.0 00 000 0000000 00.0 00 00.0 00 00.0 00 00.0 050 000 00000 00> 00.0 - 00.0 5 00.0 - 00.0 00 000 0000000 0000 0500 00 0500 00 0500 00 0500 00 0050>000 500000 mmuo< onmoz .>.m.m.z wowuumoosm oasum 000000000--.0 00000 illrlll hut. 00.593 . <\mn How owm0m>m huanoo o>0m n 0:00> uoamwa0m 00000002 I .>.m.m.z 0004 pom magmam 00000009 u <\mn monmsm @0000009 I mam ,4 2 050.0 000 050.0 000 000.0 0000 000.0 0000 0000 00000 00.0 00 00.0 - 00.0 00 00.0 000 500 00000002 00.0 000 00.0 00 00.0 0000 00.0 0000 000 030000 00.0 00 00.0 00 00.0 00 00.0 000 000 0000000 00.0 00 00.0 00 00.0 50 00.0 000 000 00000 00> 00.0 0 00.0 00 00.0 0 00.0 000 00 0000000 0000 «\00 00 0500 00 «\00 00 <\00 00 0000>000 000000 mmhofi 000002 .>.m.m.z wa0uumoo£m unsum moo>o>03m 00000009 md00> 000000000--.0 00000 25 the greatest. Shoestring has remained in low incidence through the years where M. scammelli has not been found. The fact that the disease is present in Berrien and La Porte counties, but not in large propor- tions, could be explained by the movement of symptomless diseased nur— sery stock in to the counties which later developed diseased symptoms from previous infection. Possible Virus Disease Vectors - Leafhoppers The second family of possible virus disease vectors covered in the survey are the cicadellids, or more commonly known as the leaf- hoppers. Much emphasis has been placed on this family in the thesis, since it is known to contain the vector of stunt disease of blueberries in New Jersey. This survey uncovered a greater number of genera and species (Table 4) than Tuttle obtained in 1947, a phenomenon which could be based on the superiority of the D-Vac collecting method over the sweep net method which was employed by Tuttle. Scaphytopius magdalensis (Prov.), the vector of stunt disease in New Jersey, also appears in Table 4, but was not collected during the author's original survey. Out of the 25 species of leafhoppers (Table 4), only about a dozen appear to be possible leads for further transmission tests. This conclusion is based on the relationships of the various species to virus disease incidence. If a species of leafhopper is not present on more than one farm and in more than one county its potential as being a vector is lessened due to the fact that virus diseases are widespread in at least 4 counties. 26 00 n w w a u Aum0omv mmC0oooo mamsmwoosm00w 0N . n m w w Amon0omv mooumflmw0a 0000:08000 000 000 00 00 0 000 0.0000 0000 000000000 wm0 mqm mww 00 r u A.0£>v momowu0xo 05:0000xm w n u u u m Anmamzv 000000 musoCO0nuh0m w n s u u m Ammmv mJUHHno muoooo0£uh0m mum qu mom 0000 mmw mNm Ammmv mUHE0C0 mwuwam 000.0 0000 0000 0000 0000 0000 00000000 00000 00000000 00 0 00 0 00 00 00000020 00000 000000000000000 w I n u w n A.mumv 0300000>000 mSHmnmmUOuHmn ow 000 qu 000 u u AHHDQMV wumfimcH m0moamm 00 w w u u u A.n.>v muowms0 mausaoamm on u I em I u Anou0mv 000000=o mDHHouNHnE< w u n u n w Acmeov mamasoomamm 000m00000w< m0 w 00 0N u u A.>00mv mucoHOC0swcmm 00000w00000o4 Hmuoa cowoxmsz 030000 omw000< coudm aa> cmwuuom moHUQO mam muoamo mmHucsoo mo>030 zucooono>0m 050 G0 vcsom 00000030 £000 00 ponEDC 00000 £003 .mmHUmam umaaosmmmquu.a mqmu:m umumm momfl mo Hfimm mumH CH muwnesc mwumfi :0 wcflummamm casom a .hucnoo :0 080mm nuon co vasomm mm mm 0N0 wq 00 ow omH m m 0H m I I A.Q.>V mswumowasm mDHMLQQUOummN m I Ammmv mav0MHn mdwxnofimH A.>oumv nmflmcofimwwme mswmouhsmmom I m A.n.>v mHHmucoym mdwmoumnmmom w I Ahmmv mmusum mdwmouhnmmum I I Ahmmv msumuouuw mdwmmmfizmmuwm m 0H A.>oumv mamuwaowsm msaamcuonmo m I Ammmv mvsawamm maHHHm>uoz 00H 00 A.H0umv waoum0ummm mmamumouumz m N m 0H A.Hmnv muuum mamcomhw H0000 cowmxmdz m3Muuo :mmeH< amusm cm> amwuumm mmwommm was mumcmw mmwuadoo UmSCHucooII.¢ mqm HoHoo usmflflm mnu muoz .A.>oumv mfimcoamuwma mdwmouhnmmom mo Aumoflv onEmm can Aunmwuv mama cmosuoa cowumfldmooII.¢ mumfim 32 .wmu £u03 snazc Hmumcfl wuwnu paw mwcqumE xoman £u03 zmsxc unumcH vcooom mafiaonm A.>oumv wwmcofimowme moflmouhnmmom mo mnasszI.m mumam 33 .mmmuoh 3oz c0 wcwuusooo mm Annmav downwzousm ma umcowucme uo: ucmwpw> 00000 0 00 saema uwumcw umuwm ona .caEm: umumaw suuwm vmuofloo mcoum pcmm paw msaezc umumcw cuwsu cam pcooom wowcmn vow .sashd woumcH umufim vmwcmn xomHn wa03050 A.>oumv mflmcflamvmme msflmdumnmmom mo m£mE%zII.o oumflm 0w 34 .msmmGCHA Eamonamuoo Edwcfloow> .hpuonmsfin woum>0ufiso co wcflufimomfl>o cam wcmemu wcH3ozm A.>oumv mHmeHmvmWE mswmoumnmmom mo onEomII.m mumam If .0 . '7 0 t0 . «I... o. ’ ' 00.0.1..- .0 - v 0 _' 35 .msomccflq EamonENuoo Eswcwoom> amuumnodan woum>0uaso co wchoom cam GOHumuoHoo w:030£m A.umuomv mmcfloooo mamnmooonmmuu mo mHmSII.w mumfim 36 .000000 00000 00 coflumuofioo mcw3osm A.umuomv mwcHoooo mfimcmwoonmmsw mo zaathI.m mumflm SUMMARY AND CONCLUSIONS A survey for possible virus disease vectors of stunt, shoestring, necrotic ringspot, and mosaic, which are virus diseases of the culti- vated blueberry, Vaccinium corymbosum Linnaeus, was carried out in a five-county area in Michigan (Plate 1). During the spring and summer of 1964, two farms in each of the five counties were surveyed. All the farms and counties were compared with regard to virus disease in- cidence and Arthropods present. Collection of the Arthropods present in the survey areas was completed with the aid of a vacuum-sweeper like machine called a D—Vac Model 12 Sampler. The results of this survey led to the following conclusions: 1. There is a definite correlation between virus disease in- cidence and Arthropod species present between counties, but not between farms in a county. 2. Scaphytopius magdalensis (Prov.), found for the first time in Michigan by the author, is the vector of stunt in cul- tivated blueberries in New Jersey. Therefore, it is con- cluded that S. magdalensis is the probable vector of stunt in cultivated blueberries in Michigan. 3. There are two generations, spring and fall, of S, magdalensis in New Jersey, but only a fall generation has been found in Michigan. The fact may indicate that the insect control program for cultivated blueberries now being followed in Michigan is adequate for controlling the spring generation. 4. A number of species of insects found by this survey show potential as being possible vectors of blueberry viruses. 5. Virus disease incidence is increasing annually in Michigan. 37 10. 11. 12. 13. 38 Shoestring, a virus disease of cultivated blueberries, has remained in low incidence in Berrien County. The absence of Myzus scammelli Mason, a species of aphid, in Berrien may be the controlling factor. A fall survey should be conducted to determine if S, magdalensis is present in the five counties previously sur- veyed. Since stunt incidence is high in all the counties. A more precise picture of Arthropod fauna present at this time could also be obtained. A pre-bloom,dormant and post-harvest experimental insecticide program should be initiated. The fact that some pests appear or reach greater proportions at these times, as shown in Appendix II, will test this hypothesis. Stunt disease incidence in the cultivated blueberry fields could possibly be reduced if a post-harvest insect control program were initiated. Scaphytopius magdalensis (Prov.) and Graphocephala coccinea (Forst.) can be reared on cultivated blueberry, Vaccinium corymbosum Linnaeus. Male and female adults of Scaphytopius magdalensis (Prov.) can be separated by anatomical characteristics or color variation from other species of Scaphytopius that are found in Michigan. Nymphs of Scaphytopius magdalensis (Prov.) could possibly be separated out from other leafhopper nymphs on the basis of color patterns. Extensive testing of virus transmission in cultivated blue- berries can now be shortened with the aid of the author's findings. LITERATURE CITED Delong, D. M. 1948. Leafhoppers or Cicadellidae of Illinois. Illinois Natural History Survey Bulletin, 24(2):l-376. Dorr, J. E. 1965. Pollination studies on the cultivated blueberry Vaccinium corymbosum (Linnaeus). M.S. Thesis, Mich. State Univ., East Lansing, pp. 1-59. Goheen, A. C. 1953. The cultivated highbush blueberry, pp. 784-789. I2 Plant Diseases, U.S. Dept. Agr. Yrbk. Agr. Griffin, G. D., J. E. Huguelet, and J. W. Nelson 1963. Xiphinema americanum as a vector of necrotic ringspot virus of blueberry. Plant Disease Reptr., 47(8):703-704. Holbein, Peter 1965. Personal interview concerning the economic importance of the cultivated blueberry industry in Michigan. Hutchinson, M. T. and E. H. Varney 1954. Ringspot--a virus disease of cultivated blueberry. Plant Disease Reptr., 38:260-262. Hutchinson, M. T. 1955. An ecological study of the leafhopper vectors of blueberry- stunt. J. of Econ. Entomol., 48(1):1-8. Johnston, S., D. Cation and C. A. Boyer 1945. Blueberry stunt disease. Mich. Agr. Expt. Quarterly Bu1., 27:409-412. LeClug, E. L. 1964. Crop losses due to plant diseases in the United States. Phytopathology, 54(11):1305-1319. Maki, J. R. 1965. A comparison of sampling methods used in field pesticide side effects studies. M.S. Thesis, Mich. State Univ., East Lansing, pp. 1-54. Maramorosch, K. 1955. Transmission of blueberry-stunt virus by Scaphytopius magdalensis. J. Econ. Entomol., 48:106. 39 4O Marucci, P. E. 1947. A leafhopper survey in blueberry fields, pp. 2-4. l2 Proceedings of the 16th Annual Blueberry Open House. New Jersey Agr. Exp. Stat., Rutgers. Marucci, P. E., W. E. Tomlinson, Jr., and C. A. Doehlert 1947. Cage tests for possible carriers of blueberry stunt disease, pp. 9-11. l2 Proceedings of the 16th Annual Blueberry Open House. New Jersey Agr. Exp. Stat., Rutgers. Michigan Dept. of Agriculture, Division of Plant Industry Records 1965. Number of virus diseased blueberry plants tagged per county for seven years. (Unpublished) Stretch, A. W. 1965. Ringspot virus of cultivated blueberries, pp. 3-4. £2 Proceedings of the 33rd Annual Blueberry Open House. New Jersey Agr. Exp. Stat., Rutgers. Tomlinson, W. E., Jr. 1947. The blueberry fruit fly and other pests of 1947, pp. 15. IE Proceedings of the 16th Annual Blueberry Open House. New Jersey Agr. Exp. Stat., Rutgers. Tomlinson, W. E., Jr., P. E. Marucci, and C. A. Doehlert 1951. Leafhopper transmission of blueberry stunt disease. J. of Econ. Entomol” 43(5):658. Tuttle, D. M. 1947. A study of the insect fauna of the cultivated blueberry, Vaccinium corymbosum Linnaeus. MJS. Thesis, Mich. State Univ., East Lansing, pp. 1-85. Varney, E. H. 1957. Mosaic and shoestring virus diseases of cultivated blueberry in New Jersey. Phytopathology, 47:307-309. Varney, E. H. and L. C. Raniere 1960. Necrotic ringspot, a new virus disease of cultivated blue- berry. Phytopathology, 50:241. Wilcox, R. B. 1942. Blueberry stunt, a virus disease. Plant Disease Reptr., 26(9):211-213. APPENDIX I Farms Surveyed in Michigan The specific location of each farm surveyed is listed. 42 APPENDIX I Farms Surveyed in Michigan Berrien County, Michigan Hutchinson's - W 1/4, Sec. 2, T. 78., R. 20 W. Chikaming Gardens - S 1/4, Sec. 3, T. 75, R. 20 W. Van Buren County, Michigan Hartman's - Found to be just over the line into Allegan County, SE 1/4, Sec. 36, T. 1N, R. 16 w. Wakeman's - S 1/4, Sec. 6, T. 23., R. 15 W. Allegan County, Michigan Wadsworth's - S 1/4, Sec. 31, T. 3N, R. 15 W. Double A - N 1/4, Sec. 30, T. 4N., R. 15 W. Ottawa County, Michigan DePree's - W 1/4, Sec. 13, T. 5N., R. 16W. Boo Hoot's - E 1/4, Sec. 2, T. 5N., R. 16W. Muskegon County, Michigan Paul's - W 1/4, Sec. 15, T. 9N., R. 16 W. Derkse's - E 1/4, Sec. 30, T. 9N., R. 15 W. APPENDIX II Summary of Arthropods Collected The total numbers and taxa of arthropods collected during the spring and summer of 1964 are presented in this appendix. The appendix is subdivided according to county, farms, time sample was taken, and approximate dusting dates. 44 00 00 00 00 000 «N 00 omH Nu on wqm I 00 0N \D LO l I I om NmH mmp0omsz mmcfiwuoohg mmpHHmuaonocoA mmc0a0£aomoua 00000000000000 mapwaouoHSU omeEOSOHHSU mmv0cowoaoumumo mmw0%Eowwomo ompHHOnuom mmv0N%Eouw< mumua0n mmUHHDSDcHEm mmc0zunoEoucm mfionEoHHOU oumcwwmman0sw mHEwpommwm oumfisome mHHvaEoumumo mmU0HHocwoooo 2mm mopowafihmm .fig mapwfimsomhuno 00.00% mmpwowzuq< mumumomaoo mmEHOMwanEouH mmEHOM0uaooumm . mumEmHumommz mcHumo¢ oH\m 0:0,0 00\0 00\% 000\m mmumn wc00asmm xmm\m w\m 000500000 00\0 000\00 n~\% «0\% 0m0\o0 mmumn wc00m5mm 00\m« acmc0£ousm :ome «cma .zucsoo co0uuom 45 ca 00 I I mm 0N «N Nm «OH Nm Nm ONH 00mmnuom acuwzoousma< wmwfivouhma< Hummmmm mwsmmGOmmz 000000000 mumumosom mHDHHmn mowmnuhuoo 0000000000 mamoc0mm mamzflmw mmvwwwmz mausnEHH mwnmz mmwwnmz mwauouwmsn mdfimuocowHHH mwcofluoHSU msfimuOSuuo umum mammmo mmkuwz wfiaflmx mammwmw mmwmmmu masochnuuocsomH mmvwmmwhq mnmuaflemm mmwwumahufi 000000009 mmwwumoocmumH mmnwsmuzm mmvwfifimm mmkuosm 00000000000000 00000000000002 0~\% 00\0 0m0\m 0 mmumo wc0HmEmm mN\m w\m 000500000 NN\% *o\% ¥m0\©# mmumn mafiamamm w\m an cemcfinousm come vmaaflucooIIqomH .mucsoo amfiuumm 46 I I I 000000 OH wq I I I mmvHCHUmusucmH I mmwficoEUmGSUH w 000000800m I mmvflmwcmo w mmuflowfloflmno I 0000coomum I .am 0:000H00n I mcmwummEH mDanm 000000 mumumocmE%m I mHHmucoum 030mOuN£mmom I 0500050050 05000500900 I mcoumwommm 00000000002 I 00000 mcmcomzu I mcoumwuwwc mHHmcHEmuw I 0000 mummwouom I omuuwu musmcousumum I mmwwfino musmaonsuNum I 0005050 mwuwcm I mmnmm womwomEm I 00050 mHmLQmQMHsommuQ I mumfiduomcmm 000900fifimw< mmvHHHmmeHU I mcuwdnam mflfiwNwm I mwamcum> 00>04 000080050 I mSHHmEme macumfiwnm omwflaooumo oH\m 00\00 nN\m ©\m mH\o mN\m « 0 0 0 0 « mmumn wc0HaEmm w\m 000500000 00\m *w0\w« 0N\% %o\% 0m0\o¥ mmumn wawfiaEmm 0~\m0 w\m come cowawfiuusm 00:50ucouIIqomH .hucaoo cmwuumm 47 .00000 wcwumsw mumEMxouam< 0.0 HOH oq mum mmuaumEEH 000000000 mmvwawu£uoHom< mumuaoammhsa 00000900050: 00000005030 mumumousmz mmkummwa 0000000H< mumumowflamq mmvfiumEEmuwosoHuH 00\0 0N\0 00\0 000\m 0 mmuma wcflamEmm mN\m m\m 000500000 00\m 000\m« ~N\% *o\% ¥ma\o% NN\M¥ m\m¥ mmumn wcwamamm GOmGHSUUDm 50x08 005500500II¢omH .hucsou cmwuumm 48 Nmm wq cm 00 0N 00 wq 0mm 00 wq om 0N I 00 l l oqm 0000000000000: 00000002 0000000000 000000000£0000 0000005000000 0000000000009 000000000 000000000£o 000080000050 000008000000 0000000080m 0000000 00000050008m 0000800000 .00 0500000 0000000000000 000000w00 0:00:0N3000m 0000000000 0000m 0000000000000 .00 00000u0w000 .00 g .00 00.000000. 0000008000050 0000000000 00800000008008 0080000000000m 0008w000000z 000000< 00\0 mH\w wN\n w\m ©0\© mN\m N\m 0 0 0 «0 0 0 00000 w000080m 0080003 00\m m0\w wm\m*%m\00000\o%%mw\m 00009 w000080m 008000m 00x08 0000 .000500 0005m 00> 49 N00 an n 00 00 00 00000800w 000000000 0000000000 000088000 00Nmz 0000000 0000000002 000000000 00000000 00000000000< 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