Control of Paratrichodorus allius and Corky Ringspot Disease in Potato with Shank-injected Metam Sodium

Corky ringspot disease (CRS) of potato produces necrotic areas in tubers that are considered quality defects that can lead to crop rejection. CRS is caused by tobacco rattle virus that is vectored by stubby-root nematodes (Paratrichodorus spp., Trichodorus spp.) at very low population densities, making disease management difficult and expensive. Fumigation with metam sodium (MS) is a common practice to control soil-borne fungi and increase potato yield. MS is generally applied in water via chemigation (water-run, WR) but is ineffective at controlling CRS when WR-applied, even at high rates. Therefore, WR MS is often used in combination with 1,3-dichloropropene (1,3-D), aldicarb or oxamyl to attain adequate CRS control. Between 1996 and 2000, fields with a history of CRS were treated with WR MS, shank-injected MS, and/or 1,3-D, and tubers were evaluated for symptoms of CRS. Shank injection of MS (SH MS) at depths of 41 cm, 15 and 30 cm, or 15, 30 and 45 cm controlled CRS over 3 years of testing. All rates of 280 liters/ha or greater were effective. Shank injection of metam potassium (MP) at rates of 448 liters/ha was also effective. 1,3-D controlled CRS alone or in combination with WR or SH MS. Proper shank application of MS or MP may adequately control CRS without the additional cost of other nematicides at low (<10 P. allius/250 g soil) to moderate (10 to 30 P. allius/250 g soil) populations of the nematode vector. Although SH MS was superior to WR MS, additional research is necessary to determine if this practice would be sufficient at higher CRS disease pressure or if addition of other nematicides would be necessary.     

Effects of In-furrow and Water-run Oxamyl on Paratrichodorus allius and Corky Ringspot Disease of Potato in the Klamath Basin

Corky ringspot disease (CRS) of potato (Solanum tuberosum) is caused by the tobacco rattle virus (TRV), which is vectored by stubby-root nematodes, Paratrichodorus spp. and Trichodorus spp., and is a significant threat to potato quality and production in many areas of the western United States. Between 2002 and 2005, fields with a history of CRS were planted to potato and treated with various combinations of in-furrow (IF) and chemigated (water run, WR) oxamyl [Methyl N'N'-dimethyl-N-[(methyl carbamoyl)oxy]-1-thiooxamimidate] applications. Soil samples were collected to determine how Paratrichodorus allius populations responded to the various treatment regimes (2002-2004); potato tubers were evaluated for symptoms of CRS in 2004-2005. Applications of oxamyl to potato (1.1 kg a.i./ha) did not cause significant mortality of P. allius but did prevent the populations from increasing. Oxamyl applications that began at 55 days after planting (DAP) or later did not control CRS and were not different from the untreated control. However, application schedules that began early-season, either IF at planting, early WR (33 - 41 DAP), or both, significantly reduced CRS expression in cv. Yukon Gold. Therefore, oxamyl applications must be made early in the growing season to be effective in controlling CRS. Effects of oxamyl on CRS may be due to nematostatic action that suppresses feeding activity during early field season when most virus transmission probably occurs.     

Reproduction of Belonolaimus longicaudatus,Meloidogyne javanica,Paratrichodorus minor,and Pratylenchus brachyurus on Pearl Millet (Pennisetum glaucum)

Pearl millet (Pennisetum glaucum) has potential as a grain crop for dryland crop production in the southeastern United States. Whether or not pearl millet will be compatible in rotation with cotton (Gossypium hirsutum), corn (Zea mays), and peanut (Arachis hypogaea) will depend, in part, on its host status for important plant-parasitic nematodes of these crops. The pearl millet hybrid ''TifGrain 102'' is resistant to both Meloidogyne incognita race 3 and M. arenaria race 1; however, its host status for other plant-parasitic nematodes was unknown. In this study, the reproduction of Belonolaimus longicaudatus, Paratrichodorus minor, Pratylenchus brachyurus, and Meloidogyne javanica race 3 on pearl millet (''HGM-100'' and TifGrain 102) was compared relative to cotton, corn, and peanut. Separate greenhouse experiments were conducted for each nematode species. Reproduction of B. longicaudatus was lower on peanut and the two millet hybrids than on cotton and corn. Reproduction of P. minor was lower on peanut and TifGrain 102 than on cotton, corn, and HGM-100. Reproduction of P. brachyurus was lower on both millet hybrids than on cotton, corn, and peanut. Reproduction of M. javanica race 3 was greater on peanut than on the two millet hybrids and corn. Cotton was a nonhost. TifGrain 102 was more resistant than HGM-100 to reproduction of B. longicaudatus, P. minor, and M. javanica. Our results demonstrated that TifGrain 102 was a poor host for B. longicaudatus and P. brachyurus (Rf < 1) and, relative to other crops tested, was less likely to increase densities of P. minor and M. javanica.     

Niche Distribution of Paratrichodorus minor and Belonolaimus longicaudatus Following Fumigation on Potato and Cabbage

An experiment was conducted to determine population changes and niche variation in the soil at two depths (0 to 20 cm and 20 to 40 cm) of Paratrichodorus minor and Belonolaimus longicaudatus populations following fumigation. Eight plots each of potato (Solanum tuberosum) and cabbage (Brassica oleracea var. capitata), fumigated with 1, 3-dichloropropene or nonfumigated, were established. Eight plots of sorghum-sudangrass hybrid (Sorghum bicolor × S. arundinaceum var. sudanense) were also used to monitor depth distribution (0 to 20 cm and 20 to 40 cm) of B. longicaudatus and P. minor following each cabbage/potato season. Soil samples were taken 0 to 20 cm and 20 to 40 cm deep during the potato/cabbage, and sorghum-sudangrass growing seasons. During the 1993-94 and 1994-95 potato/cabbage seasons, P. minor was found at highest numbers at 20 to 40 cm, whereas numbers of B. longicaudatus were highest at 0 to 20 cm. During the 1994 and 1995 sorghum-sudangrass growing seasons, B. longicaudatus numbers were highest at 0 to 20 cm. Paratrichodorus minor numbers were highest at 0 to 20 cm and at 20 to 40 cm deep in the 1994 and 1995 sorghum-sudangrass growing seasons, respectively. Reduction by soil fumigation of B. longicaudatus at 0 to 20 cm deep did not affect depth distribution or cause P. minor populations to increase in potato or cabbage plots. Paratrichodorus minor numbers increased at 20 to 40 cm deep in the 1994-95 cabbage season after soil fumigation.     

基于烟草脆裂病毒构建同时表达2种外源蛋白的载体及其应用

目的 构建在整个寄主植物中能同时表达两个非融合蛋白的病毒载体。方法 以烟草脆裂病毒（tobacco rattle virus，TRV）基因组RNA2的农杆菌侵染性克隆pYL156为材料，缺失TRV RNA2的2b基因5"端279 bp并将其起始密码子ATG改变为AGG、同时引入豌豆早枯病毒（pea early-browning virus，PEBV）外壳蛋白（coat protein，cp）基因启动子，获得pTRV2e²载体；在pTRV2e²载体的2b和PEBV的cp启动子下游插入不同的外源基因，测定病毒TRVe²表达外源蛋白的能力、携带外源基因后重组病毒的稳定性以及分析蛋白质的生物学功能。结果 病毒TRVe²能快速同时表达2个非融合的外源蛋白，且至少能表达70 ku外源蛋白，且该病毒携带~2.0 kb外源基因能稳定存活于寄主植物中；病毒TRVe²可用于分析蛋白质的生物学功能以及2个蛋白质间的相互作用。结论 本文构建的重组病毒TRVe²为快速有效地表达2个外源蛋白以及分析2个蛋白质间的相互作用提供技术工具。     

Identification of Sources of Resistance to Four Species of Root-knot Nematodes in Tobacco

Resistance to the southern root-knot nematode, Meloidogyne incognita races 1 and 3, has been identified, incorporated, and deployed into commercial cultivars of tobacco, Nicotiana tabacum. Cultivars with resistance to other economically important root-knot nematode species attacking tobacco, M. arenaria, M. hapla, M. javanica, and other host-specific races of M. incognita, are not available in the United States. Twenty-eight tobacco genotypes of diverse origin and two standard cultivars, NC 2326 (susceptible) and Speight G 28 (resistant to M. incognita races 1 and 3), were screened for resistance to eight root-knot nematode populations of North Carolina origin. Based on root gall indices at 8 to 12 weeks after inoculation, all genotypes except NC 2326 and Okinawa were resistant to M. arenaria race 1, and races 1 and 3 of M. incognita. Except for slight root galling, genotypes resistant to M. arenaria race 1 responded similarly to races 1 and 3 of M. incognita. All genotypes except NC 2326, Okinawa, and Speight G 28 showed resistance to M. javanica. Okinawa, while supporting lower reproduction of M. javanica than NC 2326, was rated as moderately susceptible. Tobacco breeding lines 81-R-617A, 81-RL- 2K, SA 1213, SA 1214, SA 1223, and SA 1224 were resistant to M. arenaria race 2, and thus may be used as sources of resistance to this pathogen. No resistance to M. hapla and only moderate resistance to races 2 and 4 of M. incognita were found in any of the tobacco genotypes. Under natural field infestations of M. arenaria race 2, nematode development on resistant tobacco breeding lines 81-RL-2K, SA 1214, and SA 1215 was similar to a susceptible cultivar with some nematicide treatments; however, quantity and quality of yield were inferior compared to K 326 plus nematicides.     

Genetics of Burley and Flue-Cured Tobacco Resistance to Globodera tabacum tabacum

Genotypes of burley (cultivars B-21 and B-49), flue-cured (line VA-81 and cultivar PD-4), and Connecticut broadleaf (cultivar C9) tobacco (Nicotiana tabacum) resistant (R) or susceptible (S) to the tobacco cyst nematode Globodera tabacum tabacum were crossed. F1 progeny of burley and susceptible broadleaf were selfed and backcrossed to produce additional progeny for evaluation of resistance in greenhouse experiments. Plants without adult female nematodes visible (×10 magnification) on the root surface 6 weeks after inoculation were classified as resistant, whereas those plants in which one or more females were evident were classified as susceptible. Segregation ratios for progeny of resistant and susceptible plants were not different from 3:1 and 1:1 for F2 (F1 × F1) and BC1 (F1 × S) lines, respectively, indicating that resistance in burley to G. t. tabacum is conferred by a single, dominant gene. Segregation ratios for resistance in crosses between nematode-resistant burley and flue-cured tobacco (F1 and F2 progeny) and between burley-flue-cured hybrids and broadleaf BC1 (F1 × S) and BC2 (BC1 × S) progeny were consistent with the assumption that resistance to G. t. tabacum in burley and flue-cured tobacco is conferred by the same or closely linked single, dominant gene(s).     

Vertical Distribution of Plant-parasitic Nematodes in Sandy Soil under Soybean

Vertical distribution of five plant-parasitic nematodes was examined in two north Florida soybean fields in 1987 and 1988. Soil samples were collected from 0-15 cm, 15-30 cm, and 30-45 cm deep at each site. Soil at the three depths consisted of approximately 96% sand. More than 50% of Belonolaimus longicaudatus population densities occurred in the upper 15-cm soil layer at planting, but the species became more evenly distributed through the other depths as the season progressed. Criconemella sphaerocephala was evenly distributed among the three depths in one field but was low (< 20% of the total density) in the upper 15 cm at a second site. Maximum population densities of Pratylenchus brachyurus were observed at 15-30 cm on most sampling dates. Vertical distributions of Meloidogyne incognita and Paratrichodorus minor were erratic and showed seasonal variation. A diagnostic sample from the upper 0-15 cm of these soybean fields revealed only a minority of the populations of most of the phytoparasitic species present.     

Temporal Efficacy of Selected Nematicides on Meloidogyne Species on Tobacco

Aldicarb, ethoprop, and fenamiphos were evaluated for their efficacy in controlling various species of root-knot nematodes on flue-cured tobacco and for their residual activity, as determined through periodic sampling and bioassays of soil taken from field plots. Field experiments were conducted at five locations over 2 years with flue-cured tobacco. Soil in plots treated with nematicides were formed into high, wide beds before transplanting with ''Coker 371-Gold'' or ''K 326'' tobacco. Residual control of Meloidogyne spp. was greatest (P ≤ 0.05) with fenamiphos (in some cases up to 10 weeks, as measured in tomato bioassays of infested soil and root fragments). Suppression of nematode reproduction by ethoprop was short-lived, and numbers of second-stage juveniles + eggs and numbers of galls in bioassays sometimes surpassed those of untreated plots within 4 weeks after treatment. Aldicarb gave intermediate control over time as compared to the other compounds. Although nematicidal efficacy of all compounds varied with site and season, fenamiphos and aldicarb generally produced the highest yields.     

Predisposition of Broadleaf Tobacco to Fusarium Wilt by Early Infection with Globodera tabacum tabacum or Meloidogyne hapla

In greenhouse experiments, broadleaf tobacco plants were inoculated with tobacco cyst (Globodera tabacum tabacum) or root-knot (Meloidogyne hapla) nematodes 3, 2, or 1 week before or at the same time as Fusarium oxysporum. Plants infected with nematodes prior to fungal inoculation had greater Fusarium wilt incidence and severity than those simultaneously inoculated. G. t. tabacum increased wilt incidence and severity more than did M. hapla. Mechanical root wounding within 1 week of F. oxysporum inoculation increased wilt severity. In field experiments, early-season G. t. tabacum control by preplant soil application of oxamyl indirectly limited the incidence and severity of wilt. Wilt incidence was 48%, 23%, and 8% in 1989 and 64%, 60%, and 19% in 1990 for 0.0, 2.2, and 6.7 kg oxamyl/ha, respectively. Early infection of tobacco by G. t. tabacum predisposed broadleaf tobacco to wilt by F. oxysporum.     

Structure of the Cuticle of Ceramonema carinatum (Chromadorida: Ceramonenatidae)

The cuticle of Ceramonema carinatum (Chromadorida: Ceramonematidae) is described and illustrated from scanning and transmission electron microscopy. Each of ca. 200 annules is composed of a single ring with eight external flat faces (plates), which are divided by longitudinal ridges formed by pairs of parallel upstanding vanes. Vanes and plates overlap those of the adjacent annules. Longitudinal ridges extend from the cephalic capsule to the tail spike. On the cephalic capsule a simple ridge extends each of the eight ridges to a position just anterior to the amphid. Cuticular plates are formed from the electron-dense cortical layer and contain lacunae filled with fine fibrils. The vanes are denser, with laminations on a central core. In the annular grooves between the plates there is an electron-lucent layer, which it is suggested, by comparison with other nematodes, is the basal layer. An epicuticle overlies the cortical plates, the vanes, and the interannular lucent layer. Cuficular structure is compared with that of other Ceramonematidae and related nematodes.     

Scanning Electron Microscope Studies of Steinernema anomali Kozodoi, 1984

This paper presents SEM micrographs of portions of the male, female, and infective-stage juvenile of Steinernema anomali. Included are micrographs of the cephalic and caudal region, spicules, and gubernaculum of the male, the cephalic and vulval region of the female, and the cephalic region of the infective-stage juvenile. Males have six labial and four prominent cephalic papillae and small amphids. There are 11-14 pairs and one single genital papillae; of these, 6-9 pairs are preanal and subventral, one pair preanal, lateral, one pair adanal, and three pairs postanal. Spicules have a short head, a long blade, and a reduced shaft. The distal end is enlarged and bears a dorsal aperture. Gubernaculum much shorter than spicules; cuneus of gubernaculum short and bifurcate anteriorly. Females have six labial and four cephalic papillae and small amphids. Vulva with a thickened posterior lip. Infective juveniles have a smooth head, prominent amphids, and four cephalic papillae. Labial papillae, if present, are not evident.     

Early Crop Root Destruction for Management of Tobacco Cyst Nematodes

Prompt tillage after crop harvest was investigated as a cultural control for the tobacco cyst nematode, Globodera tabacum tabacum, on stalk-cut broadleaf cigar wrapper tobacco. Stalk stumps and roots remaining after harvest were destroyed by tilling immediately or from 2 to 6 wk after harvest in field experiments over 4 yr. Cyst nematode Pf/Pi ratios ranged from 0.65 to 1.62 when plants were tilled immediately after harvest and 1.13 to 5.88 when tillage was delayed. Nematode population development was monitored by inoculating plants in pots placed in fields with J2 in eggs and sampling over time (8 to 18 wk). Three generations per year were observed, and G. t. tabacum generation time was as short as 6 wk for each generation. Destroying stalks and root systems remaining after harvesting stalk-cut broadleaf cigar wrapper tobacco removes the host to preclude development of nematodes at the end of the second and entire third generation. Early tillage resulted in consistently lower tobacco cyst nematode populations than allowing viable roots to remain in fields for an additional 8 to 18 wk. This management tactic reduces the need for nematicide application to slow nematode population increases over time and can reduce losses due to infection by G. t. tabacum.     

Enhanced Hatching of Globodera tabacum solanacearum Juveniles by Root Exudates of Flue-cured Tobacco

Stimulation of hatching of a tobacco cyst nematode (Globodera tabacum solanacearum) by root exudates from resistant NC 567 and susceptible K 326 cultivars of flue-cured tobacco, Nicotiana tabacum, was investigated. Root exudates were collected by soaking seedlings in deionized water for 2 hours at 22 °C in the dark. Fifteen mature and uniformly sized cysts were exposed at 15, 20, or 25 °C to undiluted root exudate, root exudate diluted 1:1 or 1:3 with deionized water, or deionized water alone. Hatched juveniles were counted and removed at weekly intervals during 42 and 53 days of exposure in experiments conducted in 1994 and 1995, respectively. Root exudates from both susceptible cultivar K 326 and resistant cultivar NC 567 stimulated more hatching than deionized water at 25 °C in 1994, and at all three tested temperatures in 1995. In 1994, dilution of root exudates 1:3 reduced stimulation of hatching at 25 °C compared to undiluted exudate. Hatching at 25 °C was similarly stimulated by exposure to undiluted root exudate and exudate diluted 1:1. In 1995, both dilutions reduced stimulation of hatching by root exudates at all the temperatures.     

Transmission of Nepoviruses by Xiphinema americanum-group Nematodes

The transmission of North American nepoviruses by putative species belonging to the Xiphinema americanum-group is reviewed. Xiphinema americanum sensu stricto, X. californicum, and X. rivesi each transmit cherry rasp leaf (CRLV), tobacco ringspot (TobRSV), and tomato ringspot nepovirus (TomRSV), and X. bricolensis is a vector of TomRSV. The apparent lack of specificity in the transmission of North American nepoviruses by X. americanum-group species markedly contrasts with the specific associations between European nepoviruses and their vector nematode species. Two complementary projects are described examining the taxonomic identity of putative species in the X. americanum-group, their morphological and genetic relationships, their ontogeny, and their ability to transmit viruses.     

Interaction Between Neoaplectana carpocapsae and a Granulosis Virus of the Armyworm Pseudaletia unipuncta

Neoaplectaua carpocapsae developed and reproduced in armyworm hosts infected with a granulosis virus (GV). Macerated tissues of dauer juveniles from GV-infecled hosts had sufficient GV to infect 1st and 2nd instar armyworms. Electron-microscope examination of dauer juveniles and adult female nematodes confirmed the presence of GV in the lumen of the intestine. No GV was observed in other tissues of the nematode.     

Hatch and Reproduction of Globodera tabacum tabacum in Response to Tobacco,Tomato, or Black Nightshade

The effects of broadleaf tobacco, tomato, and black nightshade on juvenile hatch and reproduction of Globodera tabacum tabacum were determined in laboratory and greenhouse experiments. Root exudates from nightshade stimulated greater egg hatch than those from either ''Rutgers'' tomato or ''86-4'' tobacco. Hatch was greater at higher proportions of root exudates for all three plant species. Root exudates from plants greater than 3 weeks old stimulated more hatch than younger plants. No regression relationships existed between plant age and nematode batch. In other experiments, hatch from eggs in cysts was higher for tomato and nightshade after 10 weeks in greenhouse pots compared to tobacco and bare soil. Numbers of second-stage juveniles in eggs in cysts produced from a previous generation on the same host were highest on nightshade and less on tomato and tobacco. Cysts of variable age recovered from field soil had increased hatch in both root exudates or water compared to recently produced cysts from plants in growth chambers. Globodera t. tabacum may be subject to both host and environmentally mediated diapause.     

Evaluation of an Antibiotic-Producing Strain of Pseudomonas fluorescens for Suppression of Plant-Parasitic Nematodes

The antibiotic 2,4-diacetylphloroglucinol (DAPG), produced by some strains of Pseudomonas spp., is involved in suppression of several fungal root pathogens as well as plant-parasitic nematodes. The primary objective of this study was to determine whether Wood1R, a D-genotype strain of DAPG-producing P. fluorescens, suppresses numbers of both sedentary and migratory plant-parasitic nematodes. An experiment was conducted in steam-heated soil and included two seed treatments (with Wood1R and a control without the bacterium) and six plant-nematode combinations which were Meloidogyne incognita on cotton, corn, and soybean; M. arenaria on peanut; Heterodera glycines on soybean; and Paratrichodorus minor on corn. Wood 1R had no effect on final numbers of M. arenaria, P. minor, or H. glycines; however, final numbers of M. incognita were lower when seeds were treated with Wood1R than left untreated, and this reduction was consistent among host plants. Population densities of Wood1R were greater on the roots of corn than on the other crops, and the bacterium was most effective in suppressing M. incognita on corn, with an average reduction of 41%. Despite high population densities of Wood1R on corn, the bacterium was not able to suppress numbers of P. minor. When comparing the suppression of M. incognita on corn in natural and steam-heated soil, egg production by the nematode was suppressed in natural compared to steamed soil, but the presence of Wood1R did not result in additional suppression of the nematodes in the natural soil. These data indicate that P. fluorescens strain Wood1R has the capacity to inhibit some populations of plant-parasitic nematodes. However, consistent suppression of nematodes in natural soils seems unlikely.     

Biotin-Avidin ELISA Detection of Grapevine Fanleaf Virus in the Vector Nematode Xiphinema index

The value of biotin-avidin (B-A) ELISA for the detection of grapevine fanleaf virus (GFLV) in Xiphinema was estimated with field populations and greenhouse subpopulations. Samples consisted of increasing numbers of adults ranging from 1 to 64 in multiples of two. Tests with virus-free X. index populations reared on grapevine and fig plants as negative controls did not reveal a noticeable effect of the host plant. ELISA absorbances of virus-free X. index samples were greater than corresponding absorbances of X. pachtaicum samples. Differences occurred between two X. index field populations from GFLV-infected grapevines in Champagne and Languedoc. In most tests, 1-, 2-, 4-, and 8-nematode samples of virus-free and virus-infected populations, respectively, could not be separated. Consequently, B-A ELISA was not a reliable method for GFLV detection in samples of less than 10 X. index adults, but comparison of the absorbances obtained with increasing numbers may allow differentiation of the viral infectious potential of several populations.