The distribution pattern of citrus tristeza virus (CTV) T-36 isolate in leaves of infected mexican lime [Citrus aurantifolia (Christm.) Swingle] plants was visualized using a whole-leaf-blot immunoassay (WLBIA) procedure in combination with a computer scanning imaging technique and CTV-specific monoclonal antibody 17G11 (CTV MAb 17G11). The distribution pattern of CTV T-36 in leaves varied with the age of the leaves and shoots of infected plants. In the young leaves, especially the about 5-day-old leaves and the completed expanded leaves, CTV T-36 was easily detected in most of the leaf veins, the main veins and the large and small primary veins. In the old leaves, CTV T-36 only was detected in the main veins, sometimes in a few of the large primary veins with weak signals, and seldom in the small primary veins. The distribution density and immunoassay reaction signals of CTV T-36 reacted to CTV MAb 17G11 in leaves from new shoots were much higher than that in leaves from old shoots. ELISA test results using leaves with different ages from different shoots of the same mexican lime plants infected with CTV T-36 supported the visualized-test results obtained by the WLBIA in combination with computer scanning imaging technique. This is the first reported visual analysis of the distribution pattern of CTV in leaves of infected citrus plants. The results indicate that the WLBIA in combination with computer scanning imaging technique is a useful tool for studying the distribution of plant viruses in leaves of virus-infected plants.
Youjian Lin and Charles A. Powell
Charles A. Powell and Youjian Lin
One hundred single brown citrus aphid (BCA) (Toxoptera citricida Kirkaldy) transmission attempts were made from each of 16 different citrus trees [8 grapefruit (Citrus paradisi Macf.) and 8 sweet orange (C. sinensis (L.) Osbeck)] previously inoculated with decline-inducing (T36-CTV), non-decline-inducing (T30-CTV), a mixture of the two Citrus tristeza virus isolate types, or no CTV. Successful CTV transmission occurred in 1.5% of attempts from grapefruit trees that had been bark-chip-inoculated with T36-CTV, 3% of attempts from orange trees inoculated with T36-CTV, 3% of attempts from grapefruit trees inoculated with both T36- and T30-CTV, 4% of attempts from orange trees inoculated with both T36- and T30-CTV, 1.5% of attempts from grapefruit trees inoculated with T30-CTV, and 3.5% of attempts from orange trees inoculated with T30-CTV. Single BCA were able to recover T30-like-CTV from trees believed to be inoculated only with T36-CTV, and T36-like-CTV from trees believed to be inoculated only with T30-CTV, suggesting that these inoculum sources were also mixtures of T36-CTV and T30-CTV. The T36-CTV was not immunologically detectable in some of the trees from which it was transmitted indicating that single BrCA can recover T36-CTV from a T36-CTV/T30-CTV mixture in which the T36-CTV is an undetectable, minority component.
Charles A. Powell and Robert R. Pelosi
Sixty-eight percent of the `Pineapple', 52% of the `Navel', 46% of the `Valencia', 38% of the `Hamlin', and 0% of the `Ambersweet' orange [Citrus sinensis (L.) Osh.] trees in five Florida citrus nurseries were infected with severe strains of citrus tristeza virus (CTV), as demonstrated by reaction with a monoclinal antibody specific for severe strains of the virus. Severe strains of CTV infected 4%, 46%, 76%, 30%, and 48% of the trees at each of the five nurseries, respectively, indicating a considerable difference in severe strain prevalence among the nurseries. Thirty-five percent of the trees in the scion blocks (budwood source) of the nurseries also contained severe strains of CTV.
Charles A. Powell and Peter J. Stoffella
Mature-green and mature-red tomato (Lycopersicon esculentum Mill.) fruit were harvested from spring- and fall-grown plants infested with sweet potato whitefly (SPWF; Bemisia tabaci Gennadins). The mature-green fruit were either ripened at 20 to 22C or cold-stored at 10 to 13C for 3 weeks and then were allowed to ripen at 20 to 22C. There was no significant difference in the appearance of either external or internal tomato irregular ripening (TIR) symptoms between the two storage–ripening regimes or in the appearance of internal TIR symptoms among the two storage regimes and vine-ripened tomatoes. Thus, removing the tomatoes from the SPWF during ripening does not reduce TIR symptoms. About half of the mature-green tomatoes, ripened with or without cold storage (10 to 13C), developed no external TIR symptoms, but about half of these tomatoes had internal TIR symptoms. About one-third of the tomatoes developed external symptoms during ripening, but these symptoms disappeared after ripening was complete. A high percentage (71%) of these tomatoes with external symptoms also had internal symptoms. The remaining tomatoes developed external TIR that did not disappear, and almost all of these tomatoes had internal symptoms. These data suggest that culling tomatoes that develop external TIR during ripening will reduce but not eliminate tomatoes with internal TIR from the fresh-fruit market.
Charles A. Powell and Peter J. Stoffella
Charles A. Powell, Robert R. Pelosi, and Phyllis A. Rundell
None of 4190 sweet orange [Citrus sinensis (L.) Osb.] nursery trees of `Hamlin', `Midsweet', `Navel', and `Valencia' sampled from five Florida citrus nurseries were infected with a decline-inducing isolate of citrus tristeza virus (CTV) as judged by enzyme-linked immunosorbent assay (ELISA) using isolate-specific monoclonal antibodies. Two of the nurseries had a relatively high level of infection (37% to 100% of composite samples containing tissue from 10 trees) with nondecline-inducing (mild) isolates of CTV, depending on the cultivar. Three of the nurseries had a lower incidence of mild CTV (0% to 22% of 10 tree composite samples). No nursery was CTV-free. ELISA of individual trees used as budwood sources by the nurseries revealed that one tree out of 260 tested contained decline-inducing CTV, and 83 contained mild CTV. These results suggest that the budwood certification program adopted in 1997 has virtually eliminated decline-inducing CTV from commercial budwood supplies.
Charles A. Powell, Phyllis A. Rundell, and Robert R. Pelosi
Bark chips from six container-grown citrus trees, infected with nondecline-inducing citrus tristeza virus (CTV) isolates and maintained in a vector-free greenhouse for 10 years, 15 commercial grapefruit (Citrus paradisi Macf.) trees, and 16 commercial sweet orange [C. sinensis (L.) Osbeck] trees were used to inoculate three indicator plants each of `Madam Vinous' sweet orange [C. sinensis (L.) Osbeck], sour orange (C. aurantium L.), `Duncan' grapefruit (C. paradisi Macf.), `Mexican' lime [C. aurantifolia (Christm.)], Swingle citrumelo [C. paradisi Macf. × Poncirus trifoliota (L.) Raf.], and sour orange grafted with `Hamlin' sweet orange [C. sinensis (L.) Osbeck]. All plants providing bark chips had repeatedly tested positive by enzyme-linked immunosorbent assay (ELISA) for CTV [reacted with monoclonal antibody (MAb) 17G11], but tested negative for Florida decline-inducing isolates of CTV (did not react with MAb MCA13). After 6 months in vector-free greenhouses, all in oculated trees (except Swingle citrumelo, which is considered CTV resistant) were positive for CTV by 17G11 ELISA. In addition, some indicator plants inoculated from nine (two container, two commercial grapefruit, and five commercial orange trees) of the 37 bark chip source trees also were positive for decline-inducing CTV by MCA13 ELISA. Some of these positive indicators also showed vein-clearing symptoms characteristic of infection with a severe isolate of CTV. No control, noninoculated indicators in the same greenhouse, became infected with either decline-inducing or nondecline-inducing CTV. These results indicate that decline-inducing isolates of CTV can be present as a minor component of a mixture at levels undetectable by ELISA, and that these decline-inducing isolates can become detectable by ELISA and sometimes by symptoms when inoculated into indicator plants.
Zhipeng Huang, Phyllis A. Rundell, Xiong Guan, and Charles A. Powell
Four field sources of citrus tristeza virus (CTV) (Y3, Y6, Y7 and Y23) collected from grapefruit trees at groves in Fort Pierce, Florida, and isolate T36 were used to evaluate the transmission and separation of different virus genotypes by single brown citrus aphids (BrCA). Analysis of the field sources of CTV by inoculation to indicator plants, ELISA and RT-PCR showed that Y6 was a decline-inducing isolate and Y23 a nondecline-inducing isolate. Assays of genotype by RT-PCR indicated that Y6 contained the T36 genotype while Y23 contained the T30 genotype. Both Y3 and Y7 were a mixture of decline-inducing and nondecline-inducing CTV isolates and were a mixture of T36 and T30 genotypes. When Y6 and Y23 were the acquisition host for single BrCA, only the T36 or T30 genotypes, respectively, were detected by RT-PCR in `Mexican' Lime receptor plants. Only the T36 genotype was transmitted to receptor plants from infected Y3 and Y7 plants although these acquisition plants contained more than one genotype. No T3 or VT genotypes were detected in any acquisition or receptor plants. CTV genotype mixtures in the various field sources were separated by single BrCA transmission and that the T36 genotype in T36/T30 mixtures was more easily transmitted than the T30 genotype when the acquisition plant was `Duncan' grapefruit and the receptor plant was `Mexican' lime.
Charles A. Powell, Mark A. Ritenour, and Robert C. Bullock
The trunk diameter of ‘Valencia’ sweet orange trees tested with seven insect control strategies was measured annually for the first 5 years after planting. Yield data (marketable fruit per tree) were collected after the fourth and fifth years. The insect control treatments were Admire (imidacloprid) applied at 12, 6, 3, or 2-month intervals; Temik (aldicarb) applied annually; Meta-Systox-R (oxydemeton-methyl) applied annually; or no insect control. Trunk diameter was significantly increased by Temik treatment at 1 and 2 years after planting. Six annual applications of Admire (at 2-month intervals) significantly increased trunk diameter 2 years after planting. None of the other treatments affected trunk diameters compared with the control. There were no trunk diameter differences among treatments at 3, 4, or 5 years after planting. Both Temik applied annually and Admire applied every other month or every 3 months significantly increased yield.
Harry S. Paris, Peter J. Stoffella, and Charles A. Powell
Summer squash (Cucurbita pepo L.) plants were grown in pots with high (290% capacity) or low (45% to 70% of capacity) soil moisture. The plants were exposed or not exposed to sweetpotato whiteflies (Bemisia tabaci Genn.). Only the plants exposed to whiteflies developed leaf silvering. Silvering was more severe in plants subjected to low soil moisture.