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Research ArticleArticles

Suppression Of Bacterial Leaf Scorch Symptoms In American Elm Through Oxytetracycline Microinjection

S.J Kostka, Tattar T.A. and Sherald J.L
Arboriculture & Urban Forestry (AUF) February 1985, 11 (2) 54-58; DOI: https://doi.org/10.48044/joa.1985.11.2.54
S.J Kostka
Shade Tree Laboratories, Department of Plant Pathology, University of Massachusetts, Amherst, Massachusetts 01003 and Ecological Services Laboratory, National Park Service, Washington, DC 20242
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Tattar T.A.
Shade Tree Laboratories, Department of Plant Pathology, University of Massachusetts, Amherst, Massachusetts 01003 and Ecological Services Laboratory, National Park Service, Washington, DC 20242
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Sherald J.L
Shade Tree Laboratories, Department of Plant Pathology, University of Massachusetts, Amherst, Massachusetts 01003 and Ecological Services Laboratory, National Park Service, Washington, DC 20242
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Abstract

American elms with bacterial-associated leaf scorch (ELS) were injected with oxytetracycline (OTC) to determine if the antibiotic would cause a delay in symptom onset or a remission of symptoms. Degree of symptom expression and presence of fastidious, xylem-inhabiting bacteria were determined in August 1982. Eleven symptomatic trees were injected in August 1982 with OTC (40 mg a.i./cm dbh) delivered via Mauget capsules. OTC was injected at 40 mg a.i./cm dbh into 14 trees and at 80 mg a.i./cm dbh into 8 trees using a pipette injection technique. Ten untreated, symptomatic trees served as controls. In June 1983, all OTC treated trees were injected using prefilled Mauget capsules at dosage levels of 50 mg a.i./cm dbh or 80 mg a.i./cm dbh. By July, symptoms appeared in 7 of 10 untreated trees but only in 2 of the remaining 29 OTC-treated trees. By late August, symptoms were absent or reduced in 22 of 23 trees treated at the low OTC levels, and 3 of 6 trees treated at the high OTC level. Symptom remission was most apparent in trees which had less than 20% of the canopy leaf area affected with ELS prior to treatment. Delay of symptom onset and symptom remission support the role of the fastidious, xylem-inhabiting bacterium as the causal agent of ELS.

Symptoms of elm leaf scorch (ELS) develop by mid-July in the Washington, DC, area (Wester and Jyllka, 1959; Hearon et al., 1980) and are characterized by an undulating, marginal necrosis bordered by a chlorotic halo. During the summer months symptoms progress acropetally, becoming more pronounced. Necrosis extends inward towards the midrib and leaves curl. Early leaf abscission commonly leaves a tuft of unaffected leaves at the branch apex. Affected trees may exhibit branch dieback (Hearon et al., 1980) and have been reported more susceptible to Ceratocystis ulmi, the Dutch elm disease fungus, than are non-scorching trees (Wester and Jyllka, 1963). Fastidious, xylem-limited bacteria have been associated with elm and oak leaf scorch (Hearon et al., 1980) and have been shown to be the cause of sycamore leaf scorch (Sherald et al., 1983) and mulberry leaf scorch (Kostka et al., 1983). Although the elm leaf scorch-associated bacterium has been cultured on artificial media, pathogenicity has not been demonstrated (Kostka et al., 1981).

Tetracycline antibiotics block protein synthesis in prokaryotes and have been used to control diseases caused by mycoplasma-like organisms (McCoy, 1982). These materials, applied as a soil drench, have been used to demonstrate the involvement of fastidious, xylem-inhabiting bacteria in Pierce ‘s disease of grape (Hopkins and Mortensen, 1971; Hopkins and Mollenhauer, 1973) and citrus young tree decline (Tucker et al., 1974). Nyland (1979) demonstrated that oxytetracycline (OTC) injected into the xylem of almonds affected by leaf scorch could serve as a therapeutic control.

The objective of this study was to determine if injections of OTC into the xylem of leaf scorch-affected American elms would cause a remission of symptoms.

Materials and Methods

Naturally infected American elms (Ulmus americana), 2 to 6 m in height, were selected for treatment. All trees were growing in naturalized sites in Washington, DC. Symptom expression was initially evaluated in August 1982. Isolations were made from all selected trees by aseptically incubating excised wood chips in a modified PW broth (Sherald et al., 1983) or by vacuum extracting bacteria from stem segments and confirming their presence using phase contrast microscopy (French et al., 1977; Hearon et al., 1980; Sherald et al., 1983).

Two microinjection techniques were selected for introduction of OTC (Terramycin Tree Injection Formula, Pfizer Chemical Company): 1) Mauget capsules (McIntyre et al, 1978; McCoy, 1982) and 2) the pipette injection technique (Lacy and McIntyre, 1978) developed for the introduction of antibiotics into pears with pear decline. OTC levels were established from the rates developed for control of elm phloem necrosis (Filer, 1976). Trees were treated in August 1982 utilizing both techniques, while in June 1983 treatments were made with Mauget capsules.

Eleven symptomatic trees (approx. 1 2 cm dbh) were injected in August 1982 with OTC, 40 mg a.i./cm dbh (low level), delivered via Mauget capsules (provided by J.J. Mauget Co.). OTC (Terramycin Tree Injection Formula) was injected at 40 mg a.i./cm dbh into 14 symptomatic trees and at 80 mg a.i./cm dbh (high level) into 8 trees using the pipette injection technique. Ten untreated, symptomatic trees served as controls. In June 1983, all OTC treated trees were injected using prefilled Mauget capsules at dosage levels of approximately 50 mg a.i./cm dbh (low level) or 80 mg a.i./cm dbh (high level).

The success or failure of treatments was based on the following observations: 1) presence or absence of symptoms in August 1983, 2) date on which first symptoms appeared, and 3) degree of symptom remission.

Results

Initial symptom development was observed in June of 1982. By August, symptoms were well developed. Symptoms in selected trees ranged from 10 to 90 percent of the total leaf area of the canopy. The presence of fastidious, xyleminhabiting bacteria in symptomatic trees was confirmed by isolation and/or buffer extraction of stem segments and microscopic observation of bacteria in extracts.

Uptake times for antibiotic solutions were approximately 24 hours for the August 1982 treatments and less than 4 hours for the June 1983 treatments. The pipette injection technique was eliminated in 1983 because of leakage around the pipette and the excessive depth required for the injection hole to securely hold the pipette.

By mid-July 1983, 7 of 10 control trees were exhibiting leaf scorch symptoms (Table 1). Symptoms were extensive in two of the trees and limited in the remaining five. Symptoms were observed in 1 of 25 trees injected at the low OTC levels (40/50 mg a.i./cm dbh) using Mauget capsules. Of the six trees injected at the high OTC level (80 mg a.i./cm dbh) symptoms had only developed in one tree. Two trees were accidently removed for right-of-way clearance. Although symptoms developed in 2 of the treated trees, the majority were symptomless in July.

Pre- and post-treatment comparison of percent elm leaf scorch-affected canopy area in American elms injected with oxytetracycline in August 1982 and June 1983.

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In August, 1983 all trees were rated for presence and severity of symptoms (Table 1). Percent total leaf area affected in untreated, control trees was equivalent to or increased above 1982 levels. Fourteen of the 23 scorch-affected elms treated at the low OTC levels showed a complete remission of symptoms. Of the remaining 9 trees, a reduction of symptoms was observed in 8, while increased symptoms were observed in 1. Two trees were removed because of Dutch elm disease. Results from trees treated at the high OTC level were inconsistent. Of the 6 trees remaining, 3 trees had substantially reduced symptoms and 3 were unchanged from 1982. A complete or near complete remission of symptoms was observed in treated trees which had 20 percent or less of the total leaf area exhibiting leaf scorch in 1982 (Table 2).

Percent leaf scorch symptoms in oxytetracycline-treated (low level) American elms before (1982) and after (1983) treatments. Trees are grouped in pre-treatment severity categories.

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Discussion

When the rules of proof (Koch ‘s postulates) cannot be fulfilled with a fastidious prokaryote (i.e., the fastidious, xylem-inhabiting bacterium), an important step in confirming the involvement of the organism in disease is demonstration of symptom remission through antibiotic treatment (Hopkins and Mortensen, 1971; Nyland, 1979; McCoy, 1982). Oxytetracycline injected into the xylem of American elms affected with bacterial leaf scorch delayed symptom onset and suppressed symptom development. Remission was most effective when 20% or less of the canopy leaf area was affected prior to treatment. Results in trees with more than 20% leaf scorch were inconsistent at either OTC dosages. The development of symptoms in treated trees was not unexpected based upon previous OTC injection studies of almond trees affected with almond leaf scorch (Nyland, 1979). Inconsistent symptom remission may be due to inadequate distribution or final concentration of the antibiotic in the xylem to inhibit bacterial growth.

The observed delay in the onset of symptoms and the complete or partial symptom remission that occurred in treated trees supports the role of an OTC-sensitive organism (i.e., the fastidious, xylem-inhabiting bacterium) as the causal agent of elm leaf scorch. Although OTC induced symptom remission, its applicability as a therapeutic control requires further study to determine long-term control and potential phytotoxicity of the antibiotic in treated trees.

Footnotes

  • ↵1. We would like to thank the J.J. Mauget Co. and the Pfizer Chemical Co. for their support in this research. Paper No. 2668, Mass. Agric. Exp. Sta. This research supported in part from Exp. Sta. Project No. 501. The current address of the senior author is Crop Genetics International N.V., 7170 Standard Drive, Dorsey, MD 21076. Mention of trademark or proprietary product does not constitute a guarantee or warranty of the product by the USDI and does not imply its approval to the exclusion of other products that may be suitable.

  • © 1985, International Society of Arboriculture. All rights reserved.

Literature Cited

  1. 1.↵
    1. Filer, T.H., Jr.
    1976. Antibiotic injections control elm phloem necrosis in the urban ecosystem. Proc. Int. Union For. Res. Org. pp. 327–333.
  2. 2.↵
    1. French, W.J.,
    2. Christie R.G., and
    3. Stassi. D.L
    . 1977. Recovery of rickettsia-like bacteria by vacuum infiltration of peach tissues affected with phony peach disease. Phytopathology 67: 945–948.
    OpenUrl
  3. 3.↵
    1. Hearon, S.S.,
    2. Sherald J.L., and
    3. Kostka. S.J.
    1980. Association of xylem-limited bacteria with elm, sycamore, and oak leaf scorch. Can. J. Bot. 58: 1986–1993.
    OpenUrlCrossRef
  4. 4.↵
    1. Hopkins, D.L., and
    2. Mollenhauer. H.H.
    1973. Rickettsia- like bacterium associated with Pierce ‘s disease of grapes. Science 179: 298–300.
    OpenUrlAbstract/FREE Full Text
  5. 5.↵
    1. Hopkins, D.L., and
    2. Mortensen. J.A.
    1971. Suppression of Pierce ‘s disease symptoms by tetracycline antibiotics. Plant Dis. Rep. 55: 610–612.
    OpenUrl
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    1. Kostka, S.J.,
    2. Sherald J.L.,
    3. Hearon S.S., and
    4. Rissler. J.F.
    1981. Cultivation of the elm leaf scorch-associated bacterium (ESB). (Abstr.) Phytopathology 71: 768.
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  7. 7.↵
    1. Kostka, S.J.,
    2. Tattar T.A., and
    3. Sherald. J.L.
    1983. Mulberry leaf scorch: Pathogenicity of the associated bacterium. (Abstr.) Phytopathology 73: 1344.
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  8. 8.↵
    1. McCoy, R.E.
    1982. Use of tetracycline antibiotics to control yellows diseases. Plant Disease 66: 539–542.
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  9. 9.↵
    1. McIntyre, J.L.,
    2. Dodds J.A.,
    3. Walton G.S., and
    4. Lacy. G.H.
    1978. Declining pear trees in Connecticut: Symptoms, distribution, symptom remission by oxytetracycline, and associated mycoplasma-like organisms. Plant Dis. Rep. 62: 503–507.
    OpenUrl
  10. 10.↵
    1. McCoy, R.E.
    1. Nyland, G.
    1979. Chemotherapy of disease of deciduous trees associated with mycoplasma and rickettsialike organisms. In: McCoy, R.E. (ed.), Proc. USROC, Taiwan pp. 139–142.
  11. 11.↵
    1. Sherald, J.L.,
    2. Hearon S.S.,
    3. Kostka S.J., and
    4. Morgan. D.L.
    1983. Sycamore leaf scorch: Culture and pathogenicity of fastidious, xylem-limited bacteria from scorch-affected trees. Plant Disease 67: 849–852.
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  12. 12.↵
    1. Tucker, D.P.H.,
    2. Bistline F.W., and
    3. Gonsalves. D.
    1974. Observations on young tree decline-affected citrus trees treated with tetracycline. Plant Dis. Rep. 58: 895–896.
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  13. 13.↵
    1. Wester, H.V., and
    2. Jyllka. E.W.
    1959. Elm scorch, graft transmissible virus of American elm. Plant Dis. Rep. 43: 519.
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  14. 14.↵
    1. Wester, H.V., and
    2. Jyllka. E.W.
    1963. High incidence of Dutch elm disease In American elms weakened by elm scorch associated with breeding attacks by Scolytus multistriatus. Plant Dis. Rep. 47: 545–547.
    OpenUrl
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Suppression Of Bacterial Leaf Scorch Symptoms In American Elm Through Oxytetracycline Microinjection
S.J Kostka, Tattar T.A., Sherald J.L
Arboriculture & Urban Forestry (AUF) Feb 1985, 11 (2) 54-58; DOI: 10.48044/joa.1985.11.2.54

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Suppression Of Bacterial Leaf Scorch Symptoms In American Elm Through Oxytetracycline Microinjection
S.J Kostka, Tattar T.A., Sherald J.L
Arboriculture & Urban Forestry (AUF) Feb 1985, 11 (2) 54-58; DOI: 10.48044/joa.1985.11.2.54
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