SUSCEPTIBILITY of SPECIES of CÜPRESSACEAE to CROWN GALL AS DETERMINED by ARTIFICIAL INOCULATION! by CLAYTON O
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SUSCEPTIBILITY OF SPECIES OF CÜPRESSACEAE TO CROWN GALL AS DETERMINED BY ARTIFICIAL INOCULATION! By CLAYTON O. SMITH Assistant plant pathologist^ California Agricultural Experiment Station INTï^ODUGTÏON The natural occurrence of crown gall on conifers has be^n reported, by Persons (2) ^ and by Barrus, Boyd, and Wood ^bn Junipenis sabina; by Brown and Evans (1) on Cupressus arizonica; hj Smith (5) on Libocedrus decurrens; by Metcalf,^ who observed large galls at and below the ground level on a street tree of Araucaria bidwillii; and by Rounds/ who found a gall 10 inches in diameter on Araucaria excelsa. Crown gall has been artificially produced on Sequoia gigantea and S, sempervirens {3), on Araucaria bidwillii (4), Libocedrus decurrens {5)f Cupressus spp. (7), and on Taxus baccata var. erecta (6), The results of artificial inoculations on species of Cupressaceae with Bacterium tumefaciens Smith and Townsend {Phytomonas turne- faciens (Smith and Town.) Bergey et al.) are reported in this paper- The cultures of the organism used in the inoculations were isolated from Prunus pérsica^ Libocedrus decurrens, and Salix sp. MATERIALS AND METHODS The Cupressaceae under investigation consisted of species of Cupressus, Juniperus, Libocedrus, Thuja, Thujopsis, and Chamae- cyparis. The plants used in this experiment were: (1) Small seedlings ob- tained from the Rancho Santa Ana Botanic Garden; (2) plants from the United States Department of Agriculture carrying the plant introduction numbers; (3) small trees purchased from a local nursery; (4) rooted cuttings from certain species; (5) plants grown from seed at. the University of California Citrus Experiment Station; and (6) large trees growing on the campus. The sources of the plants are shown in table 1. Seedlings and small trees were grown in 5-gallon tin containers in a lath house during the experiment and were in a vigor- ous growing condition. Inoculations were through wounds made by a steel needle or through deeper wounds made by cutting through the inociitem into the tissue with a scalpel. The wounds were left unprotected or were protected in various ways. In some tests the wounds were wrapped with paraffin paper. Desiccation in other tests was further checked by tying moist cotton over the wounds, which were then wrapped with paraffin paper fastened at the ends to the plant with nurseryman's 1 Received for publication June 5, 1939.^ Paper No. 404, University of California Citrus Experiment Station. 2 Italic numbers in parentheses refer to Literature Cited, p. 925. 3 BARRUS, M. F., BOYD, O. C, and WOOD, JESSIE I. DISEASES OF PLANTS IN THE UNITED STATES IN 193O. U. S. Bur. Plant Indus. Plant Dis. Rptr. Sup. 81: 133. 1931. [Mimeographed.] 4 Letter dated March 5, 1938, to the author from Woodbridge Metcalf, extension forester, University of California. 5 Oral report to the author July 8,1939, by M. B. Rounds, Associate in the Citrus Experiment Station. Journal of Agricultural Research, Vol. 59, No. 12 Washington, D. C. Dec. 15,1939 Key No. Calif .-111 210821—40 5 (919) 920 Journal oj AgricuHural Research Vol. 59, No. 12 tape. When inoculations were made near the base of the tree, a waterproof plant pot was placed around the inoculated trunk and filled with fine gravel which was kept moist. A few inoculations were made on the trunk at and below the ground level. Some of the smaller plants were inoculated and kept for a time in a moist chamber, and galls were frequently developed by the use of this method when results from unprotected inoculation wounds had failed. Positive results were obtained from all of these methods on susceptible hosts. TABLE 1.—Summary of artificial inoculations through wounds with Bacterium tumefaciens on species of Cupressaceae Host species Inocula- Gall diam- Sources of plant material i tions Galls 2 eter range 2 Number Number Millimeters Cupressus arizonica Greene Seedlings 93 18 5-20 Do s 2357, S 2358 35 7 5-15 Cupressus bakeri Jeps s 2127, S 2131 30 4 3-30 Cupressus benthami Carr SPI114036 15 7 5-20 Cupressus duttoni Jeps S 2157 . 15 7 3-25 Cupressus forbesii Jeps s 2315, s 2319, S 2335. 50 9 4-20 Cupressus glabra Sudw SPI 112084 20 O O Do - CE. S., Nursery.._. 155 O O Cupressus guadalupensis Wats s 2069 35 O O Cupressus goveniana Gord s 2181, S 2182 30 2 3-20 Cupressus lusitanica Mill SPI 73844 60 5 10-25 Cupressus knightiana Mast Nursery 10 3 2-3 Cupressus macrocarpa Hartw s 2177, S 2184 25 7 3-35 Do 165 3 2-20 Cupressus macnabiana Murr s 2118, s 2154 25 2 2-20 Cupressus montana Wigs S 2067 30 O Cupressus nevadensis Abrams S 2159 15 3 3-5 Cupressus pygmaea Sarg S 2133, S 2137 30 7 6-35 Cupressus sargenta Jeps - s 2156, s 2149, S 2185. 43 10 12-35 Cupressus sempervirens L CE. s 45 9 10-20 Cupressus thurifera Schlecht s 2356 28 4 5-25 Cupressus torulosa Don SPI 112085 15 4 3-10 Thuja plicata Don Nursery 46 9 10-20 Thuja occidentalis L do 100 5 5-25 Thuja orientalis L CE. S 30 17 10-30 Thujopsis dolabrata Sieb, and Zucc_ Nursery 45 2 10-15 Chamaecyparis lawsoniana Pari .-..-do 90 2 O Libocedrus decurrens Torr_ .__ _do 95 34 10-30 Juniperus ashei Buchbolz SPI 124965 6 3 (3) Juniperus californica Carr CE.S 95 O O Juniperus cedrus Webb and Berth.. SPI 57080 50 2 Juniperus hibernica Gord Nursery 35 4 Juniperus phoenicea L SPI 65020 20 6 5-15 Juniperus procera Höchst SPI 60553 30 3 5-10 SPI 27505 50 O O Juniperus virginiana L Nursery 35 15 5-20 1 By "seedlings" is meant plants grown from seed at the University of California Citrus Experiment Sta- tion. "S" denotes the accession number of plants from the Rancho Santa Ana Botanic Garden, and "SPI", the plant introduction number of plants from the U. S. Department of Agriculture. "C E. S." indicates large trees growing on the campus of the Citrus Experiment Station, and "Nursery" indicates trees pur- chased from a local nursery. 2 Results observed 6 months to 1 year after inoculation. 3 Knobs. Control punctures were made on the various hosts inoculated, but because of the limited plant material available, these controls were not numerous. GALLS ON CUPRESSUS The results of inoculations on species of Cupressus are listed in table 1. Galls (fig. 1) developed on all species tested, with the excep- tion of guadalupensis. The results on glabra and montana^ however, were somewhat in- conclusive. On small trees of glabra ((7. arizonica var. bonita) the inoculations caused the development of small knoblike projections Dec. 15,1939 Susceptibility of Cupressaceae to Crown Gall 921 FicuKE 1.—Artificial galls prodiiocd on species of Cwpressua by inoculation with BacUrium tumefaciens isolated from IHalix sp. for all galls except G and H, for which cultures were isolated front Prunus pérsica. The time, in months, from the date of inoculation to the date of the photograph ¡s indicated in parentheses following the description of the species: A, arizonica, a seedling (24 months); B, arizonica, S 2357 (14 months); C, macnabiana, S 2154 (18 months); D, sargenta, S 2149 (19 months); E, macrocarpa, a seedling (26 months); F, henlhami, SPl 114036 (12 months); G, lusitanica, SPI 73844 (8 5 months) ; //, same galls (24 months) ; I, Ihurifera, S 2350 (8 months) ; /, torulosa, SPI 112085 (5.5 months); K, goveniana, S 2181 (26 months); L, forbesii, S 2335 (26 months); M, pygmaea, S 2137 (14 months); A^ bakeri, S 2131 (14 months); O, duUoni, S 2157 (8.5 months). 922 Jourval of Agricultural Reftearch Vol. .TO, No. 12 that failed to become typical galls (fig. 2, F). On mature trees of glabra growing on the campus, 50 series of 3 to 5 puncture inoculations each, were all negative. (Guadalupensis and glabra are the species of Cupreasus frequently grown as ornamentals in southern California.) The galls on montana, 4 months after inoculation, were small knoblike growths 1 to 3 mm. in diameter that had grown from healed-over tissue (fig. 2, D, K). Although later observations indicated that these galls increased somewhat in size, their slow development suggests that the species has strong resistance. A few typical galls formed on Jorbesii, but most of the inoculations were negative in effect. FIGURE 2.—Artificial gaits produood on varioiiK species of Cuprossaceae. In- oculations of A, I), K, and F wore by cultures isolated from Salix sp.; inocula- tions of B were by cultures from Prunus -pérsica, and inoculations of C were by cultures from Libocedrus decurrens. The time, in months, from the date of inoculation to the date of the photograph is indicated in parentheses following the description of the species: A, Thujopsis dolabrata (9 months); D, Cha- maecyparis lawsoniana showing two overgrowths with small projections on their upper parts and between these overgrowths a small knoblike development (11 montiis); C, LiJmcudrus decurrens showing two galls, the upper gall smooth and tlie lower gall rougli (24 months) ; D, E, Cupressus montana, S 2067, small knoblike galls (4 months) ; /'', C. glabra, knoblike projections (4 months). The aerial gall on Cwpressus may first appear as a globose swelling having a smooth bark from which small projecting knobs arc often developed. This bark covering may remain smooth for about a year, but often the pressure of growth causes it to crack, and gum forms. The gall eventually becomes rougli and resembles crown gall as it forms on other susceptible hosts. The artificial galls that develop below the soil arc usually softer than aerial galls and have a smoother bark.