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Morphology and Taxonomy of Species of Phomopsis on Asparagus Author(S): F

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Morphology and Taxonomy of Species of Phomopsis on Asparagus Author(S): F Mycological Society of America Morphology and Taxonomy of Species of Phomopsis on Asparagus Author(s): F. A. Uecker and Dennis A. Johnson Source: Mycologia, Vol. 83, No. 2 (Mar. - Apr., 1991), pp. 192-199 Published by: Mycological Society of America Stable URL: http://www.jstor.org/stable/3759934 . Accessed: 05/03/2014 11:45 Your use of the JSTOR archive indicates your acceptance of the Terms & Conditions of Use, available at . http://www.jstor.org/page/info/about/policies/terms.jsp . JSTOR is a not-for-profit service that helps scholars, researchers, and students discover, use, and build upon a wide range of content in a trusted digital archive. We use information technology and tools to increase productivity and facilitate new forms of scholarship. For more information about JSTOR, please contact [email protected]. Mycological Society of America is collaborating with JSTOR to digitize, preserve and extend access to Mycologia. http://www.jstor.org This content downloaded from 134.121.161.15 on Wed, 5 Mar 2014 11:45:57 AM All use subject to JSTOR Terms and Conditions Mycologia, 83(2), 1991, pp. 192-199. ? 1991, by The New York Botanical Garden, Bronx,NY 10458-5126 MORPHOLOGY AND TAXONOMY OF SPECIES OF PHOMOPSIS ON ASPARAGUS F. A. Uecker SystematicBotany and MycologyLaboratory, USDA, AgriculturalResearch Service, BeltsvilleAgricultural Research Center,Beltsville, Maryland 20705 AND Dennis A. Johnson WashingtonState University,Irrigated Agriculture Research and Extension Center, Box 30, Prosser, Washington99350 ABSTRACT Phomopsisjavanica, a new species on Asparagus fromJava, is described and illustrated.It differs fromP. asparagi by producingparaphyses among the conidiophoresand conidiogenous cells, and is the firstPhomopsis described with these structures.Phomopsis javanica was more virulentthan P. asparagi when inoculated on asparagus foliage and stems. Phomopsis asparagicola is considered a synonymof P. asparagi. P. asparagi and P. javanica are compared in a series of photographs. Key Words: Phomopsisjavanica sp. nov.,Phomopsis asparagi, Phomopsis asparagicola, paraphyses A serious disease of asparagus (Asparagus of- Thailand (Anon., 1961). Reifschneider and Lopes ficinalis L.) plants in Java, Indonesia was found (1982) reported it in Brazil. Few of these papers to be caused by a Phomopsis. Comparison of this contain much information on the morphology fungus with the two other Phomopsis species pre? and taxonomy of the fungus and it is possible viously reported on asparagus revealed it to be that two or more fungi are involved. an undescribed species. Saccardo (1878) de? Bausa Alcalde (1952) described a second spe? scribed Phoma asparagi Sacc. on putrescent stems cies, Phomopsis asparagicola Bausa Alcalde on of A. officinalisat Padua, Italy. Although Bubak branches of Asparagus plumosus Baker. It dif? (1906) transferredthis fungus to Phomopsis, it is fered from P. asparagi because the alpha conidia still being reported as Phoma asparagi in current of P. asparagi were oblong, 7-8 x 3 /umaccord? literature (Liu and Hwang, 1988; Tanaka et al, ing to Saccardo (1878) and oblong or spindle- 1987). shaped and 5.5-9 x 2-2.5 /nm according to Phomopsis asparagi (Sacc.) Bubak is well Bubak (1906) whereas they were ellipsoid or known in Europe (da Camara and da Luz, 1943; fusiform, 7-9.5 x 2-2.5 in P. asparagicola; co? Engler and Prantl, 1900; Hruby, 1928; Lind, 1913; nidiomata were subepidermal in P. asparagi ac? Roumeguere, 1889; Solla 1915), in the Middle cording to both Saccardo and Bubak but sub? East (Engler and Prantl, 1900), Puerto Rico (Sherf epidermal then erumpent in P. asparagicola; and and MacNab, 1986) and eastern U.S. (Farr et al, the beta conidia of P. asparagi were reported to 1989) but has not been considered an important be shorter and thinner (18-25 x 1 jtm) than those pathogen in any of these places. of Phlyctaena asparagi Fautrey & Roum. (30 x The disease caused by P. asparagi on aspara? 2 jum), which was considered by Bausa Alcalde gus was first described in India by Kheswalla to be the beta form of P. asparagi. (1936) and later by Galloway (1936), Sohi et al A Phomopsis was recently collected on stems (1975), and Tripathi (1985). It attacks asparagus of asparagus in central Java in Indonesia. It dif? in China (Teng, 1932; Yao et al, 1987), Japan fers from the two previously mentioned fungi in (Tanaka et al, 1987), Korea (Nakata and Taki- that the alpha conidia are more variable in length moto, 1928; Choi et al, 1981), Taiwan (Hsu and and are consistently broader than those of the Sun, 1969; Wu, 1970; Yang et al, 1970), and other two, it produces paraphyses among the 192 This content downloaded from 134.121.161.15 on Wed, 5 Mar 2014 11:45:57 AM All use subject to JSTOR Terms and Conditions Uecker and Johnson: Phomopsis on Asparagus 193 conidiogenous cells, and it is much more virulent illuminator equipped with 4 x and 11 x objec- on inoculated asparagus than is an isolate of P. tives. asparagi. Study ofthe conidiogenous apparatus was ac- This paper describes and illustrates the new complished by using a modification ofthe phlox- fungus on asparagus from Java and compares it ine-KOH method introduced by Martin (1934). with isotypes of Phomopsis asparagi and other A single conidioma was allowed to expand in 3% herbarium specimens and with three ATCC cul? KOH. A small portion ofthe conidiogenous layer tures from Taiwan. A lectotype of P. asparagi is was stained with 1% aqueous phloxine, the designated and photographs illustrating this spe? phloxine was replaced with KOH, and the spec? cies for the firsttime are provided. Epidemiology imen was flattened. All photographs ofthe conid? of P. javanica will be discussed in a separate iogenous apparatus were made from specimens paper. stained with phloxine-KOH, which was then re? placed with 0.2% cotton blue in 85% lactic acid MATERIALSAND METHODS (LA-CB) to extend the useful life of the prepa? ration. The original collection of Phomopsis javanica Conidia were collected on cover slips by touch- was made by one of us (DAJ) in central Java in ing the cover slip to the mucilaginous drop con? January, 1989. A culture (FAU-477) from this taining conidia at the mouth of the ostiole. A collection was grown routinely on autoclaved one- small drop of water was added and the conidia inch long pieces of stem of Asparagus officinalis, were spread in an even layer across the cover alfalfa (Medicago sativa L.), soybean [Glycine max slip. In cases where conidia were not yet extruded (L.) Merr.], Stokes aster [Stokesia laevis (Hill) in a drop, conidiomata were dissected in a small Greene], kiwi (Actinidia chinensis Planchon), and drop of water on a slide and the conidia were cranberry (Vaccinium macrocarpon Ait), and on spread upon the slide. In either case, after air- grains of oat (Avena sativa L.) on water agar (WA) drying the conidia were mounted in a small drop plates. Specimens on each host material were of LA-CB. fixed in formalin-aceto-alcohol (FAA), embed? Phomopsis javanica was also grown on pieces ded in Paraplast Plus1 and sectioned at 7 jum. of stem of asparagus, alfalfa, and grape (Vitis Slides were run through a standard series of xy? labrusca L.) on WA plates and the conidiogenous lene and ethanol to water and were stained in apparatus was observed at intervals of one to methylene blue-azure II (Humphrey and Pitt- five days. Photographs were first taken on day man, 1974, omitting the basic fuchsin). Although five after inoculation, the last ones on day 40. this stain was originally applied to specimens The purpose of this series was to determine embedded in plastic, it is also useful for paraffin whether the conidiogenous apparatus in conid? sections. Used at full strength, it overstains iomata on asparagus varied morphologically at heavily. When diluted to 25% of the original any stage of development from those at other strengthand applied for 5-15 min at room tem? stages and from those on the other substrata. perature staining was more easily controlled. To compare the virulence of P. javanica with Slides were then dipped in water to remove ex- that of P. asparagi, potted asparagus plants of cess stain, blotted nearly dry on absorbent paper, cultivars Mary Washington and WSU-1 were in? and dehydrated and differentiated one to two h oculated with P. javanica and P. asparagi in three in tertiary butanol. After two 10-min baths ih separate tests. The isolate of P. asparagi (Fau- xylene, sections were mounted in Permount. 499) was derived from naturally infected aspar? Isotype specimens of Phomopsis asparagi were agus plants grown at Rutgers Research and rehydrated for several h on moist filter paper, Development Center, Bridgeton, New Jersey. In? fixed in FAA, and furthertreated as above. Co? oculum was increased on potato dextrose agar in nidiomata were measured on living or dried Petri dishes placed under continuous fluorescent specimens using a Leitz Ultropak incident light light at 20 to 23 C for 11-14 days. Conidiomata were scraped and washed from the agar, conidia 1 of a or Mention trademark,proprietary product, were filteredthrough cheesecloth and inoculated vendor does not constitutea guaranteeor warrantyof onto four to seven plants per test. One drop of theproduct by theU.S. Departmentof Agricultureand ml does not imply its approval to the exclusion of other Tween 20 was added to 500 distilled water productsthat may also be suitable. and one drop of the dilution was added to the This content downloaded from 134.121.161.15 on Wed, 5 Mar 2014 11:45:57 AM All use subject to JSTOR Terms and Conditions 194 Mycologia inoculum. Concentration of inoculum was ap? aline, short or elongate, more or less tapered to? proximately 1.8 x 106, 2.8 x 106, and 1.5 x 106 ward apex, septate both at the base and above, alpha conidia, respectively, for the three tests.
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