First Report of Seedling Blight of Eastern Poison Ivy (Toxicodendron radicans) by Colletotrichum fioriniae in Virginia.

Colletotrichum fioriniae is a member of the large cosmopolitan C. acutatum species complex (2). Known agricultural hosts of C. acutatum include apple, European blueberry, grape, olive, papaya, and strawberry (2). In contrast, the life history of C. fioriniae ranges from an epizootic of certain scale insect populations to an endophyte of plants (3,4). The present study extends the phytopathology of C. fioriniae to include poison ivy seedlings. Poison ivy (Toxicodendron radicans) drupes were collected from solitary lianas in Roanoke and Montgomery counties, Virginia. These drupes were subjected to experiments aimed at producing sterile seedlings (1); however, there was extensive blighting and wilting in the germinated seedlings. Associated with the drupes and seedlings was a fungus with white to pale olivaceous grey mycelium with orange blister-like conidiomata and sclerotial masses enclosing the drupe mesocarp as well as conidiomata emerging from blighted, necrotic leaves. Condiomata were plated onto acidified potato dextrose agar (APDA) and oatmeal agar (OA). This consistently yielded colonies identical to those described from diseased tissues and were putatively identified as C. acutatum based on the presence of acervuli containing hyaline, smooth-walled, aseptate conidia with acute ends, the absence of setae, and formation of red pigments in culture (2). Conidial dimensions of four isolates most closely aligned with reported measurements for C. fioriniae (4): mean length ± SD × width ± SD = 15.1 ± 1.7 × 4.9 ± 0.3 µm, L/W ratio = 3.04 on OA. Fungal DNA was isolated and used as template in PCR reactions using oligonucleotide primer pairs corresponding to the internal transcribed spacer (ITS) region, and a portion of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes. The resulting PCR fragments were sequenced and used as queries in BLASTN searches of the GenBank NR database. All of the amplified ITS DNA sequences (497 bp KF944356 and KF944357) were identical to Glomerella/Colletotrichum fioriniae (JN121190 and KF278459). Similarly, the amplified (672 bp) GAPDH sequences (KF944354 and KF944355) were 99.6% similar over the 254 bp overlapping with C. fioriniae (JQ948622). Pathogenicity of two randomly chosen C. fioriniae isolates, TR-123 and TR-126, was confirmed by placing 4.75 mm diam. inoculated agar plugs from 8-day-old fungal cultures or a sterile plug (negative control) at the base of an axenic young seedling ~1.5 to 6.5 cm in height with at least one set of true leaves (1). Each treatment was replicated five times. Acute wilt and blighting of leaves and production of orange acervuli on cotyledons disease symptoms developed by 3 weeks post inoculation (WPI). By 7 WPI all but one of the Colletotrichum-inoculated plants were dead, whereas all of the control plants were healthy with significantly lower area under the disease progress curve values. Colletotrichum was consistently re-isolated, and confirmed morphologically and molecularly, from six of seven diseased seedlings, whereas two of two randomly chosen control seedlings remained asymptomatic and did not yield Colletotrichum. In summary, C. fioriniae may represent a natural biocontrol agent against poison ivy and scale insect herbivores thereof. References: (1) E. Benhase and J. Jelesko. HortScience 48:1, 2013. (2) U. Damm et al. Stud. Mycol. 73:37, 2012. (3) J. Marcelino et al. J. Insect Sci. 9:25, 2009. (4) R. Shivas et al. Fungal Divers. 39:111.

First Report of Leaf Curl on Celery Caused by Colletotrichum acutatum in the United States.

In June 2010 and July 2011, celery (Apium graveolens) samples cv. Tango were submitted to the Penn State Plant Disease Clinic from Franklin and Dauphin Counties, PA, respectively. Plants exhibited curling and twisting of leaves and petioles and dark, brownish-black necrotic lesions at the base of the plant, extending up the petioles. A fungal organism with morphology consistent to Colletotrichum acutatum J.H. Simmonds was isolated from plant lesion tissue excised from the Dauphin Co. sample. Grown on half strength potato dextrose agar (PDA), the colony had gray aerial mycelium and a pink reverse. Conidia were 5.1 to 14.5 × 2.6 to 5.1 µm, aseptate, hyaline, elliptical, with one or both ends slightly pointed, and formed from the mycelium or in dense orange masses of acervuli on the aerial surface of the culture. Setae were not present. To test pathogenicity, five 23-week-old plants of the cv. Sonora and five 11-week-old plants each of the cvs. Tango and Tall Utah were sprayed until runoff with a conidial suspension (1.3 × 106 conidia/ml and 1.4 × 106 conidia/ml, respectively) and 0.025% Tween. One plant of each cv. was sprayed with milliQ water and 0.025% Tween as a control. Plastic bags were sprayed with the conidial suspension (milliQ water for the control), and secured over the individual plants for 24 h to create a humidity chamber. Plants were incubated in a growth chamber with a 16-h photoperiod, 25°C day/18°C night temperatures, and 70% humidity. Post-inoculation, all of the cv. Tango plants exhibited leaf cupping and curling after 7 days and most plants had dark stem lesions after 3 weeks, consistent with celery leaf curl symptoms. Plants of cvs. Tall Utah and Sonora developed malformed leaves and leaf curl symptoms 16 days and 10 days post-inoculation, respectively. None of the control plants developed symptoms. Infected tissue was excised from diseased plants, surface disinfested in 0.5% sodium hypochlorite for 45 s and plated on half strength PDA. Fungal colonies consistent with C. acutatum were recovered from all inoculated celery tissues (except two of the five inoculated cv. Tall Utah plants and the negative controls). To verify morphological identification, the internal transcribed spacer (ITS) rDNA region was amplified and sequenced for our original isolate and those recovered from the inoculated plants using ITS1 and ITS4 primers (2) (GenBank Accession No. JQ794875). Sequence homology revealed 99 to 100% similarity to accessioned isolates of C. acutatum, which included the holotype and a paratype of C. acutatum (Accession Nos. AF411700 and AF411701, respectively). Celery leaf curl has been reported to have caused devastating crop losses on celery in Australia (1, 3) and to our knowledge, C. acutatum causing leaf curl of celery has not been officially reported in the United States. Infected celery plants are unmarketable because of the leaf malformation and eventual plant necrosis caused by C. acutatum. As such, this disease could have serious negative implications for celery growers in the United States. References: (1) J. B. Heaton and S. R. Dullahide. Australas. Plant Pathol. 22:152, 1993. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990. (3) D. G Wright and J. B. Heaton. Australas. Plant Pathol. 20:155, 1991.