Pollok J R, Mansfield M A, Gugino B K, May S R
Department of Plant Pathology, The Pennsylvania State University, University Park 16802.
Plant Dis. 2012 Nov;96(11):1692. doi: 10.1094/PDIS-03-12-0271-PDN.
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 × 10 conidia/ml and 1.4 × 10 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.
2010年6月和2011年7月,宾夕法尼亚州富兰克林县和多芬县分别向宾夕法尼亚州立大学植物病害诊所提交了“探戈”品种的芹菜(Apium graveolens)样本。植株表现出叶片和叶柄卷曲、扭曲,植株基部出现深褐色至黑褐色坏死斑,并向上延伸至叶柄。从多芬县样本的植株病变组织中分离出一种形态与尖孢炭疽菌(Colletotrichum acutatum J.H. Simmonds)一致的真菌。在半强度马铃薯葡萄糖琼脂(PDA)上培养时,菌落有灰色气生菌丝体,背面呈粉红色。分生孢子大小为5.1至14.5×2.6至5.1μm,无隔膜,透明,椭圆形,一端或两端稍尖,由菌丝体形成或在培养物气生表面的分生孢子盘密集橙色团块中形成。没有刚毛。为测试致病性,对5株23周龄的“索诺拉”品种植株以及5株11周龄的“探戈”和“犹他高杆”品种植株分别喷施分生孢子悬浮液(浓度分别为1.3×10分生孢子/ml和1.4×10分生孢子/ml)和0.025%吐温,直至径流。每个品种的一株植株喷施超纯水和0.025%吐温作为对照。用分生孢子悬浮液(对照用超纯水)喷施塑料袋,并套在单株植株上24小时以形成湿度箱。植株在光照周期为16小时、白天温度25°C/夜间温度18°C、湿度70%的生长室中培养。接种后,所有“探戈”品种植株在7天后均表现出叶片杯状卷曲,3周后大多数植株茎部出现深色病斑,与芹菜叶卷曲症状一致。“犹他高杆”和“索诺拉”品种植株分别在接种后16天和10天出现叶片畸形和叶卷曲症状。对照植株均未出现症状。从患病植株上切下感染组织,在0.5%次氯酸钠中表面消毒45秒,然后接种在半强度PDA上。从所有接种的芹菜组织(除5株接种的“犹他高杆”品种植株中的2株和阴性对照外)中均分离出与尖孢炭疽菌一致的真菌菌落。为验证形态学鉴定结果,使用ITS1和ITS4引物(2)对原始分离株以及从接种植株中分离得到的菌株的内部转录间隔区(ITS)rDNA区域进行扩增和测序(GenBank登录号JQ794875)。序列同源性分析显示,与已登录的尖孢炭疽菌分离株的相似性为99%至100%,其中包括尖孢炭疽菌的模式菌株和一个副模式菌株(登录号分别为AF411700和AF411701)。据报道,芹菜叶卷曲病在澳大利亚已给芹菜作物造成了毁灭性损失(1, 3),据我们所知,在美国尚未正式报道过尖孢炭疽菌引起芹菜叶卷曲病。由于尖孢炭疽菌导致叶片畸形并最终使植株坏死,感染的芹菜植株无法进入市场。因此,这种病害可能会给美国的芹菜种植者带来严重的负面影响。参考文献:(1)J. B. Heaton和S. R. Dullahide。Australas. Plant Pathol. 22:152, 1993。(2)T. J. White等人。见《PCR协议:方法与应用指南》第315页。学术出版社,加利福尼亚州圣地亚哥,1990。(3)D. G Wright和J. B. Heaton。Australas. Plant Pathol. 20:155, 1991。