Patel J S, Norman D, Brennan M, Ali G S
Mid-Florida Research and Education Center and Department of Plant Pathology, University of Florida/Institute of Food and Agricultural Sciences, 2725 Binion Rd., Apopka, FL 32703.
Plant Dis. 2013 Mar;97(3):426. doi: 10.1094/PDIS-09-12-0865-PDN.
Chinese elm (Ulmus parvifolia Jacq.) is native to China, Korea, and Japan and was introduced to the U.S. to replace the American elm, which is highly susceptible to Dutch elm disease. Cultivar Emmer II trademark Allee elm (or Allee elm) is preferred by nurseries for its rich green foliage and beauty of bark. In the summer of 2011, a new disease was observed on Allee elm at a tree farm in FL. Approximately 1% of elms in the same farm and in residential areas in central Florida had similar canker-like symptoms consisting of tan to orange patches of decomposed and loose bark. These symptoms were observed on the main trunks often extending into branches of affected trees. Cankered sections of the trunk were often several feet in length and penetrated the wood to a depth just under the bark into the phloem. To isolate the causal organism, cankers were gathered from 7 trees and tissues from the margin of 1 to 3 cankers per tree were surface sterilized in 1% sodium hypochlorite, plated on PDA, and incubated at 22°C under a 16-h/8-h light/dark cycle for 7 days. Colonies displayed white, fluffy mycelium with sporadic black acervuli containing aggregated conidia. Conidia were 5-celled with two or more apical appendages or hairs; the three central cells were dark brown and the two outer cells were hyaline (1). Based on conidial morphology, these isolates were putatively identified as Pestalotiopsis spp. To identify the species, the rDNA internal transcribed spacer (ITS) region was sequenced for two field isolates (GenBank Accessions JX999998 and JX999999). A BLASTn search in GenBank revealed 100% identity to Pestalotiopsis mangiferae ITS (JX305704.1). To test Koch's postulates, experiments were performed in the field and greenhouse. A mycelial plug of isolate 11-40 was grown on PDA, inoculated on wounded trunk of healthy 18-month-old Allee elms (n = 48) in the field and 7-month-old Allee elms (n = 12) in the greenhouse. Both experiments were set up as a randomized complete block design. The trunk of each tree was wounded with a scalpel to a depth of 5 mm, the wound was inoculated with a 5-mm agar plug from a 7-day PDA culture, and the inoculated wound was wrapped with grafting tape. Plants that served as negative controls [n = 20 (field experiment) and n = 12 (greenhouse)] were mock-inoculated with a sterile PDA plug without mycelial growth. After 4 to 6 months, symptoms consisting of loose or fallen off bark developed on all pathogen-inoculated trees but not on control trees. Control wounds healed with no expansion beyond the original 5 mm inoculation zone, whereas Pestalotiopsis-inoculated cankers expanded to 3 to 8 cm in each direction in 6 months. Pestalotiopsis (confirmed by conidial morphology and ITS sequencing) was reisolated from pathogen-inoculated trees but not from control trees. These experiments were repeated with similar results at least three times, each consisting of 15 replications (greenhouse) and 12 replications (field) with additional Pestalotiopsis isolates. Many other Pestaliopsis spp. have been reported on other shade trees through the U.S. and the world. To our knowledge, this is the first report of P. mangiferae on an Ulmus species in the U.S. and the world. Since elm canker mainly affects the trunk, a featured characteristic of the Allee elm, it can potentially result in economic loss to the ornamental industry. References: (1) Y. Ko et al. Plant Dis. 91:1684, 2007. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.
榔榆(Ulmus parvifolia Jacq.)原产于中国、韩国和日本,被引入美国以取代对荷兰榆树病高度敏感的美国榆。苗圃更喜欢栽培品种Emmer II商标的林荫道榆(或林荫道榆),因其枝叶浓绿且树皮美观。2011年夏天,在佛罗里达州的一个林场,人们在林荫道榆上发现了一种新病害。同一林场以及佛罗里达州中部居民区中约1%的榆树有类似溃疡状症状,表现为浅褐色至橙色的树皮腐烂和脱落斑块。这些症状在主干上常见,常延伸至受影响树木的枝条。树干溃疡部分通常有几英尺长,深入木材至树皮下方刚好进入韧皮部的深度。为分离致病生物,从7棵树上采集溃疡部位,每棵树1至3个溃疡边缘的组织在1%次氯酸钠中进行表面消毒,接种到PDA上,并在22°C、16小时光照/8小时黑暗周期下培养7天。菌落呈现白色、蓬松的菌丝体,有散在的黑色分生孢子盘,其中含有聚集的分生孢子。分生孢子为5细胞,有两个或更多顶端附属物或毛发;三个中央细胞深褐色,两个外部细胞透明(1)。基于分生孢子形态,这些分离物初步鉴定为拟盘多毛孢属(Pestalotiopsis spp.)。为确定物种,对两个田间分离物的rDNA内部转录间隔区(ITS)进行测序(GenBank登录号JX999998和JX999999)。在GenBank中进行BLASTn搜索显示,与芒果拟盘多毛孢(Pestalotiopsis mangiferae)的ITS(JX305704.1)有100%的同一性。为验证科赫法则,在田间和温室进行了实验。分离物11 - 40的菌丝块在PDA上生长,接种到田间健康的18个月大的林荫道榆(n = 48)和温室中7个月大的林荫道榆(n = 12)的受伤树干上。两个实验均采用随机完全区组设计。用手术刀将每棵树的树干伤口切割至5毫米深,用来自7天PDA培养物的5毫米琼脂块接种伤口,接种后的伤口用嫁接带包扎。作为阴性对照的植株 [n = 20(田间实验)和n = 12(温室)] 用无菌丝生长的无菌PDA块进行假接种。4至6个月后,所有接种病原体的树木出现树皮松动或脱落的症状,而对照树木未出现。对照伤口愈合,未超出最初5毫米接种区域,而接种拟盘多毛孢的溃疡在6个月内每个方向扩展至3至8厘米。从接种病原体的树木上重新分离出拟盘多毛孢(通过分生孢子形态和ITS测序确认)但未从对照树木上分离出。这些实验至少重复了三次,结果相似,每次实验在温室中有15次重复,在田间有12次重复,并使用了其他拟盘多毛孢分离物。在美国和世界各地,许多其他拟盘多毛孢属物种已在其他遮荫树上被报道。据我们所知,这是美国和世界上首次关于芒果拟盘多毛孢在榆属物种上的报道。由于榆树溃疡主要影响主干,这是林荫道榆的一个特征特性,它可能会给观赏行业带来经济损失。参考文献:(1)Y. Ko等人,《植物病害》91:1684, 2007。(2)T.J.怀特等人,见《PCR协议:方法与应用指南》第315页。学术出版社,圣地亚哥,1990。