Shen Y M, Chao C H, Wang F C, Liu H L, Huang T C
Plant Protection Laboratory, Taichung District Agricultural Research and Extension Station, Changhua, Taiwan, R.O.C.
Plant Dis. 2012 Jun;96(6):910. doi: 10.1094/PDIS-02-12-0168-PDN.
Eustoma (Eustoma russellianum) is an economically important cut flower in Taiwan. Each year more than 1.7 million dozen flowers, mainly exported to Japan in the winter, are produced in greenhouses. In January 2011, eustoma plants with stem and leaf blight symptoms were observed in some greenhouses in Changhua County, Taiwan, at an incidence of 2%. Brown and rotten lesions were presented on the stem and nearby leaves, with white mycelia growing on the surface and black sclerotia (up to 7 mm long) produced inside the stem. Infected plants were completely blighted and eventually died. Diseased stem tissues collected from the field were surface sterilized for 3 min in 0.6% NaOCl, rinsed with sterilized distilled water, and plated on potato dextrose agar. White fungal colonies were consistently isolated. The cultures produced large sclerotia at the peripheries of the plates. Internal transcribed spacer (ITS) sequences of two voucher isolates were determined and deposited in GenBank (Accession Nos. JQ653934 and JQ653935). The sequences were 100% identical to that of Sclerotinia sclerotiorum strain ATCC MYA-4521 (Accession No. FJ810516). In addition, PCR amplified DNA fragments (approximately 630 bp) were obtained by the S. sclerotiorum specific primer pair MP_SsF and MP_UniR (1). On the basis of morphology, ITS sequence homology, and the specific PCR detection, the fungus was identified as S. sclerotiorum. The two fungal isolates (BCRC34830 and BCRC34831) were deposited in Bioresource Collection and Research Center, Hsinchu, Taiwan. Pathogenicity tests were conducted on 1-month-old, second flush eustoma cultivars Ex Rosa Pink Flash and Rosina Blue Ver. 2 after primary flowers had been harvested in the greenhouse. Fungal inoculum consisting of Tref horticultural substrate and wet sterilized rice colonized by S. sclerotiorum BCRC34830 (substrate-rice-water ratio of 2:1:1) was placed near the base of the plants. Ten plants of each cultivar were inoculated with about 800 g of the mixture. Sterile mixture applied to an equal number of plants served as negative controls. Eight plants of each cultivar showed blight symptoms after 1 month of incubation at an average temperature of 26°C. All control plants remained healthy. The pathogen reisolated from the inoculated stems produced sclerotia identical to those isolated in the field, fulfilling Koch's postulates. The pathogenicity test was repeated with similar results. S. sclerotiorum has been reported on eustoma in Argentina (2). To our knowledge, this is the first report of Sclerotinia blight on eustoma in Taiwan. Although the disease was not prevalent on eustoma, the inoculum could be dormant in the greenhouse soil. Awareness of the potential perennial problem could increase the quality of the flowers exported and benefit the flower industry. References: (1) S. Hirschhäuser and J. Fröhlich. Int. J. Food Microbiol. 118:151, 2007. (2) S. Wolcan et al. Plant Dis. 80:223, 1996.
洋桔梗(Eustoma russellianum)是台湾一种具有重要经济价值的切花。每年在温室中生产超过170万打鲜花,主要在冬季出口到日本。2011年1月,在台湾彰化县的一些温室中观察到有茎和叶枯萎症状的洋桔梗植株,发病率为2%。茎和附近叶片上出现褐色和腐烂病斑,表面生长有白色菌丝体,茎内产生黑色菌核(长达7毫米)。受感染的植株完全枯萎并最终死亡。从田间采集的患病茎组织在0.6%次氯酸钠中进行表面消毒3分钟,用无菌蒸馏水冲洗后,接种到马铃薯葡萄糖琼脂培养基上。持续分离出白色真菌菌落。培养物在平板边缘产生大型菌核。测定了两个凭证分离株的内转录间隔区(ITS)序列并保存在GenBank中(登录号JQ653934和JQ653935)。这些序列与核盘菌菌株ATCC MYA - 4521(登录号FJ810516)的序列100%相同。此外,使用核盘菌特异性引物对MP_SsF和MP_UniR通过PCR扩增获得了约630 bp的DNA片段(1)。基于形态学、ITS序列同源性和特异性PCR检测,该真菌被鉴定为核盘菌。这两个真菌分离株(BCRC34830和BCRC34831)保存在台湾新竹的生物资源保存与研究中心。对温室中已采收初花的1月龄二次抽枝洋桔梗品种Ex Rosa Pink Flash和Rosina Blue Ver. 2进行致病性测试。由核盘菌BCRC34830定殖的Tref园艺基质和湿灭菌大米组成的真菌接种物(基质 - 大米 - 水比例为2:1:1)放置在植株基部附近。每个品种的10株植株接种约800克该混合物。等量无菌混合物施用于相同数量的植株作为阴性对照。在平均温度26°C下培养1个月后,每个品种的8株植株出现枯萎症状。所有对照植株保持健康。从接种茎中重新分离出的病原菌产生的菌核与在田间分离的菌核相同,满足柯赫氏法则。重复进行致病性测试,结果相似。在阿根廷已有关于洋桔梗上核盘菌的报道(2)。据我们所知,这是台湾洋桔梗上核盘菌叶枯病的首次报道。尽管该病在洋桔梗上并不普遍,但接种体可能在温室土壤中休眠。认识到这一潜在的常年性问题可以提高出口鲜花的质量并使花卉产业受益。参考文献:(1)S. Hirschhäuser和J. Fröhlich。《国际食品微生物学杂志》118:151,2007。(2)S. Wolcan等人。《植物病害》80:223,1996。