Li Y Y, Wang Y J, Xie Z K, Wang R Y, Qiu Y, Pan H Q, Hu J C
Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou 730000, China.
Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China.
Plant Dis. 2013 Jul;97(7):993. doi: 10.1094/PDIS-11-12-1010-PDN.
Lily (Lilium spp.) is one of the most well-known horticultural crops, and plays an important economic role in China. In September 2011, wilted plants were observed on Lilium oriental hybrid cultivar 'Sorbonne' growing in Longde County, Ningxia Hui Autonomous Region, China. Disease symptoms included wilting, stem and root rot, brown spots of bulbs and then bulbs rotting and spalling from the basal disc, plus a progressive yellowing and defoliation of the leaves from the base. Diseased plants were sampled from fields. Small pieces of symptomatic bulbs, stems, and roots from 10 different plants were surface disinfected with 75% ethanol for 30 s, 3% sodium hypochlorite for 5 min, and then washed three times in sterilize distilled water. The tissues were placed onto Martin Agar (2) at 25°C for 7 days. Nine isolates with morphology similar to Fusarium were obtained from the diseased tissues. Isolates were transferred to potato dextrose agar (PDA) and carnation leaf agar (CLA) and incubated at 25°C. Seven were identified as Fusarium oxysporum and one was F. solani, which have been reported as pathogens of lily in China (1). The other isolate, when grown on PDA, rapidly produced dense, white aerial mycelium that became pink with age and formed red pigments in the medium. On CLA, macroconidia with three to five septate were abundant, relatively slender, and curved to lunate. Microconidia were abundant, oval or pyriform, and one to two celled. Chlamydospores were in chains with smooth exine. The rDNA internal transcribed spacer (ITS) region and a portion of the translation elongation factor 1-alpha (EF-1α) gene of the fungus were amplified, with universal primers ITS1/ITS4 and EF1/EF2 primers respectively (3) and sequenced. In addition, the β-tubulin gene (β-tub) of the fungus was amplified with modified primers Btu-F-F01 (5'-CAGACMGGTCAGTGCGTAA-3') and Btu-F-R01 (5'-TCTTGGGGTCGAACATCTG-3') (4). BLASTn analysis showed that the ITS sequences of the isolate (GenBank Accession No. JX989827) had 98.9% similarity with those of F. tricinctum (EF611092, JF776665, and HM776425) and the EF-1α sequences of the isolate (JX989828) had 98.1% similarity with those of F. tricinctum (EU744837 and JX397850). The β-tub sequences of the isolate (JX989829) had 99.0% similarity with those of F. tricinctum (EU490236 and AB587077). The isolate was tested for pathogenicity. Two-month-old 'Sorbonne' seedlings were inoculated by placing 5 ml of conidial suspension (about 10 conidia per ml) over the roots of plants in each pot. Control plants were treated with sterile water in the same way. Plants were placed in a greenhouse at 22 to 25°C with a 15-h photoperiod. There were eight plants per pot and three replicates for each treatment. After 3 weeks, 87.5% of the inoculated plants exhibited browning of the root tips, root rot, and yellowing of the leaves, while control plants were symptomless. The pathogen was reisolated from the infected roots and identified as F. tricinctum, thus fulfilling Koch's postulates. To our knowledge, this is the first report of Fusarium wilt of lily caused by F. tricinctum. This information will provide guidance for the control of lily wilt disease and add information useful for the production of lilies. References: (1) C. Li and J. J. Li. Acta Phytopathol. Sin. (in Chinese) 26:192, 1995. (2) J. P. Martin. Soil Sci. 38:215, 1950. (3) K. O'Donnell et al. Proc. Nat. Acad. Sci. U. S. A. 95:2044, 1998. (4) M. Watanabe et al. BMC Evol. Biol. 11:322. 2011.
百合(百合属植物)是最著名的园艺作物之一,在中国具有重要的经济地位。2011年9月,在中国宁夏回族自治区隆德县种植的东方百合杂交品种‘索邦’上观察到了萎蔫植株。病害症状包括萎蔫、茎和根腐烂、鳞茎出现褐色斑点,随后鳞茎腐烂并从基部盘脱落,以及叶片从基部开始逐渐变黄和落叶。从田间采集了患病植株样本。将来自10株不同植株的有症状的鳞茎、茎和根的小块,先用75%乙醇表面消毒30秒,再用3%次氯酸钠消毒5分钟,然后在无菌蒸馏水中冲洗三次。将这些组织置于马丁琼脂(2)上,在25°C下培养7天。从患病组织中获得了9个形态与镰刀菌相似的分离株。将分离株转移到马铃薯葡萄糖琼脂(PDA)和香石竹叶琼脂(CLA)上,在25°C下培养。其中7个被鉴定为尖孢镰刀菌,1个为茄腐镰刀菌,在中国它们已被报道为百合的病原菌(1)。另一个分离株,在PDA上生长时,迅速产生致密的白色气生菌丝体,随着时间推移变为粉红色,并在培养基中形成红色色素。在CLA上,有三到五个隔膜的大型分生孢子丰富,相对细长,弯曲呈月牙形。小型分生孢子丰富,呈椭圆形或梨形,单细胞或双细胞。厚垣孢子呈链状,外壁光滑。分别用通用引物ITS1/ITS4和EF1/EF2引物扩增该真菌的rDNA内转录间隔区(ITS)和翻译延伸因子1-α(EF-1α)基因的一部分(3)并进行测序。此外,用改良引物Btu-F-F01(5'-CAGACMGGTCAGTGCGTAA-3')和Btu-F-R01(5'-TCTTGGGGTCGAACATCTG-3')扩增该真菌的β-微管蛋白基因(β-tub)(4)。BLASTn分析表明,该分离株的ITS序列(GenBank登录号JX989827)与三线镰刀菌(EF611092、JF776665和HM776425)的ITS序列相似度为98.9%,该分离株的EF-1α序列(JX989828)与三线镰刀菌(EU744837和JX397850)的EF-1α序列相似度为98.1%。该分离株的β-tub序列(JX989829)与三线镰刀菌(EU490236和AB587077)的β-tub序列相似度为99.0%。对该分离株进行了致病性测试。将5毫升分生孢子悬浮液(约每毫升10个分生孢子)置于每盆植株的根部,对两个月大的‘索邦’幼苗进行接种。对照植株以相同方式用无菌水处理。将植株置于温度为22至25°C、光周期为15小时的温室中。每盆有8株植株,每个处理重复3次。3周后,87.5%的接种植株出现根尖褐变、根腐和叶片发黄,而对照植株无症状。从感染的根部重新分离出病原菌,并鉴定为三线镰刀菌,从而满足了柯赫氏法则。据我们所知,这是关于三线镰刀菌引起百合枯萎病的首次报道。该信息将为百合枯萎病的防治提供指导,并为百合生产增添有用信息。参考文献:(1)李超,李建强。植物病理学报(中文)26:192,1995。(2)J.P.马丁。土壤科学38:215,1950。(3)K.奥唐奈等。美国国家科学院院刊95:2044,1998。(4)M.渡边等。BMC进化生物学卷11:322,2011。