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中国烟草疫霉引起百合疫病的首次报道。

First Report of Phytophthora Blight of Lily Caused by Phytophthora nicotianae in China.

作者信息

Yang X M, Wang J H, Qu S P, Wang L H, Peng L C

机构信息

Quality Standardizing and Testing Technology Research Institute, Yunnan Academy of Agriculture Science, Kunming 650205, China.

Flower Research Institute, Yunnan Academy of Agriculture Science, Kunming 650205, China.

出版信息

Plant Dis. 2010 Jun;94(6):782. doi: 10.1094/PDIS-94-6-0782A.

Abstract

Lily (Lilium spp.) is an economically important cut flower in China. In August 2009, 30 to 40% of plants of lily cv. Siberia in a greenhouse for cut flower production in Yunnan, China were severely diseased. Infected plants developed water-soaked lesions and soft rot on the base of stems and leaves near the soil surface. As the disease progressed, stems bent and plants collapsed. Soft rot symptoms were observed on some bulbs and roots of severely diseased plants. Small, diseased tissue fragments (approximately 3 mm) were surface disinfected with 0.5% NaOCl and then plated to Phytophthora selective medium (10% V8 juice agar) (4). Inoculated dishes were incubated at 25°C in the dark. After 5 days, white colonies with abundant aerial mycelia developed from all plated tissue samples. The fungus had aseptate hyphae. Sporangia were papillate, both caducous and noncaducous, and the shape ranged from ovoid to spherical. The dimensions of sporangia were 30 to 62 × 21 to 46 μm. On the basis of morphological features, isolates were identified as Phytophthora nicotianae Breda de Haan. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS1/ITS4 and sequenced. BLAST analysis of the 835-bp fragment showed a 99% homology with the sequence of P. nicotianae AY833527. The nucleotide sequence has been assigned GenBank No. GU299778. PCR amplification of genomic DNAs using the P. nicotianae-specific primer pair ITS3-PNIC1 generated a 455-bp sequence (3). The result further confirmed the identity of P. nicotianae. Pathogenicity tests were conducted in the greenhouse on lily cv. Siberia grown in pots. Ten 3-month-old plantlets were inoculated by watering the wounded stem bases and soil surface with 30 ml of zoospore suspensions (10 spores per ml). Five uninoculated plantlets were used as controls. All plantlets were covered with plastic bags and incubated at room temperature (22 to 26°C) for 48 h. Inoculated plants developed initial symptoms of slight chlorosis and wilting of lower leaves. Within a 3-week period, all plants died due to soft rot of stem bases and leaves. The pathogen was reisolated from inoculated plants but not from control plants that were symptomless. P. nicotianae has been reported as the causal agent of Phytophthora blight on lily in Korea, Japan, and Hungary (1,2). To our knowledge, this is the first report of Phytophthora blight of lily in China. References: (1) J. Bakonyi et al. Plant Pathol. 50:795, 2001. (2) H. J. Jee and W. G. Kim. Plant Pathol. J. 14:452, 1998. (3) P. W. Tooley et al. Appl. Environ. Microbiol. 63:1467, 1997. (4) X. B. Zheng. Phytophthora and Its Research Technology. Beijing. China Agriculture Press, Beijing, 1997.

摘要

百合(百合属植物)是中国一种具有重要经济价值的切花。2009年8月,在中国云南一个用于切花生产的温室里,30%至40%的西伯利亚百合植株严重染病。染病植株在靠近土壤表面的茎基部和叶片上出现水渍状病斑和软腐症状。随着病情发展,茎弯曲,植株倒伏。在一些重病植株的鳞茎和根部也观察到软腐症状。将小块病组织(约3毫米)用0.5%次氯酸钠进行表面消毒,然后接种到疫霉属选择性培养基(10% V8汁琼脂)上(4)。接种后的培养皿在25°C黑暗条件下培养。5天后,所有接种的组织样本都长出了带有大量气生菌丝的白色菌落。该真菌具有无隔菌丝。孢子囊具乳突,有脱落型和非脱落型,形状从卵形到球形不等。孢子囊大小为30至62×21至46微米。根据形态特征,分离物被鉴定为烟草疫霉(Phytophthora nicotianae Breda de Haan)。使用引物ITS1/ITS4扩增rDNA的内部转录间隔区(ITS)并进行测序。对835碱基对片段的BLAST分析显示与烟草疫霉AY833527的序列有99%的同源性。该核苷酸序列已被赋予GenBank登录号GU299778。使用烟草疫霉特异性引物对ITS3-PNIC1对基因组DNA进行PCR扩增,产生了一个455碱基对的序列(3)。结果进一步证实了烟草疫霉的身份。在温室中对盆栽的西伯利亚百合进行致病性测试。用30毫升游动孢子悬浮液(每毫升10个孢子)浇灌受伤的茎基部和土壤表面,对10株3个月大的幼苗进行接种。5株未接种的幼苗作为对照。所有幼苗都用塑料袋覆盖,在室温(22至26°C)下培养48小时。接种的植株出现下部叶片轻微黄化和萎蔫的初始症状。在3周内,所有植株因茎基部和叶片软腐而死亡。从接种的植株中重新分离到了病原菌,但从无症状的对照植株中未分离到。在韩国、日本和匈牙利,烟草疫霉已被报道为百合疫病的病原菌(1,2)。据我们所知,这是中国百合疫病的首次报道。参考文献:(1)J. Bakonyi等人,《植物病理学》50:795,2001年。(2)H. J. Jee和W. G. Kim,《植物病理学杂志》14:452,1998年。(3)P. W. Tooley等人,《应用与环境微生物学》63:1467,1997年。(4)X. B. Zheng,《疫霉及其研究技术》,北京,中国农业出版社,北京,1997年。

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