Gao B-D, Wang X-L, Xia H
Hunan Provincial University Key Lab of Plant Disease Control and Utilization, Hunan Agricultural University, Changsha 410128, China.
Plant Dis. 2011 Aug;95(8):1026. doi: 10.1094/PDIS-04-11-0314.
A new disease on globe artichoke (Cynara scolymus L.) was observed in the springs of 2008 and 2009 and during the spring and fall seasons of 2010 in commercial fields (nearly 1,000 ha) in Changde, Hunan Province, China. Characteristic symptoms were wilting and necrosis of the outermost leaves and dark brown discoloration of the vascular tissue and pith of the stem base. Eventually, the plants wilted and died. Nearly 5, 35, and 4% (2008, 2009, and 2010, respectively) of the artichoke fields were destroyed because of the disease. Manual weeding and cuttings often led to the development of typical soft rot during propagation. To investigate the causal agent of the disease, isolations were made from rotted stems of field artichoke plants on nutrient agar (NA). Bacteria consistently isolated from the diseased tissues formed gray-white, glossy, convex, translucent, and round colonies on NA. The bacterial cells were gram-negative rods with two to eight peritrichous flagella. Ten isolates were negative for oxidase, arginine dehydrolase, HS, gelatin liquefaction, and tryptophan ammonialyase. Isolates were positive for catalase, reduced NO to NO, indole, glucuroide, galactosidase, Voges-Proskauer test, and β-galactosidase, along with being facultatively anaerobic and insensitive to erythromycin (40 μg/ml). Negative results were obtained for utilization of maltose, gluconate, and phenylacetic acid, and positive results were obtained from arabinose, glucose, mannose, N-acetyl-glucosamine, mannitol, and sodium citrate for all isolates. Acid was produced from glucose, inositol, rhamnose, melibiose, arabinose, mannitol, sucrose, and amarogentin. All test results were similar to reference strain PCC1000 (GenBank Accession No. JF721959) of Pectobacterium carotovorum subsp. carotovorum. These isolates could also cause soft rot of Chinese cabbage stem, carrot slice, pepper, lettuce and artichoke stems, and tomato and potato slices within 48 h at 28°C in an artificial inoculation test (3). PCR amplification was carried out by utilizing universal 16S rDNA primer pair 16SF/16SR and pel gene primers Y1/Y2 (1). The 16S rDNA and pel gene sequences of isolate HNXDT002 (GenBank Accession Nos. JF721958 and JF721960, respectively) had 99 and 93% nucleotide identity with strains of P. carotovorum subsp. carotovorum (GenBank Accession Nos. U80197 and CP001657, respectively). Pathogenicity was confirmed by needle-stab inoculation (1 × 10 CFU/ml) at the stem on three healthy artichoke plants held at 28°C for 48 h. Sterile distilled water was used as a negative control. Within 72 h after inoculation, water-soaking and soft-rot symptoms were observed on all inoculated artichoke plants, while controls remained healthy. The bacterium was recovered only from rotted stems of inoculated plants. In recent years, P. carotovorum was reported on such plants as Pinellia ternata (4) and Chinese cabbage (2) in China. To our knowledge, this is the first report of bacterial rot disease caused by P. carotovorum subsp. carotovorum on artichoke in China. References: (1) D. J. Brenner et al. Bergey's Manual of Systematic Bacteriology. Vol. 2. Springer, NY, 2005. (2)Y. Fang et al. Acta Microbiol. Sinica 44:136, 2004. (3) H. Yi-Bo et al. Acta Phytopathol. Sinica 37:338, 2007. (4) F. X. Ying et al. Plant Dis. 91:1359, 2007.
2008年和2009年春季以及2010年春、秋季节,在中国湖南省常德市的商业种植田(面积近1000公顷)中,人们发现了一种新的球茎甘蓝(Cynara scolymus L.)病害。其典型症状为最外层叶片萎蔫和坏死,茎基部维管束组织和髓部出现深褐色变色。最终,植株枯萎死亡。由于这种病害,2008年、2009年和2010年分别有近5%、35%和4%的球茎甘蓝种植田被毁。人工除草和扦插繁殖时,常常会引发典型的软腐病。为了探究该病害的病原菌,从田间球茎甘蓝植株的腐烂茎部在营养琼脂(NA)上进行分离培养。从患病组织中持续分离出的细菌在NA上形成灰白色、有光泽、凸起、半透明的圆形菌落。细菌细胞为革兰氏阴性杆菌,有2至8根周生鞭毛。10个分离菌株的氧化酶、精氨酸脱氨酶、硫化氢、明胶液化和色氨酸氨解酶检测均为阴性。分离菌株的过氧化氢酶、将硝酸盐还原为亚硝酸盐、吲哚、葡萄糖醛酸酶、半乳糖苷酶、Voges-Proskauer试验和β-半乳糖苷酶检测均为阳性,兼性厌氧且对红霉素(40μg/ml)不敏感。所有分离菌株利用麦芽糖、葡萄糖酸盐和苯乙酸的检测结果为阴性,利用阿拉伯糖、葡萄糖、甘露糖、N-乙酰葡糖胺、甘露醇和柠檬酸钠的检测结果为阳性。葡萄糖、肌醇、鼠李糖、蜜二糖、阿拉伯糖、甘露醇、蔗糖和苦玄参苷能产酸。所有检测结果均与胡萝卜软腐果胶杆菌胡萝卜软腐亚种的参考菌株PCC1000(GenBank登录号:JF721959)相似。在人工接种试验中,这些分离菌株在28°C条件下48小时内也能使大白菜茎、胡萝卜片、辣椒、生菜和球茎甘蓝茎以及番茄和马铃薯片发生软腐(3)。利用通用的16S rDNA引物对16SF/16SR和pel基因引物Y1/Y2进行PCR扩增(1)。分离菌株HNXDT002的16S rDNA和pel基因序列(GenBank登录号分别为JF721958和JF721960)与胡萝卜软腐果胶杆菌胡萝卜软腐亚种菌株(GenBank登录号分别为U80197和CP001657)的核苷酸同源性分别为99%和93%。通过在3株健康球茎甘蓝植株的茎部进行针刺接种(1×10CFU/ml)并在28°C下培养48小时来确认致病性。无菌蒸馏水用作阴性对照。接种后72小时内,所有接种的球茎甘蓝植株均出现水渍状和软腐症状,而对照植株保持健康。仅从接种植株的腐烂茎部重新分离到该细菌。近年来,中国报道了胡萝卜软腐果胶杆菌在半夏(4)和大白菜(2)等植物上的发生情况。据我们所知,这是中国首次报道由胡萝卜软腐果胶杆菌胡萝卜软腐亚种引起的球茎甘蓝细菌性腐烂病。参考文献:(1)D. J. Brenner等人。《伯杰氏系统细菌学手册》第2卷。施普林格出版社,纽约,2005年。(2)方勇等人。《微生物学报》第44卷:136页,2004年。(3)易波等人。《植物病理学报》第37卷:338页,2007年。(4)应福兴等人。《植物病害》第91卷:1359页,2007年。