Wei J-F, Wei J-H
Plant Protection College of Yunnan Agricultural University, Kunming 650201, China.
Foreign Languages School of Guangxi Medical University, Nanning 530021, China.
Plant Dis. 2014 May;98(5):682. doi: 10.1094/PDIS-06-13-0679-PDN.
Konnyaku (Amorphophallus rivieri Durieu) is grown in some rural areas of China as an important cash crop. In 2011, there was a serious outbreak of Konnyaku soft rot in Xuanwei District of Yunnan Province of China. The disease was characterized by partial or complete tuber rot. At its anaphase, the soft rot may move up the stem, causing the caudex to decay and the whole plant to collapse. If the stem is strong or big enough, the soft rot may develop on one side of the stem, leaving the other side healthy for several days. In this case, the stem does not collapse, and etiolation may be observed on the rotten tissue. In serious cases, up to 80% of the plants were infected. The disease is even more serious if Konnyaku is grown continuously in the same field for more than one year. At its worst, the disease can wipe out the whole crop. In 2012 and 2013, we isolated 46 strains of bacteria from 60 Konnyaku tuber samples with soft rot symptoms from Xuanwei District. All strains grew on CVP medium, and produced iridescent, cross-hatched translucent colonies in deep, cuplike depressions or pits. All strains were facultatively anaerobic, gram-negative, straight rod-shaped cells with peritrichous flagella. All strains were catalase-positive, but oxidase-negative. They were able to ferment glucose, reduce nitrate, produce β-galactosidase and HS, and they utilized L-arabinose, D-galactose, D-glucose, glycerol, D-mannose, D-ribose, and sucrose, but did not produce urease, or acid from adonitol. Pectobacterium carotovorum subsp. carotovorum (syn. Erwina carotovora subsp. carotovora) has been commonly accepted as the causal agent of Konnyaku soft rot in Japan and China (1,3). Our studies also confirmed that P. carotovorum subsp. carotovorum caused Konnyaku soft rot, but the colony morphology and physiological and biochemical characteristics of these bacteria differed greatly from those of P. carotovorum subsp. carotovorum and other pectolytic Pectobacterium species. They grew at 37°C, caused potato soft rot, produced acid from melibiose, citrate, raffinose, and lactose, but did not produce acid from sorbitol and arabitol. The strain also utilized malonate but not keto-methyl glucoside as the sole carbon source. All strains were positive for phosphatase. Forty-one of 46 strains were sensitive to erythromycin. Thirty-seven of 46 strains produced indole. All tests were conducted with P. carotovorum subsp. carotovorum standard strain C2 isolated from Chinese cabbage as a positive control. Healthy Konnyaku tubers were inoculated with suspensions of the strains with a concentration of 10 CFU/ml in sterile water to confirm pathogenicity. After ~48 h, tuber rot symptoms were observed on all inoculated Konnyaku tubers. In comparison, there were no symptoms on tubers inoculated with sterile water. The bacterium was re-isolated from the infected Konnyaku tubers and identified as Dickeya dadantii, in accordance with Koch's postulates. All strains were confirmed by using the species-specific primers ERWFOR/CHRREV (2), which amplified a 450-bp DNA fragment by PCR assay. To our knowledge, this is the first report of Konnyaku soft rot caused by D. dadantii in China. References: (1) N. Hayashi. Gunma J. Agric. Res. A (Genera1) 5:25, 1988. (2) E. J. Smid et al. Plant Pathol. 44:1058, 1995. (3) J. Y. Tang et al. J. Yunnan Agric. Univ. 16:185, 2001.
魔芋(魔芋属)在中国一些农村地区作为重要经济作物种植。2011年,中国云南省宣威地区爆发了严重的魔芋软腐病。该病的特征是块茎部分或全部腐烂。在后期,软腐病可能向上蔓延至茎部,导致茎基部腐烂,整株植物倒伏。如果茎秆粗壮或足够大,软腐病可能只在茎的一侧发展,使另一侧在几天内保持健康。在这种情况下,茎不会倒伏,腐烂组织上可能会出现黄化现象。严重时,高达80%的植株会被感染。如果魔芋在同一块田地连续种植一年以上,病情会更加严重。最严重时,该病可导致整季作物绝收。2012年和2013年,我们从云南省宣威地区60份有软腐症状的魔芋块茎样本中分离出46株细菌。所有菌株都能在CVP培养基上生长,并在深层杯状凹陷或坑中产生彩虹色、交叉状半透明菌落。所有菌株都是兼性厌氧、革兰氏阴性、直杆状细胞,有周生鞭毛。所有菌株过氧化氢酶阳性,但氧化酶阴性。它们能够发酵葡萄糖、还原硝酸盐、产生β-半乳糖苷酶和H₂S,利用L-阿拉伯糖、D-半乳糖、D-葡萄糖、甘油、D-甘露糖、D-核糖和蔗糖,但不产生脲酶,也不能从阿东糖醇产酸。胡萝卜软腐果胶杆菌胡萝卜软腐亚种(同义词:胡萝卜欧文氏菌胡萝卜软腐亚种)在日本和中国普遍被认为是魔芋软腐病的病原菌(1,3)。我们的研究也证实了胡萝卜软腐果胶杆菌胡萝卜软腐亚种会引起魔芋软腐病,但这些细菌的菌落形态以及生理生化特性与胡萝卜软腐果胶杆菌胡萝卜软腐亚种和其他果胶分解果胶杆菌属的细菌有很大差异。它们在37°C下生长,能引起马铃薯软腐病,从蜜二糖、柠檬酸盐、棉子糖和乳糖产酸,但不能从山梨醇和阿拉伯糖醇产酸。该菌株还能利用丙二酸盐作为唯一碳源,但不能利用酮甲基葡糖苷。所有菌株磷酸酶呈阳性。46株菌株中有41株对红霉素敏感。46株菌株中有37株产生吲哚。所有试验均以从大白菜中分离出的胡萝卜软腐果胶杆菌胡萝卜软腐亚种标准菌株C2作为阳性对照。用浓度为10⁸CFU/ml的菌株无菌水悬浮液接种健康魔芋块茎以确认致病性。约48小时后,所有接种的魔芋块茎上都观察到块茎腐烂症状。相比之下,接种无菌水的块茎没有症状。根据柯赫氏法则,从感染的魔芋块茎中重新分离出该细菌,并鉴定为达旦氏果胶杆菌。所有菌株均使用种特异性引物ERWFOR/CHRREV(2)进行确认,通过PCR检测扩增出一个450bp的DNA片段。据我们所知,这是中国关于达旦氏果胶杆菌引起魔芋软腐病的首次报道。参考文献:(1)N. Hayashi。群马农业研究A(综合)5:25,1988。(2)E. J. Smid等人。植物病理学44:1058,1995。(3)J. Y. Tang等人。云南农业大学学报16:185,200