Ngadze E, Coutinho T A, van der Waals J E
Department of Crop Science, University of Zimbabwe, P.O. Box Mp167, Mount Pleasant, Harare, Zimbabwe and University of Pretoria, Department of Microbiology and Plant Pathology, Pretoria 0002, South Africa.
Department of Microbiology and Plant Pathology, Forestry and Agricultural Institute (FABI), University of Pretoria, Pretoria 0002, South Africa.
Plant Dis. 2010 Oct;94(10):1263. doi: 10.1094/PDIS-05-10-0361.
A survey was carried out in the potato- (Solanum tuberosum L.) growing regions of Zimbabwe in April 2009 to assess the prevalence of bacterial soft rot. A total of 125 tubers with soft rot symptoms were collected. The disease caused severe economic losses ranging from 20 to 60% on tubers in the field and in storage. Affected tubers had symptoms that ranged from light vascular discoloration to complete seed piece decay. Infected tuber tissue was often cream colored and soft to the touch. In the field, plants showed severe wilting, often accompanied by a slimy, brown necrosis of the lower stems. Seventy-five of 125 isolations from diseased tubers yielded pectolytic bacteria on crystal violet pectate (CVP) medium and colonies were characterized after purification on King's B medium. All 75 isolates were gram-negative rods, oxidase negative, facultatively anaerobic, able to degrade pectate, and rot potato slices. They grew at 37°C, were sensitive to erythromycin, positive for phosphatase, indole production, cis-aconitate, lactose, d-arabinose, meso-tartrate, casein, d-melibiose, myo-inositol, and malonate utilization, while negative for acid production from trehalose, inuline, and α-methyl glucose. Dickeya dadantii (Erwinia chrysanthemi 3937 from the Scottish Research Institute) was included in all biochemical and pathogenicity tests. These characteristics are typical for two species, D. zeae and D. dadantii (2). Thus, the 75 isolates were further identified by PCR amplification with BOX and REP primers (3) and five isolates by gyrB sequence analysis (1). These analyses give support for the isolates being D. dadantii. Partial gyrB sequence analysis showed that the analyzed isolates had 96% sequence identity with the D. dadantii type strain Ech 586 (GenBank Accession No. CP001836.1). One-microliter suspensions (10 CFU per ml) of 20 samples were injected into the stolon end of potato tubers (S. tuberosum L.) cv. BP1. Each isolate was inoculated into three tubers, which were maintained at 25°C. Three control tubers were inoculated with sterile distilled water. Soft rot symptoms identical to those observed in the field and in storage appeared on all inoculated tubers 1 to 2 days after inoculation but not on the control tubers. A bacterium with identical characteristics to those described above was consistently reisolated from the rotted tissue of inoculated tubers. To our knowledge, this is the first report of soft rot on potato in Zimbabwe caused by D. dadantii, formerly referred to as E. chrysanthemi. This finding has implications for import and export of potato material into and out of Zimbabwe. Zimbabwe imports seed from various countries because of the current seed shortage and exports table potatoes to other African states. References: (1) C. Brady et al. Int. J. Syst. Evol. Micobiol. 59:2339, 2009. (2) R. Samson et al. Int. J. Syst. Evol. Microbiol. 55:1415, 2005. (3) J. Versalovic et al. 1991. Nucleic Acids Res. 19:6823, 1991.
2009年4月,在津巴布韦的马铃薯(Solanum tuberosum L.)种植区开展了一项调查,以评估细菌性软腐病的流行情况。共收集了125个有软腐症状的块茎。该病害在田间和储存期造成了20%至60%的严重经济损失。受影响的块茎症状从轻微的维管束变色到整个种薯腐烂不等。受感染的块茎组织通常呈奶油色,触摸起来很软。在田间,植株表现出严重萎蔫,常伴有下部茎部的黏液状褐色坏死。从125个患病块茎中分离出的75个菌株在结晶紫果胶酸盐(CVP)培养基上产生了果胶分解菌,并在King's B培养基上纯化后对菌落进行了鉴定。所有75个分离株均为革兰氏阴性杆菌,氧化酶阴性,兼性厌氧,能够降解果胶酸盐并使马铃薯切片腐烂。它们在37°C下生长,对红霉素敏感,磷酸酶、吲哚产生、顺乌头酸盐、乳糖、d -阿拉伯糖、内消旋酒石酸盐、酪蛋白、d -蜜二糖、肌醇和丙二酸盐利用呈阳性,而海藻糖、菊粉和α -甲基葡萄糖产酸呈阴性。在所有生化和致病性试验中都包括了达旦迪基氏菌(Dickeya dadantii,来自苏格兰研究所的菊花欧文氏菌3937)。这些特征是zeae达旦迪基氏菌和达旦迪基氏菌两个物种的典型特征(2)。因此,通过用BOX和REP引物进行PCR扩增(3)对75个分离株进行了进一步鉴定,并用gyrB序列分析(1)对5个分离株进行了鉴定。这些分析支持这些分离株为达旦迪基氏菌。gyrB部分序列分析表明,所分析的分离株与达旦迪基氏菌模式菌株Ech 586(GenBank登录号CP001836.1)具有96%的序列同一性。将20个样品的1微升悬浮液(每毫升10 CFU)注射到马铃薯(S. tuberosum L.)品种BP1的块茎匍匐茎端。每个分离株接种到3个块茎中,将其置于25°C下。3个对照块茎接种无菌蒸馏水。接种后1至2天,所有接种的块茎都出现了与田间和储存期观察到的相同的软腐症状,但对照块茎没有出现。从接种块茎的腐烂组织中持续重新分离出一种具有与上述相同特征的细菌。据我们所知,这是津巴布韦首次关于由达旦迪基氏菌(以前称为菊花欧文氏菌)引起的马铃薯软腐病的报道。这一发现对津巴布韦马铃薯材料的进出口有影响。由于目前种子短缺,津巴布韦从各个国家进口种子,并向其他非洲国家出口食用马铃薯。参考文献:(1)C. Brady等人,《国际系统与进化微生物学杂志》59:2339,2009年。(2)R. Samson等人,《国际系统与进化微生物学杂志》55:1415,2005年。(3)J. Versalovic等人,1991年,《核酸研究》19:6823,1991年。