Muzhinji N, Woodhall J W, Truter M, van der Waals J E
Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria 0002, South Africa and Tobacco Research Board, Zimbabwe.
The Food and Environment Research Agency, Sand Hutton, York, Y041 1LZ, UK.
Plant Dis. 2014 Apr;98(4):570. doi: 10.1094/PDIS-08-13-0815-PDN.
Rhizoctonia solani consists of 13 anastomosis groups (AGs) designated AG1 to 13. AG3-PT is considered the predominant AG in potatoes (4) and is associated with quantitative and qualitative yield losses. Qualitative losses are typically associated with the tuber blemish disease, black scurf. However, atypical tuber blemishes such as elephant hide consisting of corky lesions on the tuber surface (2) have also been attributed to Rhizoctonia. Such atypical blemishes are not considered specific to Rhizoctonia, making direct-cause effect estimates difficult (1). Koch's postulates for the elephant hide symptom and R. solani AG3-PT have not been completed. Recently, growth cracking and scab lesions were observed on potato tubers in South Africa and attributed to a new Streptomyces species (3). These lesions and cracks were similar to elephant hide symptoms attributed to R. solani AG3-PT. Therefore, the cause of the elephant hide symptom in South Africa was investigated further. Symptoms of elephant hide and cracking have been observed on tubers from the Eastern Free State, KwaZulu-Natal, Limpopo, Mpumalanga, North-Eastern Cape, Northern Cape, North West, Sandveld, and Western Free State growing regions. In 2012, three samples of potato tubers (cv. BP1) with elephant hide and cracking were selected for analysis. These samples were collected from Clanwilliam in the Sandveld potato growing region. Tubers were surface sterilized with 1% NaOCl; sections of affected tissue were excised and plated onto potato dextrose agar (PDA). Rhizoctonia-like colonies were identified and after further sub-culturing on PDA, three representative isolates (Rh3, Rh4, and Rh6) of R. solani from each sample were obtained. For each isolate, genomic DNA was extracted and the rDNA ITS region sequenced using ITS1-F and ITS4 (2). The resulting sequences (KF234142, KF234143, and KF234144) were at least 98% identical to other AG3-PT sequences on GenBank (JX27814 and KC157664). To confirm Koch's postulates, pathogenicity tests were conducted with the three isolates. PDA plugs of each isolate were added to 10 g of barley grains which were incubated for 14 days until fully colonized. The barley grains were then used to inoculate disease-free mini-tubers (cv. BP1) in 5l pots containing a sand-clay-pine bark mixture (1:1:1 ratio). Potato plants inoculated with sterile barley grains served as controls. Plants were held for 120 days in a greenhouse at 22°C with light for 12 h a day. Incidence of the elephant hide symptom for isolates Rh3, Rh4, and Rh6 was 58%, 33%, and 37.5%, respectively. Growth cracking and black scurf were also observed with each isolate. R. solani AG3-PT was successfully re-isolated from symptomatic tubers, confirming Koch's postulates. This is the first report of R. solani AG3-PT causing elephant hide in potato tubers in South Africa. Elephant hide caused by R. solani AG3-PT has been reported in tubers from France (2) and the United Kingdom (3), but Koch's postulates were not proven. In this study, Koch's postulates were proven for R. solani AG3-PT causing scab or elephant hide symptom and cracking in potato tubers. R. solani AG3-PT should thus be considered in addition to Streptomyces as a cause of this symptom and control strategies should also consider R. solani AG3-PT. References: (1) G. J. Banville et al. Pages 321-330 in: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control, B. Sneh et al., eds. Kluwer Academic Publishers, Dordrecht, The Netherlands, 1996. (2) M. Fiers et al. Eur. J. Plant. Pathol. 128:353, 2010. (3) R. Gouws and A. McLeod. Plant Dis. 96:1223, 2012. (4) J. W. Woodhall et al. Eur. J. Plant. Pathol. 136:273, 2013.
立枯丝核菌由13个融合群(AGs)组成,分别命名为AG1至AG13。AG3-PT被认为是马铃薯中的主要融合群(4),与产量的定量和定性损失有关。定性损失通常与块茎病害、黑痣病有关。然而,非典型的块茎瑕疵,如块茎表面由木栓化病斑组成的象皮状瑕疵(2),也被归因于立枯丝核菌。这种非典型瑕疵并不被认为是立枯丝核菌特有的,因此难以直接估计其致病作用(1)。针对象皮状症状和立枯丝核菌AG3-PT的柯赫氏法则尚未完成。最近,在南非的马铃薯块茎上观察到生长开裂和疮痂病斑,并归因于一种新的链霉菌物种(3)。这些病斑和裂缝与归因于立枯丝核菌AG3-PT的象皮状症状相似。因此,对南非象皮状症状的病因进行了进一步调查。在东自由邦、夸祖鲁-纳塔尔、林波波、姆普马兰加、东北开普、北开普、西北、桑德维尔德和西自由邦等种植区的块茎上都观察到了象皮状和开裂症状。2012年,选取了三个带有象皮状和开裂症状的马铃薯块茎样本(品种BP1)进行分析。这些样本采自桑德维尔德马铃薯种植区的克兰威廉。块茎用1%的次氯酸钠进行表面消毒;切除受影响组织的切片,接种到马铃薯葡萄糖琼脂(PDA)上。鉴定出类似立枯丝核菌的菌落,在PDA上进一步继代培养后,从每个样本中获得了立枯丝核菌的三个代表性分离株(Rh3、Rh4和Rh6)。对于每个分离株,提取基因组DNA,并使用ITS1-F和ITS4对rDNA ITS区域进行测序(2)。所得序列(KF234142、KF234143和KF234144)与GenBank上的其他AG3-PT序列(JX27814和KC157664)至少有98%的同一性。为了证实柯赫氏法则,对这三个分离株进行了致病性测试。将每个分离株的PDA菌块添加到10克大麦粒中,培养14天直至完全定殖。然后用这些大麦粒接种5升花盆中无菌的微型块茎(品种BP1),花盆中装有沙-粘土-松树皮混合物(比例为1:1:1)。接种无菌大麦粒的马铃薯植株作为对照。植株在22°C、每天光照12小时的温室中培养120天。分离株Rh3、Rh4和Rh6引起的象皮状症状的发病率分别为58%、33%和37.5%。每个分离株还观察到生长开裂和黑痣病。从有症状的块茎中成功重新分离出立枯丝核菌AG3-PT,证实了柯赫氏法则。这是立枯丝核菌AG3-PT在南非马铃薯块茎上引起象皮状症状的首次报道。在法国(2)和英国(3)的块茎中曾报道过由立枯丝核菌AG3-PT引起的象皮状症状,但未证实柯赫氏法则。在本研究中,证实了立枯丝核菌AG3-PT可引起马铃薯块茎的疮痂病或象皮状症状及开裂的柯赫氏法则。因此,除链霉菌外,还应考虑立枯丝核菌AG3-PT是这种症状的病因,控制策略也应考虑立枯丝核菌AG3-PT。参考文献:(1)G. J. Banville等人。载于:《立枯丝核菌物种:分类学、分子生物学、生态学、病理学和病害控制》,B. Sneh等人编。Kluwer学术出版社,荷兰多德雷赫特,1996年,第321 - 330页。(2)M. Fiers等人。《欧洲植物病理学杂志》128:353,2010年。(3)R. Gouws和A. McLeod。《植物病害》96:1223,2012年。(4)J. W. Woodhall等人。《欧洲植物病理学杂志》136:273,2013年。