Jibrin M O, Timilsina S, Potnis N, Minsavage G V, Shenge K C, Akpa A D, Alegbejo M D, Beed F, Vallad G E, Jones J B
Department of Crop Protection, Ahmadu Bello University, Zaria, Nigeria.
Plant Pathology Department, University of Florida, Gainesville 32611.
Plant Dis. 2014 Oct;98(10):1426. doi: 10.1094/PDIS-06-14-0586-PDN.
Bacterial spot (BS) has been reported as an important disease on pepper in Nigeria (4). Xanthomonas campestris pv. vesicatoria was identified as the causal agent using phenotypic and pathogenicity tests; however, X. campestris pv. vesicatoria is a synonym for two genetically distinct groups that have been elevated to the species X. euvesicatoria and X. vesicatoria (2). Furthermore, the latter two species and X. gardneri cause similar diseases on pepper (2). In order to determine the species associated with BS on pepper, leaves with irregular, dark brown lesions were collected from pepper plants in fields from northwestern Nigeria, and isolations were made on nutrient agar (NA). Yellow, mucoid colonies typical of Xanthomonas were isolated. Six strains isolated from pepper were determined to be non-amylolytic. For race determinations, bacterial suspensions of the pepper strains, derived from 24-h cultures grown on NA at 28°C, were adjusted to 10 CFU/ml and infiltrated into leaves of tomato and pepper differential genotypes (5). The six pepper strains elicited HRs on the tomato differential genotypes. The strains produced a susceptible reaction on all pepper differentials and were designated as pepper race 6 (5). Multilocus sequence analysis (MLSA) using six housekeeping genes (fusA, lacF, gyrB, gltA, gapA, and lepA) was used to further analyze the strains (1) (GenBank Accession Nos. KJ938585 to KJ938587, KJ938592 to KJ938594, KJ938599 to KJ938601, KJ938606 to KJ938608, KJ938633 to KJ938635, and KJ938640 to KJ938642). A partial sequence of hrpB2 was also sequenced since the four Xanthomonas species associated with BS can be differentiated based on sequence divergence (3) (KJ938622 to KJ938627). The housekeeping gene sequences were aligned along with other Xanthomonas sequences imported from the NCBI database using muscle tool from MEGA software, 5.2.2. Maximum likelihood phylogenetic trees constructed for the six housekeeping gene sequences individually and in concatenation revealed that the Nigerian pepper strains were identical to the X. euvesicatoria reference strain 85-10. Although BS is common in Nigeria, to our knowledge, this represents the first report for this pepper pathogen in Nigeria. References: (1) N. F. Almeida et al. Phytopathology 100:208, 2010. (3) J. B. Jones et al. System Appl. Microbiol. 27:755, 2004. (4) A. Obradovic et al. Eur. J. Plant Pathol. 88:736, 2004. (2) E. U. Opara and F. J. Odibo. J. Mol. Gen. 1:35, 2009. (5) R. E. Stall et al. Ann. Rev. Phytopathol. 47:265, 2009.
细菌性斑点病(BS)在尼日利亚被报道为辣椒上的一种重要病害(4)。通过表型和致病性测试,野油菜黄单胞菌疮痂致病变种被鉴定为致病因子;然而,野油菜黄单胞菌疮痂致病变种是两个遗传上不同的群体的同义词,这两个群体已被提升为黄单胞菌新种和疮痂黄单胞菌(2)。此外,后两个物种和加德纳黄单胞菌在辣椒上引起相似的病害(2)。为了确定与辣椒上的细菌性斑点病相关的物种,从尼日利亚西北部田间的辣椒植株上采集了带有不规则深褐色病斑的叶片,并在营养琼脂(NA)上进行分离培养。分离出了典型的黄单胞菌黄色、黏液状菌落。从辣椒中分离出的6个菌株被确定为非淀粉酶分解型。为了进行小种鉴定,将在28°C下于NA上培养24小时的辣椒菌株的细菌悬液调整至10 CFU/ml,并接种到番茄和辣椒鉴别基因型的叶片中(5)。这6个辣椒菌株在番茄鉴别基因型上引发了过敏反应。这些菌株在所有辣椒鉴别品种上产生了感病反应,并被指定为辣椒小种6(5)。使用6个管家基因(fusA、lacF、gyrB、gltA、gapA和lepA)进行多位点序列分析(MLSA)以进一步分析这些菌株(1)(GenBank登录号:KJ938585至KJ938587、KJ938592至KJ938594、KJ938599至KJ938601、KJ938606至KJ938608、KJ938633至KJ938635以及KJ938640至KJ938642)。还对hrpB2的部分序列进行了测序,因为与细菌性斑点病相关的4个黄单胞菌物种可以根据序列差异进行区分(3)(KJ938622至KJ938627)。使用MEGA软件5.2.2中的muscle工具将管家基因序列与从NCBI数据库导入的其他黄单胞菌序列进行比对。分别为6个管家基因序列单独构建以及串联构建的最大似然系统发育树表明,尼日利亚辣椒菌株与黄单胞菌新种参考菌株85 - 10相同。尽管细菌性斑点病在尼日利亚很常见,但据我们所知,这是该辣椒病原菌在尼日利亚的首次报道。参考文献:(1)N. F. Almeida等人,《植物病理学》100:208,2010。(3)J. B. Jones等人,《系统与应用微生物学》27:755,2004。(4)A. Obradovic等人,《欧洲植物病理学杂志》88:736,2004。(2)E. U. Opara和F. J. Odibo,《分子遗传学杂志》1:35,2009。(5)R. E. Stall等人,《植物病理学年度评论》47:265,2009。