Costa H, Ventura J A, Jadão A S, Rezende J A M, Mello A P O A
INCAPER, 29052-010 Vitória, ES, Brazil.
Departamento de Fitopatologia e Nematologia, ESALQ/USP, 13418-900 Piracicaba, SP, Brazil.
Plant Dis. 2010 Aug;94(8):1066. doi: 10.1094/PDIS-94-8-1066A.
Watercress (Nasturtium officinale L.), a member of the family Brassicaceae, is consumed mainly as salad. Medicinal properties have also been attributed to this species. In Brazil, watercress is grown mainly by very small farmers. The crop is primarily seed propagated and growers can harvest several times per year in an established planting. Very few diseases have been reported in this crop worldwide. In Brazil, watercress infection by Cauliflower mosaic virus (CaMV) (3), Cucumber mosaic virus (CMV) (1), and an unidentified potyvirus (2) were previously reported. In January 2009, 80% of watercress plants, cv. Gigante Redondo, exhibiting severe mosaic, leaf size reduction, and plant stunting were observed in a crop in Marechal Floriano Municipality, State of Espírito Santo, Brazil. Preliminary leaf dip analysis by transmission electron microscopy revealed the presence of potyvirus-like particles. Sap from five infected plants reacted in plate-trapped antigen (PTA)-ELISA with polyclonal antiserum against Turnip mosaic virus (TuMV), but not with antiserum against CMV. Both antisera were produced in the Plant Virology Laboratory, ESALQ/USP. Mechanically inoculated watercress plants developed similar systemic mosaic symptoms. The virus was also transmitted to Nicotiana benthamiana, which exhibited severe mosaic and stunting. The presence of TuMV on these inoculated plants was confirmed by PTA-ELISA and reverse transcription (RT)-PCR. Total RNA extracted from infected and healthy watercress and infected N. benthamiana was analyzed by RT-PCR using specific pairs of primers flanking the coat protein gene of TuMV. Degenerated anti-sense (5'-t/caacccctt/gaacgcca/cagt/ca-3') and sense (5'-gcaggtgaa/gacg/acttgat/ca/gc-3') primers were designed after analysis to an alignment of the nucleotide sequences for five isolates of TuMV available in the GenBank (Accession Nos. NC_002509, D10927, EU680574, AB362513, and D88614). One fragment of 838 bp was amplified from samples in the infected plants, but not in the healthy controls. Two amplicons were purified and directly sequenced in both directions. Comparisons of the 731-bp consensus nucleotide sequence (Accession No. HM008961) to several other isolates of TuMV revealed 94 to 95% identity in the coat protein region. To our knowledge, this is the first report of TuMV in watercress in Brazil. Management of the disease should include propagation by seeds instead of vegetative parts of the plants and rouging of diseased plants to prevent mechanical transmission during successive harvestings. References: (1) A. J. Boari et al. Fitopatol. Bras. 25:438, 2000. (2) A. J. Boari et al. Fitopatol. Bras. 27:S200, 2002. (3) M. L. R. Z. C. Lima et al. Fitopatol. Bras. 9:403, 1984.
豆瓣菜(Nasturtium officinale L.)是十字花科的一员,主要作为沙拉食用。该物种也具有药用特性。在巴西,豆瓣菜主要由非常小的农户种植。这种作物主要通过种子繁殖,种植者在已定植的地块上每年可以收获数次。全球范围内,关于这种作物的病害报道很少。在巴西,此前曾报道过豆瓣菜感染花椰菜花叶病毒(CaMV)(3)、黄瓜花叶病毒(CMV)(1)以及一种未鉴定的马铃薯Y病毒(2)。2009年1月,在巴西圣埃斯皮里图州马雷夏尔弗洛里亚诺市的一块地里,观察到80%的“Gigante Redondo”品种豆瓣菜植株出现严重花叶、叶片变小和植株矮化的症状。通过透射电子显微镜进行的初步叶片浸渍分析显示存在马铃薯Y病毒样颗粒。来自五株受感染植株的汁液在板条捕获抗原(PTA)-ELISA中与抗芜菁花叶病毒(TuMV)的多克隆抗血清发生反应,但与抗CMV的抗血清不发生反应。这两种抗血清均由ESALQ/USP植物病毒学实验室制备。机械接种的豆瓣菜植株出现了类似的系统花叶症状。该病毒也传播到了本氏烟草上,本氏烟草表现出严重花叶和矮化症状。通过PTA-ELISA和逆转录(RT)-PCR证实了这些接种植株上存在TuMV。使用位于TuMV外壳蛋白基因两侧的特异性引物对,通过RT-PCR对从受感染和健康的豆瓣菜以及受感染的本氏烟草中提取的总RNA进行了分析。在对GenBank中可获得的五个TuMV分离株(登录号分别为NC_002509、D10927、EU680574、AB362513和D88614)的核苷酸序列进行比对分析后,设计了简并反义引物(5'-t/caacccctt/gaacgcca/cagt/ca-3')和正义引物(5'-gcaggtgaa/gacg/acttgat/ca/gc-3')。从受感染植株的样本中扩增出了一个838 bp的片段,而在健康对照样本中未扩增出。对两个扩增产物进行了纯化并双向直接测序。将731 bp的共有核苷酸序列(登录号为HM008961)与其他几个TuMV分离株进行比较,发现在外壳蛋白区域的同一性为94%至95%。据我们所知,这是巴西首次报道豆瓣菜感染TuMV。该病的防治措施应包括通过种子而非植株的营养部分进行繁殖,以及拔除病株以防止在连续收获期间的机械传播。参考文献:(1)A. J. Boari等人,《巴西植物病理学》25:438,2000年。(2)A. J. Boari等人,《巴西植物病理学》27:S200,2002年。(3)M. L. R. Z. C. Lima等人,《巴西植物病理学》9:403,1984年。