Fonseca M E N, Boiteux L S, Abreu H, Nogueira I, Pereira-Carvalho R C
Embrapa Hortaliças (CNPH), CP 218, 70359-970, Brasília-DF, Brazil.
Universidade de Brasília (UnB), Departamento de Fitopatologia, 70910-900, Brasília-DF, Brazil.
Plant Dis. 2013 May;97(5):692. doi: 10.1094/PDIS-06-12-0593-PDN.
Tomato chlorosis virus (ToCV) and Tomato infectious chlorosis virus (TICV) are the two Solanaceae-infecting Crinivirus species (family Closteroviridae) of worldwide importance. In Brazil, only ToCV has been detected under natural conditions infecting tomato (Solanum lycopersicum), sweet pepper (Capsicum annuum), and potato (S. tuberosum), causing foliar chlorosis (1, 3). However, there are no formal reports of alternative weed hosts of ToCV. During crop surveys in Capão Bonito, São Paulo State, Brazil (May 2011), a high incidence (above 20%) of plants of the weed, cut leaf ground cherry (Physalis angulata L.) growing around and within a tomato (cv. Alambra) field with a high incidence of ToCV, were found displaying interveinal chlorosis on the lower leaves, similar to those induced by magnesium deficiency. The P. angulata plants also had high populations of whiteflies (Bemisia tabaci biotype B). Ten leaf samples were taken from individual symptomatic ground cherry and tomato plants for Crinivirus testing. Total nucleic acids were extracted from 2 g of symptomatic and healthy leaf tissues of both hosts using Whatman CF-11 cellulose (Sigma) as described (4). The purified double stranded RNA samples were used as a template in reverse transcription (RT)-PCR using specific primers targeting the p22 gene region in the genome of ToCV (2). Only the 566-bp ToCV-specific amplicon was detected in all field samples. Sequences of samples from the P. angulata and tomato cDNA amplicons were identical to each other (GenBank Accession No. JX187514) and they showed 99% identity with the ToCV RNA 1 from a tomato isolate from Florida (AY903447). This confirmed the initial hypothesis of Crinivirus infection. Cuttings of symptomatic P. angulata plants were also obtained and kept in a voile cage under greenhouse conditions together with healthy seedlings of P. angulata and the begomovirus-resistant inbred tomato line 'TX-468RG.' Fifty aviruliferous B. tabaci (biotype B) adults were placed in the cage. Similar symptoms were observed 50 days after exposure to whiteflies in both hosts. Transmission to P. angulata and to 'TX-468RG' was also confirmed via sequencing of ToCV-specific amplicon, demonstrating the infectivity of the isolate to both hosts. To our knowledge, this is the first report of P. angulata as a natural host of ToCV in Brazil. This weed is often present in the commercial fields because of its natural tolerance to herbicides currently used in tomato production. The ToCV-infected P. angulata plants might serve as alternative sources of inoculum for the surrounding tomato fields. The environmental persistence of P. angulata combined with its intense whitefly colonization might allow a year-round ToCV exposure for tomato plants under field conditions in this major production area of Brazil where at least 25 million tomato plants are cultivated annually. References: (1) J. C. Barbosa et al. Trop. Plant Pathol. 36: 256, 2011. (2) M. I. Font et al. Plant Dis. 86:696, 2002. (3) D. M. S. Freitas et al. Plant Dis. 96:593, 2012. (4) R. A. Valverde et al. Plant Dis. 74:285, 1990.
番茄褪绿病毒(ToCV)和番茄传染性褪绿病毒(TICV)是两种感染茄科植物的重要的长线形病毒属病毒(长线形病毒科)。在巴西,在自然条件下仅检测到ToCV感染番茄(番茄)、甜椒(辣椒)和马铃薯(马铃薯),引起叶片褪绿(1,3)。然而,尚无关于ToCV替代杂草寄主的正式报道。在巴西圣保罗州卡庞博尼托进行作物调查期间(2011年5月),发现在一个ToCV发病率很高的番茄(品种Alambra)田块周围及田内生长的杂草——截叶酸浆(酸浆)植株发病率很高(超过20%),其下部叶片出现脉间褪绿,类似于缺镁引起的症状。截叶酸浆植株上烟粉虱(烟粉虱生物型B)数量也很多。从有症状的酸浆和番茄植株上分别采集了10片叶片样本进行长线形病毒属检测。按照所述方法(4),使用Whatman CF - 11纤维素(Sigma)从两种寄主植物2克有症状和健康的叶片组织中提取总核酸。纯化的双链RNA样本用作逆转录(RT)-PCR的模板,使用针对ToCV基因组中p22基因区域的特异性引物(2)。在所有田间样本中仅检测到566 bp的ToCV特异性扩增子。酸浆和番茄cDNA扩增子样本的序列彼此相同(GenBank登录号JX187514),并且与来自佛罗里达的番茄分离株的ToCV RNA 1显示99%的同一性(AY903447)。这证实了最初关于长线形病毒属感染的假设。还获取了有症状的酸浆植株的插条,并与酸浆健康幼苗和抗双生病毒的番茄自交系“TX - 468RG”一起置于温室条件下的纱笼中。将50只无毒烟粉虱(生物型B)成虫放入笼中。在两种寄主植物中,暴露于烟粉虱50天后观察到类似症状。通过对ToCV特异性扩增子进行测序,也证实了向酸浆和“TX - 468RG”的传播,证明该分离株对两种寄主均具有感染性。据我们所知,这是截叶酸浆作为ToCV在巴西的自然寄主的首次报道。由于这种杂草对目前番茄生产中使用的除草剂具有天然耐受性,它经常出现在商业田地中。感染ToCV的截叶酸浆植株可能成为周围番茄田的替代接种源。截叶酸浆在环境中的持久性及其密集的烟粉虱定殖可能使巴西这个主要产区的番茄植株在田间条件下全年都接触到ToCV。参考文献:(1)J.C. Barbosa等人,《热带植物病理学》36:256,2011年。(2)M.I. Font等人,《植物病害》86:696,2002年。(3)D.M.S. Freitas等人,《植物病害》96:593,2012年。(4)R.A. Valverde等人,《植物病害》74:285,1990年。