Perry K L, Miller L, Williams L
Department of Plant Pathology, 334 Plant Science Bldg. Cornell University, Ithaca, NY 14853.
Plant Dis. 2005 Mar;89(3):340. doi: 10.1094/PD-89-0340C.
Impatiens necrotic spot virus (INSV; genus Tospovirus) was detected in experimental greenhouse-grown potatoes (Solanum tuberosum) and Nicotiana benthamiana in New York State in July and August of 2003 and 2004. Potato leaves exhibiting necrotic lesions with a concentric pattern similar to those induced by Tomato spotted wilt virus (1) were observed on cvs. Atlantic, Huckleberry, NY115, and Pentland Ivory. The presence of INSV was confirmed using double-antibody sandwich enzyme-linked immunosorbent assay and a rapid 'ImmunoStrip' assay (Agdia, Inc., Elkhart, IN). INSV-specific sequences were amplified from total RNA extracts using reverse transcription-polymerase chain reaction with 'Tospovirus Group' primers (Agdia, Inc.) and two independently amplified DNAs were sequenced. A common sequence of 355 nucleotides (GenBank Accession No. AY775324) showed 98% identity to coding sequences in an INSV L RNA. The virus was mechanically transmitted to potato and N. benthamiana and could be detected in asymptomatic, systemically infected potato leaves. Stems nodes and leaves were removed from infected potato plants, and sterile in vitro plantlets were established (2). None of the regenerated in vitro plantlets of cvs. Pentland Ivory (6 plantlets) or NY115 (5 plantlets) were infected with INSV. Two of ten regenerated cv. Atlantic plantlets initially tested positive, but INSV could not be detected after 6 months in tissue culture. In vitro tissue culture plantlets could not be established from infected cv. Huckleberry plants, even though they were consistently obtained from uninfected plants. Infected greenhouse plants were grown to maturity and the tubers harvested, stored for 6 months at 4°C, and replanted in the greenhouse. INSV could not be detected in plants from 26 cv. Huckleberry, 4 cv. NY115, or 4 cv. Atlantic tubers. Although this isolate of INSV was able to systemically infect potato, it was not efficiently maintained or transmitted to progeny tubers. This might explain why INSV has not been reported as a problem in potato production. Lastly, in both years, dying N. benthamiana provided the first sign of a widespread greenhouse infestation of INSV in a university facility housing ornamental and crop plants. INSV induced a systemic necrosis in N. benthamiana, and this host may be useful as a sensitive 'trap' plant indicator for natural infections in greenhouse production. References: (1) T. L. German. Tomato spotted wilt virus. Pages 72-73 in: Compendium of Potato Diseases. W. R. Stevenson et al., eds. The American Phytopathological Society, St. Paul, 2001. (2) S. A. Slack and L. A. Tufford. Meristem culture for virus elimination. Pages 117-128 in: Fundamental Methods of Plant Cell, Tissue and Organ Culture and Laboratory Operations. O. L. Gamborg and G. C. Philips, eds. Springer-Velag, Berlin, 1995.
2003年7月和8月以及2004年7月和8月,在纽约州实验温室种植的马铃薯(Solanum tuberosum)和本氏烟草(Nicotiana benthamiana)中检测到凤仙坏死斑点病毒(INSV;番茄斑萎病毒属)。在大西洋、哈克贝利、NY115和彭特兰象牙等品种的马铃薯叶片上观察到出现坏死病斑,病斑呈同心状,类似于番茄斑萎病毒诱导产生的病斑(1)。使用双抗体夹心酶联免疫吸附测定法和快速“免疫试纸”测定法(Agdia公司,印第安纳州埃尔克哈特)确认了INSV的存在。使用“番茄斑萎病毒属”引物(Agdia公司)通过逆转录 - 聚合酶链反应从总RNA提取物中扩增出INSV特异性序列,并对两个独立扩增的DNA进行了测序。一段355个核苷酸的共同序列(GenBank登录号AY775324)与INSV L RNA中的编码序列具有98%的同一性。该病毒通过机械接种传播到马铃薯和本氏烟草上,并且在无症状的、系统感染的马铃薯叶片中能够被检测到。从感染的马铃薯植株上切下茎节和叶片,并建立了无菌离体苗(2)。彭特兰象牙品种(6株)和NY115品种(5株)的再生离体苗均未感染INSV。10株再生的大西洋品种离体苗中有2株最初检测呈阳性,但在组织培养6个月后未检测到INSV。即使从未感染的哈克贝利品种植株上能持续获得离体苗,但从感染的哈克贝利品种植株上却无法建立离体组织培养苗。将感染的温室植株培育至成熟,收获块茎,在4°C下储存6个月,然后重新种植到温室中。在26个哈克贝利品种、4个NY115品种或4个大西洋品种的块茎长出的植株中均未检测到INSV。尽管这种INSV分离株能够系统感染马铃薯,但它不能有效地维持或传播给后代块茎。这可能解释了为什么INSV在马铃薯生产中未被报道为一个问题。最后,在这两年中,濒死的本氏烟草是一所种植观赏植物和农作物的大学设施中INSV在温室广泛侵染的首个迹象。INSV在本氏烟草中诱导产生系统性坏死,并且这种寄主植物可能作为温室生产中自然感染的敏感“诱捕”植物指示物。参考文献:(1)T. L. 杰曼。番茄斑萎病毒。载于《马铃薯病害汇编》第72 - 73页。W. R. 史蒂文森等人编。美国植物病理学会,圣保罗,2001年。(2)S. A. 斯拉克和L. A. 塔福德。用于病毒消除的分生组织培养。载于《植物细胞、组织和器官培养及实验室操作的基本方法》第117 - 128页。O. L. 甘伯格和G. C. 菲利普斯编。施普林格 - 维拉格出版社,柏林,1995年。