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榛树(榛子)中三叶草黄边植原体的首次报道。

First Report of Clover Yellow Edge Phytoplasma in Corylus (Hazelnut).

作者信息

Jomantiene R, Postman J D, Montano H G, Maas J L, Davis R E, Johnson K B

机构信息

USDA-ARS, Fruit Laboratory, Beltsville, MD 20705, and Institute of Botany, Vilnius, Lithuania.

USDA-ARS, National Clonal Germplasm Repository, Corvallis, OR 97333.

出版信息

Plant Dis. 2000 Jan;84(1):102. doi: 10.1094/PDIS.2000.84.1.102B.

Abstract

During investigations into the cause of a stunt syndrome affecting cultivated European hazelnut trees (Corylus avellana L.) in Oregon, the clover yellow edge (CYE) phytoplasma was detected for the first time in this crop. The cause of hazelnut stunt syndrome (HSS) is unknown, but the disease has been transmitted by grafting and apparently has moved within orchards through root grafts (1). Severely affected trees persist for many years, but their nut production is greatly reduced. Previous attempts to detect viruses, bacteria, and other pathogens have been unsuccessful. HSS has been observed only in Oregon and already had been present for more than 10 years when it was first reported in 1970 (1). In June, 1999, leaf samples were collected from two affected and two apparently healthy (symptomless) hazelnut trees in a field plot at Oregon State University, Corvallis, and from a healthy greenhouse-grown tree. Leaf samples were sent to the USDA Beltsville, MD, laboratory, where they were assessed for phytoplasma infection, using nested polymerase chain reactions (PCRs). PCRs were primed by phytoplasma universal primer pairs P1/P7 and F2n/R2 (3) for amplification of phytoplasma 16S ribosomal (r) DNA (16S rRNA gene) sequences according to the procedures of Gunderson and Lee (2). Phytoplasma-characteristic 1.2-kbp DNA sequences were amplified from all field-tree samples. No DNA sequences were amplified from samples of the greenhouse-grown tree. Restriction fragment length polymorphism patterns of rDNA digested with AluI, KpnI, HhaI, HaeIII, HpaII, MseI, RsaI, and Sau3A1 endonucleases indicated that all diseased hazelnut trees as well as symptomless field trees were infected by a phytoplasma classified in group 16SrIII (peach X-disease group), subgroup B (III-B, type strain CYE phytoplasma). No phytoplasmas were detected in samples from the greenhouse-grown tree. Nucleotide sequences were determined for 16Sr DNA fragments amplified from the hazelnut CYE phytoplasma in nested PCRs primed with F2n/R2. The sequences were deposited in GenBank under Accession no. AF189288. Sequence similarity between 16Sr DNAs of the hazelnut CYE strain (CYE-Or) and the Canadian clover yellow edge strain (CYE-C, GenBank Accession no. AF175304) phytoplasma was 99.9%. Decline and yellows disorders of hazelnut in Germany and Italy have been associated with infections by apple proliferation, pear decline, and European stone fruit yellows phytoplasmas (4). These phytoplasmas are classified in 16Sr group X, the apple proliferation group of phytoplasmas. This is the first report of the CYE phytoplasma infecting Corylus. References: (1) H. R. Cameron. Plant Dis. Rep. 54:69, 1970. (2) D. E. Gunderson and I.-M. Lee. Phytopathol. Mediterr. 35:144, 1996. (3) R. Jomantiene et al. HortScience 33:1069, 1998. (4) C. Marcone et al. Plant Pathol. 45:857,1996.

摘要

在对俄勒冈州种植的欧洲榛树(Corylus avellana L.)出现的一种矮化综合征病因进行调查期间,首次在这种作物中检测到三叶草黄边(CYE)植原体。榛树矮化综合征(HSS)的病因尚不清楚,但该病可通过嫁接传播,并且显然已在果园内通过根嫁接传播(1)。严重受影响的树木会持续多年,但坚果产量会大幅下降。此前检测病毒、细菌和其他病原体的尝试均未成功。HSS仅在俄勒冈州被观察到,1970年首次报道时它已存在了10多年(1)。1999年6月,从科瓦利斯俄勒冈州立大学一块田地中的两棵受影响和两棵明显健康(无症状)的榛树上以及一棵健康的温室培育树上采集了叶片样本。叶片样本被送往位于马里兰州贝尔茨维尔的美国农业部实验室,在那里使用巢式聚合酶链反应(PCR)评估植原体感染情况。根据冈德森和李(2)的方法,使用植原体通用引物对P1/P7和F2n/R2(3)对PCR进行引物设计,以扩增植原体16S核糖体(r)DNA(16S rRNA基因)序列。从所有田间树木样本中扩增出了具有植原体特征的1.2-kbp DNA序列。从温室培育树的样本中未扩增出DNA序列。用AluI、KpnI、HhaI、HaeIII、HpaII、MseI、RsaI和Sau3A1核酸内切酶消化rDNA后的限制性片段长度多态性模式表明,所有患病榛树以及无症状田间树均被分类为第16SrIII组(桃X病组)、亚组B(III-B,模式菌株CYE植原体)的植原体感染。在温室培育树的样本中未检测到植原体。对用F2n/R2引物进行巢式PCR扩增得到的榛树CYE植原体1.6Sr DNA片段测定了核苷酸序列。这些序列已存入GenBank,登录号为AF189288。榛树CYE菌株(CYE-Or)与加拿大三叶草黄边菌株(CYE-C,GenBank登录号为AF175304)植原体的16Sr DNA之间的序列相似性为99.9%。德国和意大利榛树的衰退和黄化病症与苹果增殖、梨衰退和欧洲核果黄化植原体感染有关(4)。这些植原体被分类为第16Sr组X,即植原体的苹果增殖组。这是关于CYE植原体感染榛树的首次报道。参考文献:(1)H. R. 卡梅隆。植物病害报告54:69,1970年。(2)D. E. 冈德森和I.-M. 李。地中海植物病理学35:144,1996年。(3)R. 乔曼蒂内等人。园艺科学33:1069,1998年。(4)C. 马尔科内等人。植物病理学45:857,1996年。

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