四种植原体病害的鉴定
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摘要
本研究采用分子生物学方法,对引起落地生根和柳树花变叶、黄栌顶枯病和榆树黄化病的植原体进行了鉴定,主要结果如下:
利用针对植原体的通用引物,通过巢式PCR从表现花变叶症状的落地生根植株中扩增到了植原体16S rRNA、核糖体蛋白基因(rp)和延伸因子Tu基因(tuf),序列分析结果表明,该植原体属于翠菊黄化组。在电镜下观察到了植原体粒子,嫁接实验证明植原体是落地生根花变叶的病原。
通过巢式PCR证明自然表现花变叶症状的柳树中含有植原体。16SrRNA序列的结果表明与柳树花变叶相关的植原体属于翠菊黄化组16SrI-B亚组,根据rp序列分析的结果,该植原体属于AY组rp I-D亚组。
证明黄栌顶枯病是由植原体引起。16S rRNA序列比较结果表明该植原体(CSDP)属于16Sr I-B亚组。通过比较CSDP与GenBank中登陆的16Sr I组12个16S rRNA序列,进一步明确CSDP株系属于16Sr I-B亚组。黄栌木虱可能是黄栌顶枯病的昆虫介体。
从榆树黄化植株中扩增到了植原体。16S rRNA序列分析的结果表明中国榆树黄化植原体属于EY组的16Sr V-B亚组。
本实验中的落地生根花变叶病、柳树花变叶病、黄栌顶枯病是国内外首次报道,榆树黄化病是国内首次报道。
The phytoplasmas associated with air-plant phyllody,willow phyllody, common smoketree dieback and Chinese elm yellows were identified at molecular levels.The main results read as follows:
The 16S rRNA gene,ribosomal protein gene(rp) and elongation factor Tu(EF-Tu) (tuf gene) of the phytoplasma associating with air-plant phyllody were amplified with normal or nested-PCR.Sequence analysis results show that the phytoplasma belongs to aster yellows group.The phytoplasma particles were observed under electron microscope.Graft experiment also conformed that air-plant phyllody is caused by phytoplasma.
Phytoplasma of aster yellows group was also detected from willow plants showing symptom of phyllody.The associating phytoplasma belongs to 16Sr I-B according to 16S rRNA sequence and belongd to rp I-D according to rp gene sequence.
Common smoketree dieback is caused by phytoplasma.Blast results of 16S rRNA sequences showed that the phytoplasma belongs to subgroup 16Sr I-B. Alignment results of this and other twelve 16S phytoplasmal rRNA that the corresponding phytoplasma belongs to subgroup 16Sr I-B.Psyllas inhabiting on common smoketree are probably the insect vector of common smoketree dieback disease.
Phytoplasmal 16S rRNA fragment is also amplified and sequence from elm plants showing yellowing.The sequence analysis results showed that the phytoplasma associating with Chinese elm yellowing belongs to subgroup16Sr V-B of elm yellows group.
利用针对植原体的通用引物,通过巢式PCR从表现花变叶症状的落地生根植株中扩增到了植原体16S rRNA、核糖体蛋白基因(rp)和延伸因子Tu基因(tuf),序列分析结果表明,该植原体属于翠菊黄化组。在电镜下观察到了植原体粒子,嫁接实验证明植原体是落地生根花变叶的病原。
通过巢式PCR证明自然表现花变叶症状的柳树中含有植原体。16SrRNA序列的结果表明与柳树花变叶相关的植原体属于翠菊黄化组16SrI-B亚组,根据rp序列分析的结果,该植原体属于AY组rp I-D亚组。
证明黄栌顶枯病是由植原体引起。16S rRNA序列比较结果表明该植原体(CSDP)属于16Sr I-B亚组。通过比较CSDP与GenBank中登陆的16Sr I组12个16S rRNA序列,进一步明确CSDP株系属于16Sr I-B亚组。黄栌木虱可能是黄栌顶枯病的昆虫介体。
从榆树黄化植株中扩增到了植原体。16S rRNA序列分析的结果表明中国榆树黄化植原体属于EY组的16Sr V-B亚组。
本实验中的落地生根花变叶病、柳树花变叶病、黄栌顶枯病是国内外首次报道,榆树黄化病是国内首次报道。
The phytoplasmas associated with air-plant phyllody,willow phyllody, common smoketree dieback and Chinese elm yellows were identified at molecular levels.The main results read as follows:
The 16S rRNA gene,ribosomal protein gene(rp) and elongation factor Tu(EF-Tu) (tuf gene) of the phytoplasma associating with air-plant phyllody were amplified with normal or nested-PCR.Sequence analysis results show that the phytoplasma belongs to aster yellows group.The phytoplasma particles were observed under electron microscope.Graft experiment also conformed that air-plant phyllody is caused by phytoplasma.
Phytoplasma of aster yellows group was also detected from willow plants showing symptom of phyllody.The associating phytoplasma belongs to 16Sr I-B according to 16S rRNA sequence and belongd to rp I-D according to rp gene sequence.
Common smoketree dieback is caused by phytoplasma.Blast results of 16S rRNA sequences showed that the phytoplasma belongs to subgroup 16Sr I-B. Alignment results of this and other twelve 16S phytoplasmal rRNA that the corresponding phytoplasma belongs to subgroup 16Sr I-B.Psyllas inhabiting on common smoketree are probably the insect vector of common smoketree dieback disease.
Phytoplasmal 16S rRNA fragment is also amplified and sequence from elm plants showing yellowing.The sequence analysis results showed that the phytoplasma associating with Chinese elm yellowing belongs to subgroup16Sr V-B of elm yellows group.
引文
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