在枣疯植原体16S rDNA片段内设计并合成引物对DZ16SF-2/DZ16SR-2（扩增190 bp片段）,用重组质粒pMD18-DZ16S为模板建立标准曲线,并建立枣疯植原体实时荧光定量PCR（SYBR Green I）检测体系。溶解曲线分析显示：该引物特异性好,该体系对...在枣疯植原体16S rDNA片段内设计并合成引物对DZ16SF-2/DZ16SR-2（扩增190 bp片段）,用重组质粒pMD18-DZ16S为模板建立标准曲线,并建立枣疯植原体实时荧光定量PCR（SYBR Green I）检测体系。溶解曲线分析显示：该引物特异性好,该体系对质粒模板的检测灵敏度达102 copies/μL。利用该体系对不同月份、不同发病级别的嫁接接种枣树接穗样品进行植原体含量测定。通过对高抗品种‘星光’,中抗品种‘黑腰子枣’、‘嘎嘎枣’和‘葫芦枣’,感病品种‘月光枣’、‘马牙枣’、‘莲蓬子’、‘北流西冬枣’和高感品种‘大红袍枣’嫁接到发病‘冬枣’砧木上所表现出的不同病级、病情指数和病原浓度比较结果发现,表现无症状的样品中植原体的含量在每克鲜重10^5～10^6个,发病Ⅰ～Ⅱ级枝条中植原体浓度在10^6～10^7个/g,Ⅲ～Ⅳ级发病枝条含量在10^7～10^8个/g鲜重,而V级发病枝条达到每克鲜重109个,表明接种发病的接穗症状严重度与其体内植原体浓度积累密切相关。高抗枣疯病‘星光’品种嫁接接穗体内病原浓度明显低于感病品种,而且有较多‘星光’接穗嫁接的发病砧木症状有所减轻,植原体浓度也有所降低。本研究可为探索枣树品种的抗病机制、抗病性鉴定及枣疯病植原体的防治提供参考。展开更多
【目的】筛查北京地区枣园感染枣疯病植原体的叶蝉种类,确定枣疯病植原体潜在介体昆虫,为预防和控制枣疯病流行提供科学依据。【方法】通过扫网法和黄板诱集法,对昌平区流村镇流村、海淀区西北旺镇唐家岭村、朝阳区奥林匹克森林公园、...【目的】筛查北京地区枣园感染枣疯病植原体的叶蝉种类,确定枣疯病植原体潜在介体昆虫,为预防和控制枣疯病流行提供科学依据。【方法】通过扫网法和黄板诱集法,对昌平区流村镇流村、海淀区西北旺镇唐家岭村、朝阳区奥林匹克森林公园、通州区永于路北京观光南瓜园4个地点枣树上叶蝉种类及枣疯病植原体感染的叶蝉进行调查和分子检测分析。【结果】北京地区枣园枣树上有13种叶蝉发生,其中包括片突菱纹叶蝉( Hishimonus lamellatus Cai et Kuoh)、凹缘菱纹叶蝉( H. sellatus (Uhler))、小绿叶蝉( Empoasca spp.)、斑叶蝉( Erythroneura sp.)、横带叶蝉( Scaphoideus festivus Matsumura)、大青叶蝉( Cicadella viridis (Linnaeus))、新县长突叶蝉( Batracomorphus xinxianensis Cai et Shen)、红闪小叶蝉( Zygina sp.)、白边大叶蝉( Kolla paulula (Walker))、桃一点叶蝉( Singapora shinshana (Matsumura))、一点木叶蝉( Phlogotettix cyclops (Mulsant et Rey))、条沙叶蝉( Psammotettix striatus (Linnaeus))和窗耳叶蝉( Ledra auditura Walker)。通过对采集的叶蝉标本采用植原体通用引物进行PCR检测,片突菱纹叶蝉( H. lamellatus )、凹缘菱纹叶蝉( H. sellatus )、大青叶蝉( C. viridis )、白边大叶蝉( K. paulula ),均发现感染枣疯病植原体,感染率分别为1.4%、3.0%、7.4%、8.3%,其他叶蝉经检测未发现感染植原体。【结论】凹缘菱纹叶蝉( H. sellatus )、片突菱纹叶蝉( H. lamellatus )、大青叶蝉( C. viridis )、白边大叶蝉( K. paulula )可能为潜在的枣疯病植原体介体昆虫。展开更多
【目的】探讨片突菱纹叶蝉Hishimonus lamellatus Cai et Kuo不同种群中枣疯植原体与沃尔巴克氏体Wolbachia的感染情况和Wolbachia在不同器官组织分布,明确枣园菱纹叶蝉中Wolbachia的感染类型和分类地位,为研究Wolbachia感染对枣疯植原...【目的】探讨片突菱纹叶蝉Hishimonus lamellatus Cai et Kuo不同种群中枣疯植原体与沃尔巴克氏体Wolbachia的感染情况和Wolbachia在不同器官组织分布,明确枣园菱纹叶蝉中Wolbachia的感染类型和分类地位,为研究Wolbachia感染对枣疯植原体潜在介体叶蝉生物学及生态学影响奠定基础。【方法】通过枣疯植原体和Wolbachia的基因特异性引物对片突菱纹叶蝉田间自然种群和实验室种群进行分子检测和鉴定。【结果】田间采集的片突菱纹叶蝉成虫植原体感染率在55%-61%之间,而Wolbachia感染率为3%-4%。田间采集的片突菱纹叶蝉自然种群经室内饲养,在1-4龄若虫中检测到Wolbachia,2-5龄若虫中检测到了植原体。片突菱纹叶蝉实验室饲养无植原体种群在其卵巢、卵和若虫中发现感染Wolbachia,在其唾液腺和消化道也检测到了Wolbachia,感染率在58%-100%之间。基于Wolbachia的wsp基因构建系统发育树,发现片突菱纹叶蝉体内的2个Wolbachia株系同属于B大组,但不同于B大组其他株系,属于新株系w Lam1和w Lam2。【结论】片突菱纹叶蝉成虫采自田间种群可以感染枣疯植原体和Wolbachia,无植原体叶蝉实验室饲养种群成虫感染Wolbachia显著高于田间种群,片突菱纹叶蝉体内2个Wolbachia株系属于B大组。这一研究结果为Wolbachia作为介体叶蝉生物防治剂进一步利用提供了基础信息。展开更多
Jujube Witches’ Broom (JWB), caused by phytoplasma, is the most serious and destructive disease of Chinese Jujube. The distribution and year_round concentration variation of JWB phytoplasma were studied under fluores...Jujube Witches’ Broom (JWB), caused by phytoplasma, is the most serious and destructive disease of Chinese Jujube. The distribution and year_round concentration variation of JWB phytoplasma were studied under fluorescence microscope using DAPI. The results showed that phytoplasma might exist in the sieve tubes of all organs, phytoplasma contents varied with organs, sides of organs and growing seasons, phytoplasma usually existed in the roots of the same direction with diseased branches. The uneven_distribution could be observed much often in lightly diseased trees than in seriously diseased one. In roots, the content of phytoplasma was highest in May, relatively low in June, July and August, and lowest in December to March. In branches, the content of phytoplasma increased gradually with the rising of the temperature after bud sprouting in April and May, then increased dramatically and reached peak in July and August, thereafter decreased in autumn. From December to February, there was still a large amount of phytoplasma in diseased branches. The content of phytoplasma in branches kept higher than in roots throughout a year.展开更多
文摘【目的】筛查北京地区枣园感染枣疯病植原体的叶蝉种类,确定枣疯病植原体潜在介体昆虫,为预防和控制枣疯病流行提供科学依据。【方法】通过扫网法和黄板诱集法,对昌平区流村镇流村、海淀区西北旺镇唐家岭村、朝阳区奥林匹克森林公园、通州区永于路北京观光南瓜园4个地点枣树上叶蝉种类及枣疯病植原体感染的叶蝉进行调查和分子检测分析。【结果】北京地区枣园枣树上有13种叶蝉发生,其中包括片突菱纹叶蝉( Hishimonus lamellatus Cai et Kuoh)、凹缘菱纹叶蝉( H. sellatus (Uhler))、小绿叶蝉( Empoasca spp.)、斑叶蝉( Erythroneura sp.)、横带叶蝉( Scaphoideus festivus Matsumura)、大青叶蝉( Cicadella viridis (Linnaeus))、新县长突叶蝉( Batracomorphus xinxianensis Cai et Shen)、红闪小叶蝉( Zygina sp.)、白边大叶蝉( Kolla paulula (Walker))、桃一点叶蝉( Singapora shinshana (Matsumura))、一点木叶蝉( Phlogotettix cyclops (Mulsant et Rey))、条沙叶蝉( Psammotettix striatus (Linnaeus))和窗耳叶蝉( Ledra auditura Walker)。通过对采集的叶蝉标本采用植原体通用引物进行PCR检测,片突菱纹叶蝉( H. lamellatus )、凹缘菱纹叶蝉( H. sellatus )、大青叶蝉( C. viridis )、白边大叶蝉( K. paulula ),均发现感染枣疯病植原体,感染率分别为1.4%、3.0%、7.4%、8.3%,其他叶蝉经检测未发现感染植原体。【结论】凹缘菱纹叶蝉( H. sellatus )、片突菱纹叶蝉( H. lamellatus )、大青叶蝉( C. viridis )、白边大叶蝉( K. paulula )可能为潜在的枣疯病植原体介体昆虫。
文摘【目的】探讨片突菱纹叶蝉Hishimonus lamellatus Cai et Kuo不同种群中枣疯植原体与沃尔巴克氏体Wolbachia的感染情况和Wolbachia在不同器官组织分布,明确枣园菱纹叶蝉中Wolbachia的感染类型和分类地位,为研究Wolbachia感染对枣疯植原体潜在介体叶蝉生物学及生态学影响奠定基础。【方法】通过枣疯植原体和Wolbachia的基因特异性引物对片突菱纹叶蝉田间自然种群和实验室种群进行分子检测和鉴定。【结果】田间采集的片突菱纹叶蝉成虫植原体感染率在55%-61%之间,而Wolbachia感染率为3%-4%。田间采集的片突菱纹叶蝉自然种群经室内饲养,在1-4龄若虫中检测到Wolbachia,2-5龄若虫中检测到了植原体。片突菱纹叶蝉实验室饲养无植原体种群在其卵巢、卵和若虫中发现感染Wolbachia,在其唾液腺和消化道也检测到了Wolbachia,感染率在58%-100%之间。基于Wolbachia的wsp基因构建系统发育树,发现片突菱纹叶蝉体内的2个Wolbachia株系同属于B大组,但不同于B大组其他株系,属于新株系w Lam1和w Lam2。【结论】片突菱纹叶蝉成虫采自田间种群可以感染枣疯植原体和Wolbachia,无植原体叶蝉实验室饲养种群成虫感染Wolbachia显著高于田间种群,片突菱纹叶蝉体内2个Wolbachia株系属于B大组。这一研究结果为Wolbachia作为介体叶蝉生物防治剂进一步利用提供了基础信息。
文摘Jujube Witches’ Broom (JWB), caused by phytoplasma, is the most serious and destructive disease of Chinese Jujube. The distribution and year_round concentration variation of JWB phytoplasma were studied under fluorescence microscope using DAPI. The results showed that phytoplasma might exist in the sieve tubes of all organs, phytoplasma contents varied with organs, sides of organs and growing seasons, phytoplasma usually existed in the roots of the same direction with diseased branches. The uneven_distribution could be observed much often in lightly diseased trees than in seriously diseased one. In roots, the content of phytoplasma was highest in May, relatively low in June, July and August, and lowest in December to March. In branches, the content of phytoplasma increased gradually with the rising of the temperature after bud sprouting in April and May, then increased dramatically and reached peak in July and August, thereafter decreased in autumn. From December to February, there was still a large amount of phytoplasma in diseased branches. The content of phytoplasma in branches kept higher than in roots throughout a year.
