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[蝴蝶兰感染建兰花叶病毒和齿舌兰环斑烟草花叶病毒复合侵染的分子鉴定及其病理超微结构变化]

[Molecular identification of cymbidium mosaic Potexvirus and Odontoglossum ringspot Tobamovirus complex infected Phalaenopsis and its pathological ultrastructural alteration].

作者信息

Shi Nong Nong, Xu Ying, Wang Hui Zhong, Xie Li, Hong Jian

机构信息

Key Laboratory of Molecular Biology and Biochemistry of Hangzhou, Hangzhou Normal University, Hangzhou 310036, China.

出版信息

Fen Zi Xi Bao Sheng Wu Xue Bao. 2007 Apr;40(2):153-63.

Abstract

Filamentous and rod-shaped virions, and aggregated crystals were observed in infected leaves of by negative staining and ultramicrotomy. Histologic study synchronously showed typical crystal forms of the two virions: the crystals from filamentous particles congregated in strips, arrayed in multilayer and piled in certain angles or helix between layers; while the crystals from rod-shaped particles arrayed in parallel, angle-layer or helix. The two kinds of crystals both presented in parenchyma cells, plasmodesma, and vascular bundles; as an evidence that indicates the existence of short distance transport of viruses between cells, plasmodesmata were produced through piercing the membrane around the reproducing viral crystals; the chloroplasts in the infected cells were hypoplastic, and the filamentous virion were observed within the chloroplasts; the mitochondrions were over-developed, swelled or even cavitated; the nucleus were also swelled and cavitated. Further multiplex RT-PCR and sequencing that the coat protein genes simultaneously expanded to Cymbidium mosaic potexvirus (CymMV) and Odontoglossum ringspot tobamovirus (ORSV) showed homology with available abstracts from GenBank, and the respective percentages of identity are 98% and 99%-100%. The instant and direct identification evidences of CymMV and ORSV complex infection on phalaenopsis are revealed at both cellular and molecular levels, and the character of its pathological ultrastructural alteration as the gist in cellular pathology for pathogenesis are also presented.

摘要

通过负染色和超薄切片技术在感染的叶片中观察到丝状和杆状病毒粒子以及聚集的晶体。组织学研究同步显示了两种病毒粒子的典型晶体形态:丝状粒子形成的晶体呈带状聚集,多层排列,层间以一定角度或螺旋状堆积;而杆状粒子形成的晶体则呈平行、角层状或螺旋状排列。两种晶体均出现在薄壁细胞、胞间连丝和维管束中;作为细胞间病毒短距离运输存在的证据,胞间连丝是通过穿透繁殖病毒晶体周围的膜而产生的;感染细胞中的叶绿体发育不全,在叶绿体内观察到丝状病毒粒子;线粒体过度发育、肿胀甚至空泡化;细胞核也肿胀并空泡化。进一步的多重RT-PCR和测序表明,外壳蛋白基因同时扩增到建兰花叶病毒(CymMV)和齿舌兰环斑病毒(ORSV),与GenBank中可用摘要显示出同源性,各自的同一性百分比分别为98%和99%-100%。在细胞和分子水平上揭示了蝴蝶兰上CymMV和ORSV复合感染的即时和直接鉴定证据,并且还呈现了其病理超微结构改变的特征作为发病机制细胞病理学的要点。

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