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类病毒在番茄叶片组织高度纯化细胞核中的亚细胞定位。

Subcellular localization of viroids in highly purified nuclei from tomato leaf tissue.

作者信息

Schumacher J, Sänger H L, Riesner D

机构信息

Institut für Physikalische Biologie, Universität Dusseldorf, FRG.

出版信息

EMBO J. 1983;2(9):1549-55. doi: 10.1002/j.1460-2075.1983.tb01622.x.

DOI:10.1002/j.1460-2075.1983.tb01622.x
PMID:11892810
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC555321/
Abstract

Approximately 95% of the viroid RNA which is present in potato spindle tuber viroid (PSTV)-infected tomato plant leaf issue, is associated with the nucleolar fraction obtained from purified nuclei. Viroids were released from the nucleolar fraction by increasing the ionic strength of the medium to 0.66 suggesting that viroid RNA is present in these subnuclear components in a protein-nucleic acid complex. A purification procedure for nuclei from leaf tissue had to be newly developed; it involves two Percoll density centrifugations as final steps. The nuclei were sonicated and the sonicate fractionated into fractions either highly enriched in nucleoli or in broken chromatin and ribonucleoprotein particles. The viroid content in the different samples was determined by gel electrophoresis. Depending upon the progress of the disease, viroid copy numbers between 200 and 10,000 per cell were observed in homogenized tissue, purified nuclei and in the nucleolar fraction. In chloroplasts, practically no viroids were detected. The results are discussed in the light of current hypotheses about the replication, pathogenicity and origin of viroids.

摘要

在感染马铃薯纺锤块茎类病毒（PSTV）的番茄植株叶片组织中，约95%的类病毒RNA与从纯化细胞核中获得的核仁部分相关联。通过将培养基的离子强度提高到0.66，类病毒从核仁部分释放出来，这表明类病毒RNA以蛋白质 - 核酸复合物的形式存在于这些亚核组分中。必须新开发一种从叶片组织中纯化细胞核的方法；该方法包括两步Percoll密度离心作为最后步骤。对细胞核进行超声处理，并将超声处理后的样品分离成高度富集核仁或破碎染色质及核糖核蛋白颗粒的组分。通过凝胶电泳测定不同样品中的类病毒含量。根据疾病进展情况，在匀浆组织、纯化细胞核和核仁部分中，观察到每个细胞的类病毒拷贝数在200到10,000之间。在叶绿体中，几乎未检测到类病毒。根据关于类病毒复制、致病性和起源的当前假说对结果进行了讨论。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/24f9/555321/693900995d47/emboj00262-0120-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/24f9/555321/4e18b3f61b8a/emboj00262-0118-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/24f9/555321/3cf81cc30dfc/emboj00262-0118-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/24f9/555321/a7b984c516ed/emboj00262-0119-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/24f9/555321/82245803603c/emboj00262-0119-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/24f9/555321/693900995d47/emboj00262-0120-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/24f9/555321/4e18b3f61b8a/emboj00262-0118-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/24f9/555321/3cf81cc30dfc/emboj00262-0118-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/24f9/555321/a7b984c516ed/emboj00262-0119-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/24f9/555321/82245803603c/emboj00262-0119-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/24f9/555321/693900995d47/emboj00262-0120-a.jpg

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本文引用的文献

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