An improved procedure for isolation of high-quality RNA from nematode-infected Arabidopsis roots through laser capture microdissection. Anjam, M. S., Ludwig, Y., Hochholdinger, F., Miyaura, C., Inada, M., Siddique, S., & Grundler, F. M. W. Plant Methods, 12(1):25, April, 2016. ![An improved procedure for isolation of high-quality RNA from nematode-infected Arabidopsis roots through laser capture microdissection [link]](https://bibbase.org/img/filetypes/link.svg "link")
Paper doi abstract bibtex Cyst nematodes are biotrophs that form specialized feeding structures in the roots of host plants, which consist of a syncytial fusion of hypertrophied cells. The formation of syncytium is accompanied by profound transcriptional changes and active metabolism in infected tissues. The challenge in gene expression studies for syncytium has always been the isolation of pure syncytial material and subsequent extraction of intact RNA. Root fragments containing syncytium had been used for microarray analyses. However, the inclusion of neighbouring cells dilutes the syncytium-specific mRNA population. Micro-sectioning coupled with laser capture microdissection (LCM) offers an opportunity for the isolation of feeding sites from heterogeneous cell populations. But recovery of intact RNA from syncytium dissected by LCM is complicated due to extended steps of fixation, tissue preparation, embedding and sectioning.
@article{anjam_improved_2016,
title = {An improved procedure for isolation of high-quality {RNA} from nematode-infected {Arabidopsis} roots through laser capture microdissection},
volume = {12},
issn = {1746-4811},
url = {https://doi.org/10.1186/s13007-016-0123-9},
doi = {10.1186/s13007-016-0123-9},
abstract = {Cyst nematodes are biotrophs that form specialized feeding structures in the roots of host plants, which consist of a syncytial fusion of hypertrophied cells. The formation of syncytium is accompanied by profound transcriptional changes and active metabolism in infected tissues. The challenge in gene expression studies for syncytium has always been the isolation of pure syncytial material and subsequent extraction of intact RNA. Root fragments containing syncytium had been used for microarray analyses. However, the inclusion of neighbouring cells dilutes the syncytium-specific mRNA population. Micro-sectioning coupled with laser capture microdissection (LCM) offers an opportunity for the isolation of feeding sites from heterogeneous cell populations. But recovery of intact RNA from syncytium dissected by LCM is complicated due to extended steps of fixation, tissue preparation, embedding and sectioning.},
number = {1},
urldate = {2024-08-30},
journal = {Plant Methods},
author = {Anjam, Muhammad Shahzad and Ludwig, Yvonne and Hochholdinger, Frank and Miyaura, Chisato and Inada, Masaki and Siddique, Shahid and Grundler, Florian M. W.},
month = apr,
year = {2016},
keywords = {Arabidopsis, Cyst nematode, LCM, Nematode, RNA degradation, Root, Syncytium},
pages = {25},
}
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