Laser Microdissection‐Mediated Isolation and In Vitro Transcriptional Amplification of Plant RNA

Scanlon, Michael J.; Ohtsu, Kazuhiro; Timmermans, Marja C.P.; Schnable, Patrick S.

doi:10.1002/0471142727.mb25a03s87

Cited by 26 publications

(29 citation statements)

References 36 publications

(62 reference statements)

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“…Hand-dissected apices of 14-d-old B73 seedlings were fixed in acetone, embedded into paraffin, and sections of 8 µm spread on 1.0 PEN membrane slides (Zeiss), as described (Scanlon et al 2009). After deparaffinization in xylene, cells of interest were captured into AdhesiveCap 500 tubes (Zeiss) using the PALM Micro-Beam system.…”

Section: Laser Microdissection and Rna-seq Library Constructionmentioning

confidence: 99%

“…Library metrics are listed in Supplemental Table S1. Gene expression levels were normalized to RPM rather than RPKM, as linearly amplified RNA captures the 3 ′ 400-500 nucleotides of transcripts (Scanlon et al 2009). Relatedness across tissue samples was determined based on the expression values of all genes expressed in each pairwise comparison using Pearson's correlation in R (R Core Team 2015).…”

Section: Gene Expression Analysesmentioning

confidence: 99%

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A high-resolution gene expression atlas links dedicated meristem genes to key architectural traits

et al. 2019

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Section: Laser Microdissection and Rna-seq Library Constructionmentioning

confidence: 99%

Section: Gene Expression Analysesmentioning

confidence: 99%

A high-resolution gene expression atlas links dedicated meristem genes to key architectural traits

et al. 2019

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“…Total above-ground plant samples were harvested for RNA at the same time that shoot apices were collected for laser microdissection. All laser-microdissected samples were fixed in ice-cold, 100% acetone and processed for RNA isolations following the procedures outlined in Scanlon et al (2009). At least 100 000 lm 2 of sectioned tissue was isolated for each biological replicate.…”

Section: Plant Tissue and Growth Conditionsmentioning

confidence: 99%

Dissecting the molecular signatures of apical cell‐type shoot meristems from two ancient land plant lineages

et al. 2015

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SummaryShoot apical meristem (SAM) structure varies markedly within the land plants. The SAMs of many seedless vascular plants contain a conspicuous inverted, pyramidal cell called the apical cell (AC), which is unidentified in angiosperms.In this study, we use transcriptomic sequencing with precise laser microdissections of meristem subdomains to define the molecular signatures of anatomically distinct zones from the AC-type SAMs of a lycophyte (Selaginella moellendorffii) and a monilophyte (Equisetum arvense). The two model species for this study represent vascular plant lineages that diverged > 400 million yr ago.Our data comprise comprehensive molecular signatures for the distinct subdomains within AC-type SAMs, an anatomical anomaly whose functional significance has been debated in the botanical literature for over two centuries.Moreover, our data provide molecular support for distinct gene expression programs between the AC-type SAMs of Selaginella and Equisetum, as compared with the SAM transcriptome of the angiosperm maize. The results are discussed in light of the functional significance and evolutionary success of the AC-type SAM within the embryophytes.

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“…Laser microdissection enables the isolation of discrete domains within microscopic samples for use in transcriptomic analyses (Nelson et al, 2006;Scanlon et al, 2009). Six samples were microdissected from developing embryos and 14-d-old seedlings, including (1) the cells comprising the embryo proper of the proembryo ( Figures 1M and 1S), (2) the organizing SAM and emerging scutellar hood of the transition-stage embryo ( Figures 1N and 1T), (3) the SAM and the initiating coleoptile of the coleoptile-stage embryo ( Figures 1O and 1U), (4) the SAM and leaf primordium of a stage 1 embryo ( Figures 1P and 1V), (5) the SAM and plastochron 1 leaf of a 14-d-old seedling (L14; Figures 1Q and 1W), and (6) the lateral meristem and the newly initiated husk leaf from the 14-d-old seedling ( Figures 1R and 1X).…”

Section: Laser Microdissection and Rna-seq Of Sam Ontogenymentioning

confidence: 99%

Ontogeny of the Maize Shoot Apical Meristem

Takacs

et al. 2012

Plant Cell

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The maize (Zea mays) shoot apical meristem (SAM) arises early in embryogenesis and functions during stem cell maintenance and organogenesis to generate all the aboveground organs of the plant. Despite its integral role in maize shoot development, little is known about the molecular mechanisms of SAM initiation. Laser microdissection of apical domains from developing maize embryos and seedlings was combined with RNA sequencing for transcriptomic analyses of SAM ontogeny. Molecular markers of key events during maize embryogenesis are described, and comprehensive transcriptional data from six stages in maize shoot development are generated. Transcriptomic profiling before and after SAM initiation indicates that organogenesis precedes stem cell maintenance in maize; analyses of the first three lateral organs elaborated from maize embryos provides insight into their homology and to the identity of the single maize cotyledon. Compared with the newly initiated SAM, the mature SAM is enriched for transcripts that function in transcriptional regulation, hormonal signaling, and transport. Comparisons of shoot meristems initiating juvenile leaves, adult leaves, and husk leaves illustrate differences in phase-specific (juvenile versus adult) and meristem-specific (SAM versus lateral meristem) transcript accumulation during maize shoot development. This study provides insight into the molecular genetics of SAM initiation and function in maize.

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Laser Microdissection‐Mediated Isolation and In Vitro Transcriptional Amplification of Plant RNA

Cited by 26 publications

References 36 publications

A high-resolution gene expression atlas links dedicated meristem genes to key architectural traits

A high-resolution gene expression atlas links dedicated meristem genes to key architectural traits

Dissecting the molecular signatures of apical cell‐type shoot meristems from two ancient land plant lineages

Ontogeny of the Maize Shoot Apical Meristem

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