Improved method for the isolation of RNA from plant tissues

Logemann, J.; Schell, Jeff; Willmitzer, Lothar

doi:10.1016/0003-2697(87)90086-8

Cited by 1,779 publications

(1,007 citation statements)

References 9 publications

Supporting

Mentioning

993

Contrasting

Unclassified

Order By: Relevance

“…RNA for real-time PCR experiments was extracted from different tissues from greenhouse grown Arabidopsis thaliana Col-0 plants as reported earlier (Logeman et al 1987). Residual genomic DNA was removed by treatment with RNAse-free DNAse (Roche, Mannheim, Germany) and approximately 2 lg of total RNA was reverse transcribed with an MMuLV reverse transcriptase (Qbiogene, Heidelberg, Gemany) using an oligo (dT) primer according to the manufacturer's suggestion, thus generating 21 ll of first-strand cDNA.…”

Section: Rna Isolation and Rt-pcrmentioning

confidence: 99%

The inositol oxygenase gene family of Arabidopsis is involved in the biosynthesis of nucleotide sugar precursors for cell-wall matrix polysaccharides

Kanter

Usadel

Guérineau

et al. 2005

Planta

137

141

View full text Add to dashboard Cite

The nucleotide sugar UDP-glucuronic acid (UDP-GlcA) is the principal precursor for galacturonic acid, xylose, apiose and arabinose residues of the plant cell-wall polymers. UDP-GlcA can be synthesized by two different functional pathways in Arabidopsis involving either UDP-glucose dehydrogenase or inositol oxygenase as the initial enzyme reaction to channel carbohydrates into a pool of UDP sugars used for cellwall biosynthesis. The genes for the enzyme myo-inositol oxygenase (MIOX) were analyzed in Arabidopsis. They represent a small gene family containing four members. The transcription of all those members indicates a transient and organ-specific gene expression pattern in growing plant tissues as analyzed by RT-PCR and in promoter::GUS reporter gene lines. Two isoforms (MIOX1, MIOX2) are expressed in almost all tissues of the plant, whereas the expression of MIOX4 and MIOX5 is largely restricted to flowers, particularly maturing pollen. T-DNA insertion lines in MIOX genes were isolated; however, single knock-outs show growth phenotypes similar to the wild type. The monosaccharide composition of the cell wall in these mutants is not significantly changed compared to wild type plants. However, the incorporation of 3 H-inositol into wall polymers of seedlings is greatly impaired in the mutant lines DMIOX1 and DMIOX2, which are the only isoforms that are expressed in seedlings.

show abstract

Section: Rna Isolation and Rt-pcrmentioning

confidence: 99%

The inositol oxygenase gene family of Arabidopsis is involved in the biosynthesis of nucleotide sugar precursors for cell-wall matrix polysaccharides

Kanter

Usadel

Guérineau

et al. 2005

Planta

137

141

View full text Add to dashboard Cite

show abstract

“…Total RNA was isolated from different stages of the moss life cycle according to Logemann et al (1987). Single stranded cDNA was subsequently produced with Superscript II reverse transcriptase (Invitrogen).…”

Section: Transcript Expression Analysismentioning

confidence: 99%

The MADS-domain protein PPM2 preferentially occurs in gametangia and sporophytes of the moss Physcomitrella patens

Quodt

Faigl

Saedler

et al. 2007

Gene

View full text Add to dashboard Cite

To date, the function of MADS-domain transcription factors in non-seed plants remains largely elusive, although a number of genes have been isolated and characterized from a variety of species. In our study we analyzed PPM2, a classical MIKC-type MADS-box gene from the moss Physcomitrella patens, taking advantage of the unique technical properties Physcomitrella offers in terms of efficient homologous recombination. We determined mRNA and protein distribution and performed targeted disruption of the genomic locus for functional analysis of PPM2. Despite weak ubiquitous expression, PPM2 protein is mostly found in male and female gametangia and basal parts of developing sporophytes. Therefore, PPM2 seems to function in both the haploid and the diploid phase of the moss life cycle. This situation reflects an evolutionary transition state of gene recruitment from an ancestral gametophytic generation into a derived sporophytic generation which became dominating in tracheophytes. However, a knock-out of the PPM2 gene did not cause visible phenotypical changes in the respective structures. The implications of our findings for the understanding of the evolutionary history of MADS-box transcription factors in plants are discussed.

show abstract

“…Total RNA was isolated from 10 g fresh weight of seed coats by the guanidine hydrochloride extraction method (Logemann et al, 1987). Poly(A) + RNA was prepared by oligo(dT)-cellulose chromatography (Sambrook et al, 1989).…”

Section: Construction Of a Seed Coat Cdna Librarymentioning

confidence: 99%

Members of the aquaporin family in the developing pea seed coat include representatives of the PIP, TIP, and NIP subfamilies

et al. 2003

View full text Add to dashboard Cite

Water and nutrients required by developing seeds are mainly supplied by the phloem and have to be released from a maternal parenchyma tissue before being utilized by the filial tissues of embryo and endosperm. To identify aquaporins that could be involved in this process four full-length cDNAs were cloned and sequenced from a cDNA library of developing seed coats of pea (Pisum sativum L.). The cDNA of PsPIP1-1 appeared to be identical to that of clone 7a/TRG-31, a turgor-responsive gene cloned previously from pea roots. PsPIP1-1, PsPIP2-1, and PsTIP1-1, or their possible close homologues, were also expressed in cotyledons of developing and germinating seeds, and in roots and shoots of seedlings, but transcripts of PsNIP-1 were only detected in the seed coat. In mature dry seeds, high hybridization signals were observed with the probe for PsPIP1-1, but transcripts of PsPIP2-1, PsTIP1-1, and PsNIP-1 were not detected. Functional characterization after heterologous expression in Xenopus oocytes showed that PsPIP2-1 and PsTIP1-1 are aquaporins whereas PsNIP-1 is an aquaglyceroporin. PsNIP-1, like several other NIPs, contains a tryptophan residue corresponding with Trp-48 in GlpF (the glycerol facilitator of Escherichia coli) that borders the selectivity filter in the permeation channel. It is suggested that PsPIP1-1 and/or its possible close homologues could play a role in water absorption during seed imbibition, and that PsPIP2-1, possibly together with PsPIP1-1, could be involved in the release of phloem water from the seed coat symplast, which is intimately connected with the release of nutrients for the embryo.Abbreviations: MIPs, major intrinsic proteins; NIPs, nodulin 26-like intrinsic proteins; PIPs, plasma membrane intrinsic proteins; SIPs, small, basic intrinsic proteins; TIPs, tonoplast intrinsic proteins

show abstract

Improved method for the isolation of RNA from plant tissues

Cited by 1,779 publications

References 9 publications

The inositol oxygenase gene family of Arabidopsis is involved in the biosynthesis of nucleotide sugar precursors for cell-wall matrix polysaccharides

The inositol oxygenase gene family of Arabidopsis is involved in the biosynthesis of nucleotide sugar precursors for cell-wall matrix polysaccharides

The MADS-domain protein PPM2 preferentially occurs in gametangia and sporophytes of the moss Physcomitrella patens

Members of the aquaporin family in the developing pea seed coat include representatives of the PIP, TIP, and NIP subfamilies

Contact Info

Product

Resources

About