A simple and rapid method for minipreparation of DNA from tissue cultured plant cells

Mettler, Irvin J.

doi:10.1007/bf02668996

Cited by 135 publications

(59 citation statements)

References 4 publications

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“…After sublimation of the nitrogen, 500 pL of buffer (100 mM Tris, 100 mM Na,EDTA, 250 mM NaC1, pH 8.0) were added and grinding was continued. Fifty microliters of 10% sarkosyl were then added, followed by 5 pL of proteinase K (10 mg/mL), and the tubes were incubated at 55°C for approximately 1 h. After centrifugation (14,000 rpm, 10 min), the supernatant was extracted twice with pheno1:chloroform:isoamyl alcohol (24:24:1, v / v), and 0.35 volumes of 5 M NaCl and 2 volumes of 100% ethanol were added to the final aqueous phase to precipitate DNA free from polysaccharides (Mettler, 1987). The pelle'ts were washed twice in 75% ethanol and resuspended in 100 pL of sterile water, and 1.5 PL were used per 2 0 -~L PCR reaction.…”

Section: Chromosome Mappingmentioning

confidence: 99%

A Brassinosteroid-Insensitive Mutant in Arabidopsis thaliana Exhibits Multiple Defects in Growth and Development

1996

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Brassinosteroids are widely distributed plant compounds that modulate cell elongation and division, but little is known about the mechanism of action of these plant growth regulators. To investigate brassinosteroids as signals influencing plant growth and development, we identified a brassinosteroid-insensitive mutant in Arabidopsis thaliana (L.) Henyh. ecotype Columbia. The mutant, termed bril, did not respond to brassinosteroids in hypocotyl elongation and primary root inhibition assays, but it did retain sensitivity to auxins, cytokinins, ethylene, abscisic acid, and gibberellins. The bril mutant showed multiple deficiencies in developmental pathways that could not be rescued by brassinosteroid treatment, including a severely dwarfed stature; dark green, thickened leaves; male sterility; reduced apical dominance; and de-etiolation of darkgrown seedlings. Cenetic analysis suggests that the Bril phenotype is caused by a recessive mutation in a single gene with pleiotropic effects that maps 1.6 centimorgans from the cleaved, amplified, polymorphic sequence marker DHSl on the bottom of chromosome IV. The multiple and dramatic effects of mutation of the BRIl locus on development suggests that the BRIl gene may play a critical role in brassinosteroid perception or signal transduction.

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Section: Chromosome Mappingmentioning

confidence: 99%

A Brassinosteroid-Insensitive Mutant in Arabidopsis thaliana Exhibits Multiple Defects in Growth and Development

1996

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“…Small amount DNA was extracted from some leaflets of regenerated plants [10]. The presence of the gus gene was detected by PCR using the following 2 primers: the forward (5 -GGGATCCATCGCAGCGTAATG-3 ) and the reverse (5 -GCCGACAGCAGCAGTTTCATC-3').…”

Section: Dna Analysismentioning

confidence: 99%

Transgenic peanut plants obtained by particle bombardment via somatic embryogenesis regeneration system

Deng

Wei

2001

Cell Res

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After pre-culture and treatment of osmosis, cotyledons of immature peanut (Arachis hypogaea L.) zygotic embryos were transformed via particle bombardment with a plasmid containing a chimeric hph gene conferring resistance to hygromycin and a chimeric intron-gus gene. Selection for hygromycin resistant calluses and somatic embryos was initiated at 10th d post-bombardment on medium containing 10-25 mg/L hygromycin. Under continuous selection, hygromycin resistant plantlets were regenerated from somatic embryos and were recovered from nearly 1.6% of the bombarded cotyledons. The presence and integration of foreign DNA in regenerated hygromycin resistant plants was confirmed by PCR (polymerase chain reaction) for the intron-gus gene and by Southern hybridization of the hph gene. GUS enzyme activity was detected in leaflets from transgenic plants but not from control, non-transformed plants. The production of transgenic plants are mainly based on a newly improved somatic embryogenesis regeneration system developed by us.

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“…Total leaf DNA (Mettler, 1987) was digested with restriction endonucleases and electrophoresed in 0.7% agarose gels (3 g per lane). For the RNA gel blots, total leaf RNA was extracted using the TRIzol reagent (GIBCO-BRL) and electrophoresed in 1% agarose/formaldehyde gels (5 g of RNA per lane).…”

Section: Dna and Rna Gel Blotsmentioning

confidence: 99%

RNA Polymerase Subunits Encoded by the Plastid rpoGenes Are Not Shared with the Nucleus-Encoded Plastid Enzyme1

1998

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Plastid genes in photosynthetic higher plants are transcribed by at least two RNA polymerases. The plastid rpoA, rpoB, rpoC1, and rpoC2 genes encode subunits of the plastid-encoded plastid RNA polymerase (PEP), an Escherichia coli-like core enzyme. The second enzyme is referred to as the nucleus-encoded plastid RNA polymerase (NEP), since its subunits are assumed to be encoded in the nucleus. Promoters for NEP have been previously characterized in tobacco plants lacking PEP due to targeted deletion of rpoB (encoding the ␤-subunit) from the plastid genome. To determine if NEP and PEP share any essential subunits, the rpoA, rpoC1, and rpoC2 genes encoding the PEP ␣-, ␤-, and ␤؆-subunits were removed by targeted gene deletion from the plastid genome. We report here that deletion of each of these genes yielded photosynthetically defective plants that lack PEP activity while maintaining transcription specificity from NEP promoters. Therefore, rpoA, rpoB, rpoC1, and rpoC2 encode PEP subunits that are not essential components of the NEP transcription machinery. Furthermore, our data indicate that no functional copy of rpoA, rpoB, rpoC1, or rpoC2 that could complement the deleted plastid rpo genes exists outside the plastids.At least two distinct RNA polymerases are involved in the transcription of plastid genes in photosynthetic higher plants. One of these contains homologs of the Escherichia coli enzyme, including the ␣-, ␤-, ␤Ј-, and ␤Љ-subunits encoded in the plastid rpoA, rpoB, rpoC1, and rpoC2 genes, and is referred to as PEP. The promoters for PEP are reminiscent of the E. coli 70 -type promoters, and have two conserved hexameric blocks of sequences (TTGACA or "-35" element; TATAAT or "Ϫ10" element) 17 to 19 nucleotides apart. Transcription from PEP promoters initiates 5 to 7 nucleotides downstream of the "Ϫ10" promoter element (Igloi and Kö ssel, 1992; Gruissem and Tonkyn, 1993;Link, 1996). Promoter specificity to PEP is conferred by nuclearencoded -like factors (Isono et al., 1997;Tanaka et al., 1997).Several reports indicate the existence of a second, NEP activity (Morden et al., 1991; Hess et al., 1993; Allison et al., 1996). A candidate for NEP is an approximately 110-kD protein that has properties similar to the mitochondrial and phage T3/T7 RNA polymerases that may be part of a larger complex (Lerbs-Mache, 1993; Hedtke et al., 1997). NEP promoters share a loose, 10-nucleotide consensus, ATA-GAATA/GAA, overlapping the transcription-initiation site, which is reminiscent of promoters recognized by the mitochondrial and phage T3/T7 RNA polymerases (Hajdukiewicz et al., 1997; Hü bschmann and Bö rner, 1998; for review, see Maliga, 1998).Plastid RNA polymerase activities with distinct sensitivities to inhibitors are present in higher plants in multisubunit complexes (Pfannschmidt and Link, 1994). Sharing of essential subunits of RNA polymerases has been reported in yeast (Sentenac et al., 1992). Therefore, plastid NEP and PEP could be part of the same complex. To test if NEP and PEP share any essential subunits,...

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A simple and rapid method for minipreparation of DNA from tissue cultured plant cells

Cited by 135 publications

References 4 publications

A Brassinosteroid-Insensitive Mutant in Arabidopsis thaliana Exhibits Multiple Defects in Growth and Development

A Brassinosteroid-Insensitive Mutant in Arabidopsis thaliana Exhibits Multiple Defects in Growth and Development

Transgenic peanut plants obtained by particle bombardment via somatic embryogenesis regeneration system

RNA Polymerase Subunits Encoded by the Plastid rpoGenes Are Not Shared with the Nucleus-Encoded Plastid Enzyme1

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