scispace - formally typeset
Search or ask a question
Journal ArticleDOI

A plant DNA minipreparation: Version II

01 Sep 1983-Plant Molecular Biology Reporter (PLANT MOLECULAR BIOLOGY REPORTER)-Vol. 1, Iss: 4, pp 19-21
TL;DR: The topic of this report is rap,d m,croscale methods for,solat,on of plant D N A without tile use of ul tracentr ,fugatlon wEth CsCI, which is of moderately high molecular weight and serves as a satisfactory substrate for most restrlctum cndonucleases and is statable for genom,c blot analys,s.
Abstract: The topic of this report is rap,d m,croscale methods for ,solat,on of plant D N A without tile use of ul tracentr ,fugatlon wEth CsCI. The D N A produced ,s of moderately high molecular weight and serves as a satisfactory substrate for most restrlctum cndonucleases and is statable for genom,c blot analys,s. In addi t ion to the rapidi ty and convenience of mlmpreps which permit a large number of samples to be processed in just a few hours, the small amount of tissue reqmred (less than 1.0 grams) allows tbr molecular analysis of plants at a very young stage Mm,prep D N A y,elds from leaf tissue of most species tested to date are typ,cally 30-100 big per gram tissue, greater than 50 kb, and remarkably uniform from sample to sample. The first mmlprep procedure we reported fi3r maize D N A isolation (Dellaporta et al , ;'*l,;tze Geneta3 Cr162162 Neu'_~letlrt. 1983) was adapted from a procedure commonly used for }'east D N A preparatmn (Dav,s et al. , 1980) Since th,s report, numerous personal commun,cat ,ons have demonstrated that the mm,prep procedure or a modification thereof, can be apphed to most plant species tested. For example, the method has been successfully used on Ntcottana hlgl~um. N. plumklgmgidtum. N. 3)/t'eJtrt~. L)s~opertcum sp.. Amar,mthm sp . Gl)~me max. Petuma h.~hra&. Several modifications have been apphed by these ,nvestlgators and in our own laboratory m order to extend the appl ,catmn of ram,prep procedures to other plant species. The select,on of a part icular protocol depends to a large degree on the plant spec,es used. However, the procedure reported here was selected to be statable for most situations.
Citations
More filters
Journal ArticleDOI
TL;DR: It was found that the gene 5 promoter is active in a tissue-specific fashion whereas this is not the case for the NOS promoter, providing the first documented instance of a gene derived from a procaryotic host the expression of which is apparently regulated by plant growth factors.
Abstract: A “plant gene vector cassette” to be used in combination with various Escherichia coli gene-cloning vectors was constructed. This cassette contains a replication and mobilization unit which allows it to be maintained and to be transferred back and forth between E. coli and Agrobacterium tumefaciens hosts provided these hosts contain plasmid RK2 replication and mobilization helper functions. The cassette also harbors a transferable DNA unit with plant selectable marker genes and cloning sites which can be combined with different bacterial replicons, thus facilitating the reisolation of transferred DNA from transformed plants in E. coli. The vector cassette contains two different promoters derived from the T-DNA-encoded genes 5 and nopaline synthase (NOS). By comparing the levels of expression of the marker enzymes linked to each of these promoter sequences, it was found that the gene 5 promoter is active in a tissue-specific fashion whereas this is not the case for the NOS promoter. This observation provides the first documented instance of a gene derived from a procaryotic host the expression of which is apparently regulated by plant growth factors.

2,064 citations

Journal ArticleDOI
TL;DR: Yields ranged from 0.3–200 nanograms of DNA per milligram of tissue, and in no case tested was inhibition observed for restriction enzymes BamHI or EcoRI.
Abstract: We have developed a DNA extraction procedure for milligram amounts of plant tissue. Yields ranged from 0.3-200 nanograms of DNA per milligram of tissue. The factors affecting yield are discussed. Fresh tissue, as well as herbarium specimens (22-118 years old) and mummified seeds and embryos (500 to greater than 44 600 years old) were used. All tissues attempted (57 types from 29 species) yielded measurable amounts of DNA. In no case tested was inhibition observed for restriction enzymes BamHI or EcoRI.

1,829 citations

Journal ArticleDOI
TL;DR: A robust CRISPR/Cas9 vector system, utilizing a plant codon optimized Cas9 gene, for convenient and high-efficiency multiplex genome editing in monocot and dicot plants and provides examples of loss-of-function gene mutations in T0 rice and Arabidopsis plants.

1,451 citations


Cites methods from "A plant DNA minipreparation: Versio..."

  • ...Mutation Detection Genomic DNA extraction from leaves of transgenic rice plants, and rosette leaves of transgenic Arabidopsis plants, was carried out using the sodium dodecyl sulfate method (Dellaporta et al., 1983)....

    [...]

  • ...Genomic DNA extraction from leaves of transgenic rice plants, and rosette leaves of transgenic Arabidopsis plants, was carried out using the sodium dodecyl sulfate method (Dellaporta et al., 1983)....

    [...]

Journal ArticleDOI
10 Jan 1997-Cell
TL;DR: Transformation of the cloned wild-type NPR1 gene into npr1 mutants not only restored the responsiveness to SAR induction with respect to PR-gene expression and resistance to infections, but also rendered the transgenic plants more resistant to infection by P. syringae in the absence of SAR induction.

1,449 citations

Journal ArticleDOI
Pbk. Kishor1, Zonglie Hong1, Guo-Hua Miao1, Caa. Hu1, D. P. S. Verma1 
TL;DR: Proline (Pro) accumulation has been correlated with tolerance to drought and salinity stresses in plants and overproduction of Pro in plants may lead to increased tolerance against these abiotic stresses, suggesting that activity of the first enzyme of the pathway is the rate-limiting factor in Pro synthesis.
Abstract: Proline (Pro) accumulation has been correlated with tolerance to drought and salinity stresses in plants. Therefore, overproduction of Pro in plants may lead to increased tolerance against these abiotic stresses. To test this possibility, we overexpressed in tobacco the mothbean [delta]-pyrroline-5-carboxylate synthetase, a bifunctional enzyme able to catalyze the conversion of glutamate to [delta]-pyrroline-5-carboxylate, which is then reduced to Pro. The transgenic plants produced a high level of the enzyme and synthesized 10- to 18-fold more Pro than control plants. These results suggest that activity of the first enzyme of the pathway is the rate-limiting factor in Pro synthesis. Exogenous supply of nitrogen further enhanced Pro production. The osmotic potentials of leaf sap from transgenic plants were less decreased under water-stress conditions compared to those of control plants. Overproduction of Pro also enhanced root biomass and flower development in transgenic plants under drought-stress conditions. These data demonstrated that Pro acts as an osmoprotectant and that overproduction of Pro results in the increased tolerance to osmotic stress in plants.

1,351 citations

References
More filters
Journal ArticleDOI
Julius Marmur1
TL;DR: A method has been described for the isolation of DNA from micro-organisms which yields stable, biologically active, highly polymerized preparations relatively free from protein and RNA, and Representative samples have been characterized for their thermal stability and sedimentation behaviour.

11,573 citations


"A plant DNA minipreparation: Versio..." refers background in this paper

  • ...6 volumes) has been reported tit separate high molecular DNA from polysaccharides (Marmur, 1961) The sodium acetate also yields ,l tight fibrous precipitate th,it is easily washed and dried The DNA will dissolve readily if allowed tit rehydrate ,it 4 ~ for one hour fi~llowed by hght vortexlng...

    [...]

  • ...…e t a t e using relatively small amounts of lsopropanol (about O. 6 volumes) has been reported tit separate high molecular DNA from polysaccharides (Marmur, 1961) The sodium acetate also yields ,l tight fibrous precipitate th,it is easily washed and dried The DNA will dissolve readily if allowed…...

    [...]

Book ChapterDOI
TL;DR: This chapter describes the rapid DNA isolations for enzymatic and hybridization analysis for identifying the size of a particular cloned DNA fragment or for surveying the genome organization in a large number of individual organisms or strains.
Abstract: Publisher Summary This chapter describes the rapid DNA isolations for enzymatic and hybridization analysis. The methods described are for the isolation of DNA from bacteria, yeast, and tissue culture cells. The ability to rapidly isolate DNA from a large number of individual organisms has facilitated the characterization of the genome of the organisms. Ability to rapidly isolate DNA from organisms containing cloned DNA segments using recombinant DNA techniques has facilitated the identification of particular clones. These methods have been used for identifying the size of a particular cloned DNA fragment or for surveying the genome organization in a large number of individual organisms or strains. The general approach is to remove enzymatically or physically any rigid cell wall and then to lyse the cells with a detergent, usually SDS. Nucleases are inactivated with diethyl oxydiformate and most of the cell debris, proteins, and SDS are precipitated by addition of potassium. The DNA is then recovered by ethanol precipitation. RNA present in the preparations interferes with most restriction endonucleases, and is eliminated by ribonuclease treatment. The digested RNA does not interfere with analysis and is not removed.

521 citations