Efficient isolation and mapping of <i>Arabidopsis thaliana</i> T‐DNA insert junctions by thermal asymmetric interlaced PCR

Liu, Yao-Guang; Mitsukawa, Norihiro; Oosumi, Teruko; Whittier, Robert F.

doi:10.1046/j.1365-313x.1995.08030457.x

Cited by 1,315 publications

(1,108 citation statements)

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“…The flanking sequence of T-DNA in rpa2c was isolated by thermal asymmetric interlaced PCR according to a previous description (Liu et al, 1995;Zhang et al, 2006). Genotyping of the T-DNA insertion in rpa2c was determined by PCR using the T-DNA left border primer NTLB5 coupled with gene-specific primers on both sides of the insertion: 2c-F/R.…”

Section: Flanking Sequence Isolation and Genotyping Of T-dna Insertiomentioning

confidence: 99%

Replication Protein A2c Coupled with Replication Protein A1c Regulates Crossover Formation during Meiosis in Rice

Chang

Xin

et al. 2013

The Plant Cell

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Replication protein A (RPA) is a conserved heterotrimeric protein complex comprising RPA1, RPA2, and RPA3 subunits involved in multiple DNA metabolism pathways attributable to its single-stranded DNA binding property. Unlike other species possessing a single RPA2 gene, rice (Oryza sativa) possesses three RPA2 paralogs, but their functions remain unclear. In this study, we identified RPA2c, a rice gene preferentially expressed during meiosis. A T-DNA insertional mutant (rpa2c) exhibited reduced bivalent formation, leading to chromosome nondisjunction. In rpa2c, chiasma frequency is reduced by ;78% compared with the wild type and is accompanied by loss of the obligate chiasma. The residual ;22% chiasmata fit a Poisson distribution, suggesting loss of crossover control. RPA2c colocalized with the meiotic cohesion subunit REC8 and the axis-associated protein PAIR2. Localization of REC8 was necessary for loading of RPA2c to the chromosomes. In addition, RPA2c partially colocalized with MER3 during late leptotene, thus indicating that RPA2c is required for class I crossover formation at a late stage of homologous recombination. Furthermore, we identified RPA1c, an RPA1 subunit with nearly overlapping distribution to RPA2c, required for ;79% of chiasmata formation. Our results demonstrate that an RPA complex comprising RPA2c and RPA1c is required to promote meiotic crossovers in rice.

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Section: Flanking Sequence Isolation and Genotyping Of T-dna Insertiomentioning

confidence: 99%

Replication Protein A2c Coupled with Replication Protein A1c Regulates Crossover Formation during Meiosis in Rice

Chang

Xin

et al. 2013

The Plant Cell

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“…To amplify genomic sequences flanking the Ds insertion in WET121, TAIL-PCR was performed essentially as described (Liu et al, 1995;Vroemen et al, 1998). In brief, two arbitrary degenerate (AD) primers, and three specific nested Ds primers were used to amplify the Ds flanking sequences in a primary, a secondary and a tertiary round of TAIL-PCR.…”

Section: Analysis Of Flanking Sequences Of the Ds Insertion In Wet121mentioning

confidence: 99%

“…To determine the location of the Ds element in the genome of WET121, genomic DNA flanking the Ds insertion was amplified by thermal asymmetric interlaced (TAIL) PCR, essentially as described previously (Liu et al, 1995;Vroemen et al, 1998). Figure 3B shows that TAIL-PCR resulted in specific PCR products in the secondary and tertiary round of TAIL-PCR.…”

Section: Molecular Analysis Of Wet121mentioning

confidence: 99%

Colonization of the Arabidopsis rhizosphere by fluorescent Pseudomonas spp. activates a root-specific, ethylene-responsive PR-5 gene in the vascular bundle

et al. 2005

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Plants of which the roots are colonized by selected strains of non-pathogenic, fluorescent Pseudomonas spp. develop an enhanced defensive capacity against a broad spectrum of foliar pathogens. In Arabidopsis thaliana, this rhizobacteria-induced systemic resistance (ISR) functions independently of salicylic acid but requires responsiveness to jasmonic acid and ethylene. In contrast to pathogen-induced systemic acquired resistance (SAR), ISR is not associated with systemic changes in the expression of genes encoding pathogenesis-related (PR) proteins. To identify genes that are specifically expressed in response to colonization of the roots by ISRinducing Pseudomonas fluorescens WCS417r bacteria, we screened a collection of Arabidopsis enhancer trap and gene trap lines containing a transposable element of the Ac/Ds system and the GUS reporter gene. We identified an enhancer trap line (WET121) that specifically showed GUS activity in the root vascular bundle upon colonization of the roots by WCS417r. Fluorescent Pseudomonas spp. strains P. fluorescens WCS374r and P. putida WCS358r triggered a similar expression pattern, whereas ISR-non-inducing Escherichia coli bacteria did not. Exogenous application of the ethylene precursor 1-aminocyclopropane-1-carboxylate (ACC) mimicked the rhizobacteria-induced GUS expression pattern in the root vascular bundle, whereas methyl jasmonic acid and salicylic acid did not, indicating that the Ds element in WET121 is inserted in the vicinity of an ethylene-responsive gene. Analysis of the expression of the genes in the close vicinity of the Ds element revealed AtTLP1 as the gene responsible for the in cis activation of the GUS reporter gene in the root vascular bundle. AtTLP1 encodes a thaumatin-like protein that belongs to the PR-5 family of PR proteins, some of which possess antimicrobial properties. AtTLP1 knockout mutant plants showed normal levels of WCS417r-mediated ISR against the bacterial leaf pathogen Pseudomonas syringae pv. tomato DC3000, suggesting that expression of AtTLP1 in the roots is not required for systemic expression of ISR in the leaves. Together, these results indicate that induction of AtTLP1 is a local response of Arabidopsis roots to colonization by non-pathogenic fluorescent Pseudomonas spp. and is unlikely to play a role in systemic resistance.

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“…To identify the Mu insertion responsible for the mto38 mutant, we used a thermal asymmetric interlaced PCR (TAIL-PCR) approach (Liu et al, 1995). DNA was extracted from 70 homozygous wild-type and 70 homozygous mto38 seedlings, and two sequential PCRs were carried out to amplify target DNA sequences with an arbitrary degenerate primer and nested Mu-specific primers that correspond to the Figure 1.…”

Section: Identification Of Mto38 and A Candidate Gene Responsible Formentioning

confidence: 99%

The Maize Zmsmu2 Gene Encodes a Putative RNA-Splicing Factor That Affects Protein Synthesis and RNA Processing during Endosperm Development

et al. 2007

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We characterized two maize (Zea mays) mutants, zmsmu2-1 and zmsmu2-3, that result from insertion of a Mutator (Mu) transposable element in the first exon of a gene homologous to the nematode gene, smu-2, which is involved in RNA splicing. In addition to having a starchy endosperm with reduced levels of zein storage proteins, homozygous zmsmu2-1 mutants manifest a number of phenotypes, including defective meristem development. The zmsmu2 mutants have poor seedling viability and surviving plants are sterile. The gene encoding ZmSMU2 is expressed in the endosperm, embryo, and shoot apex, which explains the pleiotropic nature of the mutation. We found that proper expression of Zmsmu2 is required for efficient ribosomal RNA processing, ribosome biogenesis, and protein synthesis in developing endosperm. Based on the pleiotropic nature of the mutations and the known function of animal Zmsmu2 homologs, we propose a possible role for ZmSMU2 in the development of maize endosperm, as well as a mechanism by which misregulation of zmsmu2 causes the mutant phenotypes.

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Efficient isolation and mapping of Arabidopsis thaliana T‐DNA insert junctions by thermal asymmetric interlaced PCR

Cited by 1,315 publications

References 0 publications

Replication Protein A2c Coupled with Replication Protein A1c Regulates Crossover Formation during Meiosis in Rice

Replication Protein A2c Coupled with Replication Protein A1c Regulates Crossover Formation during Meiosis in Rice

Colonization of the Arabidopsis rhizosphere by fluorescent Pseudomonas spp. activates a root-specific, ethylene-responsive PR-5 gene in the vascular bundle

The Maize Zmsmu2 Gene Encodes a Putative RNA-Splicing Factor That Affects Protein Synthesis and RNA Processing during Endosperm Development

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