Modular cloning in plant cells

Karimi, Mansour; Meyer, Björn De; Hilson, Pierre

doi:10.1016/j.tplants.2005.01.008

Cited by 549 publications

(485 citation statements)

References 15 publications

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“…Approximately 1200 bp of 59 and 39 flanking regions of the chiC2 gene were amplified from genomic DNA by PCR using the primer sequences listed in Table S2, while the hygromycin-resistance gene (hygB) was amplified from the pCT74 vector (Lorang et al, 2001) as described previously (Dubey et al, 2013). The deletion cassette was constructed using the gateway cloning technique according to the manufacturer's instructions (Invitrogen), while the pPm43GW vector (Karimi et al, 2005) was used as the destination vector in order to construct the pPm43GW-03743-ko deletion vector.…”

Section: Methodsmentioning

confidence: 99%

Identifying glycoside hydrolase family 18 genes in the mycoparasitic fungal species Clonostachys rosea

et al. 2015

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Clonostachys rosea is a mycoparasitic fungal species that is an efficient biocontrol agent against many plant diseases. During mycoparasitic interactions, one of the most crucial steps is the hydrolysis of the prey's fungal cell wall, which mainly consists of glucans, glycoproteins and chitin. Chitinases are hydrolytic enzymes responsible for chitin degradation and it is suggested that they play an important role in fungal-fungal interactions. Fungal chitinases belong exclusively to the glycoside hydrolase (GH) family 18.These GH18 proteins are categorized into three distinct phylogenetic groups (A, B and C), subdivided into several subgroups. In this study, we identified 14 GH18 genes in the C. rosea genome, which is remarkably low compared with the high numbers found in mycoparasitic Trichoderma species. Phylogenetic analysis revealed that C. rosea contains eight genes in group A, two genes in group B, two genes in group C, one gene encoding a putative ENGase (endo-b-N-acetylglucosaminidase) and the ech37 gene, which is of bacterial origin. Gene expression analysis showed that only two genes had higher transcription levels during fungal-fungal interactions, while eight out of 14 GH18 genes were triggered by chitin. Furthermore, deletion of the C group chiC2 gene decreased the growth inhibitory activity of C. rosea culture filtrates against Botrytis cinerea and Rhizoctonia solani, although the biocontrol ability of C. rosea against B. cinerea was not affected. In addition, a potential role of the CHIC2 chitinase in the sporulation process was revealed. These results provide new information about the role of GH18 proteins in mycoparasitic interactions.

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Section: Methodsmentioning

confidence: 99%

Identifying glycoside hydrolase family 18 genes in the mycoparasitic fungal species Clonostachys rosea

et al. 2015

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“…For the subcellular localization assay, the open reading frame of NFL without the stop codon was cloned into pENTR_D_TOPO vector (Invitrogen) and recombined with a destination vector pB7WGY2 (Karimi et al, 2005). This construct, named p35S:NFL-YFP, was sequenced and then transformed into wt plants using the Agrobacteriummediated transformation protocol as described (Clough and Bent, 1998).…”

Section: Subcellular Localization Of Nflmentioning

confidence: 99%

NO FLOWERING IN SHORT DAY (NFL) is a bHLH transcription factor that promotes flowering specifically under short-day in Arabidopsis

et al. 2016

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Flowering in plants is a dynamic and synchronized process where various cues including age, day length, temperature and endogenous hormones fine-tune the timing of flowering for reproductive success. Arabidopsis thaliana is a facultative long day (LD) plant where LD photoperiod promotes flowering. Arabidopsis still flowers under shortday (SD) conditions, albeit much later than in LD conditions. Although factors regulating the inductive LD pathway have been extensively investigated, the non-inductive SD pathway is much less understood. Here, we identified a key basic helix-loop-helix transcription factor called NFL (NO FLOWERING IN SHORT DAY) that is essential to induce flowering specifically under SD conditions in Arabidopsis. nfl mutants do not flower under SD conditions, but flower similar to the wild type under LD conditions. The no-flowering phenotype in SD is rescued either by exogenous application of gibberellin (GA) or by introducing della quadruple mutants in the nfl background, suggesting that NFL acts upstream of GA to promote flowering. NFL is expressed at the meristematic regions and NFL is localized to the nucleus. Quantitative RT-PCR assays using apical tissues showed that GA biosynthetic genes are downregulated and the GA catabolic and receptor genes are upregulated in the nfl mutant compared with the wild type, consistent with the perturbation of the endogenous GA biosynthetic and catabolic intermediates in the mutant. Taken together, these data suggest that NFL is a key transcription factor necessary for promotion of flowering under non-inductive SD conditions through the GA signaling pathway.

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“…The 43 Myc tag was amplified in a two-step PCR from the binary vector pGWB17 (Nakagawa et al, 2007) using the corresponding primers (Supplemental Table S1) and recombined into the entry vector pDONR P2R-P3. All three fragments (promoter, gene, and tag) were shuttled into the destination vector pB7m34GW (Karimi et al, 2005) to obtain P AHK2 :AHK2-Myc and P AHK3 :AHK3-Myc.…”

Section: Recombinant Dna Techniquesmentioning

confidence: 99%

The Cytokinin Receptors of Arabidopsis Are Located Mainly to the Endoplasmic Reticulum

et al. 2011

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The plant hormone cytokinin is perceived by membrane-located sensor histidine kinases. Arabidopsis (Arabidopsis thaliana) possesses three cytokinin receptors: ARABIDOPSIS HISTIDINE KINASE2 (AHK2), AHK3, and CYTOKININ RESPONSE1/ AHK4. The current model predicts perception of the cytokinin signal at the plasma membrane. However, cytokinin-binding studies with membrane fractions separated by two-phase partitioning showed that in the wild type, as well as in mutants retaining only single cytokinin receptors, the major part of specific cytokinin binding was associated with endomembranes. Leaf epidermal cells of tobacco (Nicotiana benthamiana) expressing receptor-green fluorescent protein fusion proteins and bimolecular fluorescence complementation analysis showed strong fluorescence of the endoplasmic reticulum (ER) network for all three receptors. Furthermore, separation of the microsomal fraction of Arabidopsis plants expressing Myc-tagged AHK2 and AHK3 receptors by sucrose gradient centrifugation followed by immunoblotting displayed the Mg 2+ -dependent density shift typical of ER membrane proteins. Cytokinin-binding assays, fluorescent fusion proteins, and biochemical fractionation all showed that the large majority of cytokinin receptors are localized to the ER, suggesting a central role of this compartment in cytokinin signaling. A modified model for cytokinin signaling is proposed.

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Modular cloning in plant cells

Cited by 549 publications

References 15 publications

Identifying glycoside hydrolase family 18 genes in the mycoparasitic fungal species Clonostachys rosea

Identifying glycoside hydrolase family 18 genes in the mycoparasitic fungal species Clonostachys rosea

NO FLOWERING IN SHORT DAY (NFL) is a bHLH transcription factor that promotes flowering specifically under short-day in Arabidopsis

The Cytokinin Receptors of Arabidopsis Are Located Mainly to the Endoplasmic Reticulum

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