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1

Improvement of an RNA purification method for grapevine (Vitis vinifera L.) suitable for cDNA library construction

100%
Daldoul S., Chenenanoui S., Mliki A., Hofer M.
Acta Physiologiae Plantarum
|
2009
|
tom 31
|
nr 4
A method is described, which consistently yields high quality total RNA from grapevine. Dissolving of crude RNA pellets in borate-containing buffer, instead of normally used water before a selective lithium chloride precipitation, was found to be a critical step, leading to a 2.5-fold increase of yield. The resulting RNA preparations were suitable for standard downstream applications and also for cDNA library construction. The method worked efficiently and reproducibly and could easily be scaled from milligram to gram quantities of plant material grown in hydroponic culture, sandy soil or Perlite. It was applied to different kinds of grapevine tissues (leaves, stem) and, after additional adaptation of the protocol, to roots.
2

In planta agro-infiltration system for transient gene expression in grapevine (Vitis spp.)

88%
Ben-Amar A., Cobanov P., Buchholz G., Mliki A., Reustle G.
Acta Physiologiae Plantarum
|
2013
|
tom 35
|
nr 11
Transient expression of foreign genes by Agrobacterium infiltration is a versatile technique that can be used as a rapid tool for functional analysis and gene silencing studies in plants. A reproducible protocol of Agrobacterium-mediated transient gene transfer was developed for gene expression analysis on greenhouse-grown grapevines, as a complementary approach towards functional genomics and alternative to transgenics. Non-detached leaves from green cuttings were used as the target organ and vacuum infiltrated for in planta inoculation with Agrobacterium tumefaciens harboring mgfp 5-ER gene construct as visual reporter gene. Step-by-step optimization was performed and showed that the quality of greenhouse material as well as agro-infiltration conditions were the major factors which influenced successful gene expression assays. Following the optimized protocol, up to half of the infiltrated leaf surface displayed green fluorescent foci found in the intercoastal areas. Monitoring of transient Green Fluorescent Protein expression daily achieved for 2 weeks post-infiltration with the highest expression level on day 6. Evidence of GFP silencing in transgenic GFP-expressing grapevine via agro-infiltration was found for the first time. The in planta infiltration system described here provides a powerful tool to explore easily gene function in grapevine avoiding tissue culture steps and the labor-intensive generation of transgenic plants.
3

Molecular cloning and characterisation of a cDNA encoding a putative alkaline alpha-galactosidase from grapevine (Vitis vinifera L.) that is differentially expressed under osmotic stress

76%
Daldoul S., Toumi I., Reustle G. M., Krczal G., Ghorbel A., Mliki A., Hofer M. U.
Acta Physiologiae Plantarum
|
2012
|
tom 34
|
nr 2
The expression of a gene originally identified by means of a salt stress-associated partial cDNA showing a genotype dependent response was studied upon exposure to salt and drought stress and during seed development of grapevines. The complete open reading frame has been cloned and the corresponding untranslated regions have been characterized by 5'- and 3'-RACE leading to a refinement and correction of the predicted gene model in the recent 12 × Vitis genome assembly. The open reading frame of the single copy gene designated Vv-α-gal/SIP encodes for a polypeptide of 774 amino acids with a calculated molecular mass of 84.9 kDa. Protein sequence comparisons suggest that the gene belongs to a growing family of plant-specific alkaline α-galactosidases. 5'-RACE identified a not yet documented transcript species derived from the same gene locus which does not lead to the formation of a full-length transcript. The ‘‘aberrant’’ transcript harbours an alternative 5'-UTR and originates from an internal transcriptional start site 1.4 kb downstream of the predominant transcriptional start site. Levels of transcripts containing the different 5'-UTRs were compared under salt stress conditions.
4

Molecular cloning and characterisation of a cDNA encoding a putative alkaline alpha-galactosidase from grapevine (Vitis vinifera L.) that is differentially expressed under osmotic stress

76%
Daldoul S., Toumi I., Reustle G. M., Krczal G., Ghorbel A., Mliki A., Hofer M. U.
Acta Physiologiae Plantarum
|
2012
|
tom 34
|
nr 3
The expression of a gene originally identified by means of a salt-stress-associated partial cDNA showing a genotype-dependent response, was studied upon exposure to salt and drought stress and during seed development of grapevines. The complete open reading frame has been cloned and the corresponding untranslated regions have been characterized by 5'- and 3'-RACE leading to a refinement and correction of the predicted gene model in the recent 12 × Vitis genome assembly. The open reading frame of the single copy gene designated Vv-α-gal/SIP encodes for a polypeptide of 774 amino acids with a calculated molecular mass of 84.9 kDa. Protein sequence comparisons suggest that the gene belongs to a growing family of plant-specific alkaline α-galactosidases. 5'-RACE identified a not yet documented transcript species derived from the same gene locus, which does not lead to the formation of a full-length transcript. The ‘‘aberrant’’ transcript harbours an alternative 5'-UTR and originates from an internal transcriptional start site 1.4 kb downstream of the predominant transcriptional start site. Levels of transcripts containing the different 5'-UTRs were compared under salt stress conditions.
5

Short-term response of wild grapevines (Vitis vinifera L. ssp. sylvestris) to NaCl salinity exposure: changes of some physiological and molecular characteristics

64%
Askri H., Daldoul S., Ammar A. B., Rejeb S., Jardak R., Rejeb M. N., Mliki A., Ghorbel A.
Acta Physiologiae Plantarum
|
2012
|
tom 34
|
nr 3
The physiological and molecular response to salt stress was studied in two wild grapevine (Vitis vinifera L. ssp. sylvestris or Vitis sylvestris) accessions ‘‘Khédhayria’’ and ‘‘Houamdia’’, previously identified as salt-tolerant and salt-sensitive pair wise. Plants from both accessions were subjected to a progressive salt stress by the use of a nutritional solution containing up to 150 mM NaCl for 2 weeks. Salt stress adversely affected growth and water potential since the first day of exposure to 150 mM NaCl. However, chlorophyll fluorescence parameters were unchanged until 14 days of salt exposure. At that time point the predawn water potential (ΨPD), the non-photochemical quenching of fluorescence (NPQ) and the coefficient of photochemical quenching (qp) were significantly less altered in the tolerant accession. At the molecular level semi-quantitative RT-PCR assays revealed a differential expression of (Vs α-gal/SIP and Vs DHN) genes within these contrasting accessions after exposure to 24 h and 14 days of salt. Comparably, the Vs RD22 gene had increased slightly after only 14 days of treatment in both accessions. These results were the first pieces of information reported on the early and late regulation of salt response genes in wild grapevines. Furthermore, genotype-dependent parameters such as NPQ, qp, mRNA levels of Vs α-gal/SIP and Vs DHN could be used to screen salt-tolerant wild grapevine genotypes.
6
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Biochemical characterization of fennel (Ferula communis L.) different parts through their essential oils, fatty acids and phenolics

64%
Rahali F. Z., Lamine M., Rebey I. B., Wannes W. A., Hammami M., Selmi S., Mliki A., Sellami I. H.
Acta Scientiarum Polonorum. Hortorum Cultus
|
2021
|
tom 20
|
nr 1
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