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1

The analysis of 14-3-3 gene family

100%
Szopa J., Czuj T., Lukaszewicz M.
Biotechnologia
|
2003
|
issue 3
95-106
EN
The 14-3-3s constitute a family of highly homologous proteins, first discovered in brain tissue and now thought to be present in all eukaryotic cells. Recently, thirteen cDNAs in Arabidopsis, seven in human cells and six in potato plant were found, all encoding highly homologous 14-3-3 protein isoforms. While there is substantial progress in the identification of diverse partners of 14-3-3 in recent years, at least two important questions need to be answered. Is there any specificity within 14-3-3 isoforms in the binding of diverse partners? Does this binding affects plant metabolism or physiology in vivo? The significance of 14-3-3 protein in potato metabolism has been shown by the use of transgenic plants in which 14-3-3 protein has been either increased by the expression of a Cucurbita pepo cDNA or decreased by an antisense RNA method. It was thus proposed that 14-3-3 protein affect the carbohydrate metabolism in potato via the regulation of catecholamine synthesis. To answer the question on isoform specificity, the isoforms gene promoters were first analysed for specific domains content by the comparison with the known sequences accumulated in database. Then, the promoter characteristic was studied in transgenic plants transformed with reporter GUS gene under the control of the 14-3-3 promoter. The data obtained strongly suggest that the function of particular isoform at least partially derives from promoter specificity.
2

Resistance gene analogues of Arabidopsis thaliana: recognition by structure

100%
Chelkowski J., Koczyk G.
Journal of Applied Genetics
|
2003
|
vol. 44
|
issue 3
311-321
EN
Following completion of Arabidopsis thaliana sequencing projects, multiple resistance gene analogues (RGAs) have been identified. In this work a review of the current state of knowledge available in protein databases and scientific articles is presented. Putative resistance genes were identified by using BLAST searches as well as HMM fingerprints (the latter to infer existence of characteristic domains). The representation of all five classes of putative resistance genes in Col-0 ecotype was examined, along with the statistics on RGAs present on all five chromosomes of Arabidopsis thaliana.
3

Transformacja Arabidopsis thaliana z udzia?em Agrobacterium tumefaciens nios?cym geny mirozynazy TGG1 lub TGG2

100%
Troczynska J., Drozdowska L.
Biotechnologia
|
2007
|
issue 4
170-179
EN
In order to obtain the plants with altered myrosinase activity, the transformation via Agrobacterium tumefaciens was performed. The myrosinase genes TGG1 or TGG2 from Arabidopsis thaliana under the control of enhanced 35S promoter and phosphotranspherase neomycin II (NPTII) as the selectable marker were introduced into Arabidopsis. The presence and expression of NPTII gene in selected plants was confirmed by ELISA and PCR analysis. The integration of T-DNA with TGG1 or TGG2 gene caused differentiated effects from a multi-fold increase of myrosinase activity to its lack. No morphological differences were observed between transgenic and control plants. All the plants were fertile and produced seeds. The majority of the plants with additional copy (copies) of TGG1 showed increased enzyme activity. Conversely, most of the plants with insertion of additional copy (copies) of TGG2 demonstrated decreased myrosinase activity. Transgenic plants may be used in further studies explaining the physiological role of glucosinolates in plant-pest interaction and the importance of the myrosinase-glucosinolates system in growth and development.
4

Effect of Fusarium culmorum infection on survivability of a T-DNA tagged mutant of Arabidopsis thaliana harboring a mutation in the peptide transporter gene At5g46050

100%
Warzecha T., Lundh D., Mandal A.
Biotechnologia
|
2011
|
issue 1
77-84
EN
Previously, we have reported a T-DNA tagged mutant (TAG_009) of Arabidopsis thaliana exhibiting a significant sensitivity to biotic stresses. We have also cloned and analyzed the tagged gene At5g46050. Based on bioinformatic and molecular characterization, we proposed that At5g46050 is involved in the transport of peptides participating in plant defense against biotic stresses. To provide further evidence for supporting our proposal, this time we exposed this mutant to Fusarium culmorum, a potential fungal pathogen. Besides TAG_009 line, in our investigations we included two SALK insertion mutants (SALK_003119 and SALK_145209), two wild-type ecotypes (WT_C24 and WT_Col-0) and an additional T-DNA tagged mutant (TAG_197-6) of A. thaliana. We have found that the highest degree of leaf damage was exhibited by TAG_009 line (damage score 4.37), whereas the lowest was observed in WT_Col-0 ecotype (damage score 3.43). The highest rate of mortality after eight weeks of inoculation with F. culmorum was also observed in TAG_009 line (85.24%), while the lowest was in WT_Col-0 line (37.22%). We have also found that plants of SALK_145209 line, despite being infected with Fusarium, produced the highest number of leaves (average 14.17 leaves per plant), whereas the lowest number of leaves was produced by plants of TAG_197-6 line ( average 9.5 leaves per plant). Statistical analyses showed that the differences between the T-DNA tagged line TAG_009 and WT_Col-0 were significant, whereas in comparison with wild-type control plants WT_C24, they were insignificant. Based on these results, we can conclude that the gene we have tagged by using T-DNA-mediated in vivo gene fusion is indeed involved in the plant defense against Fusarium infection.
5

Efficiency of direct somatic embryogenesis in Arabidopsis thaliana (L.) Heynh. under various in vitro culture conditions

88%
Gaj M. D., Czubin M.
Biotechnologia
|
2004
|
issue 1
221-235
EN
In Arabidopsis thaliana, in vitro culture of immature zygotic embryos on medium supplemented with 2,4-D results in formation of somatic embryos via direct embrogenesis (DSE). The analysis of the nature of signals/stimuli involved in determination of embryogenic response in cultured explants can reveal genetic and physiological mechanisms involved in plant embryogenesis. The key factors for DSE induction in A. thaliana are the developmental stage of the explant and the presence of 2,4-D in induction medium. The study was undertaken to analyze DSE efficiency under modified tissue culture conditions. The studied factors included: pH of induction medium, temperature during embryogenesis induction, polyamines and their precursors, genotype and origin of the explants (seed-grown or in vitro-regenerated donor plants). The significant increase of the DSE efficiency was indicated on media with higher pH (7.0-8.0) and in culture of the explants obtained from plants regenerated via secondary embryogenesis. Moreover, embryogenic potential of the regenerant-derived explants was also observed on medium lacking of 2, 4?D. Spermidine and precursors of polyamines (ornithine and arginine) included in induction medium as well as any of the tested temperatures (5,28,32C) did not stimulate the DSE efficiency in comparison to standard conditions. All 16 tested ecotypes displayed the ability for the DSE under standard culture conditions. DSE efficiency varied between the studied ecotypes, however, most of the genotypes (75%) showed high (60-100%) frequency of explants producing somatic embryos.
6

Screening for mutations affecting sexual reproduction after activation tagging in Arabidopsis thaliana

75%
Perrella G., Cremona G., Consiglio F., Errico A., Bressan R. A., Conicella C.
Journal of Applied Genetics
|
2006
|
vol. 47
|
issue 2
109-111
EN
In this work, a seed-set-based screening was performed on 70 lines of Arabidopsis thaliana after activation tagging mutagenesis to identify mutations in reproductive mechanisms. Five mutants showed significantly lower seed set than the wild type and confirmed the phenotype in the progeny. This phenotype was linked with the marker gene bar carried by T-DNA conferring glufosinate resistance. Genetic analysis revealed that the mutation inheritance was sporophytic in 3 mutants and gametophytic in 2 mutants. In addition, 2 mutants had an extra T-DNA copy. Thus activation tagging can be an effective strategy to identify new mutations affecting sporogenesis or gametogenesis.
7

Regeneration of Arabidopsis thaliana (L.) Heynh. plants via direct somatic embryogenesis

75%
Gaj M. D.
Biotechnologia
|
2001
|
issue 2
90-98
EN
In Arabidopsis biotechnology plants are regenerated in vitro via shoot organogenesis induced in callus derived from different somatic tissues. An alternative way of in vitro plant regeneration via somatic embryogenesis has not been applied in Arabidopsis so far. Recently, it was found that development of Arabidopsis somatic embryos can be induced in the culture of immature zygotic embryos and that the callus phase is not nocessary for the initiation of embryogenesis. The aim of the presented research was to determinate the in vitro culture conditions enabling high efficiency of somatic embryo induction and their conversion into plants. The influence of induction medium composition including liquid or agar medium, type and concentration of auxin, carbohydrates and ammonium sources as well as duration of auxin treatment of explants on DSE efficiency were evaluated. Advantages of described regeneration system via DSE are as follows: short time needed to induce somatic embryos (10-15 days), high efficiency of the process (up to 90% explants responded), numerous embryos produced per explant (on average 17) and high percentage of embryo conversion into fertile plants (70-80%).
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