Establishment of an Indirect Genetic Transformation Method for Arabidopsis thaliana ecotype Bangladesh
نویسندگان
چکیده
Arabidopsis thaliana is a small flowering plant belonging to the Brassicaceae family, which is adopted as a model plant for genetic research. Agrobacterium tumifaciensmediated transformation method for A. thaliana ecotype Bangladesh was established. Leaf discs of A. thaliana were incubated with A. tumefaciens strain LBA4404 containing chimeric nos. nptII. nos and intron-GUS genes. Following inoculation and co-cultivation, leaf discs were cultured on selection medium containing 50 mg/l kanamycin + 50 mg/l cefotaxime + 1.5 mg/l NAA and kanamycin resistant shoots were induced from the leaf discs after two weeks. Shoot regeneration was achieved after transferring the tissues onto fresh medium of the same combination. Finally, the shoots were rooted on MS medium containing 50 mg/l kanamycin. Incorporation and expression of the transgenes were confirmed by PCR analysis. Using this protocol, transgenic A. thaliana plants can be obtained and indicates that genomic transformation in higher plants is possible through insertion of desired gene. Although Agrobacterium mediated genetic transformation is established for A. thaliana, this study was the conducted to transform A. thaliana ecotype Bangladesh.
منابع مشابه
Differential Expression of Arabidopsis thaliana Acid Phosphatases in Response to Abiotic Stresses
The objective of this research is to identify Arabidopsis thaliana genes encoding acid phosphatases induced by phosphate starvation. Multiple alignments of eukaryotic acid phosphatase amino acid sequences led to the classification of these proteins into four groups including purple acid phosphatases (PAPs). Specific primers were degenerated and designed based on conserved sequences of PAPs isol...
متن کاملAn integrated map of Arabidopsis thaliana for functional analysis of its genome sequence.
The genome of the model plant species Arabidopsis thaliana has recently been sequenced. To accelerate its current genome research, we developed a whole-genome, BAC/BIBAC-based, integrated physical, genetic, and sequence map of the A. thaliana ecotype Columbia. This new map was constructed from the clones of a new plant-transformation-competent BIBAC library and is integrated with the existing s...
متن کاملمشکلات روشهای موجود و ارائه دو روش جدید کشت هیدروپونیک گیاه آرابیدوپسیس تالیانا
Arabidopsis thaliana is a suitable model plant for genetic and molecular biology studies in higher plants. However, its hydroponic culture for biochemical and physiological studies is a challenge due to small size, capillary roots and little biomass at maturity. Several cultural systems have been suggested for Arabidopsis thaliana hydroponic culture, each having special advantages and disadvant...
متن کاملNatural variation in expression of self-incompatibility in Arabidopsis thaliana: implications for the evolution of selfing.
The switch from an out-crossing to a self-fertilizing mating system is one of the most prevalent evolutionary trends in plant reproduction and is thought to have occurred repeatedly in flowering plants. However, little is known about the evolution of self-fertility and the genetic architecture of selfing. Here, we establish Arabidopsis thaliana as a model for genetic analysis of the switch to s...
متن کاملYeast Two Hybrid cDNA Screening of Arabidopsis thaliana for SETH4 Protein Interaction
SETH4 coding sequence with 2013 bp is a member of gene family expressed in gametophytic tissues of Arabidopsis thaliana. This fragment was PCR amplified using Kod Hi Fi DNA polymerase enzyme. This fragment was cloned into pGBKT7 bate vector and transformed E. coli DH5? cells containing vector were selected on LB medium containing Kanamycin. Finally, pGBKT7-SETH4 bate was transformed into yeast ...
متن کامل