The creation of sugar beet transgenic plants expressing bar gene

2010 ◽  
Vol 46 (1) ◽  
pp. 80-86 ◽  
Author(s):  
Ya. V. Mishutkina ◽  
A. M. Kamionskaya ◽  
K. G. Skryabin
Keyword(s):  
Transgenic Plants ◽  
Sugar Beet ◽  
Bar Gene ◽  
The Creation

scholarly journals Overexpression of a S-Adenosylmethionine Decarboxylase from Sugar Beet M14 Increased Araidopsis Salt Tolerance

2019 ◽  
Vol 20 (8) ◽  
pp. 1990 ◽  
Author(s):  
Meichao Ji ◽  
Kun Wang ◽  
Lin Wang ◽  
Sixue Chen ◽  
Haiying Li ◽  
...  
Keyword(s):  
Antioxidant Enzymes ◽  
Salt Stress ◽  
Transgenic Plants ◽  
Sugar Beet ◽  
Stress Tolerance ◽  
Membrane Damage ◽  
Pcr Analysis ◽  
Ros Generation ◽  
Salt Stress Tolerance ◽  
Adenosylmethionine Decarboxylase

Polyamines play an important role in plant growth and development, and response to abiotic stresses. Previously, differentially expressed proteins in sugar beet M14 (BvM14) under salt stress were identified by iTRAQ-based quantitative proteomics. One of the proteins was an S-adenosylmethionine decarboxylase (SAMDC), a key rate-limiting enzyme involved in the biosynthesis of polyamines. In this study, the BvM14-SAMDC gene was cloned from the sugar beet M14. The full-length BvM14-SAMDC was 1960 bp, and its ORF contained 1119 bp encoding the SAMDC of 372 amino acids. In addition, we expressed the coding sequence of BvM14-SAMDC in Escherichia coli and purified the ~40 kD BvM14-SAMDC with high enzymatic activity. Quantitative real-time PCR analysis revealed that the BvM14-SAMDC was up-regulated in the BvM14 roots and leaves under salt stress. To investigate the functions of the BvM14-SAMDC, it was constitutively expressed in Arabidopsis thaliana. The transgenic plants exhibited greater salt stress tolerance, as evidenced by longer root length and higher fresh weight and chlorophyll content than wild type (WT) under salt treatment. The levels of spermidine (Spd) and spermin (Spm) concentrations were increased in the transgenic plants as compared with the WT. Furthermore, the overexpression plants showed higher activities of antioxidant enzymes and decreased cell membrane damage. Compared with WT, they also had low expression levels of RbohD and RbohF, which are involved in reactive oxygen species (ROS) production. Together, these results suggest that the BvM14-SAMDC mediated biosynthesis of Spm and Spd contributes to plant salt stress tolerance through enhancing antioxidant enzymes and decreasing ROS generation.

Biolistic transformation of haploid isolated microspores of barley (Hordeum vulgare L.)

Genome ◽  
1997 ◽  
Vol 40 (4) ◽  
pp. 570-581 ◽  
Author(s):  
Qing A. Yao ◽  
Ecaterina Simion ◽  
Manilal William ◽  
Joan Krochko ◽  
Ken J. Kasha
Keyword(s):  
Transgenic Plants ◽  
Southern Blot ◽  
Blot Analysis ◽  
Southern Blot Analysis ◽  
Nuclear Dna ◽  
Active Agent ◽  
Biolistic Transformation ◽  
Pcr Analysis ◽  
Bar Gene ◽  
Isolated Microspores

Transgenic barley plants were produced by the direct delivery of plasmid DNA into isolated microspores of barley cv. Igri using high velocity microprojectiles. The plasmid pAHC25 contained the uidA and bar genes, each under the control of a maize Ubi1 promoter. Bombarded microspores were cultured and selected on solid medium containing varying concentrations (2–5 mg/L) of the Basta herbicide active agent bialaphos. The effectiveness of selection with bialaphos depended on its interaction with the medium component glutamine. Six transgenic plants (R0) were obtained, and the presence of the uidA and bar genes and their integration into nuclear DNA in transformed R0 plants were confirmed by PCR and Southern blot analysis. Phosphinothricin acetyltransferase activity was observed in all six R0 transgenic plants, whereas none showed β-glucuronidase (GUS) activity in histochemical GUS assays. Two of the six R0 plants were haploid and sterile; one of them was trisomic and partially sterile; the remainder were diploid, but one of them was also sterile. Inheritance of the transgenes in progeny of three seed-producing transgenic plants was investigated. Southern blot analysis of genomic DNA from R1 plants showed that the introduced bar and uidA genes were hemizygous and stably cotransmitted to the R1 progeny derived from self-pollination. Analysis of Basta resistance and the integration of the bar gene by PCR analysis in R1 plants indicated that the bar gene was being inherited and expressed as a single dominant trait. Fluorescent in situ hybridization was performed on chromosomes of the trisomic plant to confirm the presence of transgenes in the genome.Key words: barley, microspore, biolistic transformation, bialaphos, haploid, FISH.

scholarly journals Advances in development of transgenic resistance to beet necrotic yellow vein virus (BNYVV) in sugar beet

Genetika ◽  
2005 ◽  
Vol 37 (3) ◽  
pp. 181-189
Author(s):  
Nevena Nagl ◽  
Ivan Atanasov ◽  
Krasimir Rusanov ◽  
Svetlana Paunovic ◽  
Lazar Kovacev ◽  
...  
Keyword(s):  
Coat Protein ◽  
Sugar Beet ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Transformation Method ◽  
Yellow Vein ◽  
Bar Gene ◽  
Axillary Shoots ◽  
Viral Cdna

Fragments of viral cDNA containing the coat protein gene of beet necrotic yellow vein virus were cloned in plant transformation vector pCAMBIA3301M with the bar gene as selectable marker. Vector pC3301MCPL carrying coat protein gene with leader sequence, and pC3301MCPS with coat protein gene, were used in Agrobacterium - mediated transformation of sugar beet. The transformation method used was based on the fact that sugar beet develops axillary shoots in in vitro conditions, when placed on media with citokinins. Since this ability is not genotype or ploidy dependant it is widely used for sugar beet vegetative multiplication. Sterile seedlings, with removed cotyledons and lower half of hypocotyl, were used as starting material. After transformation ex-plants were put on micropropagation medium with cephotaxime and phosphinotricyn (ppt), where axillary shoots started to develop. Since concentration of ppt was not selective enough, after two subcultivations it was increased twofold. Only one sample, transformed with pC3301MCPS preserved morphogenetic potential for micropropagatio, and it was tested for presence of COS fragment and bar gene bz PCR with soecific primers.

scholarly journals BvcZR3 and BvHs1pro-1 Genes Pyramiding Enhanced Beet Cyst Nematode (Heterodera schachtii Schm.) Resistance in Oilseed Rape (Brassica napus L.)

2019 ◽  
Vol 20 (7) ◽  
pp. 1740 ◽  
Author(s):  
Xuanbo Zhong ◽  
Qizheng Zhou ◽  
Nan Cui ◽  
Daguang Cai ◽  
Guixiang Tang
Keyword(s):  
Transgenic Plants ◽  
Sugar Beet ◽  
Oilseed Rape ◽  
Gene Pyramiding ◽  
Nematode Resistance ◽  
Cyst Nematode ◽  
Heterodera Schachtii ◽  
Translocation Line ◽  
Wild Type ◽  
Beet Cyst Nematode

Beet cyst nematode (Heterodera schachtii Schm.) is one of the most damaging pests in sugar beet growing areas around the world. The Hs1pro-1 and cZR3 genes confer resistance to the beet cyst nematode, and both were cloned from sugar beet translocation line (A906001). The translocation line carried the locus from B. procumbens chromosome 1 including Hs1pro-1 gene and resistance gene analogs (RGA), which confer resistance to Heterodera schachtii. In this research, BvHs1pro-1 and BvcZR3 genes were transferred into oilseed rape to obtain different transgenic lines by A. tumefaciens mediated transformation method. The cZR3Hs1pro-1 gene was pyramided into the same plants by crossing homozygous cZR3 and Hs1pro-1 plants to identify the function and interaction of cZR3 and Hs1pro-1 genes. In vitro and in vivo cyst nematode resistance tests showed that cZR3 and Hs1pro-1 plants could be infested by beet cyst nematode (BCN) juveniles, however a large fraction of penetrated nematode juveniles was not able to develop normally and stagnated in roots of transgenic plants, consequently resulting in a significant reduction in the number of developed nematode females. A higher efficiency in inhibition of nematode females was observed in plants expressing pyramiding genes than in those only expressing a single gene. Molecular analysis demonstrated that BvHs1pro-1 and BvcZR3 gene expressions in oilseed rape constitutively activated transcription of plant-defense related genes such as NPR1 (non-expresser of PR1), SGT1b (enhanced disease resistance 1) and RAR1 (suppressor of the G2 allele of skp1). Transcript of NPR1 gene in transgenic cZR3 and Hs1pro-1 plants were slightly up-regulated, while its expression was considerably enhanced in cZR3Hs1pro-1 gene pyramiding plants. The expression of EDS1 gene did not change significantly among transgenic cZR3, Hs1pro-1 and cZR3Hs1pro-1 gene pyramiding plants and wild type. The expression of SGT1b gene was slightly up-regulated in transgenic cZR3 and Hs1pro-1 plants compared with the wild type, however, its expression was not changed in cZR3Hs1pro-1 gene pyramiding plant and had no interaction effect. RAR1 gene expression was significantly up-regulated in transgenic cZR3 and cZR3Hs1pro-1 genes pyramiding plants, but almost no expression was found in Hs1pro-1 transgenic plants. These results show that nematode resistance genes from sugar beet were functional in oilseed rape and conferred BCN resistance by activation of a CC-NBS-LRR R gene mediated resistance response. The gene pyramiding had enhanced resistance, thus offering a novel approach for the BCN control by preventing the propagation of BCN in oilseed rape. The transgenic oilseed rape could be used as a trap crop to offer an alternative method for beet cyst nematode control.

Purification and Partial Characterization of the Streptomyces viridochromogenes TÜ494 Phosphinothricin-N-acetyltransferase Mediating Resistance to the Herbicide Phosphinothricin in Transgenic Plants

1995 ◽  
Vol 50 (11-12) ◽  
pp. 796-805 ◽  
Author(s):  
Josef Vinnemeier ◽  
Wolfgang Dröge-Laser ◽  
Elfriede K Pistorius ◽  
Inge Broer
Keyword(s):  
Escherichia Coli ◽  
Transgenic Plants ◽  
Gene Product ◽  
Coli Strain ◽  
Bar Gene ◽  
Pat Gene ◽  
Sds Page ◽  
Purification Scheme ◽  
Molecular Masses

Abstract A purification scheme for the enzyme phosphinothricin-N-acetyltransferase (PAT) originating from Streptomyces viridochromogenes {pat-gene product from Streptomyces virido­chromogenes) and mediating herbicide resistance to transgenic plants was developed. The enzyme was isolated from a transformed and overproducing Escherichia coli strain. With a combination of ammonium sulfate fractionation, chromatography on DEAE-Sephadex A50-, Phenylsepharose-, Hydroxylapatite-and FPLC-Superose 12-columns it was possible to obtain PAT which was at least 90 % homogeneous on the basis of SDS-PAGE. The properties of the isolated PAT were compared with the properties of PAT from S. hygroscopicus (bar-gene product from S. hygroscopicus) previously isolated and characterisized by Botterman, J., Gossele, V., Thoen, C., Lauwereys, M. (1991), Gene 102, 33-37. Differences were observed in the molecular masses of the two native enzymes (PAT from S. viridochrogenes being a dimer of 40 kD and PAT from S. hygroscopicus being a monomer of 21 kD), and in the temperature sensitivity of the two enzymes (the PAT from S. viridochromogenes being slightly more temperature stable than PAT from S. hygroscopicus). However, since the pat and the bar-gene are to 85 % homologous, substantial similarities exist between the two enzymes especially in the kinetic values and the substrate specificity. The isolated S. virido­chromogenes PAT did not acetylate putative substrates present in the plant cell. Antibodies were raised against the isolated protein. This antiserum was able to detect PAT in transgenic plants and therefore is suitable to analyse the fate of the protein in such plants under various stress conditions.

scholarly journals Agrobacterium-Mediated Genetic Transformation of the Medicinal Plant Veratrum dahuricum

Plants ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 191 ◽  
Author(s):  
Rui Ma ◽  
Zhijing Yu ◽  
Qinan Cai ◽  
Haiyun Li ◽  
Yingshan Dong ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Anticancer Drug ◽  
Cauliflower Mosaic Virus ◽  
Changbai Mountains ◽  
35S Promoter ◽  
Bar Gene ◽  
Transformation System ◽  
Embryogenic Calli ◽  
Infection Period ◽  
Regenerated Plants

Veratrum dahuricum L. (Liliaceae), a monocotyledonous species distributed throughout the Changbai mountains of Northeast China, is pharmaceutically important, due to the capacity to produce the anticancer drug cyclopamine. An efficient transformation system of Veratrum dahuricum mediated with Agrobacterium tumefaciens is presented. Murashige and Skoog (MS) medium containing 8 mg/L picloram was used to induce embryogenic calli from immature embryos with 56% efficiency. A. tumefaciens LBA4404 carrying the bar gene driven by the cauliflower mosaic virus 35S promoter was employed for embryogenic callus inoculation. A. tumefaciens cell density OD660 = 0.8 for inoculation, half an hour infection period, and three days of co-culture duration were found to be optimal for callus transformation. Phosphinothricin (PPT, 16 mg/L) was used as the selectable agent, and a transformation efficiency of 15% (transgenic plants/100 infected calli) was obtained. The transgenic nature of the regenerated plants was confirmed by PCR and Southern blot analysis, and expression of the bar gene was detected by RT-PCR and Quick PAT/bar strips. The steroid alkaloids cyclopamine, jervine, and veratramine were detected in transgenic plants, in non-transformed and control plants collected from natural sites. The transformation system constitutes a prerequisite for the production of the pharmaceutically important anticancer drug cyclopamine by metabolic engineering of Veratrum.

Efficient production of transgenic plants using the bar gene for herbicide resistance in sweetpotato

2009 ◽  
Vol 122 (4) ◽  
pp. 649-653 ◽  
Author(s):  
Ning Zang ◽  
Hong Zhai ◽  
Shang Gao ◽  
Wei Chen ◽  
Shaozhen He ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Herbicide Resistance ◽  
Efficient Production ◽  
Bar Gene

scholarly journals Impact of soil compaction in sowing on development and crops of sugar beet

2012 ◽  
Vol 52 (No. 1) ◽  
pp. 11-16
Author(s):  
V. Uhlíř ◽  
J. Mareček ◽  
J. Červinka
Keyword(s):  
Sugar Beet ◽  
Contact Surface ◽  
Soil Compaction ◽  
Biological Characteristic ◽  
The Other ◽  
Root Crops ◽  
Other Hand ◽  
The Creation ◽  
Moisture Conditions

Putting together work operations minimizes the number of machine passes across the plot, which helps to reduce negative soil compaction and to save fuels. However, the combination of working operations also reflects in the increased weight of machines, which – on the other hand – can result exactly in soil compaction. This is why the potential adverse phenomenon can be compensated by using tyres with a larger contact surface with the base. In the case of sowing root crops, some problems may appear with the application of these tyres as a certain part of the stand has been sown in their track. The paper brings an assessment of the possibility to use twin assembly tyres on the tractor model Fendt 822 and on the sowing drill model Monosem NG plus with 18 drilling mechanisms. Parameters to be assessed were soil compaction, and the development of plants sown inside and outside the tractor track. Although the degree of soil compaction was higher in the tractor track, the biological characteristic of plants including yield reached more favourable criteria of assessment. The situation paradoxically resulted from the creation of more favourable moisture conditions in the soil.

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