scholarly journals Transformation of plants with target gene encoding glutathione S-transferase to induce stress resistance

2020 ◽  
Author(s):  
E. V. Mikhaylova ◽  
M. Yu. Shein ◽  
V. Yu. Alekseev ◽  
E. A. Baimukhametova ◽  
Kh. G. Musin ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Nicotiana Tabacum ◽  
Transgenic Plants ◽  
Stress Resistance ◽  
Target Gene ◽  
Induce Stress ◽  
Eruca Sativa ◽  
Glutathione S Transferase ◽  
Gene Encoding

Transgenic plants Nicotiana tabacum, Brassica napus, B. rapa, Eruca sativa with target gene AtGSTF11 were created. Their stress resistance is being studied.

scholarly journals Dehiscence-related expression of an Arabidopsis thaliana gene encoding a polygalacturonase in transgenic plants of Brassica napus

1999 ◽  
Vol 22 (2) ◽  
pp. 159-167 ◽  
Author(s):  
E. S. JENKINS ◽  
W. PAUL ◽  
M. CRAZE ◽  
C. A. WHITELAW ◽  
A. WEIGAND ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Brassica Napus ◽  
Transgenic Plants ◽  
Gene Encoding

scholarly journals Aktivitas ACCase mesokarp kelapa sawit dan kloning fragmen gen penyandi ACCase subunit biotin karboksilase ACCase activity of oil palm mesocarp and cloning of gene fragment encoding biotin carboxylase subunit of ACCase

2016 ◽  
Vol 74 (1) ◽  
Author(s):  
Asmini BUDIANI ◽  
Djoko SANTOSO ◽  
Hajrial ASWIDINNOOR ◽  
Antonius SUWANTO
Keyword(s):  
Arabidopsis Thaliana ◽  
Brassica Napus ◽  
Nicotiana Tabacum ◽  
Oil Palm ◽  
Annealing Temperature ◽  
Rt Pcr ◽  
Cdna Fragment ◽  
Gene Encoding ◽  
Conserved Region ◽  
Cloned Cdna

Summary Genetic engineering to produce high yielding oil palm might be done by over expressing gene encoding key enzyme for oil biosynthesis in the oil palm mesocarp, one of which is ACCase. The objective of this research was to analyze ACCase activity of mesocarp from several developmental stages of fruit and to clone conserved region cDNA of gene encoding biotin carboxylase subunit of ACCase (BC-htACCase) from oil palm mesocarp. Activity of ACCase was analyzed by HPLC. Amplification of cDNA was done by means of reverse transcriptase polymerase chain reaction (RT-PCR) using degenerate heterologous primer on several annealing temperature and MgCl2 concentration. The cDNA fragment of RT-PCR product was cloned, sequenced and analyzed to confirm that the cloned cDNA was conserved region of BC-htACCase. The result showed that ACCase activity increased from the 14 week to the 20 week-old fruit, and then decreased. Using heterologous degenerate primers, cDNA fragments of BC-htACCase conserved region (469 bp) can be specifically amplified at 60 oC annealing temperature with 2 mM MgCl2 concentration.The result of BlastX analysis showed that the sequence of cloned cDNA fragment was highly homologous with the conserved region of BC-htACCase from Glycine max, Arabidopsis thaliana, Nicotiana tabacum,  and Brassica napus with 243, 237, 236, 231 bit score, and E. value 2e-63, 1e-61, 2e-61 and 5e-60, respectively. Ringkasan Rekayasa genetika untuk menghasilkan bibit kelapa sawit berdaya hasil tinggi dapat ditempuh dengan meningkatkan ekspresi gen penyandi enzim kunci biosintesis minyak pada kelapa sawit, salah satunya adalah ACCase. Tujuan penelitian ini adalah menguji aktivitas ACCase mesokarp beberapa tahap perkem-bangan buah sawit dan mengklon fragmen cDNA daerah konservatif gen penyandi ACCase heteromerik subunit biotin karbok-silase (BC-htACCase) dari mesokarp buah sawit. Aktivitas ACCase dianalisis dengan HPLC. Amplifikasi cDNA dilakukan dengan teknik RT-PCR menggunakan primer degene-rate heterologus pada berbagai suhu penempelan dan konsentrasi MgCl2. Fragmen cDNA hasil RT-PCR diklon, disekuen dan dianalisis untuk mengkonfirmasi bahwa cDNA terklon adalah daerah konservatif BC-htACCase. Hasil penelitian menunjukkan bahwa aktivitas ACCase meningkat dari buah berumur 14 minggu hingga buah berumur  20 minggu, kemudian menurun kembali Dengan primer degenerate heterologus, fragmen cDNA daerah konservatif BC-htACCase  (469 pb) dapat diamplifikasi secara spesifik pada suhu penempelan 60 oC dan konsentrasi MgCl2 2 mM. Hasil analisis BlastX dari sekuen DNA fragmen terklon menunjuk-kan bahwa sekuen tersebut mempunyai homologi tinggi antara lain dengan gen penyandi BC-htACCase dari Glycine max, Arabidopsis thaliana, Nicotiana tabacum dan Brassica napus, masing-masing dengan skor 243, 237, 236, 231 bit, dan E. value 2e-63, 1e-61, 2e-61 dan 5e-60.

scholarly journals Study of the ascorbate-glutathione cycle in transgenic plants Nicotiana tabacum under abiotic stress conditions

2021 ◽  
pp. 11-18
Author(s):  
Kristina V. Pristupa ◽  
Tatsiana A. Kukulianskaya ◽  
Elena A. Khramtsova
Keyword(s):  
Abiotic Stress ◽  
Nicotiana Tabacum ◽  
Transgenic Plants ◽  
Biochemical Parameters ◽  
Acc Deaminase ◽  
Polluted Soils ◽  
Gene Encoding ◽  
Acds Gene ◽  
Glutathione Cycle ◽  
Transgenic Nicotiana Tabacum

We conducted a comparative analysis of biochemical parameters for the ascorbate-glutathione cycle in non-transgenic and transgenic Nicotiana tabacum, plants cultivated in heavy metal polluted soils. Transgenic plants had in their genome a bacterial acdS-gene encoding the 1-aminocyclopropane-1-carboxylate deaminase (ACC-deaminase) enzyme. The introduction of elevated concentrations of copper, chromium, and lead ions into the soil promotes induction of the acdS-gene expression and an increase in ACC-deaminase activity in transgenic plants. It was shown the content of ascorbic acid (АА), glutathione, ascorbate peroxidase (АРХ) and glutathione peroxidase (GP) and glutathione reductase (GR) increased in plants under abiotic stress.

scholarly journals Analysis of the low-molecular weight antioxidants of transgenic plants Nicotiana tabacum under abiotic stress conditions

2020 ◽  
pp. 20-26
Author(s):  
Kristina V. Pristupa ◽  
Tatsiana A. Kukulianskaya ◽  
Elena A. Khramtsova
Keyword(s):  
Abiotic Stress ◽  
Nicotiana Tabacum ◽  
Transgenic Plants ◽  
Biochemical Parameters ◽  
Acc Deaminase ◽  
Low Molecular Weight ◽  
Polluted Soils ◽  
Gene Encoding ◽  
Acds Gene ◽  
Total Antioxidant

We conducted a comparative analysis of biochemical parameters of non-transgenic and transgenic Nicotiana tabacum, plants cultivated in heavy metal polluted soils. Transgenic plants had in their genome a bacterial acdS-gene encoding the 1-aminocyclopropane-1-carboxylate deaminase (ACC-deaminase) enzyme. The introduction of elevated concentrations of copper, chromium, and lead ions into the soil promotes induction of the acdS-gene expression and an increase in ACC-deaminase activity in transgenic plants. It was shown that the content of phenolic compounds (flavonoids), ascorbic acid, the total antioxidant activity of plants increased under abiotic stress.

scholarly journals Aktivitas ACCase mesokarp kelapa sawit dan kloning fragmen gen penyandi ACCase subunit biotin karboksilase ACCase activity of oil palm mesocarp and cloning of gene fragment encoding biotin carboxylase subunit of ACCase

2016 ◽  
Vol 74 (1) ◽  
Author(s):  
Asmini BUDIANI ◽  
Djoko SANTOSO ◽  
Hajrial ASWIDINNOOR ◽  
Antonius SUWANTO
Keyword(s):  
Arabidopsis Thaliana ◽  
Brassica Napus ◽  
Nicotiana Tabacum ◽  
Oil Palm ◽  
Annealing Temperature ◽  
Rt Pcr ◽  
Cdna Fragment ◽  
Gene Encoding ◽  
Conserved Region ◽  
Cloned Cdna

Summary Genetic engineering to produce high yielding oil palm might be done by over expressing gene encoding key enzyme for oil biosynthesis in the oil palm mesocarp, one of which is ACCase. The objective of this research was to analyze ACCase activity of mesocarp from several developmental stages of fruit and to clone conserved region cDNA of gene encoding biotin carboxylase subunit of ACCase (BC-htACCase) from oil palm mesocarp. Activity of ACCase was analyzed by HPLC. Amplification of cDNA was done by means of reverse transcriptase polymerase chain reaction (RT-PCR) using degenerate heterologous primer on several annealing temperature and MgCl2 concentration. The cDNA fragment of RT-PCR product was cloned, sequenced and analyzed to confirm that the cloned cDNA was conserved region of BC-htACCase. The result showed that ACCase activity increased from the 14 week to the 20 week-old fruit, and then decreased. Using heterologous degenerate primers, cDNA fragments of BC-htACCase conserved region (469 bp) can be specifically amplified at 60 oC annealing temperature with 2 mM MgCl2 concentration.The result of BlastX analysis showed that the sequence of cloned cDNA fragment was highly homologous with the conserved region of BC-htACCase from Glycine max, Arabidopsis thaliana, Nicotiana tabacum,  and Brassica napus with 243, 237, 236, 231 bit score, and E. value 2e-63, 1e-61, 2e-61 and 5e-60, respectively. Ringkasan Rekayasa genetika untuk menghasilkan bibit kelapa sawit berdaya hasil tinggi dapat ditempuh dengan meningkatkan ekspresi gen penyandi enzim kunci biosintesis minyak pada kelapa sawit, salah satunya adalah ACCase. Tujuan penelitian ini adalah menguji aktivitas ACCase mesokarp beberapa tahap perkem-bangan buah sawit dan mengklon fragmen cDNA daerah konservatif gen penyandi ACCase heteromerik subunit biotin karbok-silase (BC-htACCase) dari mesokarp buah sawit. Aktivitas ACCase dianalisis dengan HPLC. Amplifikasi cDNA dilakukan dengan teknik RT-PCR menggunakan primer degene-rate heterologus pada berbagai suhu penempelan dan konsentrasi MgCl2. Fragmen cDNA hasil RT-PCR diklon, disekuen dan dianalisis untuk mengkonfirmasi bahwa cDNA terklon adalah daerah konservatif BC-htACCase. Hasil penelitian menunjukkan bahwa aktivitas ACCase meningkat dari buah berumur 14 minggu hingga buah berumur  20 minggu, kemudian menurun kembali Dengan primer degenerate heterologus, fragmen cDNA daerah konservatif BC-htACCase  (469 pb) dapat diamplifikasi secara spesifik pada suhu penempelan 60 oC dan konsentrasi MgCl2 2 mM. Hasil analisis BlastX dari sekuen DNA fragmen terklon menunjuk-kan bahwa sekuen tersebut mempunyai homologi tinggi antara lain dengan gen penyandi BC-htACCase dari Glycine max, Arabidopsis thaliana, Nicotiana tabacum dan Brassica napus, masing-masing dengan skor 243, 237, 236, 231 bit, dan E. value 2e-63, 1e-61, 2e-61 dan 5e-60.

Agrobacterium tumefaciens-mediated Transformation of Double Haploid Canola (Brassica napus) Lines

1997 ◽  
Vol 24 (1) ◽  
pp. 97 ◽  
Author(s):  
K. Kazan ◽  
M. D. Curtis ◽  
K. C. Goulter ◽  
J. M. Manners
Keyword(s):  
Brassica Napus ◽  
Agrobacterium Tumefaciens ◽  
Transgenic Plants ◽  
Genetic Transformation ◽  
Double Haploid ◽  
Gene Promoter ◽  
Root Explants ◽  
Hypocotyl Explants ◽  
Peroxidase Gene ◽  
Hypocotyl Segments

Double haploid (DH) genotypes of canola (Brassica napus L.) have a high level of genetic uniformity but have not been previously tested for genetic transformation. Transgenic plants from three of four DH genotypes derived from cv. Westar were obtained by inoculation of either hypocotyl segments or root explants with Agrobacterium tumefaciens. For hypocotyl transformation, A. tumefaciens strain LBA4404 containing a binary plasmid with the neomycin phosphotransferase gene (nptII) and a CaMV 35S-peroxidase gene cassette was co-cultivated with hypocotyl segments taken from the 5–6-day-old seedlings. Transformation frequencies for hypocotyl explants of two DH genotypes were 0.3–3%. Direct evidence for genetic transformation of hypocotyl explants was obtained through molecular hybridisation analysis. Using this protocol, mature transformed plants were obtained within 4–6 months of co-cultivation. A method of root transformation was successfully modified for one DH genotype of canola and transgenic plants were obtained at a frequency of 2%. Using this protocol, a peroxidase gene promoter–GUS fusion construct was introduced into a DH genotype. Tissue specific GUS expression driven by the peroxidase gene promoter in transgenic plants was analysed by GUS staining. Transformation systems for double haploid canola lines will permit the assessment of introduced genes for their effect on agronomic and physiological traits.

scholarly journals Effects of cobalt oxide nanoparticles (Co3O4 NPs) on ion leakage, total phenol, antioxidant enzymes activities and cobalt accumulation in Brassica napus L.

2020 ◽  
Vol 48 (3) ◽  
pp. 1260-1275
Author(s):  
Malihe JAHANI ◽  
Ramazan Ali KHAVARI-NEJAD ◽  
Homa MAHMOODZADEH ◽  
Sara SAADATMAND
Keyword(s):  
Brassica Napus ◽  
Cobalt Oxide ◽  
Growth Parameters ◽  
Maximum Level ◽  
Defense Responses ◽  
Guaiacol Peroxidase ◽  
Ion Leakage ◽  
Glutathione S Transferase ◽  
Brassica Napus L ◽  
Cobalt Oxide Nanoparticles

Interaction of nanoparticles (NPs) as a significant threat to ecosystems with biological processes of plants is very important. Here, the effects of cobalt oxide (Co3O4) NPs on some physio-biochemical characteristics of Brassica napus L. were investigated. The two-weeks seedlings were sprayed with different concentrations of Co3O4 NPs (0, 50, 100, 250, 500, 1000, 2000, and 4000 mg L-1). The results showed that this treatment significantly affected the fresh and dry weights, area, relative water content (RWC) and relative chlorophyll value (SPAD) of leaves. The highest reduction of growth and biomass indexes occurred at 4000 mg L-1 NPs. The content of H2O2 and electrolyte leakage (EL) increased respectively, after 100 and 250 mg L-1 of Co3O4 NPs and showed a maximum level at 4000 mg L-1. The activities of phenylalanine ammonia lyase (PAL), ascorbate peroxidase (APX) and superoxide dismutase (SOD) increased after 100 mg L-1 of Co3O4 NPs. However, tyrosine ammonia lyase (TAL) activity enhanced after 500 mg L-1. The catalase (CAT) activity and protein content decreased after 1000 mg L-1 of Co3O4 NPs. Application of concentrations higher than 500 mg L-1 of Co3O4 NPs induced polyphenol oxidase (PPO) activity but reduced glutathione reductase (GR). The activities of guaiacol peroxidase (GPX) and glutathione S-transferase (GST) increased at 250-1000 mg L-1 of Co3O4 NPs and then decreased. These results suggested that low concentrations of Co3O4 NPs induced a positive effect on growth parameters but high levels caused extensive oxidative damage and mediated defense responses by organization of phenolic compounds and antioxidative system.

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