scholarly journals Polyubiquitin Promoter-Based Binary Vectors for Overexpression and Gene Silencing in Lotus japonicus

2008 ◽  
Vol 21 (4) ◽  
pp. 375-382 ◽  
Author(s):  
Takaki Maekawa ◽  
Mitsumasa Kusakabe ◽  
Yoshikazu Shimoda ◽  
Shusei Sato ◽  
Satoshi Tabata ◽  
...  
Keyword(s):  
Rna Interference ◽  
Transgenic Plants ◽  
Lotus Japonicus ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Binary Vectors ◽  
Gus Reporter Gene ◽  
Over Expression ◽  
Gus Reporter ◽  
Alternative Choice

In this study, we compared the transcriptional activities between Cauliflower mosaic virus (CaMV)35S promoter and polyubiquitin (Ljubq1) promoter from Lotus japonicus using β-glucuronidase (gus) reporter gene in transgenic plants of L. japonicus. The promoter analysis demonstrated that the Ljubq1 promoter possessed higher activity than the CaMV35S promoter in leaves, stems, roots, nodules, and pollen. Finally, we created GATEWAY conversion technology-compatible binary vectors for over-expression and RNA interference under the Ljubq1 promoter. These materials could provide alternative choice for studies in L. japonicus.

scholarly journals An amphibian antimicrobial peptide variant expressed in Nicotiana tabacum confers resistance to phytopathogens1

2003 ◽  
Vol 370 (1) ◽  
pp. 121-127 ◽  
Author(s):  
Donatella PONTI ◽  
M. LUISA MANGONI ◽  
Giuseppina MIGNOGNA ◽  
Maurizio SIMMACO ◽  
Donatella BARRA
Keyword(s):  
Nicotiana Tabacum ◽  
Transgenic Plants ◽  
Antimicrobial Peptide ◽  
Plant Pathogens ◽  
Rana Esculenta ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Skin Secretions ◽  
Fungal Phytopathogens ◽  
Micro Organisms

Esculentin-1 is a 46-residue antimicrobial peptide present in skin secretions of Rana esculenta. It is effective against a wide variety of micro-organisms, including plant pathogens with negligible effects on eukaryotic cells. As a possible approach to enhance plant resistance, a DNA coding for esculentin-1, with the substitution Met-28Leu, was fused at the C-terminal end of the leader sequence of endopolygalacturonase-inhibiting protein, under the control of the cauliflower mosaic virus 35S promoter region, and introduced into Nicotiana tabacum. The antimicrobial peptide was isolated from the intercellular fluids of healthy leaves of transgenic plants, suggesting that it was properly processed, secreted outside cells and accumulated in the intercellular spaces. The morphology of transgenic plants was unaffected. Challenging these plants with bacterial or fungal phytopathogens demonstrated enhanced resistance up to the second generation. Moreover, transgenic plants displayed insecticidal properties.

scholarly journals Improved Cold-resistant Performance in Transgenic Grape (Vitis vinifera L.) Overexpressing Cold-inducible Transcription Factors AtDREB1b

HortScience ◽  
2009 ◽  
Vol 44 (1) ◽  
pp. 35-39 ◽  
Author(s):  
Wanmei Jin ◽  
Jing Dong ◽  
Yuanlei Hu ◽  
Zhongping Lin ◽  
Xuefeng Xu ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Vitis Vinifera ◽  
Blot Analysis ◽  
Freezing Point ◽  
Cold Treatment ◽  
Cauliflower Mosaic Virus ◽  
Vitis Vinifera L ◽  
35S Promoter ◽  
Transgenic Lines ◽  
H 26

Dehydration response element binding (DREB)1b is a cold-inducible transcription factor in Arabidopsis thaliana. DREB1b driven by cauliflower mosaic virus 35S promoter was genetically introduced into grape Vitis vinifera L. cv. Centennial Seedless through Agrobacterium-mediated transformation for improving its cold resistance and exploring new genetic breeding approaches to obtain cold-resistant cultivars. In this study, Southern blot analysis showed the DREB1b gene was integrated into the transgenic grapevines with one to two copies. Northern blot analysis showed the presence of DREB1b transcripts in the independent transgenic lines 3, 5, 6, and 7. Further characterization of transgenic grapevines confirmed that both electrolyte leakage conductivity and the freezing point of the transgenic plants were lower than those of wild-type plants. After the cold treatment at –4 °C for 12 h, 26% of transgenic plants wilted among which 95% plants recovered once being placed under the condition of temperature 22 to 25 °C. However, subjected to the same treatment, 98% of nontransgenic plants wilted and only 2% recovered. Our results lead to the conclusion that activity of DREB1b in the transgenic grape could significantly improve its resistance to cold stress.

Expression of a Chicken Ovalbumin Gene in Three Lucerne Cultivars

1991 ◽  
Vol 18 (5) ◽  
pp. 495 ◽  
Author(s):  
HE Schroeder ◽  
MRI Khan ◽  
WR Knibb ◽  
D Spencer ◽  
TJV Higgins
Keyword(s):  
Transgenic Plants ◽  
Alternative Interpretation ◽  
Cauliflower Mosaic Virus ◽  
Vector System ◽  
35S Promoter ◽  
Restriction Enzyme Digestion ◽  
Flanking Sequence ◽  
Gene Encoding ◽  
Transformed Plants ◽  
Chicken Ovalbumin

Routine procedures have been developed for the transformation of lucerne (Medicago sativa cv. Rangelander) with foreign genes using the Agrobacterium tumefaciens binary vector system and for the regeneration of transgenic plants from tissue culture, via somatic embryogenesis. Lucerne transformation was carried out with a gene encoding neomycin phosphotransferase (npt), which conferred resistance to the antibiotic kanamycin, together with a cDNA clone encoding chicken ovalbumin which was modified for expression in plant cells. The ovalbumin cDNA protein coding sequence was combined with the cauliflower mosaic virus 35S promoter and the nopaline synthase 3' flanking sequence to make a chimeric ovalbumin gene. A DNA construct containing both these genes was transferred to lucerne, and ovalbumin was detected in leaves of regenerated plants using protein immunoblots. Pulse-chase labelling experiments and analysis of leaves from the top to bottom of the transformed plants indicated that ovalbumin, once formed, was stable in the leaves of transgenic lucerne. A wide variation in ovalbumin level was frequently observed in plants regenerated from multiple embryos on a single transformed callus. This variation correlated with changes in the restriction enzyme digestion pattern of the ovalbumin DNA from the transgenic plants. These results indicate that each transformed callus may have arisen from more than one transformation event. An alternative interpretation is that the callus may have arisen from a single transformed cell but during cell proliferation the DNA in some cells may have undergone rearrangement prior to embryogenesis. Transformation and regeneration procedures were also developed for two Australian commercial cultivars of lucerne. Although the frequency of recovery of transformed plants was lower than with cv. Rangelander, these protocols open the way for a relatively rapid

scholarly journals A DNAβ Associated with Tomato Yellow Leaf Curl China Virus Is Required for Symptom Induction

2004 ◽  
Vol 78 (24) ◽  
pp. 13966-13974 ◽  
Author(s):  
Xiaofeng Cui ◽  
Xiaorong Tao ◽  
Yan Xie ◽  
Claude M. Fauquet ◽  
Xueping Zhou
Keyword(s):  
Transgenic Plants ◽  
Stop Codon ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Tomato Yellow Leaf ◽  
Cauliflower Mosaic Virus ◽  
Start Codon ◽  
35S Promoter ◽  
Yellow Leaf ◽  
Leaf Curl

ABSTRACT We report here that all 25 isolates of Tomato yellow leaf curl China virus (TYLCCNV) collected from tobacco, tomato, or Siegesbeckia orientalis plants in different regions of Yunnan Province, China, were associated with DNAβ molecules. To investigate the biological role of DNAβ, full-length infectious clones of viral DNA and DNAβ of TYLCCNV isolate Y10 (TYLCCNV-Y10) were agroinoculated into Nicotiana benthamiana, Nicotiana glutinosa, Nicotiana. tabacum Samsun (NN or nn), tomato, and petunia plants. We found that TYLCCNV-Y10 alone could systemically infect these plants, but no symptoms were induced. TYLCCNV-Y10 DNAβ was required, in addition to TYLCCNV-Y10, for induction of leaf curl disease in these hosts. Similar to TYLCCNV-Y10, DNAβ of TYLCCNV isolate Y64 was also found to be required for induction of typical leaf curl diseases in the hosts tested. When the βC1 gene of TYLCCNV-Y10 DNAβ was mutated, the mutants failed to induce leaf curl symptoms in N. benthamiana when coinoculated with TYLCCNV-Y10. However, Southern blot hybridization analyses showed that the mutated DNAβ molecules were replicated. When N. benthamiana and N. tabacum plants were transformed with a construct containing the βC1 gene under the control of the Cauliflower mosaic virus 35S promoter, many transgenic plants developed leaf curl symptoms similar to those caused by a virus, the severity of which paralleled the level of βC1 transcripts, while transgenic plants transformed with the βC1 gene containing a stop codon after the start codon remained symptomless. Thus, expression of a βC1 gene is adequate for induction of symptoms of viral infection in the absence of virus.

scholarly journals SpPKE1, a Multiple Stress-Responsive Gene Confers Salt Tolerance in Tomato and Tobacco

2019 ◽  
Vol 20 (10) ◽  
pp. 2478 ◽  
Author(s):  
Jinhua Li ◽  
Chunrui Chen ◽  
Juanjuan Wei ◽  
Yu Pan ◽  
Chenggang Su ◽  
...  
Keyword(s):  
Salt Stress ◽  
Salt Tolerance ◽  
Wild Species ◽  
Amino Acid Sequences ◽  
Cauliflower Mosaic Virus ◽  
Stress Factors ◽  
35S Promoter ◽  
Over Expression ◽  
Abiotic Tolerance ◽  
Cultivated Species

Understanding the mechanism of abiotic-tolerance and producing germplasm of abiotic tolerance are important in plant research. Wild species often show more tolerance of environmental stress factors than their cultivated counterparts. Genes from wild species show potential abilities to improve abiotic resistance in cultivated species. Here, a tomato proline-, lysine-, and glutamic-rich type gene SpPKE1 was isolated from abiotic-resistant species (Solanum pennellii LA0716) for over-expression in tomato and tobacco for salt tolerance. The protein encoded by SpPKE1 was predominantly localized in the cytoplasm in tobacco. SpPKE1 and SlPKE1 (from cultivated species S. lycopersicum cv. M82) shared 89.7% similarity in amino acid sequences and their transcripts abundance in flowers and fruits was reduced by the imposition of drought or oxidative stress and the exogenous supply of abscisic acid. The DNA of the PKE1 promoter was highly methylated in fruit and leaf, and the methylation of the coding sequence in leaf was significantly higher than that in fruit at different development stages. The over-expression of SpPKE1 under the control of a CaMV (Cauliflower Mosaic Virus) 35S promoter in transgenic tomato and tobacco plants enhanced their tolerance to salt stress. PKE1 was downregulated by abiotic stresses but enhanced the plant’s salt stress tolerance. Therefore, this gene may be involved in post-transcriptional regulation and may be an important candidate for molecular breeding of salt-tolerant plants.

Sign in / Sign up

Export Citation Format

Share Document