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.

A detoxification gene in transgenicNicotiana tabacumconfers 2,4-D tolerance

Weed Science ◽  
1999 ◽  
Vol 47 (4) ◽  
pp. 401-404 ◽  
Author(s):  
David I. Last ◽  
Danny J. Llewellyn
Keyword(s):  
Arabidopsis Thaliana ◽  
Nicotiana Tabacum ◽  
Pisum Sativum ◽  
Histone Gene ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Spray Drift ◽  
Degree Of Protection ◽  
Dioxygenase Gene ◽  
Detoxification Gene

TransgenicNicotiana tabacumwith tolerance to 2,4-D has previously been produced using a bacterial 2,4-D-dioxygenase gene (tfdA) driven by the 35S promoter of cauliflower mosaic virus. Using promoters from thePisum sativumplastocyanin gene (petE) and anArabidopsis thalianahistone gene (H4A), we demonstrate that similar protection from 2,4-D can be obtained in transgenicN. tabacumby targeting expression oftfdAto either meristematic tissues or chloroplast-containing tissues. As with the 35S promoter constructs, the plants are tolerant but not completely resistant; very young seedlings in particular are only slightly protected. However, the levels of tolerance observed could offer a useful degree of protection from accidental spray drift.

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 A Paraquat-Inducible Protein B-like (pqiB) Gene From Chromobacterium Violaceum Confers Tolerance to Paraquat in Transgenic Tobacco

2021 ◽  
Author(s):  
Lais Santos Freire ◽  
Jamilly Azevedo Leal Sena ◽  
Marcio Gilberto Costa ◽  
Fatima Alvim
Keyword(s):  
Transgenic Plants ◽  
Transgenic Tobacco ◽  
Chromobacterium Violaceum ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Tobacco Genome ◽  
Paraquat Tolerance ◽  
The Subject ◽  
Inducible Protein ◽  
Protein B

Abstract Paraquat (1,1'-dimethyl-4,4'-bipyridinium dichloride) is a contact non-selective herbicide, widely used in agriculture in several countries. Proteins induced by paraquat have been the subject of great interest because of the possibility of conferring herbicide resistance when introduced into crops. In this work, we analyzed a paraquat-inducible protein B-like ( cvpqiB ) gene, isolated from Chromobacterium violaceum, in conferring tolerance to paraquat in transgenic tobacco. A DNA fragment containing the pqiB coding sequence was isolated from the C. violaceum ATCC12472 genome, inserted into the pCAMBIA1390 vector, under the control of the cauliflower mosaic virus (CaMV) 35S promoter, and used in Agrobacterium -mediated transformation of Nicotiana tabacum cv. Havana. Analysis of the regenerants revealed the incorporation of cvpqiB into the tobacco genome and its transmission in a Mendelian fashion to the progeny of transgenic plants. Sensitivity assays using tobacco leaves demonstrated that the transgenic plants were tolerant to concentrations up to 50 µM paraquat, whereas the wild-type (WT) plants exhibited intolerance to concentrations higher than 1 μM of the herbicide. Paraquat-treated leaves of the transgenic plants also exhibited significantly reduced electrolyte leakage and their chlorophyll content was not impacted as observed in the WT plants. Besides, in contrast to the WT, negligible amounts of hydrogen peroxide (H 2 O 2 ) were detected in paraquat-treated seedlings of the transgenic plants, as revealed by 3,3’-diaminobenzidine (DAB) staining. Collectively, these results indicate that the cvpqiB gene is functional in plants and may be further used in the genetic engineering of crop plants aiming paraquat tolerance.

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.

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 Constitutive Expression of Pea Defense Gene DRR206 Confers Resistance to Blackleg (Leptosphaeria maculans) Disease in Transgenic Canola (Brassica napus)

1999 ◽  
Vol 12 (5) ◽  
pp. 410-418 ◽  
Author(s):  
Yaping Wang ◽  
Goska Nowak ◽  
David Culley ◽  
Lee A. Hadwiger ◽  
Brian Fristensky
Keyword(s):  
Brassica Napus ◽  
Transgenic Plants ◽  
Leptosphaeria Maculans ◽  
Constitutive Expression ◽  
Single Copy ◽  
Cauliflower Mosaic Virus ◽  
Adult Plant ◽  
35S Promoter ◽  
Transgenic Lines

To identify genes effective against the blackleg fungus Leptosphaeria maculans (Phoma lingam), we have transformed canola (Brassica napus) with four pea (Pisum sativum) genes under constitutive control by the cauliflower mosaic virus 35S promoter: PR10.1, chitinase, DRR206, and defensin. Transgenic lines containing single-copy T-DNA insertions for each gene were screened for both seedling (cotyledonary) and adult plant resistance. Lines for which pea DRR206 mRNA was expressed showed decreased disease scores, compared with non-expressing transgenic lines. Transgenic plants expressing pea defensin showed a slight enhancement of resistance, while for PR10 and chitinase transgenics there was little or no enhancement of resistance. Resistance to L. maculans cosegregated with DRR206 transgenes. Extracts from DRR206 and defensin transgenic plants inhibited fungal germination in vitro. DRR206 transgenic plants also demonstrated decreased hyphal growth at inoculation sites. While the precise function of DRR206 remains to be determined, these results suggest that it does play an important role in defense against fungi.

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