Transformation of potato using an antisense class I patatin gene and its effect on microtuber formation

2005 ◽  
Vol 2 (1) ◽  
pp. 7-11 ◽  
Author(s):  
Si Huai-Jun ◽  
Liu Jun ◽  
Xie Cong-Hua
Keyword(s):  
Pcr Amplification ◽  
Transgenic Potato ◽  
Class I ◽  
Total Soluble Protein ◽  
Northern Hybridization ◽  
35S Promoter ◽  
Antisense Gene ◽  
Potato Genome ◽  
Transformed Plants ◽  
Class I Patatin

AbstractAn antisense class I patatin gene under control of the CaMV 35S promoter was introduced into potato (Solanum tuberosum) cultivar E-potato 3 using the Agrobacterium tumefaciens system. PCR amplification and PCR–Southern blot analysis indicated that the antisense class I patatin gene had been integrated into the potato genome. Northern hybridization analysis showed that the antisense gene transcribed normally in the transgenic potato plants and resulted in a reduction of endogenous class I patatin mRNA. Total soluble protein content and lipid acyl hydrolase activity of microtubers, derived from transformed plants, decreased by a maximum of 36.4% and 31.4%, respectively, compared with control plants. The expression of this antisense gene also resulted in reductions of the plantlets forming tubers, tubers per plantlet and the effective tubers (≥50 mg) of the transformed plants.

Functional analysis of a class I patatin gene SK24-1 in microtuber formation of transgenic potatoes

2008 ◽  
Vol 88 (4) ◽  
pp. 593-598 ◽  
Author(s):  
Huaijun Si ◽  
Jun Liu ◽  
Jian Huang ◽  
Conghua Xie
Keyword(s):  
Escherichia Coli ◽  
Cdna Clone ◽  
Antisense Rna ◽  
Cauliflower Mosaic Virus ◽  
Class I ◽  
35S Promoter ◽  
Transformed Plants ◽  
Transgenic Potatoes ◽  
Class I Patatin

Expression of a class I patatin cDNA clone, SK24-1, in Escherichia coli revealed that the cDNA clone possessed lipid acyl hydrolase (LAH) activity. Transformed potato plants were obtained via Agrobacterium-mediated transformation using the chimeric constructs containing the sense and antisense cDNA under the control cauliflower mosaic virus 35S (CaMV 35S) promoter. In some sense transformed plants, both sense patatin RNA and LAH activity were increased and further resulted in a significant increase of percentage of plantlets that formed microtubers and numbers of microtubers per plantlet in vitro. All antisense plants displayed a reduction in LAH activity. Both sense and antisense RNA could be detected in antisense plants, but transcripts of antisense RNA resulted in a reduction of endogenous sense RNA. Moreover, expression of antisense cDNA in some antisense transformed plants led to a significant decrease in the number of microtubers formed. These results suggest that SK24-1 was involved in regulating microtuber formation. Key words: Patatin, potato, Escherichia coli, sense RNA, antisense RNA

The expression of class I patatin gene fusions in transgenic potato varies with both gene and cultivar

1991 ◽  
Vol 16 (1) ◽  
pp. 153-160 ◽  
Author(s):  
K. S. Blundy ◽  
M. A. C. Blundy ◽  
D. Carter ◽  
F. Wilson ◽  
W. D. Park ◽  
...  
Keyword(s):  
Transgenic Potato ◽  
Class I ◽  
Gene Fusions ◽  
Class I Patatin ◽  
Patatin Gene

Transgenic Potato Plants Expressing Soybean ß-1,3-Endoglucanase Gene Exhibit an Increased Resistance to Phytophthora infestans

1998 ◽  
Vol 53 (11-12) ◽  
pp. 1012-1016 ◽  
Author(s):  
Maria Borkowska ◽  
Magdalena Krzymowska ◽  
Andrzej Talarczyk ◽  
Malik F. M. Awan ◽  
Ludmila Yakovleva ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Phytophthora Infestans ◽  
Cell Walls ◽  
Fungal Pathogens ◽  
Pcr Amplification ◽  
Transgenic Potato ◽  
Fungal Cell ◽  
Potato Genome ◽  
Potato Plants ◽  
Transgenic Potato Plants

Abstract Soybean β-1,3-endoglucanase represents a model system for studies on early plant re­sponses to infection by fungal pathogens, and it has been implicated in the release of elicitors from fungal cell walls. In the present study, potato plants were transformed with the soybean β-1,3-endoglucanase cDNA via Agrobacterium delivery system. The transfer of the gene into potato genome was confirmed by (i) PCR amplification, (ii) Northern blot analyses, and (Hi) an increase in the activity of β-1,3-endoglucanase in transgenic plants. The transformation resulted in an increased resistance of selected transgenic plants to infection by Phytophthora infestans, an important pathogen.

Expression of Escherichia coli Heat-Labile Enterotoxin B Subunit in Centella (Centella asiatica (L.) Urban) via Biolistic Transformation

2020 ◽  
Vol 21 (10) ◽  
pp. 973-979
Author(s):  
Nguyen H. Loc ◽  
Nghiem V. Tung ◽  
Phan T.A. Kim ◽  
Moon S. Yang
Keyword(s):  
Escherichia Coli ◽  
Pcr Amplification ◽  
Centella Asiatica ◽  
Biolistic Transformation ◽  
Total Soluble Protein ◽  
35S Promoter ◽  
Gm1 Ganglioside ◽  
B Subunit ◽  
Heat Labile ◽  
Enterotoxin B

Background: Heat-Labile enterotoxin B subunit (LTB) produced by Escherichia coli, a non-toxic protein subunit with potential biological properties, is a powerful mucosal and parenteral adjuvant which can induce a strong immune response against co-administered antigens. Objective: In the present study, LTB protein, encoded by the optimized ltb (also known synthetic ltb, s-ltb) gene in centella plant (Centella asiatica) for use as an antigen, has been discussed. Methods: The s-ltb gene was cloned into a plant expression vector, pMYO51, adjacent to the CaMV 35S promoter and was then introduced into centella plant by biolistic transformation. PCR amplification was conducted to determine the presence of s-ltb gene in the transgenic centella plant. The expression of s-ltb gene was analyzed by immunoblotting and quantified by ELISA. In vitro activity of LTB protein was determined by GM1-ELISA. Results: PCR amplification has found seven transgenic centella individuals. However, only five of them produced LTB protein. ELISA analysis showed that the highest amount of LTB protein detected in transgenic centella leaves was about 0.8% of the total soluble protein. GM1-ELISA assay indicated that plant LTB protein bound specifically to GM1-ganglioside, suggesting that the LTB subunits formed active pentamers. Conclusion: The s-ltb gene that was successfully transformed into centella plants by the biolistic method has produced a relatively high amount of plant LTB protein in the pentameric quaternary structure that has GM1-ganglioside binding affinity, a receptor on the intestinal epithelial membrane.

scholarly journals Organ-specific gene expression in transgenic potato: the cloning a new promoter of a class I patatin gene

1995 ◽  
Vol 11 (6) ◽  
pp. 96-103 ◽  
Author(s):  
I. M. Yefimenko ◽  
T. V. Medvedeva ◽  
P. G. Kovalenko ◽  
K. G. Gazaryan ◽  
A. P. Galkin
Keyword(s):  
Gene Expression ◽  
Transgenic Potato ◽  
Class I ◽  
Specific Gene ◽  
Specific Gene Expression ◽  
Organ Specific ◽  
Class I Patatin ◽  
Patatin Gene

Localization of Ds-transposon containing T-DNA inserts in the diploid transgenic potato: linkage to the R1 resistance gene against Phytophthora infestans (Mont.) de Bary

Genome ◽  
1996 ◽  
Vol 39 (2) ◽  
pp. 249-257 ◽  
Author(s):  
A. El-Kharbotly ◽  
J. M. E. Jacobs ◽  
B. te Lintel Hekkert ◽  
W. J. Stiekema ◽  
A. Pereira ◽  
...  
Keyword(s):  
Solanum Tuberosum ◽  
Phytophthora Infestans ◽  
Resistance Gene ◽  
Dna Sequences ◽  
Transgenic Potato ◽  
Potato Genome ◽  
Rflp Map ◽  
Polymerase Chain ◽  
R1 Gene ◽  
Map Location

The Dissociation transposable element (Ds) of maize containing NPTII was introduced into the diploid potato (Solanum tuberosum) clone J91-6400-A16 through Agrobacterium tumefaciens mediated transformation. Genomic DNA sequences flanking the T-DNAs from 312 transformants were obtained with inverse polymerase chain reaction or plasmid rescue techniques and used as probes for RFLP linkage analysis. The RFLP map location of 60 T-DNAs carrying Ds–NPTII was determined. The T-DNA distribution per chromosome and the relative distance between them appeared to be random. All 12 chromosomes have been covered with Ds-containing T-DNAs, potentially enabling tagging of any gene in the potato genome. The T-DNA insertions of two transformants, BET92-Ds-A16-259 and BET92-Ds-A16-416, were linked in repulsion to the position of the resistance gene R1 against Phytophthora infestans. After crossing BET92-Ds-A16-416 with a susceptible parent, 4 desired recombinants (Ds carrying T-DNA linked in coupling phase with the R1 gene) were discovered. These will be used for tagging the R1 gene. The efficiency of the pathway from the introduction to localization of T-DNAs is discussed. Key words : Solanum tuberosum, Phytophthora infestans, Ds element, transposon tagging, R genes, euchromatin.

MYB class transcription factors bind to the tuber-specific and sucrose-response element of a class-I patatin promoter

2017 ◽  
Vol 11 (4) ◽  
pp. 239-245 ◽  
Author(s):  
Hyung-in Choi ◽  
Sun-Young Baek ◽  
Soo Young Kim
Keyword(s):  
Transcription Factors ◽  
Class I ◽  
Response Element ◽  
Patatin Promoter ◽  
Class I Patatin

The maize transcription factor Sn alters proanthocyanidin synthesis in transgenic Lotus corniculatus plants

1999 ◽  
Vol 26 (2) ◽  
pp. 159 ◽  
Author(s):  
Francesco Damiani ◽  
Francesco Paolocci ◽  
Paul D. Cluster ◽  
Sergio Arcioni ◽  
Gregory J. Tanner ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Binary Vector ◽  
Regulatory Gene ◽  
Lotus Corniculatus ◽  
35S Promoter ◽  
Gus Gene ◽  
Plant Leaves ◽  
Negative Interaction ◽  
Transformed Plants ◽  
And Control

Lotus corniculatus L. plants were transformed with Agrobacterium rhizogenes binary vector carrying the maize Sn regulatory gene driven by the 35S promoter. These plants showed modifications in the pattern of accumulation of proanthocyanidin (PA). All the transformed plants but one showed an increase in PA content in the root relative to control untransformed and control gus gene transformed plants (C). With respect to the PAaccumulation in leaves, Sn transgenic plants were grouped in two classes: suppressed (S), that showed a consistent reduction of foliar PAcontent, and unsuppressed (U) that did not differ significantly from controls. Dihydroflavanol reductase (DFR) and leucocyanidin reductase (LAR) enzyme activities in S and U plant leaves mirrored the changes seen with foliar PA accumulation. LAR activity in the roots was consistent with the root PA levels. Mature Sn mRNA accumulated in the leaves of U plants, but not in leaves of S plants; however, leaves of both S and U plants were able to initiate Sn transcription. All Sn-transformed plants accumulated Sn message in root tissue. A possible negative interaction of Sn and an unidentified homologous endogene is proposed for explaining the behaviour of S plants.

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