Phenotypic performance of transgenic potato (Solanum tuberosum L.) plants with pyramided rice cystatin genes (OCI and OCII)

The evaluation of transgenic plants which is usually carried out under controlled conditions in culture rooms and greenhouses can yield valuable information about the influence of introduced genes on a transgenic plant phenotype. However, an overall assessment of plant performance can only be made by testing transgenic plants in the field environment. Thus, the effects of pyramided rice cystatin genes OCI and OCII on morphological parameters of transgenic potato cv. Desiree, Dragacevka and Jelica lines were compared under in vitro, greenhouse, and field conditions. All analyzed OC co-expressing transgenic lines exhibited normal phenotype, both in vitro and in greenhouse conditions. In the field environment, eight of nine OCI/OCII lines were similar to the wild-type control plants in their general phenotypic appearance. Yield parameters, such as tuber number and tuber weight for these phenotypically normal OCI/OCII lines, were also comparable to the controls. Only transgenic cv. Jelica line 4 plants exhibited slightly reduced growth, atypical leaf morphology and, contrary to the plants of other transgenic lines and untransformed controls, failed to flower. However, despite the phenotypic and developmental changes under field conditions, the OCI/OCII Jelica line 4 did not exhibit a significant decrease in tuber yield. Stacking of OCI and OCII genes preserves important attributes of the parental lines, confirming that this approach could be suitable for improving agronomical traits in potato.

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Novel Approach to Parasitic Weed Control Based on Inducible Expression of Cecropin in Transgenic Plants

10.32747/2003.7586467.bard ◽

2003 ◽

Author(s):

Radi Aly ◽

James H. Westwood ◽

Carole L. Cramer

Keyword(s):

Transgenic Plants ◽

Weed Control ◽

Control Strategies ◽

Solanum Tuberosum L ◽

Transgenic Potato ◽

Inducible Expression ◽

Growth And Yield ◽

Parasitic Weeds ◽

Small Proteins ◽

Enhanced Resistance

Our overall goal was to engineer crop plants with enhanced resistance to Orobanche (broomrape) based on the inducible expression of sarcotoxin-like peptide (SLP). A secondary objective was to localize small proteins such as SLP in the host-parasite union in order to begin characterizing the mechanism of SLP toxicity to Orobanche. We have successfully accomplished both of these objectives and have demonstrated that transgenic tobacco plants expressing SLP under control of the HMG2 promoter show enhanced resistance to O. aegyptiaca and O. ramosa . Furthermore, we have shown that proteins much larger than the SLP move into Orobanche tubercles from the host root via either symplastic or apoplastic routes. This project was initiated with the finding that enhanced resistance to Orobanche could be conferred on tobacco, potato, and tomato by expression of SLP (Sarcotoxin IA is a 40-residue peptide produced as an antibiotic by the flesh fly, Sarcophaga peregrina ) under the control of a low-level, root-specific promoter. To improve the level of resistance, we linked the SLP gene to the promoter from HMG2, which is strongly inducible by Orobanche as it parasitizes the host. The resulting transgenic plants express SLP and show increased resistance to Orobanche. Resistance in this case is manifested by increased growth and yield of the host in the presence of the parasite as compared to non-transgenic plants, and decreased parasite growth. The mechanism of resistance appears to operate post-attachment as the parasite tubercles attached to the transgenic root plants turned necrotic and failed to develop normally. Studies examining the movement of GFP (approximately 6X the size of SLP) produced in tobacco roots showed accumulation of green fluorescence in tubercles growing on transformed plants but not in those growing on wild-type plants. This accumulation occurs regardless of whether the GFP is targeted to the cytoplasm (translocated symplastically) or the apoplastic space (translocated in xylem). Plants expressing SLP appear normal as compared to non-transgenic plants in the absence of Orobanche, so there is no obvious unintended impact on the host plant from SLP expression. This project required the creation of several gene constructs and generation of many transformed plant lines in order to address the research questions. The specific objectives of the project were to: 1. Make gene constructs fusing Orobanche-inducible promoter sequences to either the sarcotoxin-like peptide (SLP) gene or the GFP reporter gene. 2. Create transgenic plants containing gene constructs. 3. Characterize patterns of transgene expression and host-to-parasite movement of gene products in tobacco ( Nicotiana tabacum L.) and Arabidopsis thaliana (L.). 4. Characterize response of transgenic potato ( Solanum tuberosum L.) and tomato ( Lycopersicon esculentum Mill .) to Orobanche in lab, greenhouse, and field. Objectives 1 and 2 were largely accomplished during the first year during Dr. Aly's sabbatical visit to Virginia Tech. Transforming and analyzing plants with all the constructs has taken longer than expected, so efforts have concentrated on the most important constructs. Work on objective 4 has been delayed pending the final results of analysis on tobacco and Arabidopsis transgenic plants. The implications of this work are profound, because the Orobanche spp. is an extremely destructive weed that is not controlled effectively by traditional cultural or herbicidal weed control strategies. This is the first example of engineering resistance to parasitic weeds and represents a unique mode of action for selective control of these weeds. This research highlights the possibility of using this technique for resistance to other parasitic species and demonstrates the feasibility of developing other novel strategies for engineering resistance to parasitic weeds.

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Expression of CP:CBF3-35S:ICE1 Enhances Low Temperature Tolerance in Transgenic Arabidopsis

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.726-731.118 ◽

2013 ◽

Vol 726-731 ◽

pp. 118-121

Author(s):

Rui Mei Li ◽

Du Juan Xi ◽

Yi Meng Ji ◽

Rui Jun Duan ◽

Jiao Liu ◽

...

Keyword(s):

Low Temperature ◽

Transgenic Plants ◽

Temperature Stress ◽

Target Genes ◽

Temperature Tolerance ◽

Low Temperature Stress ◽

Stress Tests ◽

Low Temperature Tolerance ◽

Transgenic Lines

We have constructed a vector pCAMBIA1300-CP:CBF3-35S:ICE1 and transformed into Arabidopsis. Results of PCR proved that the target genes had integrated into Arabidopsis genome. Transgenic Arabidopsis showed a bit slow growth, earlier flowering, but normal at other phenotype under 22°C with 8 h daily lights. In vitro low temperature stress tests showed that the transgenic lines were survival while the wild type was nearly dead. The transgenic plants also showed an increased proline content, SOD and POD activities under low temperature stress. The phenotype and physical evidence indicated that expression of CP:CBF3-35S:ICE1 under low temperature enhances the cold tolerance in transgenic plants.

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Obtaining of transgenic potato plants (Solanum tuberosum L.) that contain antisense sequence of prolindehydrogenase gene

Faktori eksperimental'noi evolucii organizmiv ◽

10.7124/feeo.v22.965 ◽

2018 ◽

Vol 22 ◽

pp. 299-304

Author(s):

O. O. Ovcharenko ◽

V. A. Rudas ◽

N. L. Shcherbak ◽

M. V. Kuchuk

Keyword(s):

Solanum Tuberosum ◽

Transgenic Plants ◽

Genetic Transformation ◽

Solanum Tuberosum L ◽

Practical Interest ◽

Transgenic Potato ◽

Proline Dehydrogenase ◽

Potato Plants ◽

Antisense Sequence ◽

Transgenic Potato Plants

Aim. The aim of our work was to obtain transgenic potato plants of Ukrainian varieties with the expression of a double-stranded RNA suppressor of proline dehydrogenase gene. We propose the decrease of proline degradation level and increase of overall proline concentration in obtained transgenic plants. Methods. The Agrobasterium tumefaciens-mediated method of genetic transformation to obtain transgenic plants of potato was used. Internodes of aseptic potato plants were transformed with a binary vector pBi2E containing an inverted repeats of two copies of proline dehydrogenase gene’s first exon and the gene of neomycin phosphotransferase II (nptII). Results. As a result of experiments kanamycin resistant transgenic potato lines of Deseiree, Belarusian 12 and Slavianka varieties were obtained. The transgenic nature of the obtained plants was confirmed by PCR with primers specific to the first exon of proline dehydrogenase and to nptII genes. Conclusions. The optimized conditions of genetic transformation and used agrobacterial strain allow to obtain the transgenic plants of a model potato variety Désirée, as well as varieties Belorussian 12 and Slovyanka which are of practical interest for cultivation in Ukraine. Keywords: transgenic plants, potatoes (Solanum tuberosum L.), stress resistance, proline.

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Overexpression of Cytosolic Ascorbate Peroxidase in Tomato Confers Tolerance to Chilling and Salt Stress

Journal of the American Society for Horticultural Science ◽

10.21273/jashs.130.2.167 ◽

2005 ◽

Vol 130 (2) ◽

pp. 167-173 ◽

Cited By ~ 109

Author(s):

Yueju Wang ◽

Michael Wisniewski ◽

Richard Meilan ◽

Minggang Cui ◽

Robert Webb ◽

...

Keyword(s):

Salt Stress ◽

Transgenic Plants ◽

Ascorbate Peroxidase ◽

Higher Plants ◽

Visual Assessment ◽

Kanamycin Resistance ◽

Leaf Damage ◽

Cytosolic Ascorbate Peroxidase ◽

Transgenic Lines

Ascorbate peroxidase (APX) plays an important role in the metabolism of hydrogen peroxide in higher plants, affording them protection against oxidative stress. We studied the effect of overexpressing a cytosolic ascorbate peroxidase (cAPX) gene—derived from pea (Pisum sativum L.)—in transgenic tomato plants (Lycopersicon esculentum L.). Transformants were selected in vitro using kanamycin resistance and confirmed by polymerase chain reaction (PCR) and northern analyses. An APX native-gel assay indicated that, in the absence of stress, APX activity in transgenic plants was several times greater than that measured in wild-type (WT) plants. Several independently transformed lines were propagated and evaluated for resistance to chilling and salt stress. After placing seeds at 9 °C for 5 weeks, percent germination was greater for seeds obtained from transgenic lines (26% to 37%) compared to the WT (3%). Plants from transgenic lines also had lower electrolyte leakage (20% to 23%) than WT (44%) after exposure to 4 °C. Visual assessment of transgenic and WT lines exposed to salinity stress (200 or 250 mm) confirmed that overexpression of APX minimized leaf damage. Moreover, APX activity was nearly 25- and 10-fold higher in the leaves of transgenic plants in response to chilling and salt stresses, respectively. Our results substantiate that increased levels of APX activity brought about by overexpression of a cytosolic APX gene may play an important role in ameliorating oxidative injury induced by chilling and salt stress.

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

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.1999.12.5.410 ◽

1999 ◽

Vol 12 (5) ◽

pp. 410-418 ◽

Cited By ~ 92

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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Efficacy of Endogenous Satellite Expression to Confer Resistance to CMV in Satellite Transgenic Tomato under Field Conditions

HortScience ◽

10.21273/hortsci.31.4.569c ◽

1996 ◽

Vol 31 (4) ◽

pp. 569c-569

Author(s):

John R. Stommel ◽

Marie E. Tousignant ◽

Thanda Wai ◽

Jacobus M. Kaper

Keyword(s):

Transgenic Plants ◽

Field Tests ◽

Field Trials ◽

Transgenic Tomato ◽

Field Conditions ◽

Test Plot ◽

Tomato Plants ◽

Transgenic Tomato Plants ◽

Transgenic Lines ◽

Crop Disease

Viral satellite RNA associated with cucumber mosaic virus (CMV) is know to modulate CMV symptomology. Virulent CMV associated RNA 5 (CARNA 5) satellites may intensify crop disease. Naturally occurring variants of these satellites, however, attenuate CMV symptoms. Satellite transgenic tomato plants expressing the S-CARNA 5 or 1-CARNA 5 ameliorating forms of the satellite were evaluated under simulated CMV epidemic conditions in USDA–APHIS approved field trials. Trials conducted at Beltsville, Md., in 1994 and 1995 demonstrated that CMV can be effectively controlled under field conditions in satellite transgenic plants. Yields of transgenic lines infected with CMV were 50%–65% greater than that of non-transgenic infected controls. Yields of noninfected transgenic lines ranged from 5% greater than, to 33% less than, noninfected nontransgenic controls. Expression of CARNA 5 in inoculated transgenic plants greatly reduced CMV foliar symptoms and virus titers when compared to inoculated control plants. Levels of CARNA 5 were detected at varying levels in infected transgenic plants throughout the growing season. Virus or satellite was not detected in samples collected from tomato border plants and weeds growing inside and outside a nonhost crop border surrounding the test plot. Field tests conducted in 1996 will evaluate transgenic tomato plants with a double construct coding for the CMV coat protein gene and 1-CARNA 5 satellite.

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Extreme resistance to two Brazilian strains of Potato virus Y (PVY) in transgenic potato, cv. Achat, expressing the PVYº coat protein

Horticultura Brasileira ◽

10.1590/s0102-05362001000200004 ◽

2001 ◽

Vol 19 (2) ◽

pp. 118-122 ◽

Cited By ~ 3

Author(s):

Eduardo Romano ◽

Adriana T. Ferreira ◽

André N. Dusi ◽

Karina Proite ◽

Jose A. Buso ◽

...

Keyword(s):

Coat Protein ◽

Potato Virus ◽

Potato Virus Y ◽

Potato Cultivar ◽

Transgenic Potato ◽

Resistant Line ◽

Cp Gene ◽

Transgenic Lines ◽

The Stability

The coat protein (CP) gene of the potato virus Y strain "o" (PVY O) was introduced into potato, cultivar Achat, via Agrobacterium tumefaciens-mediated transformation. Sixty three putative transgenic lines were challenged against the Brazilian strains PVY-OBR and PVY-NBR. An extremely resistant phenotype, against the two strains, was observed in one line, denominated 1P. No symptoms or positive ELISA results were observed in 16 challenged plants from this line. Another clone, named as 63P, showed a lower level of resistance. Southern blot analysis showed five copies of the CP gene in the extremely resistant line and at least three copies in the other resistant line. The stability of the integrated transgenes in the extreme resistant line was examined during several in vitro multiplications over a period of three years, with no modification in the Southern pattern was observed. The stability of the transgenes, the absence of primary infections and the relatively broad spectrum of resistance suggest that the extremely resistant line obtained in this work can be useful for agricultural purposes.

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Broad-Spectrum Protection against Tombusviruses Elicited by Defective Interfering RNAs in Transgenic Plants

Journal of Virology ◽

10.1128/jvi.73.6.5070-5078.1999 ◽

1999 ◽

Vol 73 (6) ◽

pp. 5070-5078 ◽

Cited By ~ 16

Author(s):

Teresa Rubio ◽

Marisé Borja ◽

Herman B. Scholthof ◽

Paul A. Feldstein ◽

T. Jack Morris ◽

...

Keyword(s):

Transgenic Plants ◽

Broad Spectrum ◽

Nicotiana Benthamiana ◽

Tomato Bushy Stunt Virus ◽

Seed Plants ◽

Rna Sequences ◽

Transgenic Lines ◽

Negative Controls ◽

Very High

ABSTRACT We have designed a DNA cassette to transcribe defective interfering (DI) RNAs of tomato bushy stunt virus (TBSV) and have investigated their potential to protect transgenic Nicotiana benthamianaplants from tombusvirus infections. To produce RNAs with authentic 5′ and 3′ termini identical to those of the native B10 DI RNA, the DI RNA sequences were flanked by ribozymes (RzDI). When RzDI RNAs transcribed in vitro were mixed with parental TBSV transcripts and inoculated into protoplasts or plants, they became amplified, reduced the accumulation of the parental RNA, and mediated attenuation of the lethal syndrome characteristic of TBSV infections. Analysis of F1 and F2 RzDI transformants indicated that uninfected plants expressed the DI RNAs in low abundance, but these RNAs were amplified to very high levels during TBSV infection. By two weeks postinoculation with TBSV, all untransformed N. benthamiana plants and transformed negative controls died. Although infection of transgenic RzDI plants initially induced moderate to severe symptoms, these plants subsequently recovered, flowered, and set seed. Plants from the same transgenic lines also exhibited broad-spectrum protection against related tombusviruses but remained susceptible to a distantly related tombus-like virus and to unrelated viruses.

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Alleviation of Phytophthora infestans Mediated Necrotic Stress in the Transgenic Potato (Solanum tuberosum L.) with Enhanced Ascorbic acid Accumulation

Plants ◽

10.3390/plants8100365 ◽

2019 ◽

Vol 8 (10) ◽

pp. 365 ◽

Cited By ~ 2

Author(s):

Ill-Min Chung ◽

Baskar Venkidasamy ◽

Chandrama Prakash Upadhyaya ◽

Gurusaravanan Packiaraj ◽

Govindasamy Rajakumar ◽

...

Keyword(s):

Ascorbic Acid ◽

Transgenic Plants ◽

Phytophthora Infestans ◽

Solanum Tuberosum L ◽

Transgenic Potato ◽

Pr Genes ◽

Pathogen Infection ◽

Untransformed Control ◽

Expression Of Genes ◽

Pathogenesis Related

Potato is the most widely cultivated non-cereal crop in the world, and like any other crop, it is susceptible to yield losses because of various factors, including pathogen attacks. Among the various diseases of potato, late blight caused by the oomycete Phytophthora infestans is considered as the most devastating disease worldwide. In this study, transgenic potato plants overexpressing the D-galacturonic acid reductase (GalUR) gene with an enhanced level of cellular L-ascorbate (L-AsA) were challenged with Phytophthora infestans to determine the level of stress tolerance induced in those plants. With the onset of pathogen infection, necrotic lesions progressively expanded and became necrotic in the control plants. The transgenic potato lines with enhanced ascorbic acid showed reduced necrotic lesions. Hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels were relatively lower in transgenic plants compared to the untransformed control (UT) plants. The mRNA expressions of pathogenesis-related (PR) genes, such as pathogenesis related 1 (PR1) and phenylalanine ammonia-lyase (PAL) were slightly higher in GalUR overexpressing transgenic lines as compared to the untransformed control plants. Pathogen infection also altered the mRNA expression of genes associated with gibberellic acid (GA) and abscisic acid (ABA) biosynthesis. Furthermore, the increase in various antioxidant enzymes was also observed in the gene expression analysis with the transgenic plants. The complete loss of the pathogen growth and disease occurrence was not observed in our study; however, the findings indicated that an increase in the level of cellular L-ascorbate in the transgenic potato leads to enhanced cellular antioxidants, PR genes and plant defense hormones, such as GA and ABA resulting in the reduction of the disease symptoms caused by the Phytophthora infestans.

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Respon Pemberian Aspirin dan Kinetin Terhadap Pembentukan Umbi Mikro Kentang (Solanum tuberosum L.) Secara In Vitro

10.25047/agropross.2017.15 ◽

2017 ◽

Author(s):

Yunita Prameswari ◽

FNU Djenal ◽

FNU Damanhuri

Keyword(s):

Solanum Tuberosum ◽

Solanum Tuberosum L

Kebutuhan kentang yang semakin tinggi menyebabkan permintaan semakin meningkat. Rendahnya produksi kentang mengakibatkan berbagai upaya untuk peningkatan produksi terus dilakukan. Penggunaan metode kultur jaringan yaitu metode untuk mengisolasi bagian tanaman seperti protoplasma, sel, sekelompok sel, jaringan dan organ dalam kondisi aseptik, sehingga dapat diperbanyak dan beregenerasi menjadi tanaman utuh dapat dijadikan alternatif pemenuhan kebutuhan. Penelitian ini bertujuan untuk mengetahui kecepatan pembentukan umbi mikro kentang (Solanum Tuberosum L.) varietas granola kembang secarain vitro dengan menggunakan dua faktor dan 3 kali ulangan. Faktor pertama yaitu aspirin dengan tiga taraf (5,10,15) ppm. Faktor kedua yaitu kinetin dalam tiga taraf (8,10,12) ppm. Penelitian menggunakan seluruh propagul kentang yang berumur 30 hari setelah subkultur dan data yang didapat dianalisis menggunakan ANOVA. Hasil penelitian menunjukan bahwa interaksi aspirin dan kinetin tidak berpengaruh terhadap jumlah akar, kedinian umbi, dan bobot umbi. Interaksi perlakuan terbaik bagi pembentukan tunas yaitu A2K1 aspirin 10 ppm dan kinetin 8 ppm sedangkan Interaksi perlakuan terbaik pada parameter jumlah umbi yaituA3K2 aspirin 15 ppm dan kinetin 10.

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