Over-Expression of the Pikh Gene with a CaMV 35S Promoter Leads to Improved Blast Disease (Magnaporthe oryzae) Tolerance in Rice

Cotton (Gossypium hirsutumL.) was transformed with constructs for the over-expression of two enzymes involved in ethanol fermentation, alcohol dehydrogenase (ADH) and pyruvate decarboxylase (PDC), with the goal of increasing waterlogging tolerance. Four independent transgenic lines transformed with the cotton Adh2 cDNA driven by the CaMV 35S promoter showed ectopic expression of this isozyme in leaves and up to 20-fold greater in vitro ADH activity in roots. Under conditions of O2 deficiency, excised roots from these transgenic plants showed up to 80% increase in ethanol evolution compared to untransformed roots. Conversely, one line transformed with a construct containing the Adh2 coding region in the antisense orientation showed a 65% decrease in ADH activity and a 25% decrease in ethanol production from anaerobic roots relative to untransformed cotton. Lines transformed with a rice Pdc1 cDNA driven by the CaMV 35S promoter showed high levels of expression of the transgene-encoded protein in leaves, but only very low levels of protein and no in vitro enzyme activity detectable in the roots of these plants. Roots from plants transformed with the 35S-Pdc construct did not produce more ethanol than roots from untransformed controls. We tested the ability of cotton roots to withstand low O2 treatments under hydroponic conditions. Neither the ‘ADH’ nor the ‘PDC’ transgenics showed more tolerance than the wild-type on the basis of root growth under a mild stress (5% O2), a strong stress (0% O2 with or without a 5% O2 pretreatment), or in recovery growth following these treatments. Our results show that over-expression of ADH can lead to ethanol over-production (even though the activity of this enzyme by far exceeds that of PDC, its precursor in the pathway), but this is not sufficient to increase waterlogging tolerance in cotton.

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Aberrant promoter methylation occurred from multicopy transgene and SU(VAR)3 - 9 homolog 9 (SUVH9) gene in transgenic Nicotiana benthamiana

Functional Plant Biology ◽

10.1071/fp12051 ◽

2012 ◽

Vol 39 (9) ◽

pp. 764 ◽

Cited By ~ 6

Author(s):

Gi-Ho Lee ◽

Seong-Han Sohn ◽

Eun-Young Park ◽

Young-Doo Park

Keyword(s):

Gene Silencing ◽

Nicotiana Benthamiana ◽

Fluorescent Protein ◽

Cauliflower Mosaic Virus ◽

Camv 35S Promoter ◽

35S Promoter ◽

Camv 35S ◽

Histone Deacetylase 1 ◽

Transgenic Lines ◽

Green Fluorescent

The chemical modification of DNA by methylation is a heritable trait and can be subsequently reversed without altering the original DNA sequence. Methylation can reduce or silence gene expression and is a component of a host’s defence response to foreign nucleic acids. In our study, we employed a plant transformation strategy using Nicotiana benthamiana Domin to study the heritable stability of the introduced transgenes. Through the introduction of the cauliflower mosaic virus (CaMV) 35S promoter and the green fluorescent protein (GFP) reporter gene, we demonstrated that this introduced promoter often triggers a homology-dependent gene-silencing (HDGS) response. These spontaneous transgene-silencing phenomena are due to methylation of the CaMV 35S promoter CAAT box during transgenic plant growth. This process is catalysed by SU(VAR)3–9 homologue 9 (SUVH9), histone deacetylase 1 (HDA1) and domains rearranged methylase 2 (DRM2). In particular, we showed from our data that SUVH9 is the key regulator of methylation activity in epigenetically silenced GFP transgenic lines; therefore, our findings demonstrate that an introduced viral promoter and transgene can be subject to a homology-dependent gene-silencing mechanism that can downregulate its expression and negatively influence the heritable stability of the transgene.

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CaMV 35S promoter directs β-glucuronidas expression in the laticiferous system of transgenic Hevea brasiliensis (rubber tree)

Current Opinion in Plant Biology ◽

10.1016/1369-5266(88)80018-9 ◽

1998 ◽

Vol 1 (4) ◽

pp. 277

Keyword(s):

Hevea Brasiliensis ◽

Rubber Tree ◽

Camv 35S Promoter ◽

35S Promoter ◽

Camv 35S

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Multiplex, Construct-Specific, and Real-Time PCR-Based Analytical Methods for Bt Rice with cry1Ac Gene

Journal of AOAC International ◽

10.5740/jaoacint.10-429 ◽

2012 ◽

Vol 95 (1) ◽

pp. 186-194 ◽

Cited By ~ 6

Author(s):

Gurinder Jit Randhawa ◽

Monika Singh

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Analytical Methods ◽

Camv 35S Promoter ◽

35S Promoter ◽

Pcr Assay ◽

Cry1ac Gene ◽

Camv 35S ◽

Bt Rice ◽

Pcr Assays

Abstract Qualitative and quantitative analytical methods based on PCR for Bacillus thuringiensis (Bt) rice hybrid, namely, MRP 5401 Bt expressing a modified version of the Bt cry1Ac gene, are reported here. Multiplex PCR assays were developed to target the cry1Ac transgene, Cauliflower mosaic virus (CaMV) 35S promoter, Agrobacterium tumefaciens nopaline synthase (nos) terminator, the neomycin phosphotransferase II (nptII) marker gene, and an endogenous α-tubulin (TubA) gene in Bt rice. The 3.178 kb region of inserted gene construct comprising the region of the CaMV 35S promoter and cry1Ac gene was amplified, and the construct integrity was confirmed by the nested PCR. The LOD for cry1Ac gene-specific simplex PCR was 0.01%, as established using Bt rice DNA dilutions with 100, 10, 1.0, 0.1, 0.05, 0.01, and 0.001% genetically modified trait. A real-time PCR assay was also developed to quantify the cry1Ac gene. The method performance of the reported real-time PCR assay was in line with the acceptance criteria of Codex Alimentarius Commission ALINORM 10/33/23, with LOD and LOQ values of 0.05%. The reliable PCR assays prior to commercial release of Bt rice would facilitate efficient regulatory compliance for identification of genetic trait, labeling requirements, and effective risk assessment and management. They could also address consumers' concerns and legal disputes that may arise.

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A CaMV 35S promoter driven cDNA clone of tobacco mosaic virus can infect host plant tissue despite being uninfectious when manually inoculated onto leaves

Archives of Virology ◽

10.1007/s007050050069 ◽

1997 ◽

Vol 142 (1) ◽

pp. 183-191 ◽

Cited By ~ 10

Author(s):

E. M. Dagless ◽

M. H. Shintaku ◽

R. S. Nelson ◽

G. D. Foster

Keyword(s):

Tobacco Mosaic Virus ◽

Host Plant ◽

Mosaic Virus ◽

Cdna Clone ◽

Plant Tissue ◽

Camv 35S Promoter ◽

35S Promoter ◽

Camv 35S

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Parsley ubiquitin promoter displays higher activity than the CaMV 35S promoter and the chrysanthemum actin 2 promoter for productive, constitutive, and durable expression of a transgene in Chrysanthemum morifolium

Breeding Science ◽

10.1270/jsbbs.19036 ◽

2019 ◽

Vol 69 (3) ◽

pp. 536-544 ◽

Cited By ~ 1

Author(s):

Mitsuko Kishi-Kaboshi ◽

Ryutaro Aida ◽

Katsutomo Sasaki

Keyword(s):

Chrysanthemum Morifolium ◽

Camv 35S Promoter ◽

35S Promoter ◽

Camv 35S

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Evaluation of Plant-Derived Promoters for Constitutive and Tissue-Specific Gene Expression in Potato

Plants ◽

10.3390/plants9111520 ◽

2020 ◽

Vol 9 (11) ◽

pp. 1520

Author(s):

Dmitry Miroshnichenko ◽

Aleksey Firsov ◽

Vadim Timerbaev ◽

Oleg Kozlov ◽

Anna Klementyeva ◽

...

Keyword(s):

Gene Expression ◽

Specific Activity ◽

Expression Patterns ◽

Camv 35S Promoter ◽

35S Promoter ◽

Specific Gene ◽

Transcriptional Level ◽

Promoter Strength ◽

Tissue Specific ◽

Camv 35S

Various plant-derived promoters can be used to regulate ectopic gene expression in potato. In the present study, four promoters derived from the potato genome have been characterized by the expression of identical cassettes carrying the fusion with the reporter β-glucuronidase (gusA) gene. The strengths of StUbi, StGBSS, StPat, and StLhca3 promoters were compared with the conventional constitutive CaMV 35S promoter in various organs (leaves, stems, roots, and tubers) of greenhouse-grown plants. The final amount of gene product was determined at the post-transcriptional level using histochemical analysis, fluorometric measurements, and Western blot analysis. The promoter strength comparison demonstrated that the StUbi promoter generally provided a higher level of constitutive β-glucuronidase accumulation than the viral CaMV 35S promoter. Although the StLhca3 promoter was predominantly expressed in a green tissue-specific manner (leaves and stems) while StGBSS and StPat mainly provided tuber-specific activity, a “promoter leakage” was also found. However, the degree of unspecific activity depended on the particular transgenic line and tissue. According to fluorometric data, the functional activity of promoters in leaves could be arranged as follows: StLhca3 > StUbi > CaMV 35S > StPat > StGBSS (from highest to lowest). In tubers, the higher expression was detected in transgenic plants expressing StPat-gusA fusion construct, and the strength order was as follows: StPat > StGBSS > StUbi > CaMV 35S > StLhca3. The observed differences between expression patterns are discussed considering the benefits and limitations for the usage of each promoter to regulate the expression of genes in a particular potato tissue.

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A simple and accurate PCR method for detection of genetically modified rice

Journal of Environmental Health Science and Engineering ◽

10.1007/s40201-019-00401-x ◽

2019 ◽

Vol 17 (2) ◽

pp. 847-851 ◽

Cited By ~ 2

Author(s):

Payam Safaei ◽

Ebrahim Molaee Aghaee ◽

Gholamreza Jahed Khaniki ◽

Setareh Agha Kuchak Afshari ◽

Sassan Rezaie

Keyword(s):

Genetically Modified ◽

Sucrose Phosphate Synthase ◽

Cauliflower Mosaic Virus ◽

Camv 35S Promoter ◽

35S Promoter ◽

Rice Varieties ◽

Camv 35S ◽

Nos Terminator ◽

Gm Rice ◽

Pcr Method

Abstract Background Legislation regulating for labeling and use of genetically modified (GM) crops are increased considerably worldwide in order to health and safety assurance of consumers. For this purpose, a polymerase chain reaction (PCR) method has been developed for detection of GM rice in people’s food diet. Methods In this study, eighty-one non-labeled rice samples were collected randomly from different market sites of Tehran, Iran. In order to analysis, rice genomic DNA was extracted using MBST DNA extraction kit and subsequently, sucrose phosphate synthase (SPS) gene was used to confirm the quality of extracted DNA. Then, cauliflower mosaic virus (CaMV) 35S promoter and Agrobacterium nopaline synthase (NOS) terminator were selected as screening targets for detection of GM rice sequences by PCR. Results According to our results, 2 out of 81 (2.4%) samples tested were positive for CaMV 35S promoter while no positive result was detected for NOS terminator. Conclusion The obtained data indicated that this method is capable to identify the GM rice varieties. Furthermore, it can demonstrate the possibility of the presence of GM rice in Tehran’s market, thus putting emphasis on the requirement for developing a precise approach to evaluate this product.

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