Enhanced late blight resistance of transgenic potato expressing glucose oxidase under the control of pathogen-inducible promoter

Wei, Zhen; Chen, Xi; Liang, Haobo; Hu, Yuanlei; Yin, Gao; Zhang, Lin

doi:10.1007/bf02909692

Cited by 8 publications

(4 citation statements)

References 15 publications

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“…These include typical regulatory elements such as CAAT and TATA boxes, a TATTCT box that is responsive to light, a tissue-specific expression element TGHAAARK, an ACGTG responsive to dehydration and dark, a TACGTGTC responsive to ABA, a TGACG responsive to jasmonic acid, SAR-binding domains MarAbox1 (AATAAAYAAA), MarTbox (TTWTWTTWTT), and 5'-UTR (Py-rich) (Lu et al 2009;Graeber et al 2010). Also detected were elements involved in disease resistance and SA induction: the disease-resistance elements TGTCA, GAAAAA (GT1-motif), and TGAC; the SAinduction elements TGACG (AS-1 element) and GRWAAW; W-Box elements (TTGAC, TGACT, TGACY) responsive to both pathogen and SA; and a TTTGACY element (WB-box) responsive to an elicitor (Somssich, 1994;Zhen et al, 2000;Kundu et al, 2011;Maita et al, 2015). The various GUS mRNA levels generated by the different promoters (Figures 6 and 7) lead us to speculate that the high level of expression associated with P 2 might be related to its AS-1 element.…”

Section: Discussionmentioning

confidence: 99%

Cloning and functional analysis of the chitinase gene promoter in peanut

Chen¹,

Song²,

Yu³

et al. 2015

Genet. Mol. Res.

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ABSTRACT.Chitinase is an important pathogenesis-related protein in plants, and it can accumulate when induced by salicylic acid (SA) or other elicitors. Here, we found that chitinase mRNA levels were 4.5-times greater when peanut seedlings were sprayed with 1.5 mM SA, as compared to water. The upstream promoter sequence of the chitinase gene was cloned by TAIL-PCR and the potential cis-regulatory elements in this promoter were predicted by the cis-element databases PLACE and plantCARE. Elements in the promoter related to SA induction and disease resistance response included AS-1, GT1-motif, GRWAAW, TGTCA, W-box, and WB-box. The full-length promoter (P) and a series of 5'-deleted promoters (P 1 -P 5 ) were cloned and then substituted for the 35S promoter of pCAMBIA1301-xylA, which carries the xylose isomerase gene as the selectable marker and GUS as the reporter gene. Six plant expression vectors (pCAMBIA1301-xylA-P-pCAMBIA1301-xylA-P 5 ) were obtained. The six expression vectors were then transferred into onion epidermal cells and peanut plants by Agrobacterium-mediated transformation. Both the full-length and deleted promoters resulted in GUS staining of the onion epidermis cells when 12711 Characterization of peanut chitinase gene promoter ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (4): 12710-12722 (2015) induced by SA. In onion epidermis cells, GUS enzyme activity was greater after SA induction. In transgenic peanut plants, GUS mRNA levels were greater after SA induction. Consideration of the cis-regulatory elements predicted by PLACE and plantCARE suggested that AS-1, GRWAAW, and W-box are positive regulatory elements in P 2 and P 3 and that GT1-motif and TGTCA are negative regulatory elements between P and P 2 .

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Section: Discussionmentioning

confidence: 99%

Cloning and functional analysis of the chitinase gene promoter in peanut

Chen¹,

Song²,

Yu³

et al. 2015

Genet. Mol. Res.

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“…The emergence of CRISPR/Cas9 technology provides a new approach to address this problem, and this technology has worked to achieve resistance to pathogens in various crops [ 49 – 51 ]; however, to reduce the negative effects of a transgene on the whole plant, the most efficient way to confer resistance is to restrict its expression only to infected cells. Pathogen-induced promoters are unique and valuable tools for engineering resistance to plant disease [ 52 , 53 ]. Therefore, integration of the CRISPR/Cas9 system and pathogen-induced promoters (pol II promoter) to develop a pathogen-inducible gene editing system is of great significance for devising pyrethrum resistance to S. tanaceti , as this method can restrict gene editing only to infected cells and reduce metabolic disruption caused by gene editing, thus minimizing the impact on plant growth and development.…”

Section: Discussionmentioning

confidence: 99%

Ribozyme-mediated CRISPR/Cas9 gene editing in pyrethrum (Tanacetum cinerariifolium) hairy roots using a RNA polymerase II-dependent promoter

Zeng

et al. 2022

Plant Methods

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Background Traditional CRISPR/Cas9 systems that rely on U6 or U3 snRNA promoters (RNA polymerase III-dependent promoters) can only achieve constitutive gene editing in plants, hampering the functional analysis of specifically expressed genes. Ribozyme-mediated CRISPR/Cas9 systems increase the types of promoters which can be used to transcribe sgRNA. Therefore, such systems allow specific gene editing; for example, transcription of the artificial gene Ribozyme-sgRNA-Ribozyme (RGR) is initiated by an RNA polymerase II-dependent promoter. Genetic transformation is indispensable for editing plant genes. In certain plant species, including pyrethrum, genetic transformation remains challenging to do, limiting the functional verification of novel CRISPR/Cas9 systems. Thus, this study’s aim was to develop a simple Agrobacterium rhizogenes-mediated hairy root transformation system to analyze the function of a ribozyme-mediated CRISPR/Cas9 system in pyrethrum. Results A hairy root transformation system for pyrethrum is described, with a mean transformation frequency of 7%. Transgenic hairy roots transformed with the pBI121 vector exhibited significantly increased beta-glucuronidase staining as a visual marker of transgene expression. Further, a ribozyme-based CRISPR/Cas9 vector was constructed to edit the TcEbFS gene, which catalyzes synthesis of the defense-related compound (E)-β-farnesene in pyrethrum. The vector was transferred into the hairy roots of pyrethrum and two stably transformed hairy root transgenic lines obtained. Editing of the TcEbFS gene in the hairy roots was evaluated by gene sequencing, demonstrating that both hairy root transgenic lines had DNA base loss at the editing target site. Gas chromatography–mass spectrometry showed that the (E)-β-farnesene content was significantly decreased in both hairy root transgenic lines compared with the empty vector control group. Altogether, these results show that RGR can be driven by the CaMV35S promoter to realize TcEbFS gene editing in pyrethrum hairy roots. Conclusion An A. rhizogenes-mediated hairy root transformation and ribozyme-mediated CRISPR/Cas9 gene editing system in pyrethrum was established, thereby facilitating gene editing in specific organs or at a particular developmental stage in future pyrethrum research.

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“…The resulting constructs were designated as pBI-Pat, pBI-GBSS, pBI-Ubi, and pBI-Lhca ( Figure 1) and were transformed into Agrobacterium tumefaciens strain AGL0. them, such as cold-or pathogen-inducible promoters, have been isolated from the potato genome [27,28]. The aim of this work was to test and compare the efficacy of several potato-derived promoters with an aim to apply them in various functional and applied studies.…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…Alternatively, other environmentally inducible promoters with different levels of activity might be adopted to minimize the adverse effects. However, the practical use of inducible promoters is strictly limited, since only a few of them, such as cold- or pathogen-inducible promoters, have been isolated from the potato genome [ 27 , 28 ].…”

Section: Introductionmentioning

confidence: 99%

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

Miroshnichenko

Фирсов

Timerbaev

et al. 2020

Plants

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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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Enhanced late blight resistance of transgenic potato expressing glucose oxidase under the control of pathogen-inducible promoter

Cited by 8 publications

References 15 publications

Cloning and functional analysis of the chitinase gene promoter in peanut

Cloning and functional analysis of the chitinase gene promoter in peanut

Ribozyme-mediated CRISPR/Cas9 gene editing in pyrethrum (Tanacetum cinerariifolium) hairy roots using a RNA polymerase II-dependent promoter

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

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