Synthetic chitinase gene driven by root-specific LjNRT2 and AtNRT2.1 promoters confers resistance to Fusarium oxysporum in transgenic tobacco and tomato

Kong, Kynet; Makabe, So; Ntui, Valentine Otang; Khan, Raham Sher; Nakamura, Ikuo

doi:10.1007/s11816-013-0303-2

Cited by 8 publications

(5 citation statements)

References 31 publications

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“…This might be due to the high expression of the NiC gene in these two lines as compared to other transgenic lines. Similar results of antifungal activity were observed in NiC -transgenic plants against Fusarium oxysporum (Kong et al, 2014 ). Transgenic potato plants, transformed with chitinase ( ChiC ) gene also exhibited resistance against fungal pathogens, Botyrytis cinerea (gray mold), Alternaria solani and F. oxysporum (Khan et al, 2008 , 2011 ).…”

Section: Discussionsupporting

confidence: 82%

Impact of Transgenic Brassica napus Harboring the Antifungal Synthetic Chitinase (NiC) Gene on Rhizosphere Microbial Diversity and Enzyme Activities

et al. 2017

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Transgenic Brassica napus harboring the synthetic chitinase (NiC) gene exhibits broad-spectrum antifungal resistance. As the rhizosphere microorganisms play an important role in element cycling and nutrient transformation, therefore, biosafety assessment of NiC containing transgenic plants on soil ecosystem is a regulatory requirement. The current study is designed to evaluate the impact of NiC gene on the rhizosphere enzyme activities and microbial community structure. The transgenic lines with the synthetic chitinase gene (NiC) showed resistance to Alternaria brassicicola, a common disease causing fungal pathogen. The rhizosphere enzyme analysis showed no significant difference in the activities of fivesoil enzymes: alkalyine phosphomonoestarase, arylsulphatase, β-glucosidase, urease and sucrase between the transgenic and non-transgenic lines of B. napus varieties, Durr-e-NIFA (DN) and Abasyne-95 (AB-95). However, varietal differences were observed based on the analysis of molecular variance. Some individual enzymes were significantly different in the transgenic lines from those of non-transgenic but the results were not reproducible in the second trail and thus were considered as environmental effect. Genotypic diversity of soil microbes through 16S–23S rRNA intergenic spacer region amplification was conducted to evaluate the potential impact of the transgene. No significant diversity (4% for bacteria and 12% for fungal) between soil microbes of NiC B. napus and the non-transgenic lines was found. However, significant varietal differences were observed between DN and AB-95 with 79% for bacterial and 54% for fungal diversity. We conclude that the NiC B. napus lines may not affect the microbial enzyme activities and community structure of the rhizosphere soil. Varietal differences might be responsible for minor changes in the tested parameters.

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Section: Discussionsupporting

confidence: 82%

Impact of Transgenic Brassica napus Harboring the Antifungal Synthetic Chitinase (NiC) Gene on Rhizosphere Microbial Diversity and Enzyme Activities

et al. 2017

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“…Transgenic plants harboring LjNRT2-GUS showed GUS staining only in the root tissues (Kong et al 2013). In this study, those two root-specific promoters were respectively linked to wasabi defensin gene.…”

Section: Discussionmentioning

confidence: 51%

“…DNA fragments containing LjNRT2 and AtNRT2.1 promoter sequences were cloned by PCR (Kong et al 2013). The LjNRT2 (1.0 kb) or AtNRT2.1 (1.1 kb) promoter was linked to WD gene at HindIII/BamHI or HindIII/BglII restriction sites, respectively.…”

Section: Binary Vector Constructionsmentioning

confidence: 99%

Transgenic tobacco and tomato plants expressing Wasabi defensin genes driven by root-specific LjNRT2 and AtNRT2.1 promoters confer resistance against Fusarium oxysporum

Kong

Ntui

Makabe

et al. 2014

Plant Biotechnology

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Antifungal peptides are a potential group of defense molecules that have been utilized to develop resistance to various plant pathogens. Wasabi defensin (WD) gene (0.5 kb) consists of cysteine-rich peptides that show potent growth inhibition of pathogenic filamentous fungi, such as Botrytis cinerea. Under regulation by the root-specific LjNRT2 or AtNRT2.1 promoter, WD gene was expressed in the roots of transgenic tobacco and tomato plants by Agrobacteriummediated transformation. The regenerated plants showed stable integration of the transgene, with different insertion sites, and the transgene was expressed in the root tissues but not in the leaf tissues. This result confirmed that WD protein accumulated only in the roots of transgenic plants. In a bioassay for resistance to Fusarium oxysporum, all transgenic plants showed increased resistance to the fungus as compared to non-transformed plants. Protein extracts from root and leaf tissues were assayed for antifungal activity and the activity was express as the number of colonies formed per cm 2 (CFU cm −2 ). The CFU values of the root and leaf extracts of control plants did not show significant differences. In contrast, the CFU values of the root extracts of the transgenic plants were significantly lower than those of the leaf extracts and much lower than those of control. These results suggest that LjNRT2 and AtNRT2.1 promoters triggered the antifungal gene expression in the roots and conferred increased resistance to the root pathogen F. oxysporum. In the view of bio-safety, the root-specific expression of the transgene is desirable because the roots of tomato are not edible.

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“…The genes that encode chitinase from some Trichoderma species such as T. harzianum and T. virens have been also transformed into tobacco, potato, cotton and apple against some pathogens such as Alternaria alternata, A. solani, Botrytis cinerea, and Rhizoctonia solani (Lorito et al 1998;Emani et al 2003;Schäfer et al 2012). However, the expression of fungal-derived synthetic chitinase was only found in some reports such as the NiC gene from Rhizopus oligosporus for Petunia hybrida (Khan et al 2012), tobacco and tomato (Kong et al 2014) and Brassica napus (Khan et al 2017b). This study is therefore probably the first findings of the synthetic chitinase genes derived from T. asperellum that has been expressed in plants and demonstrated strong antifungal activity against S. rolfsii.…”

Section: Discussionmentioning

confidence: 99%

Transient Expression of Chi42 Genes from Trichoderma asperellum in Nicotiana benthamiana by Agroinfiltration

Tien¹

2021

IJAB

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The present study reports the transient expression of chi42 genes encoding 42 kDa chitinase from T. asperellum SH16 in N. benthamiana via agroinfiltration. The efficacy of agroinfiltration for chi42 genes including a wild-type gene (Chi42) and two synthetic genes (syncodChi42-1 and syncodChi42-2) was determined. Accordingly, coinfiltration of two vectors pMYV719 carrying one of three genes chi42 and pMYV508 carrying gene p19 expedited the higher expression of recombinant enzymes whose genes were optimized for codon usage in plant tissues. The highest chitinolytic activity of about 290 U/mL was found in plants containing the gene syncodChi42-2 after 7 days of injection, 1.7 and 2.6 times higher than that of genes syncodChi42-1 and chi42. Recombinant chitinase has also exhibited activity against the pathogenic fungus Sclerotium rolfsii under in vitro condition. A higher expression level of syncodChi42-2 gene in N. benthamiana and its antifungal activity promise potential applications in the field of transgenic crops against phytopathogenic fungi. © 2021 Friends Science Publishers

show abstract

Synthetic chitinase gene driven by root-specific LjNRT2 and AtNRT2.1 promoters confers resistance to Fusarium oxysporum in transgenic tobacco and tomato

Cited by 8 publications

References 31 publications

Impact of Transgenic Brassica napus Harboring the Antifungal Synthetic Chitinase (NiC) Gene on Rhizosphere Microbial Diversity and Enzyme Activities

Impact of Transgenic Brassica napus Harboring the Antifungal Synthetic Chitinase (NiC) Gene on Rhizosphere Microbial Diversity and Enzyme Activities

Transgenic tobacco and tomato plants expressing Wasabi defensin genes driven by root-specific LjNRT2 and AtNRT2.1 promoters confer resistance against Fusarium oxysporum

Transient Expression of Chi42 Genes from Trichoderma asperellum in Nicotiana benthamiana by Agroinfiltration

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