Dataset Information

Safety evaluation of FAD2 RNAi transgenic Brassica napus L. based on microbial diversity and metabonomic analysis.

ABSTRACT: Oleic acid desaturase (FAD2) is the key enzyme that produces polyunsaturated fatty acids in rapeseed (Brassica napus L), which is one of the main oil crops. RNA interference (RNAi) is an emerging technique that provides new opportunities for the generation of new traits in plants. To increase oleic acid content and reduce linoleic and linolenic acid content in rapeseed, we constructed an ihpRNA plant expression vector of the FAD2 gene and obtained transgenic plants for multiple generations by stable inheritance. In this study, third-generation transgenic plants (T3), seventh-generation transgenic plants (T7), and wild-type plants (WT) were used. The differences in microbial community diversity between transgenic plants and wild-type plants and the up- and downregulation of rhizosphere metabolite contents were investigated. In conclusion, the results showed that the soil microbial community structure was stable, the general microbial community structure was not changed by the transgenic rhizosphere exudates, and no significant harmful root exudate of transgenic rapeseed on the environment was found through the microbial community and metabolomics analysis. This work may provide an understanding of the impact of RNAi on plant metabolites and a safety evaluation method for transgenic plants and a reference for rapeseed breeding.

SUBMITTER: Qi Y

PROVIDER: S-EPMC9751890 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature

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Safety evaluation of FAD2 RNAi transgenic Brassica napus L. based on microbial diversity and metabonomic analysis.

Qi Yanting Y Wang Qiming Q Xie Qingxuan Q Wu Chuan C Xu Minhui M Han Shaofan S Zhou Ting T Li Juan J Xia Libing L Li Wai Chin WC Pan Weisong W

Frontiers in plant science 20221201

Oleic acid desaturase (FAD2) is the key enzyme that produces polyunsaturated fatty acids in rapeseed (Brassica napus L), which is one of the main oil crops. RNA interference (RNAi) is an emerging technique that provides new opportunities for the generation of new traits in plants. To increase oleic acid content and reduce linoleic and linolenic acid content in rapeseed, we constructed an ihpRNA plant expression vector of the FAD2 gene and obtained transgenic plants for multi ...[more]

PMID: 36531340

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Project description:BackgroundRapid-cycling Brassica napus (B. napus-RC) has potential as a rapid trait testing system for canola (B. napus) because its life cycle is completed within 2 months while canola usually takes 4 months, and it is susceptible to the same range of diseases and abiotic stress as canola. However, a rapid trait testing system for canola requires the development of an efficient transformation and tissue culture system for B. napus-RC. Furthermore, effectiveness of this system needs to be demonstrated by showing that a particular trait can be rapidly introduced into B. napus-RC plants.ResultsAn in-vitro regeneration protocol was developed for B. napus-RC using 4-day-old cotyledons as the explant. High regeneration percentages, exceeding 70%, were achieved when 1-naphthaleneacetic acid (0.10 mg/L), 6-benzylaminopurine (1.0 mg/L), gibberellic acid (0.01 mg/L) and the ethylene antagonist silver nitrate (5 mg/L) were included in the regeneration medium. An average transformation efficiency of 16.4% was obtained using Agrobacterium-mediated transformation of B. napus-RC cotyledons using Agrobacterium strain GV3101 harbouring a plasmid with an NPTII (kanamycin-selectable) marker gene and the Arabidopsis thaliana cDNA encoding ACYL-COA-BINDING PROTEIN6 (AtACBP6). Transgenic B. napus-RC overexpressing AtACBP6 displayed better tolerance to freezing/frost than the wild type, with enhanced recovery from cellular membrane damage at both vegetative and flowering stages. AtACBP6-overexpressing B. napus-RC plants also exhibited lower electrolyte leakage and improved recovery following frost treatment, resulting in higher yields than the wild type. Ovules from transgenic AtACBP6 lines were better protected from frost than those of the wild type, while the developing embryos of frost-treated AtACBP6-overexpressing plants showed less freezing injury than the wild type.ConclusionsThis study demonstrates that B. napus-RC can be successfully regenerated and transformed from cotyledon explants and has the potential to be an effective trait testing platform for canola. Additionally, AtACBP6 shows potential for enhancing cold tolerance in canola however, larger scale studies will be required to further confirm this outcome.

Project description:Abiotic stresses such as drought and salt stresses have a negative effect on the growth and productivity of plants. Improvement of stress tolerance through genetic engineering in plants has been reported in intense studies. Transcription factors play vital roles in plant adaptation to stresses by regulating expression of a great deal of target genes. A family of Arabidopsis basic region leucine zipper (bZIP) transcription factors that can recognize and bind to the abscisic acid (ABA)-responsive elements (ABREs) in promoter is named as ABRE binding factors (ABFs)/ABRE binding proteins (AREBs). They play a key role in the regulation of expression of downstream stress-responsive genes in ABA signalling. Genetic transformation of ABF/ABRE transcription factors has been suggested to be an effective approach for engineering stress-tolerant plants. However, whether the ABF/ABRE transcription factors are able to be used for generating stress-tolerant rapeseed plants has not yet been studied.BnaABF2, encoding a bZIP transcription factor, was cloned from rapeseed in this study. Subcellular localization and transactivation analyses showed that BnaABF2 was localized to the nucleus with transactivation activity in plant cells. BnaABF2 gene expression was induced by drought and salt stresses and BnaABF2 positively functions in ABA signalling during the vegetative stage. Overexpression of BnaABF2 was found to render drought and salt tolerance to Arabidopsis plants. The resistance of the BnaABF2-expressing transgenic plants to drought and salt stresses is due to reduced water-loss rate and expression of stress-responsive genes such as RD29B, RAB18 and KIN2. The expression of RD29B, RAB18 and KIN2 regulated by BnaABF2 is involved in an ABA-dependent stress signalling.Identification of the positive role of rapeseed BnaABF2 in plant tolerance to drought and salt provides evidence for ability of engineering stress-tolerant rapeseed plants by genetic transformation of BnaABF2.

Dataset Information

Safety evaluation of FAD2 RNAi transgenic Brassica napus L. based on microbial diversity and metabonomic analysis.

Publications

Safety evaluation of <i>FAD2</i> RNAi transgenic <i>Brassica napus</i> L. based on microbial diversity and metabonomic analysis.

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