Transgenic plants over-expressing insect-specific microRNA acquire insecticidal activity against Helicoverpa armigera: an alternative to Bt-toxin technology

Agrawal, Aditi; Rajamani, Vijayalakshmi; Reddy, Vanga Siva; Mukherjee, Sunil Kumar; Bhatnagar, Raj K.

doi:10.1007/s11248-015-9880-x

Cited by 71 publications

(47 citation statements)

References 40 publications

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“…The larvae of cotton bollworm, Helicoverpa armigera, that fed on bacteria expressing an artificial miRNA (amiRNA) sequence specifically targeting the ecdysone receptor (EcR) gene showed greater mortality, developmental defects, and a significant decline in reproductive ability (Yogindran and Rajam, 2016). Feeding on transgenic tobacco plants producing an amiRNA based on endogenous miR-24 against a chitinase gene significantly reduced the level of chitinase transcripts in H. armigera larvae, causing cessation of molt (Agrawal et al, 2015). Although RSB larvae that were continuously fed with transgenic Csu-novel-miR-15 rice showed a 4-day delay of pupation, no significant lethal effect was observed (Jiang et al, 2016), the target genes and biological function of this novel miRNA remain unclear.…”

Section: Introductionmentioning

confidence: 99%

Transgenic microRNA‐14 rice shows high resistance to rice stem borer

Xiao

Sun

et al. 2018

Plant Biotechnology Journal

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Rice stem borer (RSB, Chilo suppressalis) is an insect pest that causes huge economic losses every year. Control efforts rely heavily on chemical insecticides, which leads to serious problems such as insecticide resistance, environment pollution, and food safety issues. Therefore, developing alternative pest control methods is an important task. Here, we identified an insect-specific microRNA, miR-14, in RSB, which was predicted to target Spook (Spo) and Ecdysone receptor (EcR) in the ecdysone signalling network. In-vitro dual luciferase assays using HEK293T cells confirmed the interactions of Csu-miR-14 with CsSpo and with CsEcR. Csu-miR-14 exhibited high levels of expression at the end of each larval instar stage, and its expression was negatively correlated with the expression of its two target genes. Overexpression of Csu-miR-14 at the third day of the fifth instar stage led to high mortality and developmental defects in RSB individuals. We produced 35 rice transformants to express miR-14 and found that three lines had a single copy with highly abundant miR-14 mature transcripts. Feeding bioassays using both T and T generations of transgenic miR-14 rice indicated that at least one line (C#24) showed high resistance to RSB. These results indicated that the approach of miRNAs as targets has potential for improving pest control methods. Moreover, using insect-specific miRNAs rather than protein-encoding genes for pest control may prove benign to non-insect species, and thus is worthy of further exploration.

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Section: Introductionmentioning

confidence: 99%

Transgenic microRNA‐14 rice shows high resistance to rice stem borer

Xiao

Sun

et al. 2018

Plant Biotechnology Journal

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“…Furthermore, Agrawal et al. () recently reported cessation of H. armigera larval molting after feeding on transgenic tobacco engineered to express a miRNA targeting the H. armigera group I chitinase hachi . Future studies in L. hesperus will focus on using RNAi‐based methods to elucidate the roles of the 10 chitinase‐like sequences in molting and determining their potential usefulness in controlling populations of this pest species.…”

Section: Discussionmentioning

confidence: 99%

ECDYSTEROID AND CHITINASE FLUCTUATIONS IN THE WESTERN TARNISHED PLANT BUG (Lygus hesperus) PRIOR TO MOLT INDICATE ROLES IN DEVELOPMENT

Brent

Wang

Miao

et al. 2016

Arch Insect Biochem Physiol

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Vital physiological processes that drive the insect molt represent areas of interest for the development of alternative control strategies. The western tarnished plant bug (Lygus hesperus Knight) is a pest of numerous agronomic and horticultural crops but the development of novel control approaches is impeded by limited knowledge of the mechanisms regulating its molt. To address this deficiency, we examined the fundamental relationship underlying the hormonal and molecular components of ecdysis. At 27°C L. hesperus exhibits a temporally controlled nymph-adult molt that occurs about 4 days after the final nymph-nymph molt with ecdysteroid levels peaking 2 days prior to the final molt. Application of exogenous ecdysteroids when endogenous levels had decreased disrupted the nymphal-adult molt, with treated animals exhibiting an inability to escape the old exoskeleton and resulting in mortality compared to controls. Using accessible transcriptomic data, we identified 10 chitinase-like sequences (LhCht), eight of which had protein motifs consistent with chitinases. Phylogenetic analyses revealed orthologous relationships to chitinases critical to molting in other insects. RT-PCR based transcript profiling revealed that expression changes to four of the LhChts was coordinated with the molt period and ecdysteroid levels. Collectively, our results support a role for ecdysteroid regulation of the L. hesperus molt and suggest that cuticle clearance is mediated by LhCht orthologs of chitinases that are essential to the molt process. These results provide the initial hormonal and molecular basis for future studies to investigate the specific roles of these components in molting.

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“…Similarly, Arabidopsis expressing two amiRNAs targeting the mRNA of the P69 and HC-Pro proteins showed resistance to Turnip yellow mosaic virus and Turnip mosaic virus, respectively (Niu et al, 2006). Agrawal et al (2015) showed that tobacco overexpressing an engineered amiRNA was resistant to Helicoverpa armigera.…”

Section: Artificial Mir Genesmentioning

confidence: 99%

“…Greater target specificity can be achieved with overexpression driven by tissue‐specific (Niu et al ., ) or stress‐inducible (Gao et al ., ) promoters. Additional strategies include the overexpression of target mRNAs that are resistant to specific miRNAs (Guan et al ., ), the expression of artificial target mimics that can cancel out the effect of endogenous miRNA activity (Sharma et al ., ) and the expression of artificial MIR gene targeting only the desired mRNAs (Agrawal et al ., ). Jian et al .…”

Section: The Role Of Mirna In Plant Abiotic or Biotic Stress Responsesmentioning

confidence: 99%

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MicroRNAs and new biotechnological tools for its modulation and improving stress tolerance in plants

Basso

Ferreira

Kobayashi

et al. 2019

Plant Biotechnology Journal

118

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Summary Micro RNA s (mi RNA s) modulate the abundance and spatial–temporal accumulation of target mRNA s and indirectly regulate several plant processes. Transcriptional regulation of the genes encoding mi RNA s ( MIR genes) can be activated by numerous transcription factors, which themselves are regulated by other mi RNA s. Fine‐tuning of MIR genes or mi RNA s is a powerful biotechnological strategy to improve tolerance to abiotic or biotic stresses in crops of economic importance. Current approaches for mi RNA fine‐tuning are based on the down‐ or up‐regulation of MIR gene transcription and the use of genetic engineering tools to manipulate the final concentration of these mi RNA s in the cytoplasm. Transgenesis, cisgenesis, intragenesis, artificial MIR genes, endogenous and artificial target mimicry, MIR genes editing using Meganucleases, ZNF proteins, TALEN s and CRISPR /Cas9 or CRISPR /Cpf1, CRISPR / dC as9 or dC pf1, CRISPR 13a, topical delivery of mi RNA s and epigenetic memory have been successfully explored to MIR gene or mi RNA modulation and improve agronomic traits in several model or crop plants. However, advantages and drawbacks of each of these new biotechnological tools ( NBT s) are still not well understood. In this review, we provide a brief overview of the biogenesis and role of mi RNA s in response to abiotic or biotic stresses, we present critically the main NBT s used for the manipulation of MIR genes and mi RNA s, we show current efforts and findings with the MIR genes and mi RNA s modulation in plants, and we summarize the advantages and drawbacks of these NBT s and provide some alternatives to overcome. Finally, challenges and future perspectives to mi RNA modulating in important crops are also discussed.

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Transgenic plants over-expressing insect-specific microRNA acquire insecticidal activity against Helicoverpa armigera: an alternative to Bt-toxin technology

Cited by 71 publications

References 40 publications

Transgenic microRNA‐14 rice shows high resistance to rice stem borer

Transgenic microRNA‐14 rice shows high resistance to rice stem borer

ECDYSTEROID AND CHITINASE FLUCTUATIONS IN THE WESTERN TARNISHED PLANT BUG (Lygus hesperus) PRIOR TO MOLT INDICATE ROLES IN DEVELOPMENT

MicroRNAs and new biotechnological tools for its modulation and improving stress tolerance in plants

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