Proteinase inhibitors in legume herbivore defense: from natural to genetically engineered protectants

Sultana, Mst Shamira; Millwood, Reginald J.; Mazarei, Mitra; Stewart, C. Neal

doi:10.1007/s00299-021-02800-7

Cited by 9 publications

(8 citation statements)

References 108 publications

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“…Genetically modified plants and crops provide one of the solutions to increase global food production with improved gains in yield and resistance to plant diseases or insect pests. Several successful cases of genetically modified plants have conferred phytoprotection against insects, pests, and pathogens, such as overexpression of proteinase inhibitor genes from legumes [3], recombinant Bt toxic proteins from soil bacteria Bacillus thuringiensis [4], α-amylase inhibitors, and plant lectins [5]. In spite of these successful examples, there is a need to develop alternative strategies of phytoprotection.…”

Section: Introductionmentioning

confidence: 99%

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Cell-Penetrating Peptides for Use in Development of Transgenic Plants

et al. 2023

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Genetically modified plants and crops can contribute to remarkable increase in global food supply, with improved yield and resistance to plant diseases or insect pests. The development of biotechnology introducing exogenous nucleic acids in transgenic plants is important for plant health management. Different genetic engineering methods for DNA delivery, such as biolistic methods, Agrobacterium tumefaciens-mediated transformation, and other physicochemical methods have been developed to improve translocation across the plasma membrane and cell wall in plants. Recently, the peptide-based gene delivery system, mediated by cell-penetrating peptides (CPPs), has been regarded as a promising non-viral tool for efficient and stable gene transfection into both animal and plant cells. CPPs are short peptides with diverse sequences and functionalities, capable of agitating plasma membrane and entering cells. Here, we highlight recent research and ideas on diverse types of CPPs, which have been applied in DNA delivery in plants. Various basic, amphipathic, cyclic, and branched CPPs were designed, and modifications of functional groups were performed to enhance DNA interaction and stabilization in transgenesis. CPPs were able to carry cargoes in either a covalent or noncovalent manner and to internalize CPP/cargo complexes into cells by either direct membrane translocation or endocytosis. Importantly, subcellular targets of CPP-mediated nucleic acid delivery were reviewed. CPPs offer transfection strategies and influence transgene expression at subcellular localizations, such as in plastids, mitochondria, and the nucleus. In summary, the technology of CPP-mediated gene delivery provides a potent and useful tool to genetically modified plants and crops of the future.

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Section: Introductionmentioning

confidence: 99%

“…inhibitor genes from legumes [3], recombinant Bt toxic proteins from soil bacteria Bacillus thuringiensis [4], α-amylase inhibitors, and plant lectins [5]. In spite of these successful examples, there is a need to develop alternative strategies of phytoprotection.…”

Section: Introductionmentioning

confidence: 99%

Cell-Penetrating Peptides for Use in Development of Transgenic Plants

et al. 2023

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“…Most lepidopteran digestive systems are largely based on serine proteinases (trypsin and chymotrypsin), which can be deactivated by plant serine PIs (Srinivasan et al, 2006;Shamsi et al, 2016;Terra and Ferreira, 2020). The utilization of these inherent defensive genes in a genetic engineering strategy may inhibit insect herbivory through changes in expression patterns or through PI overexpression in leaves (Sultana et al, 2022). PI expression in leaves could prevent insect damage and eventual yield loss.…”

Section: Introductionmentioning

confidence: 99%

“…BBIs are composed of single-chain polypeptides with fourteen cysteine residues that form seven disulfide bridges, and two reactive sites. The structural pattern of KTI and BBI inhibitors allows them to interact with and inhibit insect serine proteinases (Sultana et al, 2022). KTIs and BBIs are commonly found in legume seeds and are known as defensive proteins (Macedo et al, 2004;Azzouz et al, 2005;Srinivasan et al, 2005;Müller et al, 2017;Sultana et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

“…The generation of TI mutants by site-directed mutagenesis could potentially led on insect resistance (Kiggundu et al, 2006;Goulet et al, 2008). Synthetic biology may someday be used to create novel TI proteins with multiple target inhibition properties, i.e., different proteinases in multiple pest species (Sultana et al, 2022). Altogether, these findings suggest that TI genes have the potential to enhance herbivore resistance.…”

Section: Introductionmentioning

confidence: 99%

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Overexpression of soybean trypsin inhibitor genes decreases defoliation by corn earworm (Helicoverpa zea) in soybean (Glycine max) and Arabidopsis thaliana

Sultana

Mazarei²,

Jurat-Fuentes³

et al. 2023

Front. Plant Sci.

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Trypsin inhibitors (TIs) are widely distributed in plants and are known to play a protective role against herbivores. TIs reduce the biological activity of trypsin, an enzyme involved in the breakdown of many different proteins, by inhibiting the activation and catalytic reactions of proteins. Soybean (Glycine max) contains two major TI classes: Kunitz trypsin inhibitor (KTI) and Bowman-Birk inhibitor (BBI). Both genes encoding TI inactivate trypsin and chymotrypsin enzymes, which are the main digestive enzymes in the gut fluids of Lepidopteran larvae feeding on soybean. In this study, the possible role of soybean TIs in plant defense against insects and nematodes was investigated. A total of six TIs were tested, including three known soybean trypsin inhibitors (KTI1, KTI2 and KTI3) and three genes encoding novel inhibitors identified in soybean (KTI5, KTI7, and BBI5). Their functional role was further examined by overexpression of the individual TI genes in soybean and Arabidopsis. The endogenous expression patterns of these TI genes varied among soybean tissues, including leaf, stem, seed, and root. In vitro enzyme inhibitory assays showed significant increase in trypsin and chymotrypsin inhibitory activities in both transgenic soybean and Arabidopsis. Detached leaf-punch feeding bioassays detected significant reduction in corn earworm (Helicoverpa zea) larval weight when larvae fed on transgenic soybean and Arabidopsis lines, with the greatest reduction observed in KTI7 and BBI5 overexpressing lines. Whole soybean plant greenhouse feeding bioassays with H. zea on KTI7 and BBI5 overexpressing lines resulted in significantly reduced leaf defoliation compared to non-transgenic plants. However, bioassays of KTI7 and BBI5 overexpressing lines with soybean cyst nematode (SCN, Heterodera glycines) showed no differences in SCN female index between transgenic and non-transgenic control plants. There were no significant differences in growth and productivity between transgenic and non-transgenic plants grown in the absence of herbivores to full maturity under greenhouse conditions. The present study provides further insight into the potential applications of TI genes for insect resistance improvement in plants.

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