Passion fruit flowers: Kunitz trypsin inhibitors and cystatin differentially accumulate in developing buds and floral tissues

Pereira, Keitty Raquel Benevides; Botelho-Júnior, Sylvio; Domingues, Dalvania Pinho; Machado, Olga Lima Tavares; Oliveira, Antônia Elenir Amâncio; Fernandes, Kátia Valevski Sales; Madureira, Hérika C.; Pereira, Telma Nair Santana; Jacinto, Tânia

doi:10.1016/j.phytochem.2011.06.009

Cited by 5 publications

(4 citation statements)

References 56 publications

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“…While the SNSL sequence in C-terminal extension of the full-length CypCl is only found in some phytocystatins, phytocystatins with C-terminal extension containing this sequence are predicted to inhibit legumain activity. Pereira et al [12] had proved that the SNSL site of the phytocystatins contained in the C-terminal extension is essential when phytocystatin with and without C-terminal extension was tested for legumain-inhibition activity; only those with C-terminal extension are able to inhibit legumain activity. Pereira et al [12] also showed the importance of amino acid N (ASN, asparagine) in SNSL sequence as when it is replaced by a K (Lys, lysine) residue, the phytocystatin is unable to inhibit legumain.…”

Section: Discussionmentioning

confidence: 99%

“…Pereira et al [12] had proved that the SNSL site of the phytocystatins contained in the C-terminal extension is essential when phytocystatin with and without C-terminal extension was tested for legumain-inhibition activity; only those with C-terminal extension are able to inhibit legumain activity. Pereira et al [12] also showed the importance of amino acid N (ASN, asparagine) in SNSL sequence as when it is replaced by a K (Lys, lysine) residue, the phytocystatin is unable to inhibit legumain. With the presences of the conserved sequences coding for the tripartite wedge responsible for papain inhibition and the C-terminal extension which is responsible for legumain inhibition, the novel CypCl could be expressed as a bifunctional protease inhibitor.…”

Section: Discussionmentioning

confidence: 99%

“…It acts as an endogenous proteolysis regulator during seed maturation and development, involves in programmed cell death, and inhibits exogenous proteases released by plant predators like insects, nematode, bacteria, and pathogens during the attack [6–12]. In recent years, however, phytocystatin has also been suggested to be involved in response to biotic and abiotic stresses as they are highly expressed during harsh condition such as cold, drought, salt stress, oxidant stress, and wounds [8, 9, 12]. All these properties encourage more research to be done on PIs, especially on phytocystatin Ayurvedic [2–4].…”

Section: Introductionmentioning

confidence: 99%

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Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric,Curcuma longa

Chan

Bakar

Mahmood

et al. 2014

BioMed Research International

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Phytocystatin, a type of protease inhibitor (PI), plays major roles in plant defense mechanisms and has been reported to show antipathogenic properties and plant stress tolerance. Recombinant plant PIs are gaining popularity as potential candidates in engineering of crop protection and in synthesizing medicine. It is therefore crucial to identify PI from novel sources like Curcuma longa as it is more effective in combating against pathogens due to its novelty. In this study, a novel cDNA fragment encoding phytocystatin was isolated using degenerate PCR primers, designed from consensus regions of phytocystatin from other plant species. A full-length cDNA of the phytocystatin gene, designated CypCl, was acquired using 5′/3′ rapid amplification of cDNA ends method and it has been deposited in NCBI database (accession number KF545954.1). It has a 687 bp long open reading frame (ORF) which encodes 228 amino acids. BLAST result indicated that CypCl is similar to cystatin protease inhibitor from Cucumis sativus with 74% max identity. Sequence analysis showed that CypCl contains most of the motifs found in a cystatin, including a G residue, LARFAV-, QxVxG sequence, PW dipeptide, and SNSL sequence at C-terminal extension. Phylogenetic studies also showed that CypCl is related to phytocystatin from Elaeis guineensis.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric,Curcuma longa

Chan

Bakar

Mahmood

et al. 2014

BioMed Research International

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“…The activity of PIs including KPIs against insect digestive proteinases has led to the promotion of their use as tolerance determinants in transgenic plants ( Green and Ryan, 1972 ; Hilder et al, 1987 ; Ryan, 1990 ; Jongsma and Bolter, 1997 ; Lee et al, 1999 ; McManus et al, 2005 ; Lima et al, 2011 ). Recent evidence also suggests that KPIs in plants are involved in programmed cell death, growth, and development ( Karrer et al, 1998 ; Yeu et al, 2007 ; Kim et al, 2009 ; Laluk and Mengiste, 2011 ; Pereira et al, 2011 ; Boex-Fontvieille et al, 2015 ; Islam et al, 2015a ). Moreover, changes in expression profile of KPIs in response to water limitation suggests a function for these proteins in abiotic stress, possibly by targeting specific proteinases and as such limiting their proteolytic activities ( Downing et al, 1992 ; Kang et al, 2001 ; Desclos et al, 2008 ; Kidric et al, 2014 ).…”

Section: Introductionmentioning

confidence: 99%

Kunitz Proteinase Inhibitors Limit Water Stress Responses in White Clover (Trifolium repens L.) Plants

et al. 2017

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The response of plants to water deficiency or drought is a complex process, the perception of which is triggered at the molecular level before any visible morphological responses are detected. It was found that different groups of plant proteinase inhibitors (PIs) are induced and play an active role during abiotic stress conditions such as drought. Our previous work with the white clover (Trifolium repens L.) Kunitz Proteinase Inhibitor (Tr-KPI) gene family showed that Tr-KPIs are differentially regulated to ontogenetic and biotic stress associated cues and that, at least some members of this gene family may be required to maintain cellular homeostasis. Altered cellular homeostasis may also affect abiotic stress responses and therefore, we aimed to understand if distinct Tr-PKI members function during drought stress. First, the expression level of three Tr-KPI genes, Tr-KPI1, Tr-KPI2, and Tr-KPI5, was measured in two cultivars and one white clover ecotype with differing capacity to tolerate drought. The expression of Tr-KPI1 and Tr-KPI5 increased in response to water deficiency and this was exaggerated when the plants were treated with a previous period of water deficiency. In contrast, proline accumulation and increased expression of Tr-NCED1, a gene encoding a protein involved in ABA biosynthesis, was delayed in plants that experienced a previous drought period. RNAi knock-down of Tr-KPI1 and Tr-KPI5 resulted in increased proline accumulation in leaf tissue of plants grown under both well-watered and water-deficit conditions. In addition, increased expression of genes involved in ethylene biosynthesis was found. The data suggests that Tr-KPIs, particularly Tr-KPI5, have an explicit function during water limitation. The results also imply that the Tr-KPI family has different in planta proteinase targets and that the functions of this protein family are not solely restricted to one of storage proteins or in response to biotic stress.

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Defense response in non-genomic model species: methyl jasmonate exposure reveals the passion fruit leaves’ ability to assemble a cocktail of functionally diversified Kunitz-type trypsin inhibitors and recruit two of them against papain

Botelho-Júnior

Machado

Fernandes

et al. 2014

Planta

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Multiplicity of protease inhibitors induced by predators may increase the understanding of a plant's intelligent behavior toward environmental challenges. Information about defense mechanisms of non-genomic model plant passion fruit (Passiflora edulis Sims) in response to predator attack is still limited. Here, via biochemical approaches, we showed its flexibility to build-up a broad repertoire of potent Kunitz-type trypsin inhibitors (KTIs) in response to methyl jasmonate. Seven inhibitors (20-25 kDa) were purified from exposed leaves by chromatographic techniques. Interestingly, the KTIs possessed truncated Kunitz motif in their N-terminus and some of them also presented non-consensus residues. Gelatin-Native-PAGE established multiple isoforms for each inhibitor. Significant differences regarding inhibitors' activity toward trypsin and chymotrypsin were observed, indicating functional polymorphism. Despite its rarity, two of them also inhibited papain, and such bifunctionality suggests a recruiting process onto another mechanistic class of target protease (cysteine-type). All inhibitors acted strongly on midgut proteases from sugarcane borer, Diatraea saccharalis (a lepidopteran insect) while in vivo assays supported their insecticide properties. Moreover, the bifunctional inhibitors displayed activity toward midgut proteases from cowpea weevil, Callosobruchus maculatus (a coleopteran insect). Unexpectedly, all inhibitors were highly effective against midgut proteases from Aedes aegypti a dipteran insect (vector of neglected tropical diseases) opening new avenues for plant-derived PIs for vector control-oriented research. Our results reflect the KTIs' complexities in passion fruit which could be wisely exploited by influencing plant defense conditions. Therefore, the potential of passion fruit as source of bioactive compounds with diversified biotechnological application was strengthened.

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Passion fruit flowers: Kunitz trypsin inhibitors and cystatin differentially accumulate in developing buds and floral tissues

Cited by 5 publications

References 56 publications

Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric,Curcuma longa

Molecular Cloning and Characterization of Novel Phytocystatin Gene from Turmeric,Curcuma longa

Kunitz Proteinase Inhibitors Limit Water Stress Responses in White Clover (Trifolium repens L.) Plants

Defense response in non-genomic model species: methyl jasmonate exposure reveals the passion fruit leaves’ ability to assemble a cocktail of functionally diversified Kunitz-type trypsin inhibitors and recruit two of them against papain

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