A biophysical and structural study of two chitinases from <i>Agave tequilana</i> and their potential role as defense proteins

Sierra-Gómez, Yusvel; Rodríguez-Hernández, Annia; Cano‐Sánchez, Patricia; Gómez-Velasco, Homero; Hernández-Santoyo, Alejandra; Siliqi, Dritan; Rodrı́guez-Romero, Adela

doi:10.1111/febs.14993

Cited by 9 publications

(5 citation statements)

References 79 publications

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“…The optimum pH and temperature ranges over which these chitinases were stable were in the range of those reported for other reported plant, fungi and bacterial chitinases ( Patel et al., 2009 ; Sukprasirt and Wititsuwannakul, 2014 ; Horiuchi et al., 2016 ; Thimoteo et al., 2017 ; Wang et al., 2019 ; Rajninec et al., 2020 , Supplementary Table S4 ). The pH optimum and pH range for Os Chi19A activity are consistent with those reported for other plant chitinases ( Patel et al., 2009 ; Sukprasirt and Wititsuwannakul, 2014 ; Horiuchi et al., 2016 ; Sierra-Gómez et al., 2019 ; Wang et al., 2019 ). However, unlike other reported plants and bacterial chitinases, all of which have reported pH optima between 4.5 and 6, Zm Chi19A and Csp Ch18A were most active at pH 8.…”

Section: Discussionsupporting

confidence: 86%

Plant root associated chitinases: structures and functions

Shobade,

Zabotina,

Nilsen-Hamilton

2024

Front. Plant Sci.

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Chitinases degrade chitin, a linear homopolymer of β-1,4-linked N-acetyl-D-glucosamine (GlcNAc) residues found in the cell walls of fungi and the exoskeletons of arthropods. They are secreted by the roots into the rhizosphere, a complex and dynamic environment where intense nutrient exchange occurs between plants and microbes. Here we modeled, expressed, purified, and characterized Zea mays and Oryza sativa root chitinases, and the chitinase of a symbiotic bacterium, Chitinophaga oryzae 1303 for their activities with chitin, di-, tri-, and tetra-saccharides and Aspergillus niger, with the goal of determining their role(s) in the rhizosphere and better understanding the molecular mechanisms underlying plant-microbe interactions. We show that Zea mays basic endochitinase (ZmChi19A) and Oryza sativa chitinase (OsChi19A) are from the GH19 chitinase family. The Chitinophaga oryzae 1303 chitinase (CspCh18A) belongs to the GH18 family. The three enzymes have similar apparent KM values of (20-40 µM) for the substrate 4-MU-GlcNAc3. They vary in their pH and temperature optima with OsChi19A activity optimal between pH 5–7 and 30–40°C while ZmChi19A and CspCh18A activities were optimal at pH 7-9 and 50–60°C. Modeling and site-directed mutation of ZmChi19A identified the catalytic cleft and the active residues E147 and E169 strategically positioned at ~8.6Å from each other in the folded protein. Cleavage of 4-MU-GlcNAc3 was unaffected by the absence of the CBD but diminished in the absence of the flexible C-terminal domain. However, unlike for the soluble substrate, the CBD and the newly identified flexible C-terminal domain were vital for inhibiting Aspergillus niger growth. The results are consistent with the involvement of the plant chitinases in defense against pathogens like fungi that have chitin exoskeletons. In summary, we have characterized the functional features and structural domains necessary for the activity of two plant root chitinases that are believed to be involved in plant defense and a bacterial chitinase that, along with the plant chitinases, may participate in nutrient recycling in the rhizosphere.

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Section: Discussionsupporting

confidence: 86%

Plant root associated chitinases: structures and functions

Shobade,

Zabotina,

Nilsen-Hamilton

2024

Front. Plant Sci.

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“…This reiterated that the change in aa residues on the surface‐exposed loop III could be a potential reason for the difference in efficacies of chitinases in our study 6 . Additionally, it has been reported that aa changes in other regions (outside of CD) also could bring conformational changes in the catalytic cleft of chitinases 47 . We therefore speculate that aa variations in CpCHI4 positively affected the (GlcNAc) 6 ligand accommodation process, thereby facilitating effective substrate binding and superior efficacy against the herbivore.…”

Section: Resultssupporting

confidence: 56%

Characterization and in planta validation of a CHI4 chitinase from Cajanus platycarpus (Benth.) Maesen for its efficacy against pod borer, Helicoverpa armigera (Hübner)

Rathinam

Marimuthu

Tyagi

et al. 2021

Pest Management Science

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BACKGROUND Pigeonpea, Cajanus cajan is one of the economically important legume food crops and a major source of dietary proteins. Management of pod borer, Helicoverpa armigera has been prominent among crop improvement programs. Lack of resistance sources in the cultivated germplasm and crossing incompatibility with pod borer‐resistant wild relatives have prompted biotechnological interventions. Identification and exploitation of genes from pigeonpea wild relatives in host plant resistance towards the pod borer assumes pertinence. Dynamic transcriptome analysis of the wild relative vis a vis cultivated pigeonpea identified a CHI4 chitinase as one of the putative insect resistance genes. RESULTS The study presents variations in important amino acids in CHI4 chitinases from C. cajan and its wild relative C. platycarpus. Comparative protein modeling and docking analysis of the two proteins demonstrated differences in substrate binding efficacy of the chitinase from C. platycarpus which resulted in a minimum binding energy of −8.7 kcal mol−1. Furthermore, we successfully evaluated the insecticidal activity of the chitinase from C. platycarpus against H. armigera challenge through heterologous expression in tobacco. Molecular characterization of transgenic plants confirmed that their efficacy against H. armigera was a result of the integration of CHI4 from C. platycarpus. CONCLUSION Docking analysis demonstrated effective substrate interaction as a possible reason for efficacy against pod borer in the chitinase from C. platycarpus. This was authenticated by successful overexpression and bioefficacy assessment against H. armigera in tobacco. The CHI4 gene from C. platycarpus can be useful in the mitigation of H. armegira in pigeonpea as well as in other crops. © 2021 Society of Chemical Industry

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“…Chitinases, also known as PR-3, are a group of proteins that protect plants against fungal diseases by cleaving the bonds between chitin molecules that are found in the fungal cell wall [50,51]. In the present study, compared to the uninfected control, a remarkable rise in PR-3, with a relative expression level of 5.76-fold at 20 dpi was reported (Figure 3).…”

Section: The Expression Levels Of Pr-related Genessupporting

confidence: 60%

Differences in Pathogenesis-Related Protein Expression and Polyphenolic Compound Accumulation Reveal Insights into Tomato–Pythium aphanidermatum Interaction

et al. 2023

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Plant diseases significantly reduce crop yields, threatening food security and agricultural sustainability. Fungi are the most destructive type of phytopathogen, and they are responsible for major yield losses in some of the most crucial crops grown across the world. In this study, a fungus isolate was detected from infected tomato plants and molecularly identified as Pythium aphanidermatum (GenBank accession number MW725032). This fungus caused damping-off disease and was shown to be pathogenic. Moreover, the expression of five pathogenesis-related genes, namely PR-1, PR-2, PR-3, PR-4, and PR-5, was quantitatively evaluated under the inoculation of tomato with P. aphanidermatum. The quantitative polymerase chain reaction (qPCR) showed that the expression levels of PR-1, PR-2, and PR-5 genes went up significantly at 5 days post-inoculation (dpi). The expression of the PR-1 gene also increased the variably, which reached its highest value at 20 dpi, with a reported relative expression level 6.34-fold higher than that of the control. At 15 dpi, PR-2 and PR-5 increased the most, while PR-1, PR-3, and PR-5 also increased noticeably at 20 dpi. On the contrary, PR-4 gene expression significantly decreased after inoculation, at all time intervals. Regarding PR-5 gene expression, the data showed a variable change in PR-5 gene expression at a different sample collection period. Still, it was highly expressed at 15 dpi and reached 3.99-fold, followed by 20 dpi, where the increasing percentage reached 3.70-fold, relative to the untreated control. The HPLC analysis indicated that the total concentration of all detected polyphenolic compounds was 3858 µg/g and 3202.2 µg/g in control and infected plant leaves, respectively. Moreover, the HPLC results concluded that Pythium infection decreased phenolic acids, such as chlorogenic and ellagic acids, which correlated with the infection–plant complex process. Based on the results, P. aphanidermatum could be a biotic stress pathogen that causes the expression of pathogen-related genes and stops the regulation of defensin phenolic compounds.

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A biophysical and structural study of two chitinases from Agave tequilana and their potential role as defense proteins

Cited by 9 publications

References 79 publications

Plant root associated chitinases: structures and functions

Plant root associated chitinases: structures and functions

Characterization and in planta validation of a CHI4 chitinase from Cajanus platycarpus (Benth.) Maesen for its efficacy against pod borer, Helicoverpa armigera (Hübner)

Differences in Pathogenesis-Related Protein Expression and Polyphenolic Compound Accumulation Reveal Insights into Tomato–Pythium aphanidermatum Interaction

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