CRISPR-Cas9-Mediated Reduced Expression of Potato Apoplastic Invertase Inhibitor Gene and Analysis of Transformation Efficiency Parameters

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dc.contributor.authorDas Dangol, Sarbesh
dc.contributor.authorCaliskan, Mehmet Emin
dc.contributor.authorBakhsh, Allah
dc.date.accessioned2024-11-07T13:32:55Z
dc.date.available2024-11-07T13:32:55Z
dc.date.issued2024
dc.departmentNiğde Ömer Halisdemir Üniversitesi
dc.description.abstractPathogen infections that affect potato yield cause severe economic losses every year. Many studies point to the role of apoplastic (cell wall) invertase (CWIN) enzyme in plant defence mechanisms. Apoplastic invertase inhibitor (INVINH1) post-translationally regulates the CWIN enzyme activity. Nevertheless, the role of INVINH1 needs to be elucidated for several effects in plant transformation parameters and its gene expression which we sought to explore using CRISPR/Cas9 technology. In this study, we sequenced the first exon of INVINH1 gene in cv. Desiree and Solanum chacoense M6. We identified in the first exon two alleles for StINVINH1 gene in cv. Desiree and one allele for ScINVINH1 gene in S. chacoense M6. We designed two single-guided RNAs (sgRNAs) to target INVINH1 gene from diploid S. chacoense M6 and tetraploid S. tuberosum cv. Desiree using CRISPR/Cas9-based technology. In our earlier study, we have already optimised the transformation protocol for M6 and cv. Desiree using Agrobacterium strains, based on which Agrobacterium strain AGL1 was chosen for the CRISPR/Cas9 experiment. Our experimentation showed that heat stress at 37 degrees C could increase the mutagenesis capability and CRISPR/Cas9 targeting affected the plant transformation parameters. It was found from the knockout experiment that the indels were present in the calli and the candidate regenerated plants showed reduced gene expression level conducted via RT-qPCR. Our study demonstrated that INVINH1 targeting affected the calli induction and regeneration rates, was effective under heat stress and reduced its gene expression level. More studies are required to comprehend the function of the INVINH1 enzyme in potato stress response and defence mechanism.
dc.description.sponsorshipTUBITAK; Graduate Scholarship Programme
dc.description.sponsorshipWe would like to thank TUBITAK for awarding the first author with the Graduate Scholarship Programme for International Students-2215 (PhD). We are thankful to Dr. Abdellah Barakate who created the CRISPR vector and designed and supervised the protocols for gene editing plant transformation.
dc.identifier.doi10.1007/s11540-024-09818-w
dc.identifier.issn0014-3065
dc.identifier.issn1871-4528
dc.identifier.scopus2-s2.0-85206619329
dc.identifier.scopusqualityQ1
dc.identifier.urihttps://doi.org/10.1007/s11540-024-09818-w
dc.identifier.urihttps://hdl.handle.net/11480/15668
dc.identifier.wosWOS:001338573000001
dc.identifier.wosqualityN/A
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.language.isoen
dc.publisherSpringer
dc.relation.ispartofPotato Research
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.snmzKA_20241106
dc.subjectApoplastic invertase inhibitor
dc.subjectCRISPR/Cas9
dc.subjectPotato
dc.subjectSolanum chacoense M6
dc.subjectSolanum tuberosum cv. Desiree
dc.titleCRISPR-Cas9-Mediated Reduced Expression of Potato Apoplastic Invertase Inhibitor Gene and Analysis of Transformation Efficiency Parameters
dc.typeArticle

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