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• W567897640 abstract "The WIP-zinc finger genes originated from the early evolutionary establishment of the Viridiplantae clade from bryophytes and pteridophytes. From the ancestral bryophytes the first terrestrial plants two main clades have evolveld, based on our cladistic analysis of the C-terminal part of the WIPs. From the in silico analysis, CmWIP1 encodes are largely bi-partite protein in nature with the N-terminal acquired for protein-protein interactions whilst the C-terminal part is possibly mainly for DNA binding and some unknown processes involved in chromatin modulation/regulation (the presence of POST-SET domains). The common theme and character demonstrated by the AtWIP-Zinc finger proteins are highlighted by their discoveries in regulatory roles of organ growth and development. However, the molecular protein interacting partners and mechanisms by which the WIP functions are orchestrated still remain unknown. Here we show molecular protein interactions and sub-cellular localization patterns of CmWIP1 and its interactors to elucidate its regulatory role in the sex determination Cucumis melo flowers. Based on the protein-protein interactions yeast two hybrid screen of the melon cDNA library generated, we report that CmWIP1 protein physically interacts very strongly with CmbZIP and CmLHP1. Also interactions were observed with the CKII complex in C. melo which is made up of CmCKIIα, CmCKIIβ1 and CmCKIIβ2 subunits. CmWIP1 also interacts with CmTHF1 and CmPTR. Reverse genetic approaches were utilized in attempts to functionally validate the key interactors in planta. The transition from male to female flowers in gynoecious lines results from epigenetic repression of the expression of a zinc finger type transcription factor, CmWIP1 located at the g loci in Cucumis melo. When expressed, CmWIP1 is responsible for carpel abortion resulting in single sexed male flowers from an initial bisexual origin. Over-expression of CmWIP1 (under 35SCaMV promoter) in Arabidopsis thaliana resulted in plants with overall growth retardation of above- and under-ground plant parts, serrated true leaves, floral and sillique growth abnormalities as well as poor seed yield. 35S:CmWIP1 insertion lines developed were continuously monitored and phenotyped over T1 to T4 generations for stability of the leaf serration phenotype. Interestingly, two other over-expression lines carrying AtWIP1 and AtWIP2 caused leaf serrations similar to that of CmWIP1 overexpression all under 35SCaMV promoters. Two stable lines homozygote for 35S:CmWIP1 insertion were selected at the T4 generation and mutagenized with 0.3 % EMS. A thousand bulks of M2 family lines (5 plants per bulk) were screened for revertant individuals with unserrated leaves and restored growth rates. Vegetative stage screens revealed both putative revertants and 7 % albinos. Subsequent screens for revertants were carried out at the reproductive stages and also for primary root length assays were carried out to validate putative revertants. Putative revertants families were further validated as candidates for suppressor mapping via Sanger sequencing of the 35S::CmWIP1 insert by screening of the sequence for mutations which could be the causative mutation." @default.
• W567897640 created "2016-06-24" @default.
• W567897640 creator A5006349885 @default.
• W567897640 date "2013-05-15" @default.
• W567897640 modified "2023-09-23" @default.
• W567897640 title "Functional charaterization of CmWIP1 gene in the sex determination pathway of Cucumis melo using Arabidopsis thaliana as a model system" @default.
• W567897640 hasPublicationYear "2013" @default.
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