Identification of CR4-regulated genes

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Date
2000
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Zhang, Yuan
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Becraft, Philip W.
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The crinkly4 (cr4) gene encodes a receptor-like protein kinase that is required for the normal epidermal differentiation in the maize leaf and endosperm. To identify genes controlled by CR4, suppression subtractive hybridization (SSH) was used to identify genes that were differentially expressed between wild type and cr4 mutant. Eighty differentially expressed clones were isolated, 50 of which are up-regulated (+) and 30 are down-regulated (-). Four plus and four minus clones were picked for Northern Blot analysis. Three of these correspond to mRNAs that are up-regulated and three correspond to mRNAs down-regulated in wild type. To facilitate the study of CR4, transgenic maize plants containing the cr4 gene under the control of a glucocorticoid-inducible promoter were generated. The system uses a commercially available glucocorticoid analog, dexamethasone (DEX), as an inducer chemical. A maximal induction level of about 100 fold of cr4 MRNA was obtained at concentrations of 10 [Mu]M and 30 [Mu]M of DEX. Treatment of mature leaf segments from transgenic maize with 30 [Mu]M DEX resulted in expression of cr4 1 h after induction with the peak at 2 h. No induction of cr4 was observed in mature leaves of untransformed controls treated with DEX. This transgenic plant will be used to study the CR4-regulated genes.
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