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Overexpression of OsERF106MZ promotes root growth in rice seedlings by relieving the ABA-mediated inhibition of root growth

ABSTRACT: Background: Roots are essential for plant growth and serve a variety of functions, such as anchoring the plant to the ground and absorbing water and nutrients. OsERF106MZ is a salinity-induced gene that is expressed in germinating seeds and the roots of rice seedlings. However, the roles of OsERF106MZ in root growth remain poorly understood. Results: Histochemical staining of GUS (β-glucuronidase) activity in transgenic rice seedlings harboring OsERF106MZp::GUS indicated that OsERF106MZ is mainly expressed in the exodermis, sclerenchyma layer, and vascular system of roots. Overexpression of OsERF106MZ in rice seedlings leads to an increase in the length of primary roots (PRs). The expression of the ABA (abscisic acid) biosynthetic gene, OsAO3, is downregulated in OsERF106MZ-overexpressing roots under normal conditions, while the expression of OsNPC3, an AtNPC4 homolog involved in ABA sensitivity, is reduced in OsERF106MZ-overexpressin roots under both normal and NaCl-treated conditions. Under normal conditions, OsERF106MZ-overexpressing roots have a significantly low level of ABA; moreover, exogenous application of 1.0 µM ABA can suppress the OsERF106MZ-mediated promotion of root growth. Meanwhile, OsERF106MZ-overexpressing roots display less sensitivity to the ABA-mediated inhibition of root growth, when they are treated with ABA at 5.0 µM under normal conditions or exposed to NaCl-treated conditions. Chromatin immunoprecipitation (ChIP)-qPCR and luciferase (LUC) reporter assays showed that OsERF106MZ can bind directly to the sequence containing the GCC-box in the promoter region of OsAO3 gene and repress its expression. Conclusion: OsERF106MZ may play a role in maintaining root growth for resource uptake when rice seeds are sprouted under salinity stress, which is arrived at by alleviating the ABA-mediated inhibition of root growth.

ORGANISM(S): Oryza sativa

PROVIDER: GSE213466 | GEO | 2023/03/22

REPOSITORIES: GEO

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Project description:For identification of genes up-regulated in abiotic stress (drought, high salinity, low temperature and ABA) treated rice, total RNA (100 ?g) was prepared from root tissues of 14-d-old rice seedlings (Oryza sativa cv Nakdong) grown under normal growth conditions. For the high salinity and ABA treatments, the 14-d-old seedlings were transferred to a nutrient solution containing 400 mM NaCl or 100 ?M ABA for 2 h in the greenhouse under continuous light of approximately 1000 ?mol m-2 s -1. For drought treatment, 14-d-old seedlings were air-dried for 2 h also under continuous light of approximately 1000 ?mol m-2 s -1. For low temperature treatments, 14-d-old seedlings were exposed at 4°C in a cold chamber for 6 h under continuous light of 150 ?mol m-2 s -1. Expression profiling was conducted using a Rice 3’-Tiling Microarray. Information on the microarray can be found at http://www.ggbio.com (GreenGene Biotech). The Rice 3’-Tiling Microarray was designed from 27,448 genes deposited at IRGSP, RAP1 database (http://rapdb.lab.nig.ac.jp). Among these, 20,507 genes were from representative RAP1 sequences with cDNA/EST supports and 6,941 genes were predicted without cDNA/EST supports. Ten 60-nt long probes were designed from each gene starting 60 bp ahead the end of stop codon with 10 bp shifts in position so that 10 probes covered 150 bp in the 3' region of the gene. In total, 270,000 probes were designed (average size, 60-nt) to have Tm values of 75 to 85 °C. The microarray was manufactured by NimbleGen Inc. (http://www.nimblegen.com/). Random GC probes (38,000) were used to monitor the hybridization efficiency and fiducial markers at the four corners (225) were included to assist with overlaying the grid on the image. The microarray was used to profile gene expression in abiotic stress treated rice. Cy3-labeled target cDNA fragments were synthesized using a Cy3-9mer primer. For normalization, data were processed with cubic alpine normalization using quartiles to adjust signal variation between chips and with Rubust Multi-Chip Analysis using a median polish algorithm implemented in NimbleScan (Workman et al., 2002; Irizarry et al., 2003). To assess the reproducibility of the microarray analysis, we repeated the experiment three times with independently prepared total RNAs.

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Overexpression of OsERF106MZ promotes root growth in rice seedlings by relieving the ABA-mediated inhibition of root growth

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