+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Characterization and functional analysis of miR166f in drought stress tolerance in mulberry (Morus multicaulis)

Characterization and functional analysis of miR166f in drought stress tolerance in mulberry (Morus multicaulis)

Molecular Breeding 38(11): 132

MicroRNAs (miRNAs) regulate gene expression accurately and effectively at the posttranscriptional level by repressing translation or directly degrading target mRNAs. Over 40 plant miRNA family genes have been associated with abiotic stress; however, few reports have identified miRNAs and their molecular functions under abiotic stress in woody plants. Mulberry trees are ecologically and economically important perennial woody plants, but few studies have examined the stress physiology, biochemistry, and molecular biology of mulberry miRNAs. Previously, we identified miR166f in Morus alba and found that its target genes encode homeobox-leucine zipper (HD-Zip) transcription factors and histone arginine demethylase, which are involved in responses to heat, cold, drought, and salt stresses. In this study, we further characterized miR166f from mulberry, and found that it targets HD-Zip induced by drought stress. We isolated, cloned, and identified pre-miR166f, the precursor of miR166f, from the leaves of mulberry (Morus multicaulis); pre-miR166f was 91bp in length with a minimum folding free energy of -51.70kcal/mol. Bioinformatics analysis showed that the 3000-bp 5 ' upstream region of miR166f contained not only TATA and CAAT boxes that functioned in transcription initiation, but also had many cis-acting elements for stress responses, such as MBS, HSE, ARE, TC-rich repeats, and several hormones response elements, such as AuxRR-core, CGTCA-motif, and P-box. This analysis suggested that miR166f might participate in abiotic and biotic stress responses by regulating the expression of stress-related genes in mulberry. We constructed a miR166f binary overexpression vector, pCAMBIA-35S-GUS-miR166f, and established a transient transformation system in mulberry. Histochemical beta-glucuronidase (GUS) staining of miR166f-overexpression in transient transgenic mulberry leaves showed the best effect and highest GUS gene expression compared to the wild-type control plants at day four posttransformation using an optimized concentration of transformation liquid (OD600=0.7). The target genes of miR166ftwo HD-Zips and one histone arginine demethylase gene (JMJD6)showed markedly lower expression levels in miR166f-overexpression transient transgenic mulberry leaves. Further investigation showed that transient transgenic mulberry trees had higher relative water content, free proline content, soluble protein content, and superoxide dismutase and peroxidase activities, and lower malondialdehyde content compared to the wild-type control plants, which suggested that overexpression of miR166f in mulberry could enhance tolerance to drought stresses in transient transgenic mulberry. The results suggested that miR166f might function as a positive regulator of drought stress tolerance in mulberry.

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 066378714

Download citation: RISBibTeXText

DOI: 10.1007/s11032-018-0886-y

Related references

Characterization of NPR1 and NPR4 genes from mulberry (Morus multicaulis) and their roles in development and stress resistance. Physiologia Plantarum 167(3): 302-316, 2019

Mulberry (Morus alba) MmSK gene enhances tolerance to drought stress in transgenic mulberry. Plant Physiology and Biochemistry 132: 603-611, 2018

Expression and functional analysis of a Pr-1 Gene, MuPr1 , involved in disease resistance response in mulberry ( Morus multicaulis ). Journal of Plant Interactions 14(1): 376-385, 2019

A Novel LncRNA, MuLnc1 , Associated With Environmental Stress in Mulberry ( Morus multicaulis ). Frontiers in Plant Science 9: 669, 2018

De novo transcriptome analysis of mulberry (Morus L.) under drought stress using RNA-seq technology. Bioorganicheskaia Khimiia 40(4): 458-467, 2014

Molecular cloning and sequence analysis of the kaurene oxidase (MmKO) gene in mulberry (Morus multicaulis) and patterns of MmKO gene expression under abiotic stress conditions. Journal of Horticultural Science and Biotechnology 91(5): 456-465, 2016

Genotypic variation in tolerance to drought stress is highly coordinated with hydraulic conductivity-photosynthesis interplay and aquaporin expression in field-grown mulberry (Morus spp.). Tree Physiology 37(7): 926-937, 2017

Characterization and expression patterns of mannose-binding lectin ( MMBL ) gene in mulberry ( Morus multicaulis ) and its prokaryotic expression in E. coli. South African Journal of Botany 113: 1-10, 2017

Mildew Resistance Locus O Gene Cloning, Characterization, and Expression Pattern in Mulberry (Morus multicaulis) and Its Prokaryotic Expression in E. coli. Russian Journal of Bioorganic Chemistry 44(1): 104-111, 2018

In vitro screening of mulberry genotypes (Morus sp.) for drought tolerance. Indian Journal of Sericulture 36(1): 60-62, 1997

Triazole induced stomatal closure and reduced transpiration in mulberry morus multicaulis perr. Geobios 19(1): 6-9, 1992

Drought Tolerance in Mulberry (Morus Spp.): a Physiological Approach WITH INSIGHTS INTO GROWTH DYNAMICS AND LEAF YIELD PRODUCTION. Experimental Agriculture 46(4): 471-488, 2010

The Latex Protein MLX56 from Mulberry ( Morus multicaulis ) Protects Plants against Insect Pests and Pathogens. Frontiers in Plant Science 8: 1475, 2017

Effects of drought stress on physiological and biochemical characteristics of mulberry (Morus alba) plants. Acta Sericologica Sinica 30(2): 117-122, 2004

Molecular cloning and induced expression of the plasma membrane intrinsic protein gene and promoter from mulberry (Morus multicaulis). Canadian Journal of Plant Science 98(6): 1245-1253, 2018