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The miRNAome of durum wheat: isolation and characterisation of conserved and novel microRNAs and their target genes.

ABSTRACT: The allotetraploid durum wheat [Triticum turgidum subsp. durum (Desf.) Husn.] is a highly economically important species especially in the Mediterranean basin. However, its genomics, transcriptomics and in particular microRNAome are still largely unknown.In the present work, two small RNA libraries from durum wheat Ciccio and Svevo cultivars were generated from different tissues at the late milk (Z77) developmental stage. A total of 167 conserved and 98 potential novel miRNAs were identified in the two libraries and interestingly, three novel miRNAs were found to be derived from ribosomal RNA. Putative target genes were predicted for conserved and novel miRNAs, the majority of which interact with nucleic acids, according to GO terms relative to molecular function. Quantitative qPCR analysis showed that several miRNAs identified were differentially expressed in the mature (Z77) developmental stage compared to young (Z14) tissues. Moreover, target gene expression analysis suggested that in roots, the putative genes encoding for the SQUAMOSA SPL2 and TGA1 proteins are regulated by ttu-miR156n, while MYB3 transcription factor by ttu-miR319f. Additionally, the Photosystem II P680 chlorophyll A apoprotein gene showed an expression level negatively correlated to that of ttu-novel-48 in leaves.Our results suggest that, in durum wheat, these genes may play important roles in root/leaf development and are subjected to miRNA regulation. The prediction of novel miRNAs putatively derived from ribosomal RNA opens new perspectives on the study of plant miRNAs.

SUBMITTER: De Paola D

PROVIDER: S-EPMC4957382 | biostudies-literature | 2016 Jul

REPOSITORIES: biostudies-literature

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The miRNAome of durum wheat: isolation and characterisation of conserved and novel microRNAs and their target genes.

De Paola Domenico D Zuluaga Diana L DL Sonnante Gabriella G

BMC genomics 20160722

<h4>Background</h4>The allotetraploid durum wheat [Triticum turgidum subsp. durum (Desf.) Husn.] is a highly economically important species especially in the Mediterranean basin. However, its genomics, transcriptomics and in particular microRNAome are still largely unknown.<h4>Results</h4>In the present work, two small RNA libraries from durum wheat Ciccio and Svevo cultivars were generated from different tissues at the late milk (Z77) developmental stage. A total of 167 conserved and 98 potenti ...[more]

PMID: 27448633

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Project description:BACKGROUND: The synaptonemal complex (SC) is a proteinaceous tripartite structure used to hold homologous chromosomes together during the early stages of meiosis. The yeast ZIP1 and its homologues in other species have previously been characterised as the transverse filament protein of the synaptonemal complex. Proper installation of ZYP1 along chromosomes has been shown to be dependent on the axial element-associated protein, ASY1 in Arabidopsis. RESULTS: Here we report the isolation of the wheat (Triticum aestivum) ZYP1 (TaZYP1) and its expression profile (during and post-meiosis) in wild-type, the ph1b deletion mutant as well as in Taasy1 RNAi knock-down mutants. TaZYP1 has a putative DNA-binding S/TPXX motif in its C-terminal region and we provide evidence that TaZYP1 interacts non-preferentially with both single- and double-stranded DNA in vitro. 3-dimensional dual immunofluorescence localisation assays conducted with an antibody raised against TaZYP1 show that TaZYP1 interacts with chromatin during meiosis but does not co-localise to regions of chromatin where TaASY1 is present. The TaZYP1 signal lengthens into regions of chromatin where TaASY1 has been removed in wild-type but this appears delayed in the ph1b mutant. The localisation profile of TaZYP1 in four Taasy1 knock-down mutants is similar to wild-type but TaZYP1 signal intensity appears weaker and more diffused. CONCLUSIONS: In contrast to previous studies performed on plant species where ZYP1 signal is sandwiched by ASY1 signal located on both axial elements of the SC, data from the 3-dimensional dual immunofluorescence localisation assays conducted in this study show that TaZYP1 signal only lengthens into regions of chromatin after TaASY1 signal is being unloaded. However, the observation that TaZYP1 loading appears delayed in both the ph1b and Taasy1 mutants suggests that TaASY1 may still be essential for TaZYP1 to play a role in SC formation during meiosis. These data further suggest that the temporal installation of ZYP1 onto pairing homologous chromosomes in wheat is different to that of other plant species and highlights the need to study this synaptonemal complex protein on a species to species basis.

Dataset Information

The miRNAome of durum wheat: isolation and characterisation of conserved and novel microRNAs and their target genes.

Publications

The miRNAome of durum wheat: isolation and characterisation of conserved and novel microRNAs and their target genes.

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