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Differential expression of miRNAs and their target genes in senescing leaves and siliques: insights from deep sequencing of small RNAs and cleaved target RNAs.

ABSTRACT: MicroRNAs (miRNAs) are a class of small RNAs, which typically function by guiding cleavage of target mRNAs. They are known to play roles in a variety of plant processes including development, responses to environmental stresses and senescence. To identify senescence regulation of miRNAs in Arabidopsis thaliana, eight small RNA libraries were constructed and sequenced at four different stages of development and senescence from both leaves and siliques, resulting in more than 200 million genome-matched sequences. Parallel analysis of RNA ends libraries, which enable the large-scale examination of miRNA-guided cleavage products, were constructed and sequenced, resulting in over 750 million genome-matched sequences. These large datasets led to the identification a new senescence-inducible small RNA locus, as well as new regulation of known miRNAs and their target genes during senescence, many of which have established roles in nutrient responsiveness and cell structural integrity. In keeping with remobilization of nutrients thought to occur during senescence, many miRNAs and targets had opposite expression pattern changes between leaf and silique tissues during the progression of senescence. Taken together, these findings highlight the integral role that miRNAs may play in the remobilization of resources and alteration of cellular structure that is known to occur in senescence.

SUBMITTER: Thatcher SR

PROVIDER: S-EPMC4304344 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature

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Differential expression of miRNAs and their target genes in senescing leaves and siliques: insights from deep sequencing of small RNAs and cleaved target RNAs.

Thatcher Shawn R SR Burd Shaul S Wright Christopher C Lers Amnon A Green Pamela J PJ

Plant, cell & environment 20140813 1

MicroRNAs (miRNAs) are a class of small RNAs, which typically function by guiding cleavage of target mRNAs. They are known to play roles in a variety of plant processes including development, responses to environmental stresses and senescence. To identify senescence regulation of miRNAs in Arabidopsis thaliana, eight small RNA libraries were constructed and sequenced at four different stages of development and senescence from both leaves and siliques, resulting in more than 200 million genome-ma ...[more]

PMID: 24965556

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Project description:BACKGROUND:MiRNAs (microRNA) are 18-24?nt endogenous noncoding RNAs that regulate gene expression at the post-transcriptional level, including tissue-specific, developmental timing and evolutionary conservation gene expression. RESULTS:This study used high-throughput sequencing technology for the first time in Larix olgensis, predicted 78 miRNAs, including 12,229,003 reads sRNA, screened differentially expressed miRNAs. Predicting target genes was helpful for understanding the miRNA regulation function and obtained 333 corresponding target genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional annotation were analysed, mostly including nucleic acid binding, plant hormone signal transduction, pantothenate and CoA biosynthesis, and cellulose synthase. This study will lay the foundation for clarifying the complex miRNA-mediated regulatory network for growth and development. In view of this, spatio-temporal expression of miR396, miR950, miR164, miR166 and miR160 were analysed in Larix olgensis during the growth stages of not lignified, beginning of lignification, and completely lignified in different tissues (root, stem, and leaf) by quantitative real-time PCR (qRT-PCR). There were differences in the expression of miRNAs in roots, stems and leaves in the same growth period. At 60?days, miR160, miR166 and miR396-2 exhibited the highest expression in leaves. At 120?days, most miRNAs in roots and stems decreased significantly. At 180?days, miRNAs were abundantly expressed in roots and stems. Meanwhile, analysis of the expression of miRNAs in leaves revealed that miR396-2 was reduced as time went on, whereas other miRNAs increased initially and then decreased. On the other hand, in the stems, miR166-1 was increase, whereas other miRNAs, especially miR160, miR164, miR396 and miR950-1, first decreased and then increased. Similarly, in the roots, miR950-2 first decreased and then increased, whereas other miRNAs exhibited a trend of continuous increase. CONCLUSIONS:The present investigation included rapid isolation and identification of miRNAs in Larix olgensis through construction of a sRNA library using Solexa and predicted 78 novel miRNAs, which showed differential expression levels in different tissues and stages. These results provided a theoretical basis for further revealing the genetic regulation mechanism of miRNA in the growth and development of conifers and the verification of function in target genes.

Project description:BackgroundMost studies of photosynthesis in chlorenchymas under potassium (K) deficiency focus exclusively on leaves; however, little information is available on the physiological role of K on reproductive structures, which play a critical role in plant carbon gain. Brassica napus L., a natural organ-succession species, was used to compare the morphological, anatomical and photo-physiological differences between leaves and siliques exposed to K-deficiency.ResultsCompared to leaves, siliques displayed considerably lower CO2 assimilation rates (A) under K-deficient (-K) or sufficient conditions (+K), limited by decreased stomatal conductance (g s), apparent quantum yield (α) and carboxylation efficiency (CE), as well as the ratio of the maximum rate of electron transport (J max) and the maximum rate of ribulose 1,5-bisphosphate (RuBP) carboxylation (V cmax). The estimated J max, V cmax and α of siliques were considerably lower than the theoretical value calculated on the basis of a similar ratio between these parameters and chlorophyll concentration (i.e. J max/Chl, V cmax/Chl and α/Chl) to leaves, of which the gaps between estimated- and theoretical-J max was the largest. In addition, the average ratio of J max to V cmax was 16.1% lower than that of leaves, indicating that the weakened electron transport was insufficient to meet the requirements for carbon assimilation. Siliques contained larger but fewer stoma, tightly packed cross-section with larger cells and fewer intercellular air spaces, fewer and smaller chloroplasts and thin grana lamellae, which might be linked to the reduction in light capture and CO2 diffusion. K-deficiency significantly decreased leaf and silique A under the combination of down-regulated stomatal size and g s, chloroplast number, α, V cmax and J max, while the CO2 diffusion distance between chloroplast and cell wall (D chl-cw) was enhanced. Siliques were more sensitive than leaves to K-starvation, exhibiting smaller reductions in tissue K and parameters such as g s, V cmax, J max and D chl-cw.ConclusionSiliques had substantially smaller A than leaves, which was attributed to less efficient functioning of the photosynthetic apparatus, especially the integrated limitations of biochemical processes (J max and V cmax) and α; however, siliques were slightly less sensitive to K deficiency.

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Differential expression of miRNAs and their target genes in senescing leaves and siliques: insights from deep sequencing of small RNAs and cleaved target RNAs.

Publications

Differential expression of miRNAs and their target genes in senescing leaves and siliques: insights from deep sequencing of small RNAs and cleaved target RNAs.

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