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Clustering and evolutionary analysis of small RNAs identify regulatory siRNA clusters induced under drought stress in rice.

ABSTRACT: Drought tolerance is an important trait related to growth and yield in crop. Until now, drought related research has focused on coding genes. However, non-coding RNAs also respond significantly to environmental stimuli such as drought stress. Unfortunately, characterizing the role of siRNAs under drought stress is difficult since a large number of heterogenous siRNA species are expressed under drought stress and non-coding RNAs have very weak evolutionary conservation. Thus, to characterize the role of siRNAs, we need a well designed biological and bioinformatics strategy. In this paper, to characterize the function of siRNAs we developed and used a bioinformatics pipeline that includes a genomic-location based clustering technique and an evolutionary conservation tool.By comparing the wild type Nipponbare and two drought resistant rice varities, we found that 21 nt and 24 nt siRNAs are significantly expressed in the three rice plants but at different time points under a short-term (0, 1, and 6 hrs) drought treatment. siRNAs were up-regulated in the wild type at an early stage while the up-regulation was delayed in the two drought tolerant plants. Genes targeted by up-regulated siRNAs were related to oxidation reduction and proteolysis, which are well known to be associated with water deficit phenotypes. More interestingly, we found that siRNAs were located in intronic regions as clusters and were of high evolutionary conservation among monocot grass plants. In summary, we show that siRNAs are important respondents to drought stress and regulate genes related to the drought tolerance in water deficit conditions.

SUBMITTER: Jung I

PROVIDER: S-EPMC5260113 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature

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Clustering and evolutionary analysis of small RNAs identify regulatory siRNA clusters induced under drought stress in rice.

Jung Inuk I Ahn Hongryul H Shin Seon-Ju SJ Kim Jukon J Kwon Hawk-Bin HB Jung Woosuk W Kim Sun S

BMC systems biology 20161223 Suppl 4

<h4>Motivation</h4>Drought tolerance is an important trait related to growth and yield in crop. Until now, drought related research has focused on coding genes. However, non-coding RNAs also respond significantly to environmental stimuli such as drought stress. Unfortunately, characterizing the role of siRNAs under drought stress is difficult since a large number of heterogenous siRNA species are expressed under drought stress and non-coding RNAs have very weak evolutionary conservation. Thus, t ...[more]

PMID: 28155667

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Project description:BACKGROUND: Selection for grain yield under drought is an efficient criterion for improving the drought tolerance of rice. Recently, some drought-tolerant rice varieties have been developed using this selection criterion and successfully released for cultivation in drought-prone target environments. The process can be made more efficient and rapid through marker-assisted breeding, a well-known fast-track approach in crop improvement. QTLs have been identified for grain yield under drought with large effects against drought-susceptible varieties. Most of the identified QTLs show large QTL?×?environment or QTL?×?genetic background interactions. The development of mapping populations in the background of popular high-yielding varieties, screening across environments, including the target environments, and the identification of QTLs with a consistent effect across environments can be a suitable alternative marker-assisted breeding strategy. An IR74371-46-1-1?×?Sabitri backcross inbred line population was screened for reproductive-stage drought stress at the International Rice Research Institute, Philippines, and Regional Agricultural Research Station, Nepalgunj, Nepal, in the dry and wet seasons of 2011, respectively. A bulk segregant analysis approach was used to identify markers associated with high grain yield under drought. RESULTS: A QTL, qDTY12.1, significantly associated with grain yield under reproductive-stage drought stress was identified on chromosome 12 with a consistent effect in two environments: IRRI, Philippines, and RARS, Nepalgunj, Nepal. This QTL explained phenotypic variance of 23.8% and contributed an additive effect of 45.3% for grain yield under drought. The positive QTL allele for qDTY12.1 was contributed by tolerant parent IR74371-46-1-1. CONCLUSIONS: In this study, qDTY12.1 showed a consistent effect across environments for high grain yield under lowland reproductive-stage drought stress in the background of popular high-yielding but drought-susceptible recipient variety Sabitri. qDTY12.1 was also reported previously [Crop Sci 47:507-516, 2007] to increase grain yield under upland reproductive-stage drought stress situations. qDTY12.1 is the only QTL reported so far in rice to have shown a large effect against multiple recipient genetic backgrounds as well as under highly diverse upland and lowland rice ecosystems. qDTY12.1 can be successfully introgressed to improve grain yield under drought of popular high-yielding but drought-susceptible lowland as well as upland adapted varieties following marker-assisted breeding.

Project description:BackgroundPlant transcriptome profiling has provided a tool for understanding the mechanisms by which plants respond to stress conditions. Analysis of genome-wide transcriptome will provides a useful dataset of drought responsive noncoding RNAs and their candidate target genes that may be involved in drought stress responses.ResultsHere RNA-seq analyses of leaves from drought stressed rice plants was performed, producing differential expression profiles of noncoding RNAs. We found that the transcript levels of 66 miRNAs changed significantly in response to drought conditions and that they were negatively correlated with putative target genes during the treatments. The negative correlations were further validated by qRT-PCR using total RNAs from both drought-treated leaves and various tissues at different developmental stages. The drought responsive miRNA/target pairs were confirmed by the presence of decay intermediates generated by miRNA-guided cleavages in Parallel Analysis of RNA Ends (PARE) libraries. We observed that the precursor miR171f produced two different mature miRNAs, miR171f-5p and miR171f-3p with 4 candidate target genes, the former of which was responsive to drought conditions. We found that the expression levels of the miR171f precursor negatively correlated with those of one candidate target gene, but not with the others, suggesting that miR171f-5p was drought-responsive, with Os03g0828701-00 being a likely target. Pre-miRNA expression profiling indicated that miR171f is involved in the progression of rice root development and growth, as well as the response to drought stress. Ninety-eight lncRNAs were also identified, together with their corresponding antisense transcripts, some of which were responsive to drought conditions.ConclusionsWe identified rice noncoding RNAs (66 miRNAs and 98 lncRNAs), whose expression was highly regulated by drought stress conditions, and whose transcript levels negatively correlated with putative target genes.

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Clustering and evolutionary analysis of small RNAs identify regulatory siRNA clusters induced under drought stress in rice.

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Clustering and evolutionary analysis of small RNAs identify regulatory siRNA clusters induced under drought stress in rice.

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