title: Excess fertilizer responsive miRNAs revealed in Linum usitatissimum L.
abstract: Effective fertilizer application is necessary to increase crop yields and reduce risk of plant overdosing. It is known that expression level of microRNAs (miRNAs) alters in plants under different nutrient concentrations in soil. The aim of our study was to identify and characterize miRNAs with expression alterations under excessive fertilizer in agriculturally important crop - flax (Linum usitatissimum L.). We have sequenced small RNAs in flax grown under normal and excessive fertilizer using Illumina GAIIx. Over 14 million raw reads was obtained for two small RNA libraries. 84 conserved miRNAs from 20 families were identified. Differential expression was revealed for several flax miRNAs under excessive fertilizer according to high-throughput sequencing data. For 6 miRNA families (miR395, miR169, miR408, miR399, miR398 and miR168) expression level alterations were evaluated on the extended sampling using qPCR. Statistically significant up-regulation was revealed for miR395 under excessive fertilizer. It is known that target genes of miR395 are involved in sulfate uptake and assimilation. However, according to our data alterations of the expression level of miR395 could be associated not only with excess sulfur application, but also with redundancy of other macro- and micronutrients. Furthermore expression level was evaluated for miRNAs and their predicted targets. The negative correlation between miR399 expression and expression of its predicted target ubiquitin-conjugating enzyme E2 gene was shown in flax for the first time. So we suggested miR399 involvement in phosphate regulation in L. usitatissimum. Revealed in our study expression alterations contribute to miRNA role in flax response to excessive fertilizer.
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia. Electronic address: mnv-4529264@yandex.ru.
authors.forename: Nataliya V
authors.initials: NV
authors.lastname: Melnikova
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Alexey A
authors.initials: AA
authors.lastname: Dmitriev
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia; Research Institute of Physicochemical Medicine, Malaya Pirogovskaya Str. 1a, 119992 Moscow, Russia.
authors.forename: Maxim S
authors.initials: MS
authors.lastname: Belenikin
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia; Department of Higher Plants, Lomonosov Moscow State University, Lenin Hills 1, 119234 Moscow, Russia.
authors.forename: Anna S
authors.initials: AS
authors.lastname: Speranskaya
authors.affiliation: Department of Higher Plants, Lomonosov Moscow State University, Lenin Hills 1, 119234 Moscow, Russia.
authors.forename: Anastasia A
authors.initials: AA
authors.lastname: Krinitsina
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Olga A
authors.initials: OA
authors.lastname: Rachinskaia
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Valentina A
authors.initials: VA
authors.lastname: Lakunina
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: George S
authors.initials: GS
authors.lastname: Krasnov
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Anastasiya V
authors.initials: AV
authors.lastname: Snezhkina
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Asiya F
authors.initials: AF
authors.lastname: Sadritdinova
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Leonid A
authors.initials: LA
authors.lastname: Uroshlev
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Nadezda V
authors.initials: NV
authors.lastname: Koroban
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Tatiana E
authors.initials: TE
authors.lastname: Samatadze
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Alexandra V
authors.initials: AV
authors.lastname: Amosova
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Zelenin
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Olga V
authors.initials: OV
authors.lastname: Muravenko
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Nadezhda L
authors.initials: NL
authors.lastname: Bolsheva
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilov Str. 32, 119991 Moscow, Russia.
authors.forename: Anna V
authors.initials: AV
authors.lastname: Kudryavtseva
mesh_terms: D005308:Fertilizers; D019597:Flax / Q000235:genetics*; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D059014:High-Throughput Nucleotide Sequencing; D035683:MicroRNAs / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics
chemical_list: D005308:Fertilizers; D035683:MicroRNAs; D010940:Plant Proteins; D018749:RNA, Plant; D044763:Ubiquitin-Conjugating Enzymes
keywords: Excess fertilizer; Flax; High-throughput sequencing; MicroRNA; miR395