e-journal

Combined small RNA and degradome sequencing reveals novel miRNAs and their targets in response to low nitrate availability in maize

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Background and Aims:
MicroRNAs (miRNAs) play an important role in the responses and adaptation of plants to many stresses including low nitrogen (LN). Characterizing relevant miRNAs will improve our understanding of nitrogen
(N) use efficiency andLN tolerance and thus contribute to sustainable maize production. The objective of this
study was to identify novel and known miRNAs and their targets involved in the response and adaptation of maize (Zea mays) to LN stress.
Methods:
MiRNAs and their targets were identified by combined analysis of deep sequencing of small RNA and degradome libraries. The identity of target genes was confirmed by gene-specific RNA ligase-mediated rapid amplification of 5′ cDNA ends (RLM-RACE) and by quantitative expression analysis.
Key Results:
Over 150 million raw reads of small RNA and degradome sequence datawere generated. A total of 46 unique mature miRNA sequences belonging to 23 maize miRNA families were sequenced. Eighty-five potentially newmiRNAswere identified, with corresponding miRNA* also identified for 65 of them. Twenty-five new miRNAs
showed.2-fold relative change in response to LN. In addition to known miR169 species, two novel putative miR169 species were identified. Deep sequencing of miRNAs and the degradome, and RLM-RACE and quantitative polymerase chain reaction (PCR) analyses of their targets showed that miRC10- and miRC68-mediated target cleavage may play a major role among miR169 families in the adaptation to LN by maize seedlings.
Conclusions:
Small RNA and degradome sequencing combined with quantitative reverse transcription–PCR and RLM-RACE verification enabled the efficient identification of miRNAs and their target genes. The generated data
sets and the two novel miR169 species that were identified will contribute to our understanding of the physiological basis of adaptation to LN stress in maize plants.

Key words: Maize, Zea mays, low-nitrate stress, adaptation, miRNA, degradome, combined analysis.

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Informasi Detail

Judul Seri

Annals of Botany

No. Panggil

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Penerbit

Oxford : Oxford University Press on behalf of the Annals of Botany Company., 2013

Deskripsi Fisik

Annals of Botany 112: 633–642, 2013

Bahasa

English

ISBN/ISSN

10958290

Klasifikasi

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Tipe Isi

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Tipe Media

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Tipe Pembawa

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Edisi

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Subjek

PERTANIAN.

Info Detail Spesifik

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Pernyataan Tanggungjawab

agus

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