Identification and characterization of tissue specific microRNAs in chickpea

Chickpea (Cicer arietinum) is the most important pulse crop in the Indian subcontinent. MicroRNAs (miRNAs) are essential components of complex gene regulatory networks that are involved in plant development. Several conserved and plant-specific miRNAs have been characterized to date, predominantly from the plant species whose genomes are sequenced. In the current study, miRNAs from leaf, root and flower tissues were identified from chickpea. Computational analysis of small RNA sequences obtained using Illumina Genome analyser led to identification of total of 368 miRNAs. 24 families consisting of 97 conserved miRNAs and 73 novel miRNAs were identified in root and leaf tissues after extensive sequence analysis. Likewise, 25 families consisting of 112 conserved miRNAs and 86 novel miRNAs were identified in desi and kabuli flower tissues. Poly(A)-based qRT-PCR (Quantitative real-time PCR) expression analysis of several miRNAs was done for leaf, root and flower tissues of chickpea. Nine conserved and 3 novel miRNA were found to be differentially expressed between leaf and root tissues. Nine conserved and 4 novel miRNAs were found to be differentially expressed between desi and kabuli flower tissues. Both miR159 and miR172, were up-regulated in desi flower and target six APETALA genes which are repressors of flowering. In root tissue, mir390 and mir169 were found to be up-regulated and down regulated in leaf. miR156 and 159 were found to be up-regulated in leaf tissue and down-regulated in root. The target genes of all the identified miRNAs were predicted by psRNATarget web server using chickpea transcriptome database. The predicted target genes (approximately 358 different transcripts) were extensively involved in different biological processes involving a large number of gene families. Some of these genes encoded Transcription Factors, DNA replication proteins and those that are involved in cellular metabolism in addition to a variety of stress response-associated proteins. This study has identified several conserved and novel miRNAs along with their expression profiling and has predicted several target genes and thereby lays the foundation for studies on gene regulation in response to chickpea development. चने एम. आई. आर. एन. एज़ पहचान तथा उनका सार चना भारतीय दलहन फ़सल है। पादप से जुड़े जीन आर. एन. ए. (एम.आई.आर.एन.एज़) घटक है। अब तक कई एवं पादप एम. आई. आर. एन. एज़ का जा चकुा है, उन पादप से, िजनके का गया । चने , जड़ एवं से एम. आई. आर. एन. एज़ गयी है । जीनोम अनालाइसर कुल 368 एम. आई. आर. एन. ए. पहचान हो । सघन के जड़ एवं के 97 एम. आई. आर. एन. एज़ एवं 73 नवीन एम. आई. आर. एन. एज़ वाले 24 पहचान गयी । इसी से देशी एवं चने के 112 एम. आई. आर. एन. एज़ एवं 86 नवीन एम. आई. आर. एन. एज़ वाले 25 पहचान गयी । चने के , जड़ एवं हेतु कई एम. आई. आर. एन. एज़ का (ए) – पी॰सी॰आर॰ ( टाइम पी॰सी॰आर॰) गया । एवं जड़ के बीच 9 एवं 3 नवीन एम. आई. आर. एन. ए. देखी गयी । एम. आई. आर. 159 एवं एम. आई. आर. 172, देशी एवं छह , जो का दमन करते , अपहुआ । जड़ ,एम. आई. आर. 390 एवं एम. आई. आर. 169 का अपहुआ और इनका डाउनहुआ । एम. आई. आर. 156 एवं 159 का अपएवं जड़ डाउनपाया गया । चना डटेाबेस का उपयोग कर पी॰एस॰आर॰एन॰ए॰ वेब पहचान गए सभी एम. आई. आर. एन. ए. का गया । (लगभग 358 ) संबंधी से पाये गए, बहुत बड़ी जीन का समावेश था । से कुछ , डी॰एन॰ए॰ तथा उपपाचयन से एवं इसके से को करता है । इस कई एवं एम. आई. आर. एन. एज़ पहचान गयी है तथा गयी है तथा कई का गया है, और इस से, इस से चने के जीन संबंधी हेतु आधार तयैार है ।