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Citation:

Expression Analysis of 12 miRNA Families Specific to Conifers during Somatic Embryogenesis of Larch

  • Received Date: 2012-03-13
  • In order to identify miRNA and investigate it’s expression profile during somatic embryogenesis (SE) of larch, a small RNA library was constructed and sequenced by Solexa technique, and expression analysis of miRNAs and four precursors during eight developmental stages of SE was evaluated by qRT-PCR. The results demonstrated that a total of 87 miRNAs was identified, of which 29 miRNAs from 12 families were specific to conifers, including miR946, miR947, miR950, miR951, miR1311, miR1312, miR1313, miR1314, miR1316, miR3701, miR3702 and miR3704, which could be used for further analysis and next experiment. Of these, 25 miRNAs were 22nt in length, while 4 miRNAs were 21nt. The abundance of miRNAs varied largely in the small RNA library, such as miR1311 was detected by 18 copies, while miR950 with 33 009 copies. 34 potential target genes were predicted for 12 miRNA families, which were linked to many respects of regulation, including plant growth and development, disease resistance, AGO protein feedback regulation. qRT-PCR analysis showed that, for 72% miRNAs, the minor expression peak was at late single embryo or early cotyledonary embryo, the lowest level was at middle cotyledonary embryo, while the major peak was at late cotyledonary embryo, which was consistent with the physiology events of cotyledon formation, embryo pre-mature, and embryo dormancy. However, the expression model of miRNA precursor was differed from that of miRNA, and precursor expression with much smaller extent than that of miRNA. The discovery of miRNAs specific to conifers and their targets using embryogenic tissue could provide references for the study on the growth and development of gymnosperms.
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Expression Analysis of 12 miRNA Families Specific to Conifers during Somatic Embryogenesis of Larch

  • 1. Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China
  • 2. Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, Beijing 100091, China

Abstract: In order to identify miRNA and investigate it’s expression profile during somatic embryogenesis (SE) of larch, a small RNA library was constructed and sequenced by Solexa technique, and expression analysis of miRNAs and four precursors during eight developmental stages of SE was evaluated by qRT-PCR. The results demonstrated that a total of 87 miRNAs was identified, of which 29 miRNAs from 12 families were specific to conifers, including miR946, miR947, miR950, miR951, miR1311, miR1312, miR1313, miR1314, miR1316, miR3701, miR3702 and miR3704, which could be used for further analysis and next experiment. Of these, 25 miRNAs were 22nt in length, while 4 miRNAs were 21nt. The abundance of miRNAs varied largely in the small RNA library, such as miR1311 was detected by 18 copies, while miR950 with 33 009 copies. 34 potential target genes were predicted for 12 miRNA families, which were linked to many respects of regulation, including plant growth and development, disease resistance, AGO protein feedback regulation. qRT-PCR analysis showed that, for 72% miRNAs, the minor expression peak was at late single embryo or early cotyledonary embryo, the lowest level was at middle cotyledonary embryo, while the major peak was at late cotyledonary embryo, which was consistent with the physiology events of cotyledon formation, embryo pre-mature, and embryo dormancy. However, the expression model of miRNA precursor was differed from that of miRNA, and precursor expression with much smaller extent than that of miRNA. The discovery of miRNAs specific to conifers and their targets using embryogenic tissue could provide references for the study on the growth and development of gymnosperms.

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