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Bioinformatics and expression pattern analysis of miR156 family and its regulatory target SPL in Dimocarpus longan*(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

2020 03
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Bioinformatics and expression pattern analysis of miR156 family and its regulatory target SPL in Dimocarpus longan*
JIANG Mengqi SU Liyao HUANG Shuqi CHEN Xu XU Xiaoping Zhang ZihaoLAI Zhongxiong & LIN Yuling**
Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou 350002, China
Dimocarpus longanMiR156Evolutionary characteristicsSomatic embryogenesisSPL

MiR156 gene family is one of the ancient and conservative families and it involved in the regulation of plant growth and development. However, the miR156 gene family evolutionary characteristics and its function in Dimocarpus longan somatic embryos have not been reported yet. Therefore we investigated the evolution characteristics of miR156 gene family and its regulatory role in the early somatic embryogenesis of longan. we analyzed the mature and precursor sequences, and the phylogenetic of longon and other plants , predicted the secondary structure of precursors miR156 (pre-miR156) family members , analyzed the regulatory patterns of miR156a and its target genes SPL6/9/16 during the early somatic embryogenesis of longan. The longan miR156 family contain ed 13 mature sequences and 6 precursor sequences. Mfold prediction showed that the sequences of the pre-mir156 family members could form a typical stable secondary stem-loop structures , and their minimum folding free energy (ΔG) ranged from -43.30 kal·mol-1 to - 62.60kal·mol-1 ; Multiple sequences alignment of these pre-miR1 56 members showed that the regions generating mature miRNA were highly conservative. Phylogenetic tree analysis showed that miR156 family of longan was more closely related to those from rice and Arabidopsis thaliana. According to prediction of target gene, the target genes of miR156a were SPL2, SPL6, SPL9, SPL16 and SPL18. We detected SPL6/9/16 fragments in longan callus using an improved RLM-RACE, and miR156a cleaved the target mRNA at the 10th nucleotide. Real-time fluorescence quantitative PCR showed that in the early somatic embryos of longan, the expression of miR156a was stable at day 0-9th, but only decreased significantly in globular embryos (GE) , while SPL6/9/16 was increased significantly. These results showed that the mature sequences of miR156 families in plants are highly conserved , the decreased miR156 and the accumulation of SPL6/9/16 may be great significance to the construction of longan globular embryos.




Last Update: 2019-09-04