Cloning and expression analysis of <i>PeLAC</i> in <i>Phyllostachys edulis</i>

Li Li-Chao; Sun Hua-Yu; Lou Yong-Feng; Yang Yi-Hong; Zhao Han-Sheng; Gao Zhi-Min

doi:10.11913/PSJ.2095-0837.2017.20252

Plant Science Journal > 2017 > 35(2): 252-259. > DOI: 10.11913/PSJ.2095-0837.2017.20252 CSTR: 32231.14.PSJ.2095-0837.2017.20252

Li Li-Chao, Sun Hua-Yu, Lou Yong-Feng, Yang Yi-Hong, Zhao Han-Sheng, Gao Zhi-Min. Cloning and expression analysis of PeLAC in Phyllostachys edulis[J]. Plant Science Journal, 2017, 35(2): 252-259. DOI: 10.11913/PSJ.2095-0837.2017.20252

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Cloning and expression analysis of PeLAC in Phyllostachys edulis

International Center for Bamboo and Rattan, State Forestry Administration Key Laboratory on the Science and Technology of Bamboo and Rattan, Beijing 100102

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This work was supported by grants from the Special Fund for Forest Scientific Research in the Public Welfare from State Forestry Administration of China (201504106), and National Sci-Tech Support Plan of the Twelfth Five-Year (2015BAD04B01)

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Received Date: July 14, 2016
Revised Date: September 13, 2016
Available Online: October 31, 2022
Published Date: April 27, 2017

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Abstract

Abstract

Laccase (LAC) has many functions in plants, playing important roles in both growth and development. The cDNA, genomic DNA, and promoter sequences of PeLAC were isolated from Moso bamboo (Phyllostachys edulis(Carr.) Lehaie) using PCR techniques. The open reading frame (ORF) and genomic DNA sequences of PeLAC in Ph. edulis were 1692 bp and 2785 bp, respectively. There were six exons separated by five introns in the PeLAC genomic sequence, which were in full compliance with the intron splicing principle of GT-AG. PeLAC encoded 563 amino acid residues with a calculated molecular weight of 62.3 kD and theoretical isoelectric point of 9.056. Phylogenetic analysis demonstrated that PeLAC had high identities with LACs from other plants. Prediction showed that a putative miR397-targeted site was found in the coding region of PeLAC, which was further validated by RLM-5' RACE. The sequencing result of cleavage products showed that the cleavage site was at the 10th base of miR397 complementary to PeLAC in Ph. edulis, confirming that miR397 can guide mRNA cleavage of PeLAC. Tissue specific expression analysis showed that PeLAC was expressed at the highest level in the stem, followed by the root and leaf sheath, but hardly at all in the leaf blade. With increasing shoot height, the expression abundance of PeLAC increased to a maximum at 15 cm, and then declined at 30 cm; whereas the expression of miR397 showed the opposite pattern. The promoter sequence of PeLAC was also isolated, and was 2000 bp and included many responsive elements such as ABRE and MBS related to abiotic stresses, indicating that PeLAC might be involved in responses to hormones and drought. Further experiments confirmed that PeLAC was obviously upregulated in roots under ABA (100 μmol/L for 24 h) and NaCl (400 mmol/L for 4 h) treatment, but was downregulated after treatment with GA₃ (100 μmol/L for 3 h). The isolation and expression analysis of PeLAC presented in this research has laid a foundation for further study on the biological functions of LACs in Ph. edulis.
- Phyllostachys edulis,
- LAC,
- miR397,
- Expression analysis

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References (32)

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Cloning and expression analysis of PeLAC in Phyllostachys edulis

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