Identification and functional analysis of CLE gene family in Dendrobium officinale Kimura et Migo

doi:10.3969/j.issn.1004-1524.20231020

Acta Agriculturae Zhejiangensis ›› 2024, Vol. 36 ›› Issue (7): 1583-1590.DOI: 10.3969/j.issn.1004-1524.20231020

• Horticultural Science • Previous Articles Next Articles

Identification and functional analysis of CLE gene family in Dendrobium officinale Kimura et Migo

ZHU Yan¹(), DING Lan², CHEN Yiqian¹, HUANG Xiujing¹, JIANG Weiwei¹, CHEN Donghong¹^,^*()

1. The State Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300 China
2. Agricultural and Forestry Technology Extension Center of Lin’an District, Hangzhou 311300, China

Received:2023-08-28 Online:2024-07-25 Published:2024-08-05

Abstract

Abstract:

CLE peptide are a class of small secreted proteins that widely participate in plant development and cell-to-cell communication, playing a key regulatory role in maintaining the proliferation and differentiation of stem cells. In order to study the function of CLE family member in Dendrobium officinale Kimura et Migo, RNA was extracted from the leaves, roots, stems, buds and fruits of D. officinale and reverse transcribed into cDNA, and the expression of CLE family genes in each tissue was detected by semi-quantitative RT-PCR. The results showed that 17 CLE members were identified in D. officinale, all of which conserved 12-amino acid CLE motifs. Semi-quantitative RT-PCR performed on roots, stems, leaves, flowers and fruits showed that these CLE members had different tissue expression profiles, particularly CLE19635 with specifical expression in flower buds and fruits, and CLE22175 with specifical expression in fruits. In vitro synthesis of CLE peptides for treatment of Arabidopsis thaliana revealed that CLE peptides impacted the growth of A. thaliana roots, CLE05351 promoted root elongation, CLE18468 had no obvious effect, but the other CLE peptides all inhibited root elongation. mPS-PI staining did not find that the cell pattern of root-tip meristem was affected by CLE peptides. In addition, T₂ homozygotes of A. thaliana overexpressing CLE02038 showed a short root phenotype, which was consistent with the result of CLE peptides treatment. The results laid a foundation for the further exploring the role of CLE family in the epiphytic root of D. officinale and its signal regulation network, and provided potential target genes for breeding of ideal root systems in D. officinale.

Key words: Dendrobium officinale, CLE gene family, peptide hormone, root development, Arabidopsis

CLC Number:

S567.239

ZHU Yan, DING Lan, CHEN Yiqian, HUANG Xiujing, JIANG Weiwei, CHEN Donghong. Identification and functional analysis of CLE gene family in Dendrobium officinale Kimura et Migo[J]. Acta Agriculturae Zhejiangensis, 2024, 36(7): 1583-1590.

Figures/Tables 6

References 27

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基因名称 Gene name	正向引物 Forward primer (5'→3')	反向引物 Reverse primer (5'→3')	基因ID Gene ID	产物长度 Product length/bp
CLE109525	CTCTTACAGTACCTGATCATACAC	TTATGCCTGTCCAGGCAA	LOC110109525	525
CLE01792	ATGTCTCTCCAATTGCTCTTG	TTAATGGTGGAGTGGGTTGG	PKU85961	372
CLE02038	GACTTCATGAGGAAGAATAGAA	TCAGTTATGAAGAGGATTAGGTC	PKU86207	228
CLE04631	ATGGCTAAACTAAAAGAGATAGG	TCATCTGTTATGCAGAGGATTAG	PKU65016	267
CLE05351	ATGGCTTCTGCTGAAGTCTC	CTACCTATTGGAGTCTGGGTT	PKU82346	267
CLE11641	ATGGCATTCTCTTCAAGAGC	TCAATGGTGTTGAGGGTCTG	PKU78021	231
CLE5097	ATGGCAAGAGAAGTAGTGAGG	TCAATTGTGTAGCGGGTTTG	PKU64567	171
CLE15729	ATGGTTGGGCAAAGAGAGATG	TCAGTTGTGCAAAGGATTAGG	PKU75207	264
CLE18468	ATGGACACTGAGCTGATCAG	TCACCTATTTGAATCAGGATTC	PKU83835	336
CLE19635	ATGCTGCCCACTCTGCCG	CTAGTGATGGAGGGGATTCG	PKU59933	177
CLE20689	ATGCAAGGAATTAGAAGGACT	TCAGTTGTGAAGTGGATTGG	PKU74917	249
CLE22175	ATGGGGAGAATCAATATTCT	TTAATGGTGTTCAGGATCTG	PKU68119	210
CLE22943	ATGGTTGGGCTAAGAGAGA	CTATCTGTTGTGCAAAGGATT	PKU86730	267
CLE29071	ATGCTTCCCACTCTGCCG	CTAGTGATGGAGGGGATTCGC	PKU62687	177
CLE79016	ATGGCGTTCACATACTCAAGAG	TCAATGGTGTTGGGGGTCTG	PKU79016	231
CLE82107	ATGCCAACTTCAACGAGGC	TTACCGGTTGTGAAGGCGG	PKU82107	234
CLE66413	ATGGGGCTGCGTAAAAGAG	TCACAGCTCTTTTACGCAGC	PKU66413	237
DcACTIN1	GATTTGCTGGTGACGATGC	AGGATACCTCTTTTGGACTGG	LOC110111141	136

基因名称 Gene name	正向引物 Forward primer (5'→3')	反向引物 Reverse primer (5'→3')	基因ID Gene ID	产物长度 Product length/bp
CLE109525	CTCTTACAGTACCTGATCATACAC	TTATGCCTGTCCAGGCAA	LOC110109525	525
CLE01792	ATGTCTCTCCAATTGCTCTTG	TTAATGGTGGAGTGGGTTGG	PKU85961	372
CLE02038	GACTTCATGAGGAAGAATAGAA	TCAGTTATGAAGAGGATTAGGTC	PKU86207	228
CLE04631	ATGGCTAAACTAAAAGAGATAGG	TCATCTGTTATGCAGAGGATTAG	PKU65016	267
CLE05351	ATGGCTTCTGCTGAAGTCTC	CTACCTATTGGAGTCTGGGTT	PKU82346	267
CLE11641	ATGGCATTCTCTTCAAGAGC	TCAATGGTGTTGAGGGTCTG	PKU78021	231
CLE5097	ATGGCAAGAGAAGTAGTGAGG	TCAATTGTGTAGCGGGTTTG	PKU64567	171
CLE15729	ATGGTTGGGCAAAGAGAGATG	TCAGTTGTGCAAAGGATTAGG	PKU75207	264
CLE18468	ATGGACACTGAGCTGATCAG	TCACCTATTTGAATCAGGATTC	PKU83835	336
CLE19635	ATGCTGCCCACTCTGCCG	CTAGTGATGGAGGGGATTCG	PKU59933	177
CLE20689	ATGCAAGGAATTAGAAGGACT	TCAGTTGTGAAGTGGATTGG	PKU74917	249
CLE22175	ATGGGGAGAATCAATATTCT	TTAATGGTGTTCAGGATCTG	PKU68119	210
CLE22943	ATGGTTGGGCTAAGAGAGA	CTATCTGTTGTGCAAAGGATT	PKU86730	267
CLE29071	ATGCTTCCCACTCTGCCG	CTAGTGATGGAGGGGATTCGC	PKU62687	177
CLE79016	ATGGCGTTCACATACTCAAGAG	TCAATGGTGTTGGGGGTCTG	PKU79016	231
CLE82107	ATGCCAACTTCAACGAGGC	TTACCGGTTGTGAAGGCGG	PKU82107	234
CLE66413	ATGGGGCTGCGTAAAAGAG	TCACAGCTCTTTTACGCAGC	PKU66413	237
DcACTIN1	GATTTGCTGGTGACGATGC	AGGATACCTCTTTTGGACTGG	LOC110111141	136

基因名(CLE多肽名称) Gene name (Name of CLE peptide)	保守区段基序 Motif sequence	基因名(CLE多肽名称) Gene name (Name of CLE peptide)	保守区段基序 Motif sequence	基因名(CLE多肽名称) Gene name (Name of CLE peptide)	保守区段基序 Motif sequence
CLE66413 (PKU66413)	RKVPNASDPLHN	CLE05351 (PKU82346)	HEVPNGPNPDSN	CLE82107 (PKU82107)	RPVPSCPDRLHN
CLE109525 (LOC110109525)	RKVPKGPDPIHN	CLE18468 (PKU83835)	HEVPSGPNPDSN	CLE19635	RLVPSGANPLHH
				(PKU59933/PKU62687)
CLE79016 (PKU79016)	RLSPGGSDPQHH	CLE04631 (PKU65016)	RNVYTGPNPLHN	CLE20689 (PKU74917)	RLVPSGPNPLHN
CLE22175 (PKU68119)	RLSPGGPDPEHH	CLE15729	RTVYTGPNPLHN	CLE01792 (PKU85961)	RFVPCGPNPLHH
		(PKU75207/PKU86730)
CLE11641 (PKU78021)	RLSPGGPDPQHH	CLE02038 (PKU86207)	RIIYAGPNPLHN	CLE5097 (PKU64567)	RFVPQGPNPLHN

基因名(CLE多肽名称) Gene name (Name of CLE peptide)	保守区段基序 Motif sequence	基因名(CLE多肽名称) Gene name (Name of CLE peptide)	保守区段基序 Motif sequence	基因名(CLE多肽名称) Gene name (Name of CLE peptide)	保守区段基序 Motif sequence
CLE66413 (PKU66413)	RKVPNASDPLHN	CLE05351 (PKU82346)	HEVPNGPNPDSN	CLE82107 (PKU82107)	RPVPSCPDRLHN
CLE109525 (LOC110109525)	RKVPKGPDPIHN	CLE18468 (PKU83835)	HEVPSGPNPDSN	CLE19635	RLVPSGANPLHH
				(PKU59933/PKU62687)
CLE79016 (PKU79016)	RLSPGGSDPQHH	CLE04631 (PKU65016)	RNVYTGPNPLHN	CLE20689 (PKU74917)	RLVPSGPNPLHN
CLE22175 (PKU68119)	RLSPGGPDPEHH	CLE15729	RTVYTGPNPLHN	CLE01792 (PKU85961)	RFVPCGPNPLHH
		(PKU75207/PKU86730)
CLE11641 (PKU78021)	RLSPGGPDPQHH	CLE02038 (PKU86207)	RIIYAGPNPLHN	CLE5097 (PKU64567)	RFVPQGPNPLHN

Identification and functional analysis of CLE gene family in Dendrobium officinale Kimura et Migo

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