铁皮石斛CCO基因家族鉴定及其在花发育中的表达分析

doi:10.16420/j.issn.0513-353x.2023-0566

摘要/Abstract

摘要：

为了解铁皮石斛（Dendrobium officinale）中类胡萝卜素裂解氧化酶（carotenoid cleavage oxygenase，CCO）基因家族的功能，通过生物信息的方法鉴定铁皮石斛CCO家族成员，同时分析它们在不同组织、不同发育时期的花及茉莉酸甲酯（methyl jasmonate，MeJA）处理后幼苗叶片中的表达量。结果表明，铁皮石斛有12个CCO家族成员，10个属于CCD亚家族，2个属于NCED亚家族，不均匀定位于7条染色体上，且存在串联重复基因对。该家族基因启动子上存在大量激素响应和逆境响应元件。DoCCO家族基因在铁皮石斛各组织中均有表达，多数在花粉和根中高表达，它们在不同发育时期的花和MeJA处理后叶片中呈现差异化表达，可能参与不同组织的生长发育及MeJA响应，其中DoCCD1b可能在花发育中发挥重要作用。

关键词: 铁皮石斛, 类胡萝卜素裂解氧化酶, 基因家族, 花发育, 表达分析

Abstract:

To investigate the functions of carotenoid cleavage oxygenase（CCO）gene family in Dendrobium officinale Kimura et Migo，bioinformatics methods were employed to identify the CCO family members in D. officinale. Simultaneously，their expression levels in different tissues，different stages of flower development，and after the application of methyl jasmonate（MeJA）. The results revealed that a total of 12 CCO family members were excavated，with ten belonging to the carotenoid cleavage dioxygenase（CCD）subfamily and two belonging to the 9-cis-epoxycarotenoid dioxygenase（NCED）subfamily. They were unevenly distributed on 7 chromosomes，and harbored tandem duplications. The promoter regions of these DoCCO genes contained numerous hormone and stress response elements. The expression of DoCCO genes varied across different tissues of D. officinale，with higher expression observed in pollen and roots. They were differentially expressed during flower development and in response to MeJA treatment，suggesting that their potential roles in growth，development and the MeJA response. Specifically，DoCCD1b，may play a crucial role during flower development.

Key words: Dendrobium officinale, carotenoid cleavage oxygenase, gene family, flower development, expression analysis

赵佳莹, 曾周婷, 岑欣颖, 施姣淇, 李效贤, 沈晓霞, 俞振明. 铁皮石斛CCO基因家族鉴定及其在花发育中的表达分析[J]. 园艺学报, 2024, 51(9): 2075-2088.

ZHAO Jiaying, ZENG Zhouting, CEN Xinying, SHI Jiaoqi, LI Xiaoxian, SHEN Xiaoxia, YU Zhenming. Genome-Wide Identification and Expression Analysis of CCO Gene Family in Dendrobium officinale During Flower Development[J]. Acta Horticulturae Sinica, 2024, 51(9): 2075-2088.

图/表 11

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引物名称 Primer name	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse primer
EF-1α	GCTTGAGAAGGAGCCCAAGT	CCAACAGCCACAGTTTGTCG
DoCCD1a	CCGACGGAGCTAGCAATTTAT	CTGTTTCCTGCCAGTGTAGTT
DoCCD1b	GCACAATCCAGTGCCAATAAC	CCATTTCCTTTGGTCGGAAATAC
DoCCD4	CCTCAGACGAAGCCTCAATATC	GTGTGGTGGGTCGATGAAA
DoCCD7a	CCGGCCAAAGAAACAAGTTC	GTTGACCTTCACCACAGAGT
DoCCD7b	CCATTGGCCTCACAGACATTA	CTTAGTGATTCCGGCGATGAA
DoCCD7c	GAGCACCGGGTTTGAAATTG	GCCAATAACGTCGGCTCTAA
DoCCD7d	CTTGTGCTCCATTGTTGATTCC	GAAGTCGAGCTAGCGTACATAAA
DoCCD8a	GCTCCAACTACAGGGAGAAATC	AAAGGTGGCGGAAATAGTAGTC
DoCCD8b	ATGCCGCTGGAGATAGATTTG	TCTTGGGATGAGCCATGAAAG
DoCCD8c	CGATCCTTCCTCTCTTGGATTT	CGGAGTGAATGGAATGAGTTTG
DoNCED1	CGATACGCCTACCTTGCAATAG	CGAATTTCCCTTTCCCGTACTC
DoNCED2	CTCCAGGCCATTCAATCTCATC	GGACGGAATTAGGAGTGGTAGA

引物名称 Primer name	上游引物（5′-3′） Forward primer	下游引物（5′-3′） Reverse primer
EF-1α	GCTTGAGAAGGAGCCCAAGT	CCAACAGCCACAGTTTGTCG
DoCCD1a	CCGACGGAGCTAGCAATTTAT	CTGTTTCCTGCCAGTGTAGTT
DoCCD1b	GCACAATCCAGTGCCAATAAC	CCATTTCCTTTGGTCGGAAATAC
DoCCD4	CCTCAGACGAAGCCTCAATATC	GTGTGGTGGGTCGATGAAA
DoCCD7a	CCGGCCAAAGAAACAAGTTC	GTTGACCTTCACCACAGAGT
DoCCD7b	CCATTGGCCTCACAGACATTA	CTTAGTGATTCCGGCGATGAA
DoCCD7c	GAGCACCGGGTTTGAAATTG	GCCAATAACGTCGGCTCTAA
DoCCD7d	CTTGTGCTCCATTGTTGATTCC	GAAGTCGAGCTAGCGTACATAAA
DoCCD8a	GCTCCAACTACAGGGAGAAATC	AAAGGTGGCGGAAATAGTAGTC
DoCCD8b	ATGCCGCTGGAGATAGATTTG	TCTTGGGATGAGCCATGAAAG
DoCCD8c	CGATCCTTCCTCTCTTGGATTT	CGGAGTGAATGGAATGAGTTTG
DoNCED1	CGATACGCCTACCTTGCAATAG	CGAATTTCCCTTTCCCGTACTC
DoNCED2	CTCCAGGCCATTCAATCTCATC	GGACGGAATTAGGAGTGGTAGA

基因ID Gene ID	氨基酸数 Number of amino acid	分子量/kD Molecular weight	等电点 pI	不稳定系数 Instability index	脂肪系数 Aliphatic index	总平均亲水系数 Grand average of hydropathicity	RPE65保守域 RPE65 domain	亚细胞定位 Subcellular localization
DoCCD1a	512	57.63	9.12	30.53	89.30	0	1 ~ 319	叶绿体、细胞质 Chloroplast，cytoplasm
DoCCD1b	501	56.47	9.19	35.52	83.05	-0.278	17 ~ 384	叶绿体、细胞质 Chloroplast，cytoplasm
DoCCD4	633	68.58	6.30	45.29	81.39	-0.163	148 ~ 622	叶绿体、细胞质 Chloroplast，cytoplasm
DoCCD7a	194	21.94	9.25	51.25	68.81	-0.236	26 ~ 167	叶绿体、细胞质 Chloroplast，cytoplasm
DoCCD7b	261	28.58	7.72	41.14	96.82	0.153	66 ~ 216	叶绿体 Chloroplast
DoCCD7c	600	67.57	6.65	37.66	79.65	-0.218	65 ~ 597	细胞质 Cytoplasm
DoCCD7d	296	33.84	8.10	50.41	85.30	-0.136	112 ~ 222	叶绿体、细胞质 Chloroplast，cytoplasm
DoCCD8a	769	85.68	5.05	42.49	80.01	-0.332	298 ~ 764	叶绿体、细胞质 Chloroplast，cytoplasm
DoCCD8b	559	61.50	6.04	36.29	87.73	-0.110	88 ~ 394；424 ~ 554	细胞质 Cytoplasm
DoCCD8c	333	37.94	6.42	48.06	67.39	-0.425	195 ~ 281	叶绿体、细胞质、细胞核Chloroplast，cytoplasm，nucleus
DoNCED1	592	65.40	5.98	42.95	79.71	-0.279	121 ~ 584	细胞质 Cytoplasm
DoNCED2	610	67.31	6.27	50.40	79.26	-0.300	137 ~ 600	细胞质 Cytoplasm

基因ID Gene ID	氨基酸数 Number of amino acid	分子量/kD Molecular weight	等电点 pI	不稳定系数 Instability index	脂肪系数 Aliphatic index	总平均亲水系数 Grand average of hydropathicity	RPE65保守域 RPE65 domain	亚细胞定位 Subcellular localization
DoCCD1a	512	57.63	9.12	30.53	89.30	0	1 ~ 319	叶绿体、细胞质 Chloroplast，cytoplasm
DoCCD1b	501	56.47	9.19	35.52	83.05	-0.278	17 ~ 384	叶绿体、细胞质 Chloroplast，cytoplasm
DoCCD4	633	68.58	6.30	45.29	81.39	-0.163	148 ~ 622	叶绿体、细胞质 Chloroplast，cytoplasm
DoCCD7a	194	21.94	9.25	51.25	68.81	-0.236	26 ~ 167	叶绿体、细胞质 Chloroplast，cytoplasm
DoCCD7b	261	28.58	7.72	41.14	96.82	0.153	66 ~ 216	叶绿体 Chloroplast
DoCCD7c	600	67.57	6.65	37.66	79.65	-0.218	65 ~ 597	细胞质 Cytoplasm
DoCCD7d	296	33.84	8.10	50.41	85.30	-0.136	112 ~ 222	叶绿体、细胞质 Chloroplast，cytoplasm
DoCCD8a	769	85.68	5.05	42.49	80.01	-0.332	298 ~ 764	叶绿体、细胞质 Chloroplast，cytoplasm
DoCCD8b	559	61.50	6.04	36.29	87.73	-0.110	88 ~ 394；424 ~ 554	细胞质 Cytoplasm
DoCCD8c	333	37.94	6.42	48.06	67.39	-0.425	195 ~ 281	叶绿体、细胞质、细胞核Chloroplast，cytoplasm，nucleus
DoNCED1	592	65.40	5.98	42.95	79.71	-0.279	121 ~ 584	细胞质 Cytoplasm
DoNCED2	610	67.31	6.27	50.40	79.26	-0.300	137 ~ 600	细胞质 Cytoplasm