基于比较转录组的延胡索组织差异性表达分析

doi:10.16420/j.issn.0513-353x.2021-0822

摘要/Abstract

摘要：

延胡索（Corydalis yanhusuo）干燥块茎为传统中药，异喹啉生物碱类化合物是其主要的活性成分。采用Illumina高通量测序技术对延胡索块茎、根茎和叶片组织进行转录组测序，鉴定异喹啉生物碱生物合成的相关酶基因，并分析3种组织间的差异表达基因。在块茎vs.根茎、块茎vs.叶、根茎vs.叶比对中分别鉴别到6 161、8 852和5 772个差异表达基因。差异表达基因的KEGG富集分析表明，核糖体、淀粉和蔗糖代谢途径，苯丙素生物合成途径，植物激素信号转导途径以及异喹啉生物碱生物合成途径是主要的富集途径。其中47个转录本与延胡索异喹啉生物碱生物合成相关，其在块茎中的表达量最高，根茎次之，叶中的表达量最低。

关键词: 延胡索, 异喹啉生物碱, 转录组, 表达分析

Abstract:

The dried tubers of Corydalis yanhusuo can be used as the traditional Chinese medicine，and isoquinoline alkaloid compounds are the main active ingredient. To identify genes related to isoquinoline alkaloid biosynthesis and analyze the differentially expressed genes of tuber，rhizome and leaf tissues of C. yanhusuo，transcriptome sequencing of these tissues using Illumina high-throughput sequencing technology were performed. A total of 6 161，8 852 and 5 772 differentially expressed genes were identified in comparisons of tuber vs. rhizome，tuber vs. leaf and rhizome vs. leaf，respectively．KEGG enrichment analysis of differentially expressed genes indicated that major enrichment pathways were starch and sucrose metabolism，phenylpropanine biosynthesis，plant hormone signal transduction and isoquinoline alkaloid biosynthesis pathways. Fourty-seven transcripts were identified to be related to isoquinoline alkaloid biosynthesis and the highest level of gene expression was found in tuber，following by rhizome and leaf.

Key words: Corydalis yanhusuo, isoquinoline alkaloids, transcriptome, expression analysis

中图分类号:

S567

赵雪艳, 王琪, 王莉, 王方圆, 王庆, 李艳. 基于比较转录组的延胡索组织差异性表达分析[J]. 园艺学报, 2023, 50(1): 177-187.

ZHAO Xueyan, WANG Qi, WANG Li, WANG Fangyuan, WANG Qing, LI Yan. Comparative Transcriptome Analysis of Differential Expression in Different Tissues of Corydalis yanhusuo [J]. Acta Horticulturae Sinica, 2023, 50(1): 177-187.

图/表 8

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基因名称（编号） Gene name（ID）	正向引物序列（5′-3′） Forward primer sequence	反向引物序列（5′-3′） Reverse primer sequence
Actin (AT1G49240)	GGTAACATTGTGCTCAGTGGTGG	AACGACCTTAATCTTCATGCTGC
NCS（transcript_36281）	TAGCAAGCATGGCAGCAGTCATC	TAGCCACCTCCACCTCTGTTGTC
6OMT（transcript_4831）	AGGGACTGGTCACTTGGGATCG	AAAAGGCGTCGGTGCATCGG
CNMT（transcript_29289）	AGTGGGAAGCATTTCTCACGTACC	TTCCTCCATGATTGCCATCACTGC
NMCH（transcript_4028）	GCCAAAGCCCAATCAAGGAATCAG	TGGGGTTGGAGGGTGAAGTCTTAG
4’OMT（transcript_4823）	CGTAGCACTGGAGGAGGGATCTG	CCTTTCTCTCCCACCTGTGTTCAC
SOMT（transcript_19389）	TTGCTGGGAAGGAGCCAAAAGTG	ACCACCACCAACATCAACCAACTC

基因名称（编号） Gene name（ID）	正向引物序列（5′-3′） Forward primer sequence	反向引物序列（5′-3′） Reverse primer sequence
Actin (AT1G49240)	GGTAACATTGTGCTCAGTGGTGG	AACGACCTTAATCTTCATGCTGC
NCS（transcript_36281）	TAGCAAGCATGGCAGCAGTCATC	TAGCCACCTCCACCTCTGTTGTC
6OMT（transcript_4831）	AGGGACTGGTCACTTGGGATCG	AAAAGGCGTCGGTGCATCGG
CNMT（transcript_29289）	AGTGGGAAGCATTTCTCACGTACC	TTCCTCCATGATTGCCATCACTGC
NMCH（transcript_4028）	GCCAAAGCCCAATCAAGGAATCAG	TGGGGTTGGAGGGTGAAGTCTTAG
4’OMT（transcript_4823）	CGTAGCACTGGAGGAGGGATCTG	CCTTTCTCTCCCACCTGTGTTCAC
SOMT（transcript_19389）	TTGCTGGGAAGGAGCCAAAAGTG	ACCACCACCAACATCAACCAACTC

样品 Sample	Clean read 数量 Clean read number	碱基数 Base number	GC/%	% ≥ Q30	比对率/% Mapped ratio
块茎 Tuber	20 214 211	6 055 395 267	43.43	93.21	86.72
根茎 Rhizome	21 439 743	6 422 326 271	43.24	93.17	72.78
叶 Leaf	24 116 972	7 222 883 792	43.40	93.40	71.96

样品 Sample	Clean read 数量 Clean read number	碱基数 Base number	GC/%	% ≥ Q30	比对率/% Mapped ratio
块茎 Tuber	20 214 211	6 055 395 267	43.43	93.21	86.72
根茎 Rhizome	21 439 743	6 422 326 271	43.24	93.17	72.78
叶 Leaf	24 116 972	7 222 883 792	43.40	93.40	71.96

样品对比 Sample pair	总数 All	COG	GO	KEGG	KOG	Pfam	Swiss-Prot	eggNOG	Nr
块茎 vs. 根茎 Tuber vs. Rhizome	5 869	2 735	4 201	2 420	3 341	4 995	4.667	5 723	5 846
块茎 vs. 叶 Tuber vs. Leaf	8 487	3 989	6 126	3 937	5 234	7 270	6.629	8 261	8 458
根茎 vs. 叶 Rhizome vs. Leaf	5 529	2 674	3 989	2 557	3 465	4 741	4.340	5 361	5 508