转录组测序法研究草珊瑚叶和根的基因差异表达

车苏容; 张家源; 卢伟; 祁克明; 魏艺聪

doi:10.19303/j.issn.1008-0384.2020.06.005

转录组测序法研究草珊瑚叶和根的基因差异表达

doi: 10.19303/j.issn.1008-0384.2020.06.005

福建中医药大学，福建　福州　350122

基金项目: 福建省自然科学基金项目（2017J01841）

详细信息

作者简介:
车苏容（1974−），男，硕士生导师，副教授，研究方向：中药品种鉴定与质量评价研究（E-mail8612040@qq.com）

通讯作者:
魏艺聪（1981−），男，博士，副教授，研究方向：中药资源及其药效物质作用机制研究（E-mailyicongwei@126.com）

中图分类号: R 932
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出版历程
- 收稿日期: 2019-11-09
- 修回日期: 2020-06-19
- 刊出日期: 2020-08-10

Profiling Differential Gene Expressions in Leaves and Roots of Sarcandra glabra Based on Transcriptome

Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian　350122, China

摘要

摘要: 目的从基因表达的水平，初步分析草珊瑚叶和根之间次生代谢差异的分子机制，为二者之间临床疗效差异形成的分子机制分析提供信息。方法以福建省福州市的草珊瑚作为样品，采用Illumina HiSeq^TM高通量测序技术测定草珊瑚叶和根的转录组，然后经过滤和Trinity组装，得到的unigenes再通过blast与Nr、Nt、Pfam、KOG、Swiss-Prot、Kegg和GO进行比对注释，并对叶和根的基因差异表达进行分析，尤其是对KEGG代谢通路富集的差异基因进行分析。结果转录组测序结果共获得0.4亿多个clean reads，经Trinity组装后共得到508 271个unigenes，其平均长度为740 bp，最大长度为17.3 kb。基于blast分析，共有148 561个unigenes在七大功能注释数据库中得到成功注释，占总基因数的58.80%。在分析基因表达水平差异时，发现草珊瑚叶和根的共同基因有93 127个，叶和根的差异基因分别为36 327个和52 268个；同时还发现在29 732种不同表达的unigenes中，有12 511个上调基因和17 221个下调基因；代谢相关KEGG具有显著差异的通路有淀粉和蔗糖代谢、苯丙烷类生物合成、乙醛酸和二羧酸代谢、光合生物的固碳作用、吞噬体、谷胱甘肽代谢、光合作用、丙氨酸、天冬氨酸和谷氨酸代谢、倍半萜类和三萜类生物合成、卟啉和叶绿素代谢、氮素代谢、昼夜节律-植物、光合作用-天线蛋白、芪类、二芳基庚酸和姜酚生物合成、不饱和脂肪酸生物合成、柠檬烯和蒎烯降解、类胡萝卜素生物合成、二萜类生物合成、类黄酮生物合成、脂肪酸延伸等。其中与药效密切相关的次生代谢通路苯丙烷类、倍半萜类和三萜类、二萜类、类黄酮类生物合成等途径分别有193个、82个、40个、35个差异表达基因，而上调倍半萜合酶、ent-kaur-16-烯合酶、黄酮醇合酶/黄烷酮3-羟化酶等基因和下调8-羟基香叶醇脱氢酶、vinorine合酶、角鲨烯合酶等关键酶基因差异显著。结论草珊瑚叶和根中苯丙烷类、倍半萜类和三萜类、二萜类、类黄酮次生代谢途径的相关基因差异最为显著，其中差异显著的关键酶基因可为分析其叶和根之间次生代谢差异的分子机制提供重要信息。
- 草珊瑚 /
- 叶 /
- 根 /
- Illumina测序 /
- 基因差异表达分析 /
- 次生代谢
Abstract: Objective Base on transcriptome sequencing, the molecular mechanisms that caused the secondary metabolic differences between the leaves and roots of Sarcandra glabra were studied for clinic applications of the two parts, as well as for determination of the effective components in the medicinal herb. Method Specimens of S. glabra were collected from Fuzhou, Fujian for a transcriptome analysis on the leaves and roots using the Illumina HiSeq platform. After filtration and the Trinity assembly, the unigenes were compared with Nr, Nt, Pfam, KOG, Swiss-Prot, KEGG, and GO by BLAST, and the differentially expressed genes analyzed. A special attention was paid on the differentially enriched genes in the KEGG metabolic pathway. Result More than 40 million clean reads were obtained from the sequencing. The Trinity assembly yielded 508 271 unigenes with an average length of 740 bp. Based on BLAST, 148 561 unigenes, accounting for 58.80% of the total, were successfully annotated using 7 functional annotation databases. There were 29 732 unigenes identified with differential gene expressions between the leaves and the roots. Of which, 12 511 were up-regulated and 17 221 down-regulated. Dozens of significantly different KEGG metabolic pathways were found that associated with functions such as starch/sucrose metabolism, phenylpropanoid biosynthesis, glyoxylate/dicarboxylate metabolism, carbon fixation in photosynthetic organisms, phagosome/glutathione metabolism, photosynthesis, alanine/aspartate/glutamate metabolism, sesquiterpenoid/triterpenoid biosynthesis, porphyrin/chlorophyll metabolism, nitrogen metabolism, circadian rhythm-plant/photosynthesis-antenna proteins, stilbenoid/diarylheptanoid/gingerol biosynthesis, unsaturated fatty acids biosynthesis, limonene/pinene degradation, carotenoid biosynthesis, diterpenoid biosynthesis, flavonoid biosynthesis, fatty acid elongation, etc. Insofar as pharmacodynamics is concerned, the secondary metabolic pathway of the phenylpropanoid biosynthesis had 193 differentially expressed genes between the leaves and the roots, that of the sesquiterpene/triterpene biosynthesis 82, that of the diterpene biosynthesis 40, and that of the flavonoid biosynthesis 35. In addition, the up-regulated genes of sesquiterpene synthase, ent-kaur-16-ene synthase, and flavonol synthase/flavanone 3-hydroxylase, as well as the down-regulated genes of 8-hydroxygeraniol dehydrogenase, vinornine synthase, and squalene synthase were found significantly different between the two parts. Conclusion The genes related to the secondary metabolic pathways of phenylalanine sesquiterpenoid and triterpenoid, diterpenes, and flavonoids most significantly differed in leaves and roots of S. glabra. The significantly differentiated genes associated with the key enzymes provided important information for analyzing the molecular mechanisms of the medicinal herb.
- Sarcandra glabra (Thunb.) Nakai /
- leaves /
- roots /
- illumina sequencing /
- differential gene expression analysis /
- secondary metabolism

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图 1 拼接转录本与基因序列长度分布

Figure 1. Distribution of lengths of spliced transcripts and gene sequences

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图 2 GO分类

Figure 2. GO classification

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图 3 KEGG分类

Figure 3. KEGG classification

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图 4 利用维恩图分析草珊瑚叶和根的基因差异表达

Figure 4. Gene expression by Venn diagram

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表 1 样品RNA测序质量情况

Table 1. Quality of RNA sequencing on samples

样品名 Sample	原始序列数据量 Number of Raw Reads	过滤后的测序数据量 Number of Clean Reads	碱基错误率 Error/%	Q20/%	Q30/%	GC/%
L1	48 191 554	47 613 498	0.02	97.69	93.16	45.60
L2	49 296 702	48 604 312	0.02	97.83	93.51	45.70
L3	44 518 042	43 929 418	0.02	97.85	93.48	45.65
R1	48 079 532	47 367 140	0.02	97.77	93.33	45.22
R2	40 631 848	40 030 256	0.02	97.77	93.33	45.31
R3	41 827 608	41 131 944	0.02	97.86	93.71	45.65
注：Q20、Q30：指的是Phred值大于20、30的碱基占总体碱基的百分比；GC：指的是碱基G和C的数量和占总的碱基数量的百分比。 Note: Q20, Q30: the percentage of bases with Phred values greater than 20,30 in total base; GC: the number of bases G and C and the percentage of the total bases number.

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表 2 转录本拼装长度分布情况

Table 2. Distribution of lengths of transcript assembly

项目 Project	最小长度 Min length/bp	平均长度 Mean length/bp	中位数长度 Median length/bp	最大长度 Max length/bp	N50/bp	N90/bp	总核苷酸 Total nucleotides/个
转录本 Transcripts	201	740	418	17 345	1 178	298	376 095 538
基因 Genes	201	1 138	806	17 345	1 627	537	287 368 837
注：N50/N90：指的是将拼接转录本从长到短排序后累加转录本的长度，直到不小于总长50%/90%的拼接转录本的长度。 Note: N50/N90: the length of the spliced transcript that is accumulated after sequencing from long to short until the length of the spliced transcripts is not less than 50/90% of the total length.

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表 3 基因注释成功率

Table 3. Success rate of gene annotation

数据库 Database	基因数目 Number of Genes	占比 Percentage/%
在NR数据库有中得到注释 Annotated in NR	124 514	49.28
在NT数据库有中得到注释 Annotated in NT	62 462	24.72
在PFAM数据库有中得到注释 Annotated in PFAM	91 970	36.40
在KOG数据库有中得到注释 Annotated in KOG	33 736	13.35
在SwissPro数据库有中得到注释 Annotated in SwissProt	87 941	34.81
在KO数据库有中得到注释 Annotated in KO	43 227	17.11
在GO数据库有中得到注释 Annotated in GO	92 738	36.70
在所有数据库中得到注释 Annotated in all Databases	14 603	5.78
至少在一个数据库中得到注释 Annotated in at least one Database	148 561	58.80
总基因 Total Unigenes	252 630	100.00

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表 4 样品差异倍数前10名基因

Table 4. Top 10 genes with multi-fold differentiation in specimens

部位 Position	序号 Number	基因编号 Gene id	读数 Readcount	差异倍数 Log2 fold change	基因表达说明 NR Description
叶 Leaf	1	Cluster-32258.95760	2 209.65	13.54	抗坏血酸过氧化酶 L-ascorbate peroxidase
	2	Cluster-32258.103145	3 524.70	13.25	2-甲基-6-植酰苯醌甲基转移酶 2-methyl-6-phytylbenzoquinone methyltranferase
	3	Cluster-32258.100314	8 286.64	12.59	5′-腺苷硫酸盐还原酶 5′-adenylylsulfate reductase
	4	Cluster-32258.72246	957.15	12.33	CBL相互作用蛋白激酶 CBL-interacting protein kinase
	5	Cluster-32258.93899	937.73	12.30	叶绿体茎环结合蛋白 chloroplast stem-loop binding protein
	6	Cluster-32258.93433	858.84	12.17	逆转录转座子多蛋白 retrotransposon polyprotein
	7	Cluster-32258.97994	7 598.24	12.14	细胞质同工酶 cytoplasmic isozyme
	8	Cluster-32258.102576	825.51	12.12	淀粉合成酶 stachyose synthase
	9	Cluster-32258.52249	796.50	12.07	钾转运体 potassium transporter
	10	Cluster-32258.89876	707.29	11.89	赤霉素2-β-双加氧酶 gibberellin 2-beta-dioxygenase
根 Root	1	Cluster-13209.0	29.70	−20.77	葡萄糖6-脱氢酶 glucose 6-dehydrogenase
	2	Cluster-32258.15742	11 132.11	−15.97	枯草芽孢杆菌类蛋白酶 subtilisin-like protease
	3	Cluster-32258.155788	3 560.27	−14.32	阳离子氨基酸转运体 cationic amino acid transporter
	4	Cluster-32258.178815	3 308.58	−14.22	谷胱甘肽S-转移酶 glutathione S-transferase
	5	Cluster-32258.178517	2 443.78	−13.78	膜联蛋白样蛋白 annexin-like protein
	6	Cluster-32258.184073	2 002.57	−13.49	甘油-3-磷酸2-O-酰基转移酶 glycerol-3-phosphate 2-O-acyltransferase
	7	Cluster-32258.15882	1 941.66	−13.45	细胞色素 Cytochrome
	8	Cluster-32258.15168	1 825.90	−13.36	过氧化物酶超家族蛋白 Peroxidase superfamily protein
	9	Cluster-32258.11664	1 596.82	−13.17	黄瓜素 Cucumisin
	10	Cluster-32258.15447	1 461.26	−13.04	DNA结合蛋白 DNA-binding protein

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表 5 草珊瑚叶和根KEGG富集前20的代谢通路

Table 5. Metabolic pathways of S. glabra leaves and roots with 20 highest KEGG enrichment

序号 Number	通路名称 Pathway term	富集度 Rich factor	q值 q value	基因数目 Gene number/个
1	淀粉和蔗糖代谢 Starch and sucrose metabolism	0.28	0.004	213
2	苯丙素类生物合成 Phenylpropanoid biosynthesis	0.38	0.000	193
3	乙醛酸和二羧酸代谢 Glyoxylate and dicarboxylate metabolism	0.33	0.000	161
4	光合生物的固碳作用 Carbon fixation in photosynthetic organisms	0.34	0.000	153
5	吞噬体 Phagosome	0.30	0.007	121
6	谷胱甘肽代谢 Glutathione metabolism	0.29	0.020	119
7	光合作用 Photosynthesis	0.38	0.000	94
8	丙氨酸，天冬氨酸和谷氨酸代谢 Alanine, aspartate and glutamate metabolism	0.29	0.034	86
9	倍半萜类和三萜类生物合成 Sesquiterpenoid and triterpenoid biosynthesis	0.51	0.000	82
10	卟啉和叶绿素代谢 Porphyrin and chlorophyll metabolism	0.32	0.006	82
11	氮素代谢 Nitrogen metabolism	0.36	0.004	64
12	昼夜节律-植物 Circadian rhythm - plant	0.35	0.004	64
13	光合作用-天线蛋白 Photosynthesis - antenna proteins	0.64	0.000	63
14	芪类，二芳基庚酸和姜酚生物合成 Stilbenoid, diarylheptanoid and gingerol biosynthesis	0.45	0.000	55
15	不饱和脂肪酸生物合成 Biosynthesis of unsaturated fatty acids	0.32	0.029	55
16	柠檬烯和蒎烯降解 Limonene and pinene degradation	0.33	0.022	53
17	类胡萝卜素生物合成 Carotenoid biosynthesis	0.34	0.022	51
18	二萜类生物合成 Diterpenoid biosynthesis	0.37	0.022	40
19	类黄酮生物合成 Flavonoid biosynthesis	0.46	0.003	35
20	脂肪酸延伸 Fatty acid elongation	0.39	0.024	31

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表 6 苯丙烷类代谢途径上差异倍数前10名基因

Table 6. Top 10 genes with multi-fold differentiation on phenylalanine metabolic pathway

部位 Position	序号 Number	基因编号 Gene id	叶中的读数 Leaf readcount	根中的读数 Root readcount	差异倍数 Log2 fold change	基因表达说明 NR Description
叶 Leaf	1	Cluster-32258.118654	359.81	1.66	7.64	第三类过氧化物酶 class III peroxidase
	2	Cluster-32258.57844	670.29	4.06	7.38	莽草酸氧-羟基肉桂酰基转移酶 shikimate O-hydroxycinnamoyltransferase
	3	Cluster-32258.75671	22.25	0.00	6.90	溶酶体β葡萄糖苷酶样异构体 X1 lysosomal beta glucosidase-like isoform X1
	4	Cluster-21604.0	18.74	0.00	6.66	假设的蛋白质 VITISV 011546 hypothetical protein VITISV 011546
	5	Cluster-22722.0	13.73	0.00	6.21	过氧化物酶P7样 peroxidase P7-like
	6	Cluster-32258.162333	12.17	0.00	6.03	假设的蛋白质 SPRG_01919 hypothetical protein SPRG 01919
	7	Cluster-32258.93656	10.98	0.00	5.89	过氧化物酶 N1 Peroxidase N1
	8	Cluster-32258.59976	181.06	3.49	5.56	假设的蛋白质 PHAVU 010G162700g hypothetical protein PHAVU 010G162700g
	9	Cluster-32258.92806	15.42	0.27	5.41	β-葡萄糖苷酶31样 beta-glucosidase 31-like
	10	Cluster-32258.105029	15.42	0.27	5.41	β-葡萄糖苷酶31样 beta-glucosidase 31-like
根 Root	1	Cluster-32258.15168	0.00	1 825.90	−13.36	过氧化物酶超家族蛋白 Peroxidase superfamily protein
	2	Cluster-32258.179987	0.61	2 036.76	−11.67	8-羟基香叶醇脱氢酶 8-hydroxygeraniol dehydrogenase
	3	Cluster-32258.183657	0.00	294.75	−10.73	PRX3过氧氧还蛋白 PRX3 peroxiredoxin
	4	Cluster-32258.8233	0.00	275.66	−10.63	过氧化物酶10样 peroxidase 10-like
	5	Cluster-10163.0	0.00	208.35	−10.23	未命名蛋白质产品 unnamed protein product
	6	Cluster-32258.172620	0.98	798.80	−9.72	可能的甘露醇脱氢酶 probable mannitol dehydrogenase
	7	Cluster-32258.23969	9.49	7 453.68	−9.62	可能的甘露醇脱氢酶亚型 X2 probable mannitol dehydrogenase isoform X2
	8	Cluster-32258.874	0.00	131.92	−9.57	过氧化物酶9样 peroxidase 9-like
	9	Cluster-32258.179634	0.00	111.64	−9.33	过氧化物酶17样 peroxidase 17-like
	10	Cluster-32258.179997	0.32	210.46	−9.28	过氧化物酶7样 peroxidase 7-like

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表 7 倍半萜类和三萜类代谢途径上差异倍数前10名基因

Table 7. Top 10 genes with multi-fold differentiation on sesquiterpenoid and triterpenoid metabolic pathways

部位 Position	序号 Number	基因编号 Gene id	叶中的读数 Leaf readcount	根中的读数 Root readcount	差异倍数 Log2 fold change	基因表达说明 NR Description
叶 Leaf	1	Cluster-32258.64073	623.05	0.00	11.71	假设的蛋白质 VITISV 030783 hypothetical protein VITISV 030783
	2	Cluster-32258.111485	2 274.22	2.03	10.17	假设的蛋白质 VITISV 000109 hypothetical protein VITISV 000109
	3	Cluster-32258.111497	1 356.27	1.76	9.63	未命名蛋白质产品 unnamed protein product
	4	Cluster-32258.97968	192.97	0.33	9.06	卤酸脱卤酶样 haloacid dehalogenase-like
	5	Cluster-32258.108941	189.67	0.33	9.03	倍半萜合酶 sesquiterpene synthase
	6	Cluster-32258.97970	73.89	0.00	8.64	锗烷烯-D合酶 germacrene-D synthase
	7	Cluster-32258.70247	66.66	0.00	8.49	倍半萜合酶 sesquiterpene synthase
	8	Cluster-32258.102983	1 747.64	5.94	8.13	倍半萜合酶 sesquiterpene synthase
	9	Cluster-32258.98251	461.98	1.61	8.12	锗烷烯-D合酶 germacrene-D synthase
	10	Cluster-32258.102455	79.05	0.27	7.77	锗烷烯-D合酶 germacrene-D synthase
根 Root	1	Cluster-32258.180183	0.00	548.81	−11.62	细胞色素P45071D11样 cytochrome P450 71D11-like
	2	Cluster-32258.5750	0.00	177.43	−9.99	（-）-锗烷烯D合酶（-）-germacrene D synthase
	3	Cluster-32258.177549	0.00	78.25	−8.82	Valencene合酶样 valencene synthase-like
	4	Cluster-32258.5749	0.00	73.25	−8.72	（-）-锗烷烯D合酶（-）-germacrene D synthase
	5	Cluster-32258.181501	0.00	61.55	−8.47	未命名蛋白质产物 unnamed protein product
	6	Cluster-32258.177548	0.30	67.20	−7.63	Valencene合酶样 valencene synthase-like
	7	Cluster-32258.5751	0.00	17.33	−6.64	δ-镉烯合酶同工酶 Delta-cadinene synthase isozyme A
	8	Cluster-32258.178134	0.00	16.37	−6.56	Valencene合酶样 valencene synthase-like
	9	Cluster-32258.7220	0.00	14.12	−6.35	（-）-锗烷烯D合酶（-）-germacrene D synthase-like
	10	Cluster-7136.0	0.00	12.04	−6.11	鲨烯合酶 squalene synthase

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表 8 二萜类代谢途径上差异倍数前10名基因

Table 8. Top 10 genes with multi-fold differentiation on diterpenes metabolic pathway

部位 Position	序号 Number	基因编号 Gene id	叶中的读数 Leaf readcount	根中的读数 Root readcount	差异倍数 log2 Fold Change	基因表达说明 NR Description
Leaf	1	Cluster-32258.89876	707.29	0.00	11.89	赤霉素2-β-双加氧酶8-样 gibberellin 2-beta-dioxygenase 8-like
	2	Cluster-32258.89871	257.30	0.00	10.44	赤霉素2-β-双加氧酶8-样 gibberellin 2-beta-dioxygenase 8-like
	3	Cluster-32258.89879	124.88	0.00	9.39	赤霉素2-β-双加氧酶8-样 gibberellin 2-beta-dioxygenase 8-like
	4	Cluster-32258.89883	86.18	0.00	8.86	赤霉素2-β-双加氧酶8-样 gibberellin 2-beta-dioxygenase 8-like
	5	Cluster-32258.98988	1 138.80	3.04	8.50	ent-Kaur-16-烯合成酶 ent-kaur-16-ene synthase
	6	Cluster-32258.89886	62.52	0.00	8.39	赤霉素2-β-双加氧酶8-样 gibberellin 2-beta-dioxygenase 8-like
	7	Cluster-32258.121945	48.27	0.00	8.02	ent-Kaur-16-烯合成酶 ent-kaur-16-ene synthase
	8	Cluster-32258.89870	175.69	1.67	6.70	赤霉素2-β-双加氧酶8-样 gibberellin 2-beta-dioxygenase 8-like
	9	Cluster-32258.89878	1 829.05	22.15	6.39	赤霉素2-β-双加氧酶8-样 gibberellin 2-beta-dioxygenase 8-like
	10	Cluster-32258.89868	72.51	0.83	6.24	赤霉素2-β-双加氧酶8-样 gibberellin 2-beta-dioxygenase 8-like
Root	1	Cluster-32258.187014	0.00	377.06	−11.08	en-kaurene氧化酶 ent-kaurene oxidase
	2	Cluster-32258.8593	0.00	322.60	−10.86	戊二烯丙基二磷酸合酶 ent-copalyl diphosphate synthase
	3	Cluster-32258.177819	0.00	186.82	−10.07	ent-贝壳杉酸氧化酶2样 ent-kaurenoic acid oxidase 2-like
	4	Cluster-32258.187433	0.00	176.53	−9.99	ent-贝壳杉酸氧化酶1样 ent-kaurenoic acid oxidase 1-like
	5	Cluster-32258.187015	0.00	172.68	−9.96	en-kaurene氧化酶2样 ent-kaurene oxidase 2-like
	6	Cluster-32258.13043	0.00	126.65	−9.51	假设的蛋白质F775 52245 hypothetical protein F775 52245
	7	Cluster-32258.44968	2.83	1 419.27	−8.97	ent-贝壳杉酸氧化酶1样 ent-kaurenoic acid oxidase 1-like
	8	Cluster-32258.187016	0.00	71.95	−8.69	en-kaurene氧化酶2样 ent-kaurene oxidase 2-like
	9	Cluster-32258.12198	0.00	53.27	−8.26	ent-贝壳杉酸氧化酶2样 ent-kaurenoic acid oxidase 2-like
	10	Cluster-32258.185460	0.00	51.04	−8.19	赤霉素2-β-双加氧酶2-样 gibberellin 2-beta-dioxygenase 2-like

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表 9 类黄酮代谢途径上差异倍数前10名基因

Table 9. Top 10 genes with multi-fold differentiation on flavonoids metabolic pathway

部位 Position	序号 Number	基因编号 Gene id	叶中的读数 Leaf readcount	根中的读数 Root readcount	差异倍数 Log2 fold change	基因表达说明 NR Description
叶 Leaf	1	Cluster-32258.71949	86.75	0.00	8.87	黄酮醇合成酶/黄酮3-羟化酶 flavonol synthase/flavanone 3-hydroxylase
	2	Cluster-32258.115774	82.00	0.00	8.79	黄酮醇合成酶/黄酮3-羟化酶 flavonol synthase/flavanone 3-hydroxylase
	3	Cluster-32258.115780	2 057.23	7.28	8.14	黄酮醇合成酶/黄酮3-羟化酶 flavonol synthase/flavanone 3-hydroxylase
	4	Cluster-32258.57844	670.29	4.06	7.38	莽草酸氧-羟基肉桂酰基转移酶 shikimate O-hydroxycinnamoyltransferase
	5	Cluster-32258.90886	7.93	0.00	5.42	查耳酮合酶 chalcone synthase
	6	Cluster-32258.71390	6.28	0.00	5.08	莽草酸氧-羟基肉桂酰基转移酶 shikimate O-hydroxycinnamoyltransferase
	7	Cluster-32258.90882	1 753.10	56.76	4.94	查耳酮合酶 chalcone synthase
	8	Cluster-32258.142410	4 158.40	206.09	4.33	查尔酮和二苯乙烯合成酶家族蛋白 Chalcone and stilbene synthase family protein
	9	Cluster-32258.106648	42.80	2.31	4.20	黄烷酮3-羟化酶 flavanone 3-hidroxylase
	10	Cluster-32258.87019	32.35	1.98	4.02	查耳酮合酶 chalcone synthase
根 Root	1	Cluster-32258.10764	0.00	20.79	−6.90	长春瑞碱合成酶 vinorine synthase
	2	Cluster-1755.1	0.00	7.72	−5.46	细胞色素 P450CYP75B79样 cytochrome P450 CYP75B79-like
	3	Cluster-32258.10891	7.38	278.86	−5.25	细胞色素 P450CYP73A100样 cytochrome P450 CYP73A100-like
	4	Cluster-32258.57819	948.36	9 832.09	−3.37	咖啡酰辅酶-CoAO-甲基转移酶 caffeoyl-CoA O-methyltransferase
	5	Cluster-32258.152146	295.63	2 811.13	−3.24	4-香豆酸3-羟化酶 5-4-coumarate 3-hydroxylase
	6	Cluster-32258.66450	15.59	78.37	−2.33	咖啡酰辅酶-CoAO-甲基转移酶 caffeoyl-CoA O-methyltransferase
	7	Cluster-32258.96920	700.62	2 261.42	−1.69	莽草酸氧-羟基肉桂酰基转移酶 shikimate O-hydroxycinnamoyltransferase
	8	Cluster-32258.72425	372.27	1 064.20	−1.51	咖啡酰辅酶-CoA3-O-甲基转移酶 caffeoyl-CoA 3-O-methyltransferase
	9	Cluster-32258.124422	229.83	480.10	−1.06	咖啡酰辅酶-CoAO-甲基转移酶 caffeoyl-CoA O-methyltransferase
	10	—	—	—	—	—

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