花生油脂合成相关基因的表达谱分析

doi:10.3724/SP.J.1006.2021.04150

Abstract

Abstract:

In order to survey the regulation patterns of genes expression in the synthesis of oil at different developmental stages in peanut seed, F18 (high-oleic medium oil peanut variety) and ‘Luhua 6’ (low-oleic acid low oil variety) were used as research materials. The expression pattern analysis of genes was performed for peanut seeds on the 10, 30, 40, and 60 DAP, respectively. The results indicated that 130, 3556, and 2783 genes were significantly differentially expressed in the two varieties (lines) on the 30, 40, and 60 DAP, respectively. GO annotation and KEGG enrichment showed that DEGs were mainly enriched in the fatty acid synthesis and photosynthetic pathways, such as FAB2, FAD2, WRI1 genes, which were involved in the accumulation of oleic acid. All the genes involved in photosynthesis pathway were photochlorophyll binding proteins and all of them were up-regulated. KEGG pathway indicated that all the genes involved in fatty acid biosynthesis pathway on the 40 DAP and 60 DAP were up-regulated. In summary, these results provide a theoretical basis for molecular study of fatty acid synthesis in the development stage of peanut seeds and offer some candidate genes as gene resources of quality improvement in peanut breeding.

Key words: peanut, microarray, differentially expressed genes, genes involved in lipid synthesis

XU Jing, PAN Li-Juan, LI Hao-Yuan, WANG Tong, CHEN Na, CHEN Ming-Na, WANG Mian, YU Shan-Lin, HOU Yan-Hua, CHI Xiao-Yuan. Expression pattern analysis of genes related to lipid synthesis in peanut[J].Acta Agronomica Sinica, 2021, 47(6): 1124-1137.

Figures/Tables 12

Table 1

Table 2

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Table 3

Differentially expressed genes involved in the lipid metabolism on 40 DAP"

基因编号 Gene ID		基因注释 Gene annotation			GO富集 GO enrichment		调控^a Regulation^a	差异表达倍数 Fold change
AHTC1031703		移酶1-酰基-sn-甘油-3-磷酸酰基转			磷脂生物合成		上调	4.19
AHTC1031703		1-acyl-sn-glycerol-3-phosphate acyltransferase-like			Phospholipid biosynthetic process		Up	4.19
基因编号 Gene ID	基因注释 Gene annotation			GO富集 GO enrichment			调控^a Regulation^a	差异表达倍数 Fold change
AHTC1022314	3-酮脂酰-合酶			脂肪酸生物合成			上调	2.88
AHTC1022314	3-ketoacyl-synthase			Fatty acid biosynthetic process			Up	2.88
AHTC1029564	类3-酮脂酰-合酶10			脂肪酸生物合成			上调	2.30
AHTC1029564	3-ketoacyl-synthase 10-like			Fatty acid biosynthetic process			Up	2.30
AHTC1007527	3-酮脂酰-合酶11			脂肪酸生物合成			下调	3.28
AHTC1007527	3-ketoacyl-synthase 11			Fatty acid biosynthetic process			Down	3.28
AHTC1008395	类3-酮脂酰-合酶11			脂肪酸生物合成			下调	2.55
AHTC1008395	3-ketoacyl-synthase 11-like			Fatty acid biosynthetic process			Down	2.55
AHTC1016489	3-酮脂酰-合酶21			脂质生物合成			上调	5.25
AHTC1016489	3-ketoacyl-synthase 21			Lipid biosynthetic process			Up	5.25
AHTC1003640	类3-酮脂酰-合酶21			脂肪酸生物合成			上调	3.51
AHTC1003640	3-ketoacyl-synthase 21-like			Fatty acid biosynthetic process			Up	3.51
AHTC1003641	类3-酮脂酰-合酶21			脂肪酸生物合成			上调	6.43
AHTC1003641	3-ketoacyl-synthase 21-like			Fatty acid biosynthetic process			Up	6.43
AHTC1004544	3-酮脂酰-合酶6			脂肪酸生物合成			上调	2.86
AHTC1004544	3-ketoacyl-synthase 6			Fatty acid biosynthetic process			Up	2.86
AHTC1019082	α-乙酰羧化酶亚基			脂肪酸生物合成; 丙二酰辅酶A生物合成			下调	2.07
AHTC1019082	Alpha-acetyl-carboxylase subunit			Fatty acid biosynthetic process; malonyl-CoA biosynthetic process			Down	2.07
AHTC1001812	β-酮酰基-ACP还原酶			脂肪酸生物合成; 氧化还原			上调	2.05
AHTC1001812	Beta-ketoacyl-acp reductase 1-2			Fatty acid biosynthetic process; oxidation-reduction process			Up	2.05
AHTC1001844	β-酮酰基-ACP合酶			脂肪酸生物合成			上调	2.17
AHTC1001844	Beta-ketoacyl-acp synthase i-2			Fatty acid biosynthetic process			Up	2.17
AHTC1005915	叶绿体ω-6 脂肪酸去饱和酶			二磷酸生物合成			上调	2.06
AHTC1005915	Chloroplast omega-6 fatty acid desaturase			Diphosphate biosynthetic process			Up	2.06
AHTC1032453	酰基-载体-蛋白合酶			脂肪酸生物合成			上调	2.16
AHTC1032453	Fatty acid biosynthesis 1 isoform 1			Fatty acid biosynthetic process			Up	2.16
AHTC1032454	酰基-载体-蛋白合酶			脂肪酸生物合成			上调	2.57
AHTC1032454	Fatty acid biosynthesis 1 isoform 1			Fatty acid biosynthetic process			Up	2.57
AHTC1020107	鞘脂Δ8 去饱和酶			脂质代谢; 氧化还原			上调	2.04
AHTC1020107	Fatty acid sphingolipid desaturase			Lipid metabolic process; oxidation- reduction process			Up	2.04
AHTC1035094	脂质转移蛋白			脂质转移			上调	14.59
AHTC1035094	Lipid transfer protein gpi-anchored			Lipid transport			Up	14.59
AHTC1009675	长链酰基合成酶			长链脂肪酸代谢			上调	2.44
AHTC1009675	Long chain acyl-synthetase 1-like isoform×1			Long-chain fatty acid metabolic process			Up	2.44
AHTC1026013	长链酰基合成酶			长链脂肪酸代谢			上调	3.03
AHTC1026013	Long chain acyl-synthetase 1-like isoform×1			Long-chain fatty acid metabolic process			up	3.03
AHTC1017689	长链酰基合酶2			长链脂肪酸代谢			上调	2.65
AHTC1017689	Long chain acyl-synthetase 2			Long-chain fatty acid metabolic process			up	2.65
AHTC1006429	长链酰基合成酶2异构体			长链脂肪酸代谢			上调	3.44
AHTC1006429	Long chain acyl-synthetase 2-like isoform×2			Long-chain fatty acid metabolic process			Up	3.44
AHTC1034617	微粒体ω-3脂肪酸去饱和酶			脂质代谢; 氧化还原			上调	5.40
AHTC1034617	Microsomal omega-3 fatty acid desaturase			Lipid metabolic process; oxidation- reduction process			Up	5.40
基因编号 Gene ID	基因注释 Gene annotation		GO富集 GO enrichment			调控^a Regulation^a		差异表达倍数 Fold change
AHTC1020991	非特异性脂质转移酶蛋白		脂质转移			上调		3.92
AHTC1020991	Non-specific lipid-transfer protein		Lipid transport			Up		3.92
AHTC1017861	ω-3去饱和酶家族蛋白		不饱和脂肪酸生物合成; 氧化还原			下调		2.55
AHTC1017861	Omega-3 desaturase family protein		Unsaturated fatty acid biosynthetic process; oxidation-reduction process			Down		2.55
AHTC1008662	硬脂酰ACP去饱和酶		脂肪酸生物合成			上调		3.48
AHTC1008662	Stearoyl-acyl-carrier desaturase		Fatty acid biosynthetic process; oxidation-reduction process			Up		3.48

Table 3

Table 4

Differentially expressed genes involved in the lipid metabolism on 60 DAP"

基因编号 Gene ID	基因注释 Gene annotation	GO富集 GO enrichment	调控^a Regulation^a	差异表达倍数 Fold change
AHTC1005431	类3-酮脂酰-合酶11	脂肪酸生物合成	上调	7.27
AHTC1005431	3-ketoacyl-synthase 11-like	Fatty acid biosynthetic process	Up	7.27
AHTC1000075	酰基载体蛋白	脂肪酸生物合成; 氧化还原	上调	3.10
AHTC1000075	Acyl carrier protein 1	Fatty acid biosynthetic process; oxidation- reduction process	Up	3.10
AHTC1000128	酰基载体蛋白	脂肪酸生物合成; 氧化还原	上调	2.72
AHTC1000128	Acyl carrier protein 1	Fatty acid biosynthetic process; oxidation- reduction process	Up	2.72
AHTC1028756	α-羧基转移酶前体	脂肪酸生物合成	上调	4.68
AHTC1028756	Alpha-carboxyltransferase precursor	Fatty acid biosynthetic process	Up	4.68
AHTC1005278	α-羧基转移酶	脂肪酸生物合成; 泛醌生物合成	上调	4.26
AHTC1005278	Alpha-carboxyl transferase 2	Fatty acid biosynthetic process; coenzyme biosynthetic process	Up	4.26
AHTC1000881	β-羟烷基-ACP脱氢酶	脂肪酸生物合成	上调	18.60
AHTC1000881	Beta-hydroxyacyl-acp dehydratase	Fatty acid biosynthetic process	Up	18.60
AHTC1001812	β-酮酰基-ACP还原酶	脂肪酸生物合成; 氧化还原	上调	7.89
AHTC1001812	Beta-ketoacyl-acp reductase 1-2	Fatty acid biosynthetic process; oxidation- reduction process	Up	7.89
AHTC1001844	β-酮酰基-ACP合酶	脂肪酸生物合成	上调	4.49
AHTC1001844	Beta-ketoacyl-acp synthase i-2	Fatty acid biosynthetic process	Up	4.49
AHTC1007010	β-酮酰基-ACP合酶	脂肪酸生物合成	上调	3.25
AHTC1007010	Beta-ketoacyl-acp synthase ii-1	Fatty acid biosynthetic process	Up	3.25
AHTC1002452	生物素羧基载体蛋白	脂肪酸生物合成	上调	6.29
AHTC1002452	Biotin carboxyl carrier protein 2-2	Fatty acid biosynthetic process	Up	6.29
AHTC1014935	生物素羧基载体蛋白	脂肪酸生物合成	上调	2.83
AHTC1014935	Biotin carboxyl carrier protein 2-2	Fatty acid biosynthetic process	Up	2.83
AHTC1022457	生物素羧基载体蛋白	脂肪酸生物合成	上调	4.01
AHTC1022457	Biotin carboxyl carrier protein 2-2	Fatty acid biosynthetic process	Up	4.01
AHTC1032453	酰基-载体-蛋白合酶	脂肪酸生物合成	上调	3.90
AHTC1032453	Fatty acid biosynthesis 1 isoform 1	Fatty acid biosynthetic process	Up	3.90
AHTC1009675	长链酰基合成酶	长链脂肪酸代谢	上调	7.57
AHTC1009675	Long chain acyl-synthetase 1-like isoform×1	Long-chain fatty acid metabolic process	Up	7.57
基因编号 Gene ID	基因注释 Gene annotation	GO富集 GO enrichment	调控^a Regulation^a	差异表达倍数 Fold change
AHTC1026013	长链酰基合成酶	长链脂肪酸代谢	上调	9.64
AHTC1026013	Long chain acyl-synthetase 1-like isoform×1	Long-chain fatty acid metabolic process	Up	9.64
AHTC1017689	长链酰基合酶2	长链脂肪酸代谢	上调	6.75
AHTC1017689	Long chain acyl-synthetase 2	Long-chain fatty acid metabolic process	Up	6.75
AHTC1020991	非特异性脂质转移酶蛋白	脂质转移	上调	3.89
AHTC1020991	Non-specific lipid-transfer protein	Lipid transport	Up	3.89
AHTC1002315	非特异性脂质转移酶蛋白	脂质转移	上调	3.73
AHTC1002315	Non-specific lipid-transfer protein at5g64080-like	Lipid transport	Up	3.73
AHTC1015374	非特异性脂质转移酶蛋白	脂质转移	上调	3.13
AHTC1015374	Non-specific lipid-transfer protein at5g64080-like	Lipid transport	Up	3.13
AHTC1019533	酰基转移酶	甘油酯生物合成	下调	6.23
AHTC1019533	O-acyltransferase wsd1-like	Glycerolipid biosynthetic process	Down	6.23
AHTC1005424	油酰-酰基载体蛋白硫酯酶	脂肪酸生物合成	上调	2.72
AHTC1005424	Oleoyl-acyl carrier protein thioesterase chloroplastic-like	Fatty acid biosynthetic process	Up	2.72
AHTC1000715	ω6 脂肪酸去饱和酶	脂质代谢; 氧化还原	上调	3.82
AHTC1000715	Omega 6 fatty acid desaturase	Lipid metabolic process; oxidation-reduction process	Up	3.82
AHTC1021791	过氧化物酶体脂肪酸-β氧化多功能蛋白	脂肪酸生物合成; 氧化还原	下调	2.62
AHTC1021791	Peroxisomal fatty acid beta-oxidation multifunctional protein mfp2-like	Fatty acid metabolic process; oxidation- reduction process	Down	2.62
AHTC1001749	硬脂酰-ACP去饱和酶	脂肪酸生物合成; 氧化还原	上调	29.89
AHTC1001749	Searoyl-acp desaturase	Fatty acid biosynthetic process; oxidation- reduction process	Up	29.89
AHTC1003174	超长链3-酰基还原酶	超长链脂肪酸生物合成	上调	3.92
AHTC1003174	Very-long-chain 3-oxoacyl-reductase 1-like	Very long-chain fatty acid biosynthetic process	Up	3.92
AHTC1001068	类超长链烯酰还原酶异构体	脂质代谢; 氧化还原	上调	3.97
AHTC1001068	Very-long-chain enoyl-reductase-like isoformx1	Lipid metabolic process; oxidation-reduction process	Up	3.97
AHTC1001069	类超长链烯酰还原酶异构体	脂质代谢; 氧化还原	上调	3.80
AHTC1001069	Very-long-chain enoyl-reductase-like isoformx1	Lipid metabolic process; oxidation-reduction process	Up	3.80

Table 4

Fig. 6

Fig. 7

Table 5

RT-PCR validation results of the differentially expressed genes"

基因编号 Gnen ID	基因注释 Gene annotation	发育时期 Development stage	调控^a Regulation ^a	芯片数据(倍数) Microarray data (fold change)	RT-PCR验证结果(倍数) RT-PCR result (fold change)
AHTC1032413	黄酮醇合酶	DAP30	上调	6.05	13.64
AHTC1032413	Flavonol synthase flavanone 3-hydroxylase-like	DAP30	Up	6.05	13.64
AHTC1030930	锌指蛋白	DAP30	下调	-3.78	-4.55
AHTC1030930	Dof zinc finger protein	DAP30	Down	-3.78	-4.55
AHTC1005724	非特异性脂质转移酶	DAP40	上调	4.04	3.25
AHTC1005724	Non-specific lipid-transfer protein 3-like	DAP40	Up	4.04	3.25
AHTC1030772	β-羟酰基-ACP脱水酶	DAP40	上调	2.86	2.11
AHTC1030772	Beta-hydroxyacyl-acp dehydrase 1	DAP40	Up	2.86	2.11
AHTC1020107	鞘脂Δ8 去饱和酶	DAP40	上调	2.04	3.48
AHTC1020107	Fatty acid sphingolipid desaturase	DAP40	Up	2.04	3.48
AHTC1012845	WRI1 转录因子	DAP40	上调	2.15	2.31
AHTC1012845	Ethylene-responsive transcription factor wri1-like	DAP40	Up	2.15	2.31
AHTC1030930	锌指蛋白	DAP40	下调	-5.30	-10.24
AHTC1030930	Dof zinc finger protein	DAP40	down	-5.30	-10.24
AHTC1014100	肌醇半乳糖醇合酶	DAP40	下调	-3.09	-4.07
AHTC1014100	Galactinol synthase 8	DAP40	Down	-3.09	-4.07
AHTC1001812	β-酮酰基-ACP还原酶	DAP60	上调	7.89	5.09
AHTC1001812	Beta-ketoacyl-acp reductase 1-2	DAP60	Up	7.89	5.09
AHTC1001844	β-酮酰基-ACP合酶	DAP60	上调	4.49	4.83
AHTC1001844	Beta-ketoacyl-acp synthase i-2	DAP60	Up	4.49	4.83
AHTC1012845	WRI1 转录因子	DAP60	上调	5.83	21.21
AHTC1012845	Ethylene-responsive transcription factor wri1-like	DAP60	Up	5.83	21.21
AHTC1005424	酰基载体蛋白硫酯酶	DAP60	上调	2.72	3.47
AHTC1005424	Oleoyl-acyl carrier protein thioesterase chloroplastic-like	DAP60	Up	2.72	3.47
基因编号 Gnen ID	基因注释 Gene annotation	发育时期 Development stage	调控^a Regulation^a	芯片数据(倍数) Microarray data (fold change)	RT-PCR验证结果(倍数) RT-PCR result (fold change)
AHTC1001749	硬脂酰-ACP去饱和酶	DAP60	上调	29.89	16.26
AHTC1001749	Stearoyl-acp desaturase	DAP60	Up	29.89	16.26
AHTC1000715	ω6 脂肪酸去饱和酶	DAP60	上调	3.82	1.55
AHTC1000715	Omega 6 fatty acid desaturase	DAP60	Up	3.82	1.55
AHTC1002482	天冬酰胺合成酶	DAP60	下调	-4.87	-6.12
AHTC1002482	Asparagine synthetase 2	DAP60	Down	-4.87	-6.12
AHTC1027178	Hsf 转录因子	DAP60	下调	-4.39	-9.85
AHTC1027178	Heat stress transcription factor a-2-like	DAP60	Down	-4.39	-9.85

Table 5

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基因编号 Gene ID	正向引物 Forward primer (5°-3°)	反向引物 Reverse primer (5°-3°)
AHTC1032413	ACGAAGGTTATGGAAGTAG	AGGAAGGTAGTGAAGGAA
AHTC1030930	AGGTGAAGAATCTGTTAGTC	CTTATCTTGTGAATTACTTGTCT
AHTC1005724	GATAAGTGGAGCAGTAGC	CAAGAAGACGATGATATAAGC
AHTC1030772	TCAATCAGATTCGTAACATTC	ACTCAATAACTCTATCCACTAG
AHTC1020107	CTACTACTCATCATTCATTGTC	CTTAACTCACCCACACAT
AHTC1012845	TCCTCTTCTTCATCTTCATC	CCTGGTTACTCCTCTGTA
AHTC1014100	ACTCCTTCTCCTCTTCAT	ATATTAGGCAGCAGATACC
AHTC1001812	AATGGCAGGAGGTTATTG	TTCTTCTTCATCATAATCTTAGC
AHTC1001844	GTGACTTGGCAGAGATTA	AATTGATTGATTGACGGATG
AHTC1001749	TCTTGGAGTTCCTTGTTG	GCTATGCGACTTCTTCAT
AHTC1005424	GAGTGATGTAGTTGAGATAGA	CTCTTCCGATGACTTGAT
AHTC1000715	TGGTTAAGAGGAGCATTG	ATGAGGCATTGTTGAGAA
AHTC1002482	GTTGCTCTGATGACTCTC	TATGGCTAACCGTTGATG
AHTC1027178	TCTCACTCACTCTCATCA	CCACCACCATCTTAGAAG
Actin11	TTGGAATGGGTCAGAAGGATGC	AGTGGTGCCTCAGTAAGAAGC

样品名称 Name	发育时期 Development stage	含油量 Oil content (%)	油酸含量 Oleic content (%)	亚油酸含量 Linoleic content (%)	油亚比 O/L
F18	10 DAP	13.86	58.91	15.39	3.83
	30 DAP	35.21	74.01	5.61	13.19
	40 DAP	46.82	76.06	2.43	31.30
	60 DAP	50.81	70.75	1.55	45.65
鲁花6号 Luhua 6	10 DAP	19.67	31.63	37.14	0.85
	30 DAP	36.95	34.96	41.28	0.85
	40 DAP	44.44	37.67	39.39	0.96
	60 DAP	45.44	43.98	34.28	1.28

Expression pattern analysis of genes related to lipid synthesis in peanut

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