向日葵DGATs基因家族的鉴定及表达分析

doi:10.3724/SP.J.1006.2023.14217

Abstract

Abstract:

Diacylglycerol acyltransferase (DGAT) plays an important role in plant oil metabolism and stress resistance. To explore the evolution of DGAT gene in sunflower (Helianthus annuus L.) and its function in oil biosynthesis and response to abiotic stresses, 18 homologous gene sequences of DGAT were obtained from the sunflower genome database by alignment with Arabidopsis thaliana AtDGATs genes. Then, chromosome distribution, gene structure, conserved protein motifs, phylogenetic relationship, tissue-specific expression of HaDGATs, and their expressing patterns under abiotic stresses were systematically investigated. The results showed that sunflower DGATs genes were divided into four subfamilies (DGAT1, DGAT2, DGAT3, and WSD), and the members of the same subfamily shared similar gene structures and conserved motifs. The promoter regions of HaDGATs harbored multiple cis-elements related to environmental stresses and plant hormone response. The main factor for HaDGATs amplification was fragment duplication. The qRT-PCR indicated that HaDGAT1, HaDGAT2, and HaDGAT3 were mainly expressed in the early and middle stages of sunflower seed development, which coincided with the rapid accumulation period of oil in seeds, while HaWSD subfamily genes were mainly expressed in stems, leaves, and petals. Most of HaWSD genes could be induced by NaCl, low temperature, drought, and ABA treatments in sunflower roots, stems, and leaves, indicating that they might play essential roles in dealing with various abiotic stresses. The results revealed that HaDGATs had functional differentiation in regulating lipid biosynthesis and abiotic stresses. This study provides an important foundation for further understanding the function of sunflower DGAT genes.

Key words: sunflower, DGAT gene family, abiotic stresses, gene expression

ZHANG Cheng, ZHANG Zhan, YANG Jia-Bao, MENG Wan-Qiu, ZENG Ling-Lu, SUN Li. Genome-wide identification and relative expression analysis of DGATs gene family in sunflower[J].Acta Agronomica Sinica, 2023, 49(1): 73-85.

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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°)
HaDGAT1-1	TACGTAGGCGTCCGATCACA	AATAGGCCTGCATGACTCTGTTT
HaDGAT1-2	GAGGAGCGTGAAGCCTGATG	ATGCGCTGGAGAAGAAGCAC
HaDGAT1-3	GCCTAACATCGCCGAATCCA	AACGCAACACACTCGTCTGA
HaDGAT2-1	AGTACGGGCGTGGCTATTTT	GCCAATGACAGTCCCATCCA
HaDGAT2-2	TCTGGCCGATTGGAGTTGTC	TGAACGCCACCTGGTATGAC
HaDGAT3	GTGGGTGTTAAGCAGCGGAA	TTGACTTGCTGACCCAACCC
HaWSD1	GAGGATACCGAAACGCCGAT	ACCTTCGTCTTTTTGCCCCT
HaWSD2	TGGATGCCGCTAAACACGTC	CGGCGATGAAACCGTGAGAT
HaWSD3	TGGAGGATAGGCCAAGACGA	TTGTTTCCCCATGTGCCATCT
HaWSD4	GAAAGGAGAGGACCACGACC	GCTCCCGTTTTCCTTGTCCA
HaWSD5	GTTGGTCACGGACAAACACG	ACGATAGAAGCGAAACGCCA
HaWSD6	GGGGCCAAAGATGACCAAGA	CCACCACCGTTGCTTCTAGT
HaWSD7	TTCGTTAGGCGACGGTTTGT	GCCCATGCTGCCTTTTAACG
HaWSD8	TGTGTGCACTGGGCTGTTTC	ACACCCGCGATTTCACTGTATC
HaWSD9	CACTAAAAGGGTCCGCAGGT	GTCGAACGCCTTTGGGAATG
HaWSD10	GCACGACGAACCACTATCCC	TCCGCCAATGTTCATGTCCG
HaWSD11	AGCGTGTTTGGCCTCGAAC	GTGAGGTCGGCTAGGTATTCG
HaWSD12	TGTGTGGAGGATGTGACCATGT	GGTCTTCCCCCTTCACTTCAAC
18S rRNA	CTACCACATCCAAGGAAGGCAG	CGACAGAAGGGACGAGTAAACC

转录本ID Transcript ID	基因名称 Gene name	氨基酸数量 Amino acid amounts (aa)	分子量 Molecular weight (kD)	等电点 pI	亲水性系数 Hydrophilicity coefficient	跨膜结构域数量Transmembrane domain amounts
OTG30061	HaDGAT1-1	479	55.74	9.24	0.243	8
OTG13343	HaDGAT1-2	517	59.81	8.56	0.166	9
OTG03739	HaDGAT1-3	507	58.52	9.11	0.219	9
OTF98266	HaDGAT2-1	330	37.47	9.12	0.294	2
OTF96040	HaDGAT2-2	336	38.09	9.85	0.191	2
OTG25557	HaDGAT3	329	35.59	8.63	-0.416	0
OTG29248	HaWSD1	666	74.73	6.84	-0.249	0
OTG29986	HaWSD2	455	50.68	8.62	-0.037	0
OTG23138	HaWSD3	476	54.00	8.95	-0.241	0
OTG23434	HaWSD4	486	55.54	9.20	-0.244	0
OTG11715	HaWSD5	474	52.63	8.55	-0.003	0
OTG11716	HaWSD6	461	51.91	8.77	-0.025	0
OTG12880	HaWSD7	491	56.20	9.00	-0.221	1
OTG03949	HaWSD8	508	57.33	9.21	-0.176	1
OTG03950	HaWSD9	509	57.11	9.21	-0.190	1
OTF91362	HaWSD10	458	51.51	8.41	-0.079	0
OTF91364	HaWSD11	456	50.79	8.65	-0.057	0
OTF86778	HaWSD12	475	53.16	7.11	-0.111	0

重复基因对 Duplicated gene pairs	非同义替换率 Ka	同义替换率 Ks	Ka/Ks	重复类型 Duplicated type	选择类型 Selective type	分化时间 Differentiation time (Mya)
HaDGAT1-1/HaDGAT1-2	0.1424	1.1312	0.1259	片段重复Segmental	纯化Purifying	37.70
HaDGAT1-1/HaDGAT1-3	0.1334	1.0506	0.1270	片段重复Segmental	纯化Purifying	35.02
HaDGAT1-2/HaDGAT1-3	0.0906	0.4019	0.2255	片段重复Segmental	纯化Purifying	13.39
HaWSD3/HaWSD4	0.1488	0.4223	0.3523	串联重复Tandem	纯化Purifying	14.07
HaWSD7/HaWSD8	0.1271	0.5899	0.2155	片段重复Segmental	纯化Purifying	19.66
HaWSD7/HaWSD9	0.1349	0.5677	0.2377	片段重复Segmental	纯化Purifying	18.92
HaWSD8/HaWSD9	0.0635	0.3272	0.1942	串联重复Tandem	纯化Purifying	10.90
HaWSD10/HaWSD11	0.1251	0.3299	0.3793	串联重复Tandem	纯化Purifying	10.99

Genome-wide identification and relative expression analysis of DGATs gene family in sunflower

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