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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (1): 73-85.doi: 10.3724/SP.J.1006.2023.14217

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

ZHANG Cheng1(), ZHANG Zhan2, YANG Jia-Bao1, MENG Wan-Qiu1, ZENG Ling-Lu1, SUN Li1,*()   

  1. 1College of Life Science, Shihezi University, Shihezi 832003, Xinjiang, China
    2Bingtuan Xingxin Vocational and Technical College, Bazhou 841007, Xinjiang, China
  • Received:2021-11-23 Accepted:2022-05-05 Online:2023-01-12 Published:2022-05-24
  • Contact: SUN Li E-mail:2020002371@qq.com;sunlishz@126.com
  • Supported by:
    National Natural Science Foundation of China(31760064);National Natural Science Foundation of China(31360052)

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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

Table 1

Primers for qRT-PCR"

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

Table 2

Detailed information of HaDGATs genes in sunflower"

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

Fig. 1

Phylogenetic tree of DGAT family proteins in sunflower and other plants Ha: Helianthus annuus; At: Arabidopsis thaliana; Gm: Glycine max; Ah: Arachis hypogaea; Bn: Brassica napus; Sb: Sorghum bicolor; Os: Oryza sativa; Eg: Elaeis guineensis; Aa: Artemisia annua; Ct: Carthamus tinctorius; Zm: Zea mays."

Fig. 2

Gene structures of HaDGATs genes in sunflower"

Fig. 3

Conserved motifs of HaDGATs proteins in sunflower"

Fig. 4

Chromosome distribution (A) and cis-acting elements in sunflower HaDGATs promoters (B)"

Table 3

Gene duplication events and divergence time of HaDGATs family in sunflower"

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

Fig. 5

Collinearity analysis A: collinearity analysis of HaDGATs in sunflower; B: collinearity analysis of DGATs among sunflower, Arabidopsis, and safflower."

Fig. 6

Relative expression patterns of HaDGATs in different tissues and seed development stages of sunflower DAF: days after flowering. The relative expression levels of HaDGATs genes are compared with the relative expression in roots."

Fig. 7

Oil contents in sunflower seeds at different developmental stages Different lowercase letters indicate there are significantly different at P < 0.05."

Fig. 8

Relative expression levels of sunflower HaDGATs under salt (A) and low temperature (B) stresses"

Fig. 9

Relative expression levels of sunflower HaDGATs under ABA (A) and drought (B) treatments"

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