Cloning and expression analysis of the CiDGAT1 from Carya illinoinensis

doi:10.3969/j.issn.1000-2006.201905023

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2020, Vol. 44 ›› Issue (4): 55-62.doi: 10.3969/j.issn.1000-2006.201905023

Cloning and expression analysis of the CiDGAT1 from Carya illinoinensis

ZHOU Xiaofeng¹(), MO Zhenghai¹, SHANG Yangjuan¹, LUO Min¹, SU Wenchuan¹, TAN Pengpeng¹, HAN Minghui¹, DENG Qiuju¹, TIAN Zhaoxia²(), PENG Fangren¹()

^1.Co -Innovation Center for the Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, China
^2.School of Life Sciences, University of Science and Technology of China, Hefei 230027, China

Received:2019-05-23 Revised:2020-03-07 Online:2020-07-22 Published:2020-08-13
Contact: TIAN Zhaoxia,PENG Fangren E-mail:zhxf032302@163.com;zxtian@ustc.edu.cn;frpeng@ njfu.edu.cn

Abstract

Abstract: Objective

The purpose of the study was to elucidate the structure and expression patterns of the CiDGAT1 to provide a theoretical reference for a further analysis of the function of CiDGAT1 gene.

Method

A full-length CiDGAT1 gene was isolated from the fruit of Carya illinoinensis using rapid amplification of cDNA ends with specific primers according to the conserved DGAT1 fragment sequence. The genetic structure , protein character, and evolutionary relationship were analyzed using bioinformatics. In addition, the CiDGAT1 temporal expression pattern was analyzed in different varieties using quantitative real-time PCR.

Result

CiDGAT1 has an open reading frame with 1 617 bp and encodes 538 amino acids. The gene structure analysis showed that the CiDGAT1 gene consists of 15 exons and 14 introns. The phylogenetic analysis revealed that the CiDGAT1 protein is clustered in the dicotyledonous branch with a close relationship to oak DGAT1 and filbert DGAT1. The qRT-PCR analysis showed that the expression of CiDGAT1 is higher in the growth process of ‘Pawnee’ and peaks in the mature stage at 145 d. Whereas, CiDGAT1 expression is lower during the growth process of ‘Mahan’ and ‘Jinhua’ varieties; it rapidly increased after 152 d, reaching the highest level at final maturity.

Conclusion

CiDGAT1 belongs to the MBOAT gene family, and its expression tends to increase during the growth process of the fruit of C. illinoinensis, which is a key regulator in the process of oil synthesis in plants.

Key words: Carya illinoinensis, lipids synthesis, CiDGAT1, gene cloning, gene expression patterns

CLC Number:

S664.1

ZHOU Xiaofeng, MO Zhenghai, SHANG Yangjuan, LUO Min, SU Wenchuan, TAN Pengpeng, HAN Minghui, DENG Qiuju, TIAN Zhaoxia, PENG Fangren. Cloning and expression analysis of the CiDGAT1 from Carya illinoinensis[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2020, 44(4): 55-62.

Figures/Tables 6

Table 1

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

References 30

1	ZHANG C Y，ISKANDAROV U，KLOTZ E T，et al．A thraustochytrid diacylglycerol acyltransferase 2 with broad substrate specificity strongly increases oleic acid content in engineered Arabidopsis thaliana seeds[J]．J Exp Bot，2013，64(11)：3189-3200．DOI:10.1093/jxb/ert156.
2	LUS P，STURTEVANT D，AZIZ M，et al．Spatial analysis of lipid metabolites and expressed genes reveals tissue⁃specific heterogeneity of lipid metabolism in high⁃and low⁃oil Brassicanapus L．seeds[J]．Plant J，2018，94(6)：915-932．DOI:10.1111/tpj.13959.
3	郑玲.花生二酰甘油酰基转移酶(AhDGAT)基因家族的功能与调控研究[D]．济南：山东大学，2018．
	ZHENG L.Function and regulation research of peanut diacylgycerolacyl transferase (AhDGAT) gene family[D]．Jinan：Shandong University，2018.
4	TURCHETTO⁃ZOLET A C，CHRISTOFF A P，KULCHESKI F R，et al．Diversity and evolution of plant diacylglycerolacyltransferase (DGATs) unveiled by phylogenetic，gene structure and expression analyses[J]．Genet Mol Biol，2016，39(4)：524-538．DOI:10.1590/1678-4685-GMB-2016-0024.
5	HERNÁNDEZ M L，WHITEHEAD L，HE Z S，et al. A cytosolic acyltransferase contributes to triacylglycerol synthesis in sucrose⁃rescued Arabidopsis seed oil catabolism mutants[J].Plant Physiol，2012，160(1)：215-225．DOI:10.1104/pp.112.201541．
6	LI F L，WU X M，LAM P，et al．Identification of the wax ester synthase/acyl⁃coenzyme A：diacylglycerolacyltransferase WSD1 required for stem wax ester biosynthesis in Arabidopsis[J]．Plant Physiol，2008，148(1)：97-107．DOI:10.1104/pp.108.123471.
7	ROSLI R，CHAN P L，CHAN K L，et al．In silico characterization and expression profiling of the diacylglycerolacyl transferase gene family (DGAT1，DGAT2，DGAT3 and WS/DGAT) from oil palm，Elaeisguineensis[J]．Plant Science，2018，275：84-96．DOI:10.1016/j.plantsci.2018.07.011.
8	彭方仁，李永荣，郝明灼.我国薄壳山核桃生产现状与产业化发展策略[J].林业科技开发，2012，26（4）：1-4. DOI: 10.3969/j.issn.1000-8101.2012.04.00. PENG F R,
	LI Y R, HAO M Z, et al. Production status and industrialization development strategy of pecan in China[J]. China Forestry Science and Technology, 2012, 26（4）: 1-4.
9	NEIL C E O，KEAST D R，NICKLAS T A，et al．Out⁃of⁃hand nut consumption is associated with improved nutrient intake and health risk markers in US children and adults：National Health and Nutrition Examination Survey 1999-2004[J]．Nutrition Research，2012，32(3)：185-194．DOI：10.1016/j.nutres.2012.01.005．
10	HUANG J Q，ZHANG T，ZHANG Q X，et al．The mechanism of high contents of oil and oleic acid revealed by transcriptomic and lipidomic analysis during embryogenesis in Carya cathayensis Sarg[J]．BMC Genomics，2016，17：113．DOI:10.1186/s12864-016-2434-7.
11	李晖，张瑞，彭方仁，等．美国山核桃种质资源遗传多样性ISSR分析[J]．南京林业大学学报(自然科学版)，2015，39(4)：7-12．
	LI H，ZHANG R，PENG F R，et al．Genetic diversity and identification of pecan by ISSR markers[J]．J Nanjing For Univ （Nat Sci Ed），2015，39(4)：7-12．DOI:10.3969/j.issn.1000-2006.2015.04.002.
12	周丹，赵江哲，柏杨，等．植物油脂合成代谢及调控的研究进展[J]．南京农业大学学报，2012，35(5)：77-86．
	ZHOU D，ZHAO J Z，BAI Y，et al．Research advance in triacylglycerol synthesis，metabolism，and regulation in plants[J]．J Nanjing Agric Univ，2012，35(5)：77-86．
13	ZABOROWSKA Z，STARZYCKI M，FEMIAK I，et al．Yellow lupine gene encoding stearoyl⁃ACP desaturase：organization，expression and potential application[J]．Acta Biochim Pol，2002，49(1)：29-42．DOI:10.18388/abp.2002_3818.
14	ZOU J，WEI Y，JAKO C，et al．The Arabidopsis thaliana TAG1 mutant has a mutation in a diacylglycerolacyltransferase gene[J]．Plant J，1999，19(6)：645-653．DOI:10.1046/j.1365-313x.1999.00555.x.
15	刘凯，梁国校，杨麒，等．油桐DGAT1基因在不同品种及种子发育阶段的表达模式[J]．中南林业科技大学学报，2014，34(10)：61-64．
	LIU K，LIANG G X，YANG Q，et al．Expression patterns of DGAT1 gene in different varieties and seeds developmental stages of Vernicia fordii[J]．J Central South Univ For Technol，2014，34(10)：61-64．DOI:10.14067/j.cnki.1673-923x.2014.10.030.
16	HE X H，TURNER C，CHEN G Q，et al．Cloning and characterization of a cDNA encoding diacylglycerolacyltransferase from Castor bean[J]．Lipids，2004，39(4)：311-318．DOI：10.1007/s11745-004-1234-2.
17	NYKIFORUK C．Characterization of cDNAs encoding diacylglycerolacyltransferase from cultures of Brassica napus and sucrose⁃mediated induction of enzyme biosynthesis[J]．Biochimicaet Biophysica Acta⁃Molecular and Cell Biology Lipids，2002，1580(2/3)：95-109．DOI：10.1016/s1388-1981(01)00200-1.
18	NYKIFORUK C L，FURUKAWA⁃STOFFER T L，HUFF P W，et al．Characterization of cDNAs encoding diacylglycerolacyltransferase from cultures of Brassicanapus and sucrose⁃mediated induction of enzyme biosynthesis[J].BBA⁃Mol Cell Biol Lipids，2002，1580(2/3)：95-109．DOI:10.1016/S1388-1981(01)00200-1.
19	CHI X Y，HU R B，ZHANG X W，et al．Cloning and functional analysis of three diacylglycerolacyltransferase genes from peanut (Arachishy pogaea L．)[J].PLoS One，2014，9(9)：e105834．DOI:10.1371/journal.pone.0105834.
20	ZHENG P Z，ALLEN W B，ROESLER K，et al．A phenylalanine in DGAT is a key determinant of oil content and composition in maize[J]．Nat Genet，2008，40(3)：367-372．DOI:10.1038/ng.85.
21	朱红霞，胡利宗，邓小莉，等．三种豆科植物DGAT1基因家族的分子特征与进化分析[J]．生物技术通报，2011(10)：163-166．
	ZHU H X，HU L Z，DENG X L，et al．Molecular characterization and evolutionary analysis of DGAT1 gene family in three Fabaceae plants[J]．Biotechnol Bull，2011(10)：163-166． DOI:10.13560/j.cnki.biotech.bull.1985.2011.10.034.
22	LU C L，DE NOYER S B，HOBBS D H，et al．Expression pattern of diacylglycerol acyltransferase⁃1，an enzyme involved in triacylglycerol biosynthesis，in Arabidopsis thaliana[J]．Plant Mol Biol，2003，52(1)：31-41.DOI:10.1023/a:1023935605864.
23	陈义挺，冯新，赖瑞联，等．猕猴桃AdCCS1基因的克隆及其表达调控[J]．园艺学报，2018，45(12)：2371-2382．
	CHEN Y T，FENG X，LAI R L，et al.Cloning and expression regulation of the AdCCS1 from Actinidia chinensis var.deliciosa[J]．Acta Hortic Sin，2018，45(12)：2371-2382．DOI:10.16420/j.issn.0513-353x.2018-0433.
24	刘翔如. 薄壳山核桃果实发育及脂肪酸累积规律的研究[D]. 南京：南京林业大学， 2013.
	LIU X R. The study of pecan fruit development and the law of fatty acid accumulation[D]. Nanjing： Nanjing Forestry University, 2013.
25	卢翔，李效文，夏海涛，等．薄壳山核桃胚发育过程中主要营养成分的变化[J]．林业科技，2018，43(3)：40-44．
	LU X，LI X W，XIA H T，et al．Embryonic developmental and main nutritional components of Carya illinoinensis[J]．For Sci Technol，2018，43(3)：40-44．
26	张玲慧．紫苏种子油脂代谢及限速酶DGAT1基因的表达分析[D]．太谷：山西农业大学，2014．ZHANG L H，Study on the lipid metabolism in seeds and expression pattern of the rate⁃limiting enzyme gene DGAT1 of Perilla[D]．Taigu：Shanxi Agricultural University，2014.
27	TAYLOR D C，ZHANG Y，KUMAR A，et al. Molecular modification of triacylglycerol accumulation by over⁃expression of DGAT1 to produce canola with increased seed oil content under field conditions[J]. Botany，2009，87(6): 533–543. DOI：10.1139/B08-101.
28	蓝碧秀，吴子恺，王凛，等. DGAT1-2基因在超高油玉米中的转化、表达及对油脂含量的影响[J]. 华南农业大学学报，2015，36(5): 43-47.
	LAN B X,WU Z K，WANG L，et al. Transformation and expression of DGAT1⁃2 gene and its effect on the oil content of super⁃high oil corn[J]. Journal of South China Agricultural University，2015，36(5): 43-47．DOI: 671/j.issn.1001-411X.2015.05.008.
29	张飞，高秀清，张靖洁，等．种子特异表达异源DGAT1基因提高大豆种子含油量和营养品质[J]．生物工程学报，2018，34(9)：1478-1490．
	ZHANG F，GAO X Q，ZHANG J J，et al．Seed⁃specific expression of heterologous gene DGAT1 increase soybean seed oil content and nutritional quality[J]．Chin J Biotechnol，2018，34(9)：1478-1490． DOI:10.13345/j.cjb.180236.
30	LU C F，HILLS M J．Arabidopsis mutants deficient in diacylglycerolacyltransferase display increased sensitivity to abscisic acid，sugars，and osmotic stress during germination and seedling development[J]．Plant Physiol，2002，129(3)：1352-1358．DOI:10.1104/pp.006122.

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引物 primers	引物序列5'→3' sequences	作用function
3'RACE	ATACGACTCACTATAGGGTTGTAAAACGACGGC CAGTGTTTTTTTTTTTTTTTTTT	反转3'RACE使用的cDNA
GSP1	ATTTGGCGTTGTGTTGATGC	3'RACE巢式PCR第1轮PCR
T7	TAATACGACTCACTATAGGG	3'RACE巢式PCR第1轮PCR
GSP2	ATGACATGAGAGCACTTACC	3'RACE巢式PCR第2轮PCR
M13	GTAAAACGACGGCCAGTG	3'RACE巢式PCR第2轮PCR
GSP3	TCTGTATTCTGGGTCAACCC	3'RACE巢式PCR第3轮PCR，克隆到PolyA
CiDGAT1-F	CCCAAGCTTTGCAATGGCGATTTCGGATATG	开放读码框（ORF）
CiDGAT1-R	CGCGGATCCTGATAGTTTATTCAGTCTGCCC	开放读码框（ORF）

Cloning and expression analysis of the CiDGAT1 from Carya illinoinensis

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