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

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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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