Characterization and Ectopic Expression ofCoWRI1, an AP2/EREBP Domain-Containing Transcription Factor from Coconut (Cocos nuciferaL.) Endosperm, Changes the Seeds Oil Content in TransgenicArabidopsis thalianaand Rice (Oryza sativaL.).
Coconut (Cocos nuciferaL.) is a key tropical crop and a member of the monocotyledonous family Arecaceae (Palmaceae). Few genes and related metabolic processes involved in coconut endosperm development have been investigated. In this study, a new member of theWRI1gene family was isolated from coconut endosperm and was namedCoWRI1. Its transcriptional activities and interactions with the acetyl-CoA carboxylase (BCCP2) promoter ofCoWRI1were confirmed by the yeast two-hybrid and yeast one-hybrid approaches, respectively. Functional characterization was carried out through seed-specific expression inArabidopsisand endosperm-specific expression in rice. In transgenicArabidopsis, high over-expressions ofCoWRI1in seven independent T2 lines were detected by quantitative real-time PCR. The relative mRNA accumulation of genes encoding enzymes involved in either fatty acid biosynthesis or triacylglycerols assembly (BCCP2, KASI, MAT, ENR, FATA, and GPDH) were also assayed in mature seeds. Furthermore, lipid and fatty acids C16:0 and C18:0 significantly increased. In two homozygous T2 transgenic rice lines (G5 and G2), differentCoWRI1expression levels were detected, but no CoWRI1 transcripts were detected in the wild type. Analyses of the seed oil content, starch content, and total protein content indicated that the two T2 transgenic lines showed a significant increase (P< 0.05) in seed oil content. The transgenic lines also showed a significant increase in starch content, whereas total protein content decreased significantly. Further analysis of the fatty acid composition revealed that palmitic acid (C16:0) and linolenic acid (C18:3) increased significantly in the seeds of the transgenic rice lines, but oleic acid (C18:1) levels significantly declined.
Department of Bioengineering, College of Material and Chemical Engineering, Hainan University Haikou, China.
RuHao
R
Sun
National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University Wuhan, China.
Rongjian
R
Ye
Department of Bioengineering, College of Material and Chemical Engineering, Hainan University Haikou, China.
Lingchao
L
Gao
Department of Bioengineering, College of Material and Chemical Engineering, Hainan University Haikou, China.
Lin
L
Zhang
National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University Wuhan, China.
Rui
R
Wang
Department of Bioengineering, College of Material and Chemical Engineering, Hainan University Haikou, China.
Ting
T
Mao
Department of Bioengineering, College of Material and Chemical Engineering, Hainan University Haikou, China.
Yusheng
Y
Zheng
Department of Bioengineering, College of Material and Chemical Engineering, Hainan University Haikou, China.
Dongdong
D
Li
National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University Wuhan, China.
Yongjun
Y
Lin
CoWRI1; coconut; endosperm; lipid; rice (Oryza sativa L.); seed-specific expression