Thambugala Dinushika, Cloutier Sylvie
Department of Plant Science, University of Manitoba, 66 Dafoe Rd, Winnipeg, MB, Canada, R3T 2N2.
J Appl Genet. 2014 Nov;55(4):423-32. doi: 10.1007/s13353-014-0222-0. Epub 2014 May 29.
Little is known about the relationship between expression levels of fatty acid desaturase genes during seed development and fatty acid (FA) composition in flax. In the present study, we looked at promoter structural variations of six FA desaturase genes and their relative expression throughout seed development. Computational analysis of the nucleotide sequences of the sad1, sad2, fad2a, fad2b, fad3a and fad3b promoters showed several basic transcriptional elements including CAAT and TATA boxes, and several putative target-binding sites for transcription factors, which have been reported to be involved in the regulation of lipid metabolism. Using semi-quantitative reverse transcriptase PCR, the expression patterns throughout seed development of the six FA desaturase genes were measured in six flax genotypes that differed for FA composition but that carried the same desaturase isoforms. FA composition data were determined by phenotyping the field grown genotypes over four years in two environments. All six genes displayed a bell-shaped pattern of expression peaking at 20 or 24 days after anthesis. Sad2 was the most highly expressed. The expression of all six desaturase genes did not differ significantly between genotypes (P = 0.1400), hence there were no correlations between FA desaturase gene expression and variations in FA composition in relatively low, intermediate and high linolenic acid genotypes expressing identical isoforms for all six desaturases. These results provide further clues towards understanding the genetic factors responsible for FA composition in flax.
关于亚麻种子发育过程中脂肪酸去饱和酶基因的表达水平与脂肪酸(FA)组成之间的关系,目前所知甚少。在本研究中,我们研究了六个脂肪酸去饱和酶基因的启动子结构变异及其在种子发育过程中的相对表达。对sad1、sad2、fad2a、fad2b、fad3a和fad3b启动子的核苷酸序列进行计算分析,发现了几个基本转录元件,包括CAAT盒和TATA盒,以及几个转录因子的假定靶结合位点,据报道这些位点参与脂质代谢的调控。使用半定量逆转录PCR,在六种亚麻基因型中测量了六个脂肪酸去饱和酶基因在种子发育过程中的表达模式,这些基因型的脂肪酸组成不同,但携带相同的去饱和酶同工型。通过对在两种环境中四年田间种植的基因型进行表型分析来确定脂肪酸组成数据。所有六个基因均呈现出钟形表达模式,在开花后20或24天达到峰值。Sad2表达量最高。六个去饱和酶基因在不同基因型之间的表达没有显著差异(P = 0.1400),因此,对于所有六种去饱和酶表达相同同工型的低、中、高亚麻酸基因型,脂肪酸去饱和酶基因表达与脂肪酸组成变化之间没有相关性。这些结果为理解亚麻脂肪酸组成的遗传因素提供了进一步的线索。