Bryant Fiona M, Munoz-Azcarate Olaya, Kelly Amélie A, Beaudoin Frédéric, Kurup Smita, Eastmond Peter J
Department of Plant Biology and Crop Science, Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, United Kingdom (F.M.B., O.M.-A., F.B., S.K., P.J.E.); andSchool of Life Sciences, University of Warwick, Coventry, CV4 7AL, United Kingdom (A.A.K.).
Department of Plant Biology and Crop Science, Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, United Kingdom (F.M.B., O.M.-A., F.B., S.K., P.J.E.); andSchool of Life Sciences, University of Warwick, Coventry, CV4 7AL, United Kingdom (A.A.K.)
Plant Physiol. 2016 Sep;172(1):154-62. doi: 10.1104/pp.16.00836. Epub 2016 Jul 26.
Omega-7 monounsaturated fatty acids (ω-7s) are specifically enriched in the aleurone of Arabidopsis (Arabidopsis thaliana) seeds. We found significant natural variation in seed ω-7 content and used a Multiparent Advanced Generation Inter-Cross population to fine-map a major quantitative trait loci to a region containing ACYL-ACYL CARRIER PROTEIN DESATURASE1 (AAD1) and AAD3 We found that AAD3 expression is localized to the aleurone where mutants show an approximately 50% reduction in ω-7 content. By contrast, AAD1 is localized to the embryo where mutants show a small reduction in ω-9 content. Enzymatic analysis has previously shown that AAD family members possess both stearoyl- and palmitoyl-ACP Δ(9) desaturase activity, including the predominant isoform SUPPRESSOR OF SALICYLIC ACID INSENSITIVE2. However, aad3 ssi2 aleurone contained the same amount of ω-7s as aad3 Within the AAD family, AAD3 shares the highest degree of sequence similarity with AAD2 and AAD4. Mutant analysis showed that AAD2 also contributes to ω-7 production in the aleurone, and aad3 aad2 exhibits an approximately 85% reduction in ω-7s Mutant analysis also showed that FATTY ACID ELONGASE1 is required for the production of very long chain ω-7s in the aleurone. Together, these data provide genetic evidence that the ω-7 pathway proceeds via Δ(9) desaturation of palmitoyl-ACP followed by elongation of the product. Interestingly, significant variation was also identified in the ω-7 content of Brassica napus aleurone, with the highest level detected being approximately 47% of total fatty acids.
ω-7单不饱和脂肪酸(ω-7s)在拟南芥种子的糊粉层中特异性富集。我们发现种子ω-7含量存在显著的自然变异,并利用多亲代高级杂交群体将一个主要数量性状位点精细定位到包含酰基-酰基载体蛋白去饱和酶1(AAD1)和AAD3的区域。我们发现AAD3的表达定位于糊粉层,其突变体的ω-7含量降低了约50%。相比之下,AAD1定位于胚,其突变体的ω-9含量略有降低。酶学分析先前表明,AAD家族成员具有硬脂酰-和棕榈酰-ACP Δ(9)去饱和酶活性,包括水杨酸不敏感2的主要异构体。然而,aad3 ssi2糊粉层中ω-7s的含量与aad3相同。在AAD家族中,AAD3与AAD2和AAD4的序列相似性最高。突变体分析表明,AAD2也有助于糊粉层中ω-7的产生,aad3 aad2的ω-7s含量降低了约85%。突变体分析还表明,脂肪酸延长酶1是糊粉层中极长链ω-7产生所必需的。这些数据共同提供了遗传证据,表明ω-7途径是通过棕榈酰-ACP的Δ(9)去饱和作用,随后产物延长而进行的。有趣的是,在甘蓝型油菜糊粉层的ω-7含量中也发现了显著变异,检测到的最高水平约为总脂肪酸的47%。