Cahoon E B, Shanklin J, Ohlrogge J B
Department of Botany and Plant Pathology, Michigan State University, East Lansing 48824.
Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11184-8. doi: 10.1073/pnas.89.23.11184.
Little is known about the metabolic origin of petroselinic acid (18:1 delta 6cis), the principal fatty acid of the seed oil of most Umbelliferae, Araliaceae, and Garryaceae species. To examine the possibility that petroselinic acid is the product of an acyl-acyl carrier protein (ACP) desaturase, Western blots of coriander and other Umbelliferae seed extracts were probed with antibodies against the delta 9-stearoyl-ACP desaturase of avocado. In these extracts, proteins of 39 and 36 kDa were detected. Of these, only the 36-kDa peptide was specific to tissues which synthesize petroselinic acid. A cDNA encoding the 36-kDa peptide was isolated from a coriander endosperm cDNA library, placed under control of the cauliflower mosaic virus 35S promoter, and introduced into tobacco by Agrobacterium tumefaciens-mediated transformation. Expression of this cDNA in transgenic tobacco callus was accompanied by the accumulation of petroselinic acid and delta 4-hexadecenoic acid, both of which were absent from control callus. These results demonstrate the involvement of a 36-kDa putative acyl-ACP desaturase in the biosynthetic pathway of petroselinic acid and the ability to produce fatty acids of unusual structure in transgenic plants by the expression of the gene for this desaturase.
对于岩芹酸(18:1 Δ6顺式)的代谢起源知之甚少,岩芹酸是大多数伞形科、五加科和鞘柄木科植物种子油中的主要脂肪酸。为了研究岩芹酸是酰基 - 酰基载体蛋白(ACP)去饱和酶产物的可能性,用抗鳄梨Δ9 - 硬脂酰 - ACP去饱和酶的抗体对芫荽和其他伞形科种子提取物进行蛋白质免疫印迹分析。在这些提取物中,检测到了39 kDa和36 kDa的蛋白质。其中,只有36 kDa的肽段对合成岩芹酸的组织具有特异性。从芫荽胚乳cDNA文库中分离出编码36 kDa肽段的cDNA,将其置于花椰菜花叶病毒35S启动子的控制下,并通过根癌农杆菌介导的转化引入烟草。该cDNA在转基因烟草愈伤组织中的表达伴随着岩芹酸和Δ4 - 十六碳烯酸的积累,而对照愈伤组织中则没有这两种物质。这些结果证明了一种36 kDa的假定酰基 - ACP去饱和酶参与了岩芹酸的生物合成途径,并且通过表达该去饱和酶基因能够在转基因植物中产生结构异常的脂肪酸。