植物脂肪酸不饱和性对植物抗寒性影响的研究

对植物脂肪酸不饱和性与植物抗寒性关系、催化不饱和脂肪酸形成的酶特性和转化膜质脱饱和酶和酰基转移酶基因提高植物抗寒性的研究进展进行了分析论述。膜是植物低温伤害的首要发生地,细胞膜的稳定性主要决定于膜质脂肪酸不饱和性,不饱和度越高,膜稳定性越强,抗寒性越强。膜质脂肪酸不饱和度主要由膜质脱饱和酶活性和酰基转移酶的特异性不同所致,目前克隆和研究了10多个与植物细胞膜质不饱和性相关的基因,通过高效表达这些基因可以明显提高冷敏感植物的抗寒性。因此转化膜脂不饱和脂肪酸合成酶基因是提高植物抗寒性的重要途径之一。通过分析近年来的研究进展认为,今后植物脂肪酸不饱和性的研究应向农业生产应用转化,同时要加强脂肪酸脱饱和酶表达调控的研究。     

Expression of a cDNA encoding palmitoyl-acyl carrier protein desaturase from cat's claw (Doxantha unguis-cati L.) in Arabidopsis thaliana and Brassica napus leads to accumulation of unusual unsaturated fatty acids and increased stearic acid content in the seed oil

A cDNA encoding palmitoyl‐acyl carrier protein (ACP)‐desaturase from cat's claw (Doxantha unguis‐cati L.) was expressed in Arabidopsis thaliana and Brassica napus L. with the goal of decreasing the saturated fatty acid (FA) content of the seed oil. In general, transformation of Arabidopsis resulted in a greater change in the FA composition of the seed oil than for B. napus. An increase in palmitoleic acid (16:1cisΔ⁹) was obtained in transgenic lines, suggesting that the 16:0‐ACP‐desaturase cDNA was expressed in the manner originally intended. Other effects on lipid metabolism, however, were observed in the seed of transgenic plants. In Arabidopsis, there was a large increase in the proportions of cis‐vaccenic acid (18:1cisΔ¹¹) and cis‐13‐eicosenoic acid (20:1cisΔ¹³), possibly generated through elongation of 16:1cisΔ⁹. Elongation of 18:1cisΔ¹¹ to 20:1cisΔ¹³, however, was not observed in B. napus indicating that certain aspects of lipid metabolism in the model plant, Arabidopsis, may not apply to B. napus. As well, the appearance of 18:1cisΔ¹¹ was accompanied by a decrease in the proportion of oleic acid (18:1cisΔ⁹). Although the introduced ACP‐desaturase resulted in synthesis of some unsaturated FAs, the overall saturated FA content was maintained at similar levels to the control or was enhanced. Increased levels of saturation were mainly associated with an increase in stearic acid, which unlike 16:0, is considered non‐atherogenic. The results suggest that a mechanism exists further downstream in oil biosynthesis to counteract the decrease in saturation brought about by the 16:0‐ACP‐desaturase action.     

甘薯zds基因的克隆与植物表达载体构建

β-胡萝卜素是维生素A原,对人体健康具有重要作用,可降低多种癌症特别是肺癌的发病率.ζ-胡萝卜素脱氢酶是β-胡萝卜素生物合成途径的关键酶之一.根据其它植物ζ-胡萝卜素脱氢酶基因(zds)的保守序列,设计简并引物,从甘薯块根总RNA逆转录的cDNA中扩增出608 bp的zds保守序列,然后用RACE技术扩增出该基因的全长cDNA.zds全长cDNA共2057 bp,有一1773 bp的开放阅读框,编码591个氨基酸,相对分子质量为65 ku.并成功构建了zds植物表达载体p2300-zds,可进一步用于提高作物β-胡萝卜素含量的遗传转化研究.     

Physiological responses, desaturase activity, and fatty acid composition in milkfish (Chanos chanos) under cold acclimation

Physiological responses of milkfish (Chanos chanos) under cold shock and acclimation were investigated. The experimental milkfish, a warm-water teleost, were initially acclimated at 25 °C and then transferred directly to 15 °C; stress responses of this species were monitored for 1 week. Parameters monitored included plasma glucose, lactate, and lipids, as well as stearoyl-CoA desaturase activity and fatty acid compositions of hepatic membranes. All parameters showed significant changes in the process of cold acclimation. A hyperglycemic response indicated by a notable and steady increase in plasma glucose levels from 85 mg dl⁻¹ to the highest level of 458.2 mg dl⁻¹ in 24 h was followed by a rapid decline thereafter. The elevation in plasma glucose content under cold shock resulted from gluconeogenesis and glycogenolysis, indicated by correlated changes in plasma glucose with fructose-1,6-biphosphatase and phosphorylase a activities. Plasma lactate concentrations remarkably increased from 47 mg dl⁻¹ on day 0 to 149.6 and 120.4 mg dl⁻¹ on days 1 and 2, respectively, and then rapidly declined to the same level as the control thereafter. In contrast, plasma lipids increased gradually from 44.8 mg dl⁻¹ to 191 mg dl⁻¹ over the 5-day acclimation period, followed by a declining trend from day 6 on. Furthermore, changes in monounsaturated fatty acids were highly correlated with those of stearoyl-CoA desaturase activities in hepatic microsomes of milkfish during cold acclimation. Results indicate that in milkfish subjected to cold stress, plasma hyperglycemic and hyperlactemic responses can be used as acute stress indicators, and plasma lipids can be used as a chronic stress indicator.     

花生中两个硬脂酰-ACP去饱和酶基因的克隆与序列分析

脂肪酸去饱和酶是不饱和脂肪酸合成途径的关键酶,催化脂肪酸链特定位置上脱氢形成双键.本研究通过构建花生种子全长cDNA文库,结合大规模EST测序和RACE克隆等方法,从花生中克隆得到了两个FAB2基因,分别命名为AhFAB2-2和AhFAB2-3.其中AhFAB2-2全长为1387bp,ORF为1158bp,理论分子量为43.7 kD,理论等电点为5.69;AhFAB2-3全长为1452bp,ORF为1188bp,理论分子量为44.9 kD,理论等电点为6.37.它们推导的氨基酸序列与拟南芥的相似性依次为60.8％和57.2％;与大豆的相似性分别是62.3％和59.3％.这两个基因在GenBank上的登录号分别为KF358459和KF358460.     

花生油酸脱氢酶基因RNAi表达载体的构建

根据克隆的花生油酸脱氢酶基因序列,选取高度保守的260 bp片段,插入植物双元表达栽体pFGC1008中,构建完成具有反向重复序列的pFGC/2nd表达栽体,利用RNAi以抑制内源目的基因表达,为获得高油酸亚油酸比值的花生种质奠定基础.     

ORIGINAL ARTICLE: Intravenous injections of cobalt reduce fatty acid desaturation products in milk and blood of lactating cows

The objective of this study was to determine whether intravenous infusion of Co affects levels of fatty acid desaturation products in bovine milk. Six cows were assigned to two replicated 3 × 3 Latin squares with 14‐day periods. Treatment occurred on days 1 to 5 and depuration occurred on days 6–14. Two treatments were tested, the first consisting of per os supplementation of 3.5 g Co daily in the form of Co acetate and the second consisting of intravenous injection of 175 mg Co daily in the form of Co acetate diluted in saline solution. The third treatment was a control. Both Co treatments decreased cis‐9 18:1 levels from approximately 18 to 14 g/100 g fatty acids, and increased 18:0 levels from 11 to 17 g/100 g fatty acids in milk fat (p < 0.001). The proportions of cis‐9 10:1, cis‐9 12:1, cis‐9 14:1, cis‐9 16:1 and cis‐9 17:1 decreased (p < 0.001), whereas 17:0 and trans‐11 18:1 increased (p < 0.001). In blood plasma, levels of cis (6,9,12) 18:3 (p < 0.001) and cis (8,11,14,17) 20:4 (p = 0.008) decreased after both the Co treatments. It is concluded that intravenous supply of Co reduces levels of fatty acid desaturation products in both bovine milk and blood.     

奶山羊SCD基因CDS区的克隆、序列分析及过表达

【目的】研究奶山羊硬脂酰辅酶A去饱和酶基因（stearoyl-CoA desaturase,SCD）在乳腺上皮细胞中对不饱和脂肪酸合成过程相关基因的调控及对细胞脂肪酸组成的影响。【方法】用RT-PCR方法从西农萨能羊乳腺组织中扩增SCD基因,进行序列分析和组织表达谱分析;构建重组腺病毒质粒,进行包装和扩增后,感染体外培养的奶山羊原代乳腺上皮细胞,实时定量检测相关基因的表达,western blotting检测蛋白变化,气相色谱-质谱联用分析细胞内脂肪酸组成变化。【结果】①获得了全长1 080 bp的山羊乳腺SCD基因CDS（GenBank登录号：GU947654）并对其进行了生物信息学分析;②奶山羊SCD基因在乳腺、肺和皮下脂肪组织中高表达,而在心脏、瘤胃和卵巢组织中的表达量较低;③成功构建了奶山羊SCD基因的重组腺病毒载体并制备出高滴度的重组病毒液,重组腺病毒感染原代乳腺上皮细胞后,检测结果表明细胞中SCD基因过表达极显著,FASN、H-FABP、LEPR和LXRα等基因表达下调,而PPARγ和LPL基因的表达量显著性增加;④棕榈酸和硬脂酸的含量下降,不饱和脂肪酸含量上升。【结论】SCD基因在奶山羊乳腺中发挥着重要作用,FASN、H-FABP、LEPR、LXRα、PPARγ和LPL基因的表达与其相关,这些基因共同影响乳脂中脂肪酸的组成。     

TaULP5 contributes to the compatible interaction of adult plant resistance wheat seedlings-stripe rust pathogen

Adult plant resistance indicates that plant is susceptible to pathogen at seedling stage, but resistant at adult stage. Understanding the mechanism of the interactions between APR wheat plants and Puccinia striiformis f. sp. tritici (Pst) is important for the creation of strategies to improve cultivar disease resistance. In this study, a full-length cDNA was isolated from APR wheat cultivar Xingzi 9104 (XZ), and was designated as ubiquitin-like protein 5 (TaULP5). TaULP5 was likely to be located in the cytoplasm, with a percentage of 75.9% Arabidopsis protoplasts number. The expression of TaULP5 was largely induced in the compatible interaction of wheat seedlings to Pst, while no obvious change was found in the incompatible interaction of wheat adult plants to Pst. Moreover, when TaULP5 was knocked down, the wheat resistance at seedling stage to Pst was improved. In addition, knockdown of TaULP5 increased the expression levels of some biotic stress-related genes, such as PR1 and PR2. It is the first time to confirm that ubiquitin-like protein could contribute to the compatible interaction of XZ to Pst, and the results will lay a foundation for understanding the mechanisms of different interactions between APR wheat plants and Pst at post-translational level.