植物表皮蜡质相关文献阅读

植物 表 关于同志近三年现实表现材料材料类招标技术评分表图表与交易pdf视力表打印pdf用图表说话 pdf 皮蜡质相关文献阅读 蜡质相关文献阅读 (1) (2) 菊科千里光蜡质合成相关蛋白 [The major cuticular components have been shown to be synthesized in the epidermis. Therefore, cloning of epidermis-specific genes could yield information to be used to isolate and characterize the enzymes involved in the cuticle biosynthesis. A subtractive cDNA library was prepared from Senecio odorus in which epidermis-specific cDNAs were enriched. Differential screening of the library using epidermal and non-epidermal probes revealed two cDNAs. One of them designated epi425 was identified, based on the sequence homology, as a member of a new class in the LTP gene family and the other clone designated epi23 as a gene encoding an aldehyde decarbonylase. Northern blot analyses showed that epi425 and epi23 cDNAs hybridized with a transcript of about 600 and 2,100 nucleotides, respectively, from the epidermis but not from the non-epidermal tissues. Further characterization of these clones will provide more information on the mechanism of the cuticle biosynthesis.](3) Senecio odorus lipid transfer protein mRNA, 3' end GenBank: L33792.1 lipid transfer protein [Senecio odorus] GenBank: AAA33934.1 千里光多年生草本。茎木质细长，高约2,5米，曲折呈攀援状，上部多分枝， 有脱落性的毛。 叶互生;椭圆状三角形，或卵状披针形，长7,10厘米，宽3.5,4.5厘米，先 端渐尖，基部戟形至截形，边缘具不规则缺刻状的齿牙，或呈微波状，或近于全 缘，有时基部稍有深裂，两面均有细软毛。 头状花序顶生，排列成伞房花序状，头状花序径约1厘米;总苞圆筒形，苞片 10,12片，披针形或狭椭圆形，长5,6毫米，宽2毫米，先端尖，无毛或 少 有细毛;周围舌状花黄色，雌性，约8朵，长约9毫米，宽约2毫米，先端3 齿裂;中央管状花，黄色，两性，长约6.5毫米，先端5裂。瘦果圆筒形，长约 3 毫米，有细毛;冠毛长约7毫米，白色。 花期10月到翌年3月。果期2,5月。 生于路旁及旷野间。 可否从这个蛋白入手? 测蜡质的方法: , 3.1. Collection of epicuticular wax from raspberry plants, sample preparation and analysis by GC and GC?MS and conduct of biosassay with aphids Details of plant growth, sample collection, analytical instrumentation, chromatographic conditions and ana-lytical methodology for chemical analysis of the wax by GC and GC?MS, are given in the preceding paper and by Shepherd et al. (1995a).(4) , , Surface wax from the top 3 cm of bolting stems, following removal of buds, flowers and siliques, was extracted in hexane for 30 s. Total [intracellular and cuticular] wax components were extracted from epidermal peels by immersion of epidermal peels in hexane. To all extracts, 10 μg of 17:1 FAME internal standard were added and the extracts were then dried under a stream of nitrogen gas. N,Obis (trimethylsilyl) trifluoroacetamide with 1% trimethylchlorosilane was added and derivatization of samples proceeded at 80oC for 90 min, followed by analysis on an HP6890 GC system. The GC column was an HP-5 (30 m length, 0.32 mm capillary diameter) with helium as carrier gas. The wax program used an initial temperature of 140oC, increasing at 4oC min-1 to 320oC where it remained for 10 min. Wax components were identified by retention times, compared to known standards, and quantified based on flame ionization detector peak areas, compared to the internal standard. Wax loads were expressed per unit surface area, which was calculated from stem diameters (measured microscopically on free hand sections) and 3 cm length. Results were confirmed by independent GC and GC-MS analyses using the methods as outlined in R. Jetter, S.Schäffer Plant Physiol. 126, 1725 (2001)(5). Analysis of CuticularWax Composition and Loads Cuticular waxes were extracted from the leaves (200 to 1000 mg) andstems (200 mg) of 4-week-old plants in chloroform for 30 s at room temperature. n-Octacosane, docosanoic acid, and 1-tricosanol were added to the extracted chloroform solvent as internal standards. The solvent was subsequently evaporated under a gentle stream of nitrogen and redissolved in a mixture of 100 mL of pyridine and 100 mL of bis-N,N-(trimethylsilyl)trifluoroacetamide. The wax mixtures were heated at 908C for 30 min to convert waxes into trimethylsilyl derivatives. Qualitative and quantitative composition analyses were conducted as described previously (Lee et al., 2009a, 2009b). The P values from each comparisonwere corrected for multiple tests using FDR control (Benjamini and Hochberg,1995). Analysis of Cutin Polyester Monomers Rosette leaves of 4-week-old plants grown in soil were used to quantify cutin polyester monomers. Methyl heptadecanoate and v-pentadecalactone were added as internal standards into the delipidated and dried leaves and then depolymerized by hydrogenolysis with LiAlH4 or by methanolysis with NaOCH3. Cutin polyesters were analyzed by gaschromatography–mass spectrometry (GCMS-QP2010; Shimazu) with aHP-5 column (60 m, 0.32 mm inner diameter, film thickness 0.1 mm;Agilent). The analysis system was maintained at 1108C. The temperature was increased to 3008C at a rate of 2.58Cmin21 and maintained at 3008C for 3 min. [The MYB96 Transcription Factor Regulates Cuticular Wax Biosynthesis under Drought] 蜡质成分分析: GC-MS[气象色谱-质谱联用技术] 取第二至三片全展叶叶片，计算其表面积后，立即进行蜡质的提取。将叶片置于室温状态下 30mL氯仿中，30s后取出，再将处理后的叶片置于60?氯仿中萃取30s，将两次萃取的氯仿合并 后，用氮吹仪吹干氯仿，称取蜡质质量，计算单位面积蜡质含量。在取样品中加入5μg正二十四 烷作为内参，然后将样品转入GC瓶，加入10μl BSTFA和10μl吡啶，70?衍生1小时。液氮吹去 BSTFA和吡啶。每个样品加200μl氯仿溶解。GC-MS分析。蜡质组分用通用型VF-17MS毛细管 柱，规格30m×0.25mm×0.25μm,GC-MS仪为GC–MS-QP2010。进样量1μl,载气为氦气，柱流 速为2ml/min恒流。进样温度为280?，50?保持2min后，以40?/min升温至200?，保持2min; 再以3?/min升温到320?，该温度保持25min。蜡质组分可以通过离子峰出峰时间，从质谱库中 检索判定蜡质成分。蜡质含量依据峰面积与内参比较进行计算。单位内蜡质含量依据叶表蜡质抽 提面积进行计算。(6) 蜡质合成路径: Pathway was created on Thu Jun 2, 2011. Contributed by aracyc: Above-ground epidermal surfaces of vascular plants are covered by a lipophilic layer known as the cuticle. Plant cuticles are composed of cutin (cutin biosynthesis) and cuticular wax. The major components of cuticular wax are very long chain fatty acids (chain length is greater than 18 carbon) and very long chain fatty acid derived aldehydes, alkanes, secondary alcohols, ketones, primary alcohols, and wax esters. The composition of cuticular wax varies among species and even within species in different tissues and at different developmental stages.【成分:长链脂肪酸，醛，烷烃，二级醇，酮，一级醇，蜡酯。】 Parts of this pathway occur in : cytosol细胞质中 nucleus细胞核中 Symbol colors - regulation Legend: Symbols cytosol enzymatic gene or metabolite nucleus composition RNA Interaction types other + regulation protein (complex) 蜡质合成相关基因: (7) (8) compounds (Aarts et al. 1995; Chen et al. 2003; Fiebig et al.2000; Hansen et al. 1997; Millar et al. 1999; Negruk et al. 1996; Pruitt et al. 2000; St-Pierre et al. 1998; Toddet al. 1999; Xia et al. 1996, 1997; Xu et al. 1997; Zhanget al. 2005), whereas CER3, GL2, GL15, and WIN1/SHN1encode regulatory proteins (Aharoni et al. 2004; Broun et al.2004; Hannoufa et al. 1996; Moose and Sisco 1996; Tackeet al. 1995). Mutation in most of these genes showed alteredwax accumulation (Jenks et al. 2002). Co-suppression ofsome of the genes in Arabidopsis resulted in reduced wax onstems (Millar et al. 1999; Todd et al. 1999), and overexpressionof some of these genes in the Arabidopsismutants complemented the mutant phenotypes (Fiebig et al.2000; Hannoufa et al. 1996). However, only a few reportsdiscuss the effect of over-expression of these genes in thewild-type background. Over-expression of the condensingenzyme gene CER6/CUT1 under the control of theCaMV35S promoter failed to promote wax deposition(Millar et al. 1999), whereas under the control of the epidermis-specific CER6 promoter, CER6/CUT1 overexpressionled to increased wax load in stems of Arabidopsis(Hooker et al. 2002). The only report on increased wax accumulation in leaf tissues of Arabidopsis was on theover-expression of AP2/EREBP transcriptional activator(Aharoni et al. 2004; Broun et al. 2004). Over-expression of WXP1 under the control of the CaMV35S promoter led toincreased cuticular wax loading on the leaf surfaces, reducedwater loss, and enhanced drought tolerance of transgenicalfalfa (Zhang et al. 2005). Transgenic expression of WXP1or of its paralog WXP2 in Arabidopsis also leads to increased wax deposition and enhanced drought tolerance(Zhang et al. 2007). CER1; 一些可能是蜡质合成的调节基因, 如CER3。CER6 是目前唯一研究得较为清楚且功能明确的蜡质基因, 它是延长C24 超长链脂肪酸必需的基因。CER6 在拟南芥整个生育期都有很高的表达量, 而且仅在表皮细胞中 表达, 唯一例外的是将要成熟的花粉中CER6 的mRNA 是在绒毡层中表达的。一些植株CER6 的过量表达导致拟 南芥茎表皮蜡质含量增加, 所以可以判断CER6 的表达水平是拟南芥茎表皮蜡质积累的控制因素之一。而在 WIN1 过表的植株中, 这些蜡质合成基因被诱导, 其中CER1 的变化最明显, KCS1 和CER2 也显著增加。(9) 参考文献: 1. Sturaro M, et al. (2005) Cloning and characterization of GLOSSY1, a maize gene involved in cuticle membrane and wax production. Plant Physiology 138(1):478. 2. 向建华 (2006) 水稻 WAX2 同源基因的克隆及遗传转化的研究. (湖南农业大学硕士学位论文). 3. Pyee J (1996) Cloning of epidermis-specific cDNAs encoding a lipid transfer protein and an aldehyde decarbonylase from Senecio odorus. Journal of Plant Biology (Korea Republic). 4. Shepherd T, Robertson G, Griffiths D, & Birch A (1999) Epicuticular wax ester and triacylglycerol composition in relation to aphid infestation and resistance in red raspberry (Rubus idaeus L.). Phytochemistry 52(7):1255-1267. 5. Pighin JA, et al. (2004) Plant cuticular lipid export requires an ABC transporter. Science 306(5696):702. 6. 王友华 (2010) 水稻 ERF 转录激活子 DRF2 调控叶表蜡质合成. (中国农业科学院). 7. Samuels L, Kunst L, & Jetter R (2008) Sealing plant surfaces: cuticular wax formation by epidermal cells. Plant Biology 59(1):683. 8. Islam MA, Du H, Ning J, Ye H, & Xiong L (2009) Characterization of Glossy1-homologous genes in rice involved in leaf wax accumulation and drought resistance. Plant Molecular Biology 70(4):443-456. 9. 李法莲, 张淼, 朱彩霞, & 邵群 (2008) 植物表皮蜡质的研究进展及 WIN1 对植物表皮蜡质的影响. 现代农业科 11. 技 10. Zhang JY, Broeckling CD, Sumner LW, & Wang ZY (2007) Heterologous expression of two Medicago truncatula putative ERF transcription factor genes, WXP1 and WXP2, in Arabidopsis led to increased leaf wax accumulation and improved drought tolerance, but differential response in freezing tolerance. Plant Molecular Biology 64(3):265-278. 11. Leide J, Hildebrandt U, Reussing K, Riederer M, & Vogg G (2007) The developmental pattern of tomato fruit wax accumulation and its impact on cuticular transpiration barrier properties: Effects of a deficiency in a beta-ketoacyl-coenzyme A synthase (LeCER6). Plant Physiology 144(3):1667-1679. 12. Bergman D, Dillwith J, Zarrabi A, Caddel J, & Berberet R (1991) Epicuticular lipids of alfalfa relative to its susceptibility to spotted alfalfa aphids (Homoptera: Aphididae). Environmental entomology 20(3):781-785. Here we report the functional characterization of two putative ERF transcription factor genes WXP1and its paralog WXP2 from Medicago truncatula. Transgenic expression of WXP1 and WXP2 in Arabidopsis (ecotype Columbia) led to significantly increased cuticular wax deposition on leaves of 4-week-old and 6-week-old transgenic plants, assessed based on fresh weight or based on surface area. Both WXP1 and WXP2 transgenic plants showed significantly enhanced whole plant drought tolerance. the WXP1 plants had increased freezing tolerance while the WXP2 plants were more sensitive to low temperature when compared to the control.(10) 候选基因:CER6(11) WSD1 WSD1 384bp BLAST结果:相似性56% ACCGGGACACCTGCAATCATGAACTCCTCTTTCGGGCCAACCACATTTGAGAATGTGAAACTAGTATTACATACTATCCTGTAATTAAGCAAGCTTGCAAANNNNNNACTTTCCTCCTAATGTCTTAGTAACAAAGTATGCGAGTTTGTATACCAAAAAAGCCTCACAAGAAAGCTTCTTTTTATCGATCATCGTTTTGGCTCTCTTCAAATACTGCAGAGGATCGGAGCCTTTTTTATGGTAGTATACTGGTAATAGCATCATACCAAATTTGTTTCCCCACTTTGATTTAGAATTTTTCTTCATTAAT TCTTCGATTTCCTGGAGTCCTAATGATGGTCTCAGGTTAACCATGGCTGCACCAGTGATCCGAAGCCCTTCTTTAAACGCCATTGAAAGGTTCACTAGGTGTCGAGAATAGGCCTAGGCGTTTTGTTCGTAGAAGTGTAAGCTTAGCCGATATTAAGGCACTGAAGAATGCTATGAATGTGACCTTGAACGACGTTGTGCTTGGAGTTACTCAGGCAGGTTTATCGTGCTACCTGAATCGCAGATACAGTAAAATCAGAGGACTAGACTATAACAGTAAAAAGGATGTTATCCCCAAAAACATTCGTCTTCGTGCCACGTTCTTTTTCAACCTAAGAGCAACCACCAGGATTGACACTCTTGTCGAAACAATGAAAACTGGAAAAATGGGTCAATGGGGCAACAAGATCGGGTATGTGCTTCTCCCATTTACAATCGGACTCAAAAGCAACCCCTTAGACTACGTAAAAGAAGCTAAGGCAGTCATTGATCGAAAGAAAGCATCGTTGGAACCTCTGTACACTTATTTTGTTCTTTACGTGGTCCTTAAGCTGTTCGGGATTAAGGCTGCAGGGAAACTAAACCACAAGG 【58%】 蜡质与抗蚜虫 小红莓(4)紫花苜蓿(12)豌豆(13)