UV-B辐射增强对拟南芥表皮蜡质的影响

以野生型拟南芥、蜡质不同程度缺失突变体CER1、CER3、CER4、CER6、CER10、CER20及KCS1为试验材料,通过施加50μW/cm2、长达10 d的UV-B辐射,研究了拟南芥表皮蜡质晶体结构、组分及蜡质基因对UV-B辐射的响应机制。结果表明:UVB辐射增强改变了拟南芥表皮蜡质晶体结构,表皮蜡质松针状(CER1)、柱状、杆状(CER3、CER10与KCS1)晶体结构显著减少,球状蜡质晶体类型出现在CER6表面,无规则片状、膜状结构覆盖在KCS1与CER10茎表面。野生型拟南芥蜡质晶体结构类型无明显变化,但在部分区域积累了大量水平杆状、管状结构,增加了蜡质层厚度。UV-B辐射增强也改变了拟南芥表皮蜡质组分的分泌量。野生型在UV-B处理后一级醇、酸、醛含量显著上升,烷、次级醇及酮含量显著下降,蜡质总量增加不显著。一级醇含量的增加及酮和次级醇含量的减少在拟南芥各材料响应UV-B辐射中具有普遍性。UV-B辐射增强诱导了野生型CER3、CER4、KCS1基因表达的上调,其中CER4大量表达,促进了蜡质组分中一级醇、酸和醛含量的积累;CER1在UV-B处理后表达量下调,可能导致烷合成下游分支途径相关产物(烷类、次级醇及酮类)的减少。WIN1表达量的下调对蜡质总量没有显著影响。UV-B辐射增强使蜡质前体从烷合成分支途径更多地转向一级醇分支途径。     

甘蓝型油菜叶表皮蜡质组分及结构与菌核病抗性关系

本试验选用6个抗病性不同的甘蓝型油莱品种,研究其叶表皮蜡质组成及结构与菌核病抗性的关系。结果表明,抗病品种在去除叶表皮蜡质后病情指数显著增加;感病品种无显著变化。不同抗性品种（系）间除酯类组分含量无显著差异外,其余蜡质组分含量差异显著。相关分析表明,蜡质组分中酯类含量与病情指数呈显著负相关关系,醇类、酮类含量与病情指数呈显著正相关,其余组分和蜡质总量与病情指数无显著相关关系。抗性品种叶表皮蜡质中烷类及酯类所占比重较高,而易感品种酮类比重较高。扫描电镜结果显示,抗病品种（系）的蜡质晶体主要为颗粒状、杆状、丝状;而感病品种（系）的蜡质晶体中不规则片状晶体所占比例较大。这些结果说明油菜叶表皮蜡质的组分及结构可能是抗病品种抵抗和延迟病原菌侵入的机制之一。     

空气湿度与土壤水分胁迫对紫花苜蓿叶表皮蜡质特性的影响

选用2个抗旱性不同的紫花苜蓿品种,敖汉(强抗旱)和三得利(弱抗旱),设置空气湿度(45%-55%和75%-85%)和土壤水分胁迫(75%和35%田间持水量)处理,分析紫花苜蓿叶表皮蜡质含量、组分及晶体结构、气体交换参数、水势及脯氨酸含量的变化规律。结果表明,单独土壤水分胁迫时,紫花苜蓿叶表皮蜡质晶体结构及蜡质总量无显著变化;敖汉蜡质组分中烷类、酯类含量增加,醇类含量下降;三得利醇类含量下降,烷类、酯类含量变化不显著。低空气湿度胁迫时,两品种蜡质总量无显著变化,烷类和酯类含量显著增加,醇类含量显著下降,叶表皮片状蜡质晶体结构熔融呈弥漫性,扩大了对叶表面积的覆盖,其蒸腾速率显著低于正常湿度。复合胁迫处理时,叶表皮片状蜡质晶体结构继续呈弥漫性,烷类、酯类、未知蜡质组分含量均高于单独胁迫处理,醇类含量最低,而蜡质总量除三得利显著高于对照外,其余均无显著差异。紫花苜蓿叶表皮蜡质各组分含量(除醇类)及蜡质总量与光合速率呈显著负相关,与蒸腾速率无显著相关关系。蜡质总量与叶水势呈显著正相关。总体上,敖汉蜡质总量显著高于三得利,蜡质组分中烷类物质的增加有助于提高植株的抗旱性。在复合胁迫下,强抗旱品种主要通过气孔因素控制水分散失,而弱抗旱品种通过气孔和非气孔因素共同控制植物水分散失。     

小麦品系CP98(11)不同器官表皮蜡质组分和蜡质晶体结构的差异分析

为明确小麦不同器官表皮蜡质晶体结构和蜡质组分的差异,该研究以小麦品系CP98(11)为材料,在小麦扬花期分别取小麦的旗叶、叶鞘、穗下茎、花药和颖壳,利用气相色谱技术对各器官表皮蜡质组分进行鉴定,并通过扫描电镜观察其蜡质晶体结构。结果表明:(1)小麦不同器官的蜡质成分共鉴定出30种,主要为初级醇、二酮、烷烃、脂肪醛、脂肪酸、酯。(2)叶鞘、穗下茎、颖壳的蜡质中二酮含量最高,分别占蜡质总量的78.96%、67.03%和68.6%;花药的蜡质中烷烃含量最高(75.82%);旗叶的蜡质中初级醇含量最高(45.91%),其次为烷烃33.19%。(3)扫描电镜观察显示,旗叶正面的蜡质晶体呈片状结构,旗叶反面和颖壳的蜡质晶体结构呈片状与柱状混合的结构,花药的蜡质呈明显的波浪状结构,穗下茎和叶鞘的蜡质晶体呈柱状结构。     

拟南芥雄性不育突变体ms214的遗传与功能分析

经过EMS诱变、背景纯化以及遗传分析,得到一株隐性核基因控制的拟南芥雄性不育突变体ms214,用图位克隆的方法将突变基因MS214定位于拟南芥第一条染色体上顶端700kb的区间内。生物信息学分析发现,该区间内有一个与蜡质合成有关的基因CERl;测序分析表明在突变体ms214中,CERl基因第一个外显子上碱基由C^509变成了u^509的突变,导致CER蛋白在该处的氨基酸由脯氨酸^170变成了亮氨酸^170;等位实验结果表明ms214和cerl是等位突变体。ms214突变体的茎和果荚呈现出与野生型不同的亮绿色;组织切片观察结果表明,突变体花药发育各个时期无异常变化;扫描电镜观察发现ms214的茎和果荚的表皮没有蜡质的形成,但是突变体成熟花粉粒表面含油层异常,具有许多小的脂肪小滴。这些结果揭示了MS214蛋白质参与蜡质合成过程,而且脯氨酸^170是该蛋白质行使功能所必需的。     

高粱表皮蜡质缺失突变体sb1抗旱生理生化分析

植物抗旱性与体表蜡质积累有关，高粱是抗旱性极强的作物，茎秆和叶片表面覆盖一层厚厚的蜡质，开展高粱体表蜡质层对高粱抗旱能力的研究，为高粱抗旱品种选育和抗旱分子机制提供理论依据。以高粱品种BTx623和表皮蜡质缺失突变体sb1为试验材料，统计农艺性状以及扫描电镜观察叶片表面蜡质形态结构；分析表皮蜡质的有无对离体叶片失水速率和叶绿素浸提率的影响；检测在干旱胁迫处理下植株的表型变化以及超氧化物歧化酶（SOD）、过氧化物酶（POD）的活性变化。结果表明，与对照BTx623相比，表皮蜡质缺失突变体sb1株高明显降低、抽穗开花期延后1周、叶片远轴面片状蜡质缺失，其他农艺性状变化不明显。突变体sb1离体叶片失水速率和叶绿素浸提率显著高于对照。在干旱胁迫处理条件下，随着干旱胁迫时间的增长，对照BTx623和突变体sb1的SOD和POD活性均增强，但对照SOD和POD活性均显著高于突变体sb1，且干旱处理96 h和复水后突变体sb1都造成叶片大面积干枯。突变体sb1表皮蜡质缺失后叶片渗透性增强，清除活性氧的能力减弱，抗旱性降低。     

植物蜡质及其与环境的关系

陆生植物的地上部分如叶、茎、花、果实等的表面覆盖着一层蜡质,它是由一系列复杂化合物组成的具有三维微结构的疏水层,在植物生长和发育过程中起着不可或缺的作用,具有很好的生物学功能。作为植物与环境的第一接触面,蜡质对外界环境因子的响应较敏感,当植物受到外界不利环境因子胁迫时,蜡质会改变自身晶体结构形态或化学组分构建防御机制以减少胁迫因子的作用,有效地协调植物与环境的关系。综述了近年来国内外关于植物蜡质的研究进展,在阐述蜡质层结构及其化学组分的基础上,着重介绍植物与环境因子的作用,包括非生物环境因子如水分、温度、光照、环境污染等以及植食性昆虫和病原菌等生物环境因子的作用。研究显示,胁迫环境下植物蜡质化学组分的变化,是由于不利环境因子的作用足以改变蜡质各产物的合成途径,从而影响蜡质产物。植物蜡质利用各种生理、化学机制对胁迫环境因子的适应以及响应,是植物适应各种生境的基础,因此通过对植物蜡质与环境关系的研究为进一步解析植物与环境关系提供证据。     

宁夏枸杞果皮蜡质微形态及蜡质组分研究北大核心CSCD

果实制干是宁夏枸杞炮制的主要过程,宁夏枸杞不同品种在制干特性方面存在一定的差异。该研究以宁夏枸杞栽培中制干差异较大的品种‘宁杞1号’(易制干)和‘宁杞5号’(不易制干)果实为材料,采用扫描电镜技术和GC-MS技术对2个宁夏枸杞品种不同发育时期(青果期、色变期、成熟期)的果实果皮结构以及果皮蜡质微形态、含量和组分进行了观测,从果皮蜡质微形态及组分的积累变化初步揭示枸杞果皮蜡质的积累规律,以明确不同枸杞品种果皮蜡质组分差异,为不同品种适宜促干剂的筛选以及促干剂的合理使用提供理论依据。结果表明:(1)‘宁杞1号’和‘宁杞5号’枸杞表皮细胞外侧细胞壁均呈现脊状突起的结构,在果实青果期脊状突起不连续,脊和脊之间排列紧密;随着发育时期的延后,脊状突起的连续性逐渐增强,且脊与脊之间的间距逐渐变宽,蜡质呈膜状覆盖于凸起的脊和两脊之间的沟内。(2)‘宁杞1号’和‘宁杞5号’两个枸杞品种在果实发育过程中,果皮单位面积蜡质含量均呈先下降后上升的趋势,在成熟期单位面积蜡质含量最高,且‘宁杞1号’3个发育时期果皮单位面积蜡质含量均高于同期‘宁杞5号’,呈现出与扫描电镜观察到的蜡质分布基本一致的变化趋势。(3)2个品种3个时期的果实表皮蜡质组分均由烷烃类、酮类、醇类、酸类、醛类、酯类和碘代烷烃类组成,两品种青果期和色变期果皮蜡质组分相同,主要由烷烃类、醇类和碘代烷烃类组成,它们成熟期的果皮蜡质组分主要由烷烃类、醇类和酯类组成。(4)主成分分析结果显示,3个生育时期‘宁杞5号’果皮蜡质组分多为烷烃类物质,较‘宁杞1号’更利于阻挡果实水分的散失。研究发现,影响枸杞果实制干的原因在于枸杞果皮蜡质含量、结构和蜡质组分,烷烃类组分能够有效阻止果实体内水分的散失,‘宁杞5号’果皮蜡质中烷烃组分含量更高,果皮保水性更强,致果实不易制干。     

cAMP介导的梨果表皮物化信号对链格孢侵染的调控

采用药理学方法,用cAMP抑制剂阿托品（atropine）处理链格孢Alternaria alternata孢子悬浮液,通过体外试验分析cAMP信号级联通路在链格孢响应梨果皮蜡质疏水性、化学组分和外源乙烯利等刺激后启动孢子萌发、附着胞形成的调控作用,并通过体内试验研究其对链格孢致病性的调控。结果表明,高疏水性表面和梨果蜡涂膜表面及1μmol/L的乙烯利均可显著促进链格孢的孢子萌发和附着胞形成。cAMP信号级联通路抑制剂atropine处理后显著抑制了表皮疏水性、蜡质和外源乙烯介导的链格孢的孢子萌发和附着胞形成,其中抑制剂处理后4h,链格孢在疏水性、果蜡涂膜表面和乙烯等处理中附着胞形成率分别较对照降低了75.3%、63.7%和74.3%,同时抑制剂处理还可抑制损伤接种链格孢早酥梨黑斑病的扩展。外源cAMP可以部分恢复抑制剂的作用,外源cAMP+atropine处理后4h,在高疏水性（108°）和果蜡涂膜表面,链格孢附着胞形成率为抑制剂atropine单独处理的2.4倍和1.6倍,表明cAMP信号级联通路可通过调控侵染结构的形成而影响链格孢对梨果表皮物理化学信号的识别和应答。     

UV-B辐射增强对马铃薯叶片结构及光合参数的影响

叶片作为植物进行光合作用的主要器官,在长期进化过程中形成了对不同光照环境条件的形态可塑性和相应的适应机制,以保证植物能在变化的、非适宜环境下的生存与繁衍。随着大气臭氧层衰减引起地表UV-B辐射增强,其对植物叶片结构和光合作用的影响显著,但这种气候变化趋势对马铃薯叶片形态结构、光合作用的影响尚不明确。设置增强UV-B辐射2.5 kJm~(-2)d~(-1)(T1)、5.0 kJm~(-2)d~(-1)(T2)、自然光(CK)3个处理,以6个马铃薯品种(系)为材料,研究增强辐射对不同基因型马铃薯叶片结构和光合参数的影响。结果表明:增强的UV-B辐射使马铃薯叶片解剖结构不同程度增厚,叶片厚度增加;叶片气孔和非腺毛的密度增加明显,腺毛有增多倾向。扫描电镜显示处理后的近轴面叶片角质层厚度增加,蜡质晶体增多,但表皮细胞变小且失水萎缩,细胞轮廓模糊;气孔、腺毛及非腺毛附属结构受胁迫影响呈萎缩状态。透射电镜显示处理后的叶肉细胞中基粒类囊体肿胀,结构层次紊乱,胁迫引起细胞质壁分离,细胞壁扭曲并有较多的沉淀物;部分品种过氧化物酶体可见清晰的过氧化氢酶晶体。叶片缩小增厚、腺毛增多、角质层和蜡质增厚、胞内积累过氧化氢酶的形态适应和生理响应并未能有效减少UV-B辐射对光合参数和光合效率的影响,合作88、丽薯6号、师大6号的净光合速率、气孔导度等参数均受到抑制,光能利用效率明显降低,属于UV-B辐射敏感型品种;剑川红21-3、21-1和转心乌3个品种(系)的相关光合特性几乎不受影响,显示云南地方品种具有较强的UV-B辐射耐受性,有待于进一步从生理生化和分子水平探究更多的适应机制。     

植物角质层蜡质的化学组成研究综述

角质层是植物与外界的第一接触面,而角质层蜡质则是由位于角质层外的外层蜡质和深嵌在角质层中的内层蜡质两部分构成。植物角质层蜡质成分极其复杂,具有重要的生理功能。综述了有关植物角质层蜡质的化学组成信息,探讨了目前植物角质层蜡质化学成分研究中存在的一些问题,展望了角质层蜡质成分的研究前景。     

Ontogenetic variation in chemical and physical characteristics of adaxial apple leaf surfaces

The reaction of plants to environmental factors often varies with developmental stage. It was hypothesized, that also the cuticle, the outer surface layer of plants is modified during ontogenesis. Apple plantlets, cv. Golden Delicious, were grown under controlled conditions avoiding biotic and abiotic stress factors. The cuticular wax surface of adaxial apple leaves was analyzed for its chemical composition as well as for its micromorphology and hydrophobicity just after unfolding of leaves ending in the seventh leaf insertion. The outer surface of apple leaves was formed by a thin amorphous layer of epicuticular waxes. Epidermal cells of young leaves exhibited a distinctive curvature of the periclinal cell walls resulting in an undulated surface of the cuticle including pronounced lamellae, with the highest density at the centre of cells. As epidermal cells expanded during ontogenesis, the upper surface showed only minor surface sculpturing and a decrease in lamellae. With increasing leaf age the hydrophobicity of adaxial leaf side decreased significantly indicated by a decrease in contact angle. Extracted from plants, the amount of apolar cuticular wax per area unit ranged from only 0.9 microgcm(-2) for the oldest studied leaf to 1.5 microgcm(-2) for the youngest studied leaf. Differences in the total amount of cuticular waxes per leaf were not significant for older leaves. For young leaves, triterpenes (ursolic acid and oleanolic acid), esters and alcohols were the main wax components. During ontogenesis, the proportion of triterpenes in total mass of apolar waxes decreased from 32% (leaf 1) to 13% (leaf 7); absolute amounts decreased by more than 50%. The proportion of wax alcohols and esters, and alkanes to a lesser degree, increased with leaf age, whereas the proportion of acids decreased. The epicuticular wax layer also contained alpha-tocopherol described for the first time to be present also in the epicuticular wax. The modifications in the chemical composition of cuticular waxes are discussed in relation to the varying physical characteristics of the cuticle during ontogenesis of apple leaves.     

Nanotubules on plant surfaces: chemical composition of epicuticular wax crystals on needles of Taxus baccata L

Needles of Taxus baccata L. were covered with tubular epicuticular wax crystals varying in diameters (100 and 250 nm) and lengths (300-500 and 500-1000 nm) on the abaxial and adaxial surfaces, respectively. Various sampling protocols were employed to study the chemical composition of the needle waxes on three different levels of spatial resolution. First, a dipping extraction of whole needles yielded the total cuticular wax mixture consisting of very long chain fatty acids (21%), alkanediols (19%), phenyl esters (15%), and secondary alcohols (9%) together with small amounts of aldehydes, primary alcohols, alkanes, alkyl esters, and tocopherols. Second, waxes from both sides of the needle were sampled separately by brushing with CHCl3-soaked fabric glass. Both sides showed very similar qualitative composition, but differed drastically in quantitative aspects, with nonacosan-10-ol (18%) and alkanediols (33%) dominating the abaxial and adaxial waxes, respectively. Third, the epi- and intracuticular wax layers were selectively sampled by a combination of mechanical wax removal and brushing extraction. This provided direct evidence that the tubular wax crystals contained high percentages of nonacosane-4,10-diol and nonacosane-5,10-diol on the abaxial surface, and nonacosan-10-ol on the adaxial surface of the needles. Together with these compounds, relatively large amounts of fatty acids and smaller percentages of aldehydes, primary alcohols, alkyl esters, and alkanes co-crystallized in the epicuticular layer. In comparison, the intracuticular wax consisted of higher portions of cyclic constituents and aliphatics with relatively high polarity. The formation of the tubular crystals is discussed as a spontaneous physico-chemical process, involving the establishment of gradients between the epi- and intracuticular wax layers and local phase separation.     

Composition of epicuticular wax layer of two species of Mandevilla (Apocynoideae,Apocynaceae) from Rio de Janeiro,Brazil

The chemical composition of epicuticular waxes of Mandevilla guanabarica and Mandevilla moricandiana was comparatively analyzed by extraction in n-hexane and chloroform. The mean wax content per unit of leaf area in the n-hexane extract was about 13–30 μg cm⁻² for M. guanabarica, containing 20–28% n-alkanes and 55–63% triterpenes; for M. mori-candiana, the mean content was 19 μg cm⁻², containing 73% n-alkanes and 14% triterpenes. In the chloroform extract, the wax yield was 40–80 μg cm⁻² for M. guanabarica, with about 9–11% n-alkanes and 75–82% triterpenes; while for M. moricandiana, the wax yield was 110 μg cm⁻², with 52% n-alkanes and 14% triterpenes. The major compounds identified were lupeol, pentacyclic triterpenes of the α- and β-amyrin class, and n-alkanes such as nonacosane, hentriacontane and tritriacontane. These results indicate that the quantitative chemical profiles of epicuticular waxes of M. guanabarica and M. moricandiana are distinct and could be used as an additional feature in taxonomic identification.     

Surface wax of Andreaea and Pogonatum species

The mosses Andreaea rupestris, Pogonatum aloides and P. urnigerum contain surface waxes in amounts of 0.05–0.12% dry wt. The waxes consisted of esters (C₃₈-C₅₄), primary alcohols (C₂₀-C₃₂), free fatty acids (C₁₆-C₃₀), and alkanes (C₂₁-C₃₁). Additionally, aldehydes (C₂₂-C₃₀) were major constituents in the wax of P. urnigerum. The classes and their chain length distributions in the surface waxes of these mosses are comparable to those of epicuticular waxes of higher plants.     

Composition of the epicuticular and intracuticular wax layers on Kalanchoe daigremontiana (Hamet et Perr. de la Bathie) leaves

Epicuticular and intracuticular waxes from both adaxial and abaxial surfaces of the leaves of Kalanchoe daigremontiana were analyzed. All wax mixtures were found to contain approximately equal amounts of triterpenoids and very long chain fatty acid (VLCFA) derivatives. The triterpenoid fraction consisted of glutinol (8-19% of the total wax) and friedelin (4-9%), together with smaller amounts of glutanol, glutinol acetate, epifriedelanol, germanicol and β-amyrin. The VLCFA derivatives comprised C₂₇-C₃₅ alkanes (19-37% of the total wax), C₃₂-C₃₄ aldehydes (3-7%), C₃₂ and C₃₄ fatty acids (0.2-3%), C₂₆-C₃₆ primary alcohols (4-8%), and C₄₂-C₅₂ alkyl esters (2-9%). The wax layers were found to differ in triterpenoid amounts, with the intracuticular wax containing higher percentages of most triterpenoids than the epicuticular wax. Friedelin, the only triterpenoid ketone present, showed the opposite distribution with higher proportions in the epicuticular wax. VLCFA derivatives also accumulated to higher percentages in the epicuticular than in the intracuticular wax layer. Epicuticular wax crystals were observed on both the adaxial and abaxial leaf surfaces.     

Epicuticular waxes of Sorghum and some compositional changes with plant age

Epicuticular wax from mature plants of Sorghum bicolor SD-102 was compared with that from panicles and seedlings of the same variety at the fourth-fifth leaf stage of growth. The composition of wax from SD-102 panicles was quite different from that of mature leaf blades and sheaths. Free fatty alcohols were the dominant class of wax from SD-102 seedlings and C₃₂ was the major homologue of alcohols and aldehydes. For comparison purposes, the epicuticular waxes from whole plants of two other S. bicolor varieties, Alliance A and Martin A, grown up to the tasseling stage, have been analysed. The major wax components were free fatty acids. The typical chain lengths of aldehydes, free alcohols and free fatty acids were C₂₈ and C_30.p-Hydroxybenzaldehyde was not a wax component of the studied varieties of sorghum.     

Intraspecific Variation in the Epicuticular Wax Composition of Four Species of Chionochloa

Intraspecific variation in four New Zealand species of Chionochloa, C. flavescens, C. pallens, C. rigida; and C. rubra, was investigated by examining the major carbon chain lengths of fatty acids, alcohols, aldehydes, wax esters and alkanes of the epicuticular waxes. The major even-carbon chain lengths ranged generally from C₂₄ to C₃₂ in the acids, alcohols and aldehydes; C₂₉ to C₃₃ in the alkanes; and even-carbon chains between C₃₆ and C₅₂ in the wax esters. A computer program was used to calculate the degree of similarity between samples in terms of chain length distribution. In C. rigida eastern and western South Island localities were identified; in C. flavescens Canterbury and Nelson, western South Island and southern North Island regions were recognized; and C. pallens and C. rubra were divisible into four regions; Canterbury, Nelson, western South Island and southern North Island. The possible elongation-decarboxylation pathways and the specificity of the enzymes in the biosynthetic pathways of epicuticular wax synthesis suggest the possibility that the northwest Nelson region could be a biogenetic centre from which wax synthesis has diversified along three routes, one to the western South Island, another to eastern South Island and the third to southern North Island. Identification of each of the four species based on the distribution of the carbon chain lengths in the individual lipid fractions is impossible unless the locality of collection is known. Intraspecific variation in lipid composition is not coincident with patterns of variation already reported.