水稻响应缺铁的韧皮部汁液蛋白质组学分析

为鉴定水稻(Oryza sativa)响应缺铁的根冠长距离信号转导物质, 采用TMT标记技术分析了不同浓度铁处理下水稻韧皮部汁液的蛋白质组学变化, 共鉴定出206个差异蛋白, 其中54个蛋白表达丰度上调, 152个蛋白表达丰度下调。差异蛋白的KEGG通路分类主要包括激素信号代谢、谷胱甘肽代谢、碳代谢以及mRNA转运等代谢途径。此外, 对差异蛋白对应的生理指标进行测定, 发现激素、蔗糖、谷胱甘肽和转运蛋白等在缺铁条件下变化显著, 后续对这些差异蛋白的功能研究有助于揭示水稻响应铁素营养的长距离信号途径。     

木薯叶的成分

木薯(Manihot esculenta Crantz)是一种热带作物,木薯根富含淀粉,世界上约有5亿人把它作为主食。木薯的叶子长势茂盛并有极高的蛋白含量。尽管木薯叶可以在改善热带地区居民的食品营养结构方面起到重要的作用,但是,人们对叶的食用远不如对根的食用普遍,其原因在于木薯叶中含有氢氰酸。     

VIGS技术鉴定木薯糖转运蛋白Mesweet18的功能研究

糖转运蛋白（sugar will eventually be exported transporter,SWEET）在植物运输糖类、生殖和发育、逆境性、与病原体互作等方面发挥着重要作用。选择木薯糖转运蛋白Mesweet18基因沉默的靶基因区域,通过病毒诱导的基因沉默(virus-induced gene silencing,VIGS)技术注射木薯SC9的盆栽苗叶片。qRT-PCR结果表明,Mesweet18在沉默植株中的表达量显著下调,分别是对照的46.80%、30.23%、21.12%。叶片叶绿素和可溶性糖含量检测结果表明,与对照相比,叶绿素a、b和总含量均出现不同程度的下降,蔗糖和果糖含量显著增加,而葡萄糖含量出现轻微下降。研究Mesweet18在木薯中的分子功能,为深入研究糖转运蛋白SWEET在木薯中的分子机制奠定了基础。     

HPLC-ELSD法在刺五加药材糖类成分分析中的应用

建立一种高效、快速的高效液相色谱—蒸发光散射检测法(HPLC-ELSD)分析刺五加药材中糖类成分—葡萄糖及蔗糖组分。采用CHROMATOREX NH2柱为固定相;以乙腈：水为84.5：15.5(V/V)为流动相,流速1.0 mL·min^-1;漂移管温度为100℃;氮气流速为2.5 L·min^-1。在上述色谱条件下,葡萄糖和蔗糖的线性范围分别为0.023 2～0.116 0 mg·mL^-1及0.099 6～0.496 5 mg·mL^-1;且该二种糖的检测限分别（信/噪=3）为3.87 ng和3.31 ng。本方法无须进行糖衍生化而直接用于刺五加药材中糖类成分的分离分析。     

大气CO_2浓度倍增对油菜韧皮部汁液成分及根部氮素积累的影响

以甘蓝型欧洲油菜(Brassica napus)‘814’和‘湘油15’两个品种为研究对象,探寻在两个CO2浓度(自然CO2浓度:400μmol·mol–1和高CO2浓度:(800±20)μmol·mol–1)和两个氮素水平(低氮和常氮)处理下,欧洲油菜韧皮部汁液中可溶性糖和游离氨基酸含量,以及根部干物质量和氮素累积量的变化。结果表明:1)CO2浓度升高提高欧洲油菜韧皮部汁液可溶性糖含量,施氮条件下‘814’在抽薹期达到0.29%,‘湘油15’在盛花期达到0.25%,其含量均显著高于自然CO2浓度处理。2)CO2浓度对韧皮部汁液游离氨基酸含量的影响因品种而异,无论施氮与否,CO2浓度升高使‘814’的游离氨基含量降低;而‘湘油15’CO2浓度升高促使其在低氮条件下含量升高,在常氮条件下则降低。3)根部干物质量和氮素累积量均随CO2浓度升高而增加,且欧洲油菜韧皮部汁液中可溶性糖含量与游离氨基酸含量与根部干物质量和氮素累积量呈显著正相关。     

大气CO2浓度倍增对油菜韧皮部汁液成分及根部氮素积累的影响

以甘蓝型欧洲油菜(Brassica napus) ‘814’和‘湘油15’两个品种为研究对象, 探寻在两个CO₂浓度(自然CO₂浓度: 400 μmol·mol^-1和高CO₂浓度: (800 ± 20) μmol·mol^-1)和两个氮素水平(低氮和常氮)处理下, 欧洲油菜韧皮部汁液中可溶性糖和游离氨基酸含量, 以及根部干物质量和氮素累积量的变化。结果表明: 1) CO₂浓度升高提高欧洲油菜韧皮部汁液可溶性糖含量, 施氮条件下‘814’在抽薹期达到0.29%, ‘湘油15’在盛花期达到0.25%, 其含量均显著高于自然CO₂浓度处理。2) CO₂浓度对韧皮部汁液游离氨基酸含量的影响因品种而异, 无论施氮与否, CO₂浓度升高使‘814’的游离氨基含量降低; 而‘湘油15’ CO₂浓度升高促使其在低氮条件下含量升高, 在常氮条件下则降低。3)根部干物质量和氮素累积量均随CO₂浓度升高而增加, 且欧洲油菜韧皮部汁液中可溶性糖含量与游离氨基酸含量与根部干物质量和氮素累积量呈显著正相关。     

马尾松和杉木树干韧皮部水溶性糖 δ13C值对气象因子的响应

通过测定亚热带马尾松和杉木树干韧皮部水溶性糖δ¹³C值的连日变化,及其对天气变化过程的响应,研究δ¹³C值对短期天气变化动态的响应特征。结果显示,春季马尾松和杉木树干韧皮部水溶性糖δ¹³C日均值分别介于-26.81‰到-26.49‰之间,以及-29.26‰到-27.47‰之间,平均值分别为(-26.58±0.12)‰和(-28.67±0.65)‰。进一步分析表明,马尾松树干韧皮部水溶性糖δ¹³C值与取样之前第4天的太阳辐射、水气压亏缺、相对湿度和空气温度显著相关(P≤0.05),杉木树干韧皮部水溶性糖δ¹³C值取样之前第3天的太阳辐射、水气压亏缺和相对湿度显著相关(P≤0.05),但与空气温度的相关性不显著(P≤0.05)。在所测定的环境因子中,太阳辐射是影响马尾松和杉木树干韧皮部水溶性糖δ¹³C值的首要因素。当天降水事件可能导致马尾松和杉木树干韧皮部水溶性糖δ¹³C值连日变化出现异常波动。马尾松和杉木韧皮部水溶性糖δ¹³C值可以敏感记录短期天气变化动态。     

土壤干旱条件下木质部汁液成分变化及其在根冠信号传递中的作用

介绍了植物木质部汁液收集方法和干旱条件下 ,蒸腾流中碳水化合物、蛋白质、pH等的变化 ,并对这些成分变化在根冠信号传递中的可能作用进行了讨论     

木薯高、低粉品种贮藏根生长关键期的生理生化特性比较

通过研究木薯Manihot esculenta高粉和低粉品种贮藏根在发育关键期(形成–膨大期)主要生理生化性质的差异及其动态变化,为木薯栽培管理和育种改良提供理论基础。对高粉木薯品种‘辐选01’(FX01)和‘Kasetsart 50’(KU50)以及低粉品种‘华南124’(SC124)和‘9I’于种植后120、130、140、151和165 d五个时期分别取贮藏根样品,测定淀粉、可溶性糖、脱落酸(ABA)、保护酶、活性氧(ROS)、丙二醛(MDA)等生理生化指标并分析变化规律。结果表明,可溶性糖含量在贮藏根皮层中始终高于淀粉储藏区;高粉品种中总可溶性糖含量在贮藏根膨大期快速增加,且表现出较高的抗氧化酶活性和较低的ROS及MDA含量;ABA含量呈现出高粉品种比低粉品种低的趋势。木薯低粉和高粉品种中可溶性糖的分布与其转运后合成淀粉的规律相一致,高粉品种中淀粉的积累速率始终高于低粉品种,其较高的抗氧化酶活性和较低的ROS及MDA含量有利于木薯贮藏根中淀粉的积累。     

基于HPLC-ELSD指纹图谱和多成分定量的浙贝母与湖北贝母质量差异研究CSCD

建立浙贝母、湖北贝母HPLC-ELSD指纹图谱,结合多成分定量分析,比较两种贝母属药材的差异。采用Waters ACQUITY HSS T3(4.6 mm×250 mm,5μm)色谱柱,以乙腈-0.1%三乙胺溶液为流动相,流速为1.1 mL/min,梯度洗脱;柱温为38℃;蒸发光散射检测;建立浙贝母和湖北贝母HPLC-ELSD指纹图谱,通过化学计量学方法和5种生物碱类成分的含量测定比较浙贝母和湖北贝母的差异。结果显示,浙贝母指纹图谱标定7个共有峰,而湖北贝母有8个;指认出其中6个峰,分别为伊贝辛、贝母辛、贝母素甲、贝母素乙、异贝母甲素、湖贝甲素,其中湖贝甲素为湖北贝母的专属性成分;HCA和PCA均能很好地区分浙贝母和湖北贝母,OPLS-DA共找到4个差异性标志物,含测结果显示,浙贝母中贝母素甲的含量明显高于湖北贝母,而贝母辛、贝母素乙和异贝母甲素的含量则明显低于湖北贝母。该方法可以有效鉴别浙贝母和湖北贝母质量的差异性,为其质量控制提供参考。     

Measurements of pH, sucrose and potassium ions in the phloem sap of castor bean (Ricinus communis) plants

The composition of phloem sap, sampled at different heights along, the stem of castor bean ( Ricinus communis L. cv. Gibsonii) plants, was determined. A gradient in pH was observed; the highest pH values occurred near the shoot apex, decreasing towards the base of the stem. The sucrose content of the exudate exhibited a similar gradient. The concentration of potassium ions was highest near the uppermost, full-grown leaves, decreasing towards the apex and the base of the stem. The importance of these findings for the understanding of phloem translocation and unloading is discussed.     

Influence of secondary compounds in the phloem sap of cassava on expression of antibiosis towards the mealybugPhenacoccus manihoti

Identification and assay of cyanogenic and phenolic compounds in phloem sap of cassava (Manihot esculenta Crantz, Euphorbiaceae) and in honeydew of the cassava mealybugPhenacoccus manihoti Matt. Ferr. (Homoptera, Pseudococcidae) were realised. Cyanogenic glucosides and three flavonoid glycosides (rutin, kaempferol glycoside-1 and kaempferol glycoside-2) were found to be translocated in cassava phloem sap and consumed by the mealygug. Differences in profiles of secondary compounds of phloem sap and honeydew samples, characterised mainly by the appearance of free cyanide and of a free flavonoid, suggest the metabolic processing of at least some of the ingested compounds. The relationship between foliar concentrations of these different compounds and expression of the antibiotic resistance of cassava towardsP. manihoti was also investigated in 7 varieties of cassava and in the ‘faux-caoutchouc’ hybrid. Infestation by mealybug was followed by a clear increase in levels of both rutin and kaempferol glycoside-2, while no modification in cyanide contents was noted. The best rank correlation between antibiotic resistance (measured by the intrinsic rate of increase r _c ) and secondary compounds analyzed is observed with rutin contents of infested plants (ρ=−0.73; p=0.05). The possible implication of this compound in the biochemical mechanisms accompanying cassava defence reaction to mealybug attack is discussed.

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Résumé Ce travail présente l'identification et le dosage des substances secondaires (composés cyanés et phénoliques) présentes dans la sève phloémienne du manioc (Manihot esculenta Crantz, Euphabiaceae) et dans le miellat de la cochenille farineusePhenacoccus manihoti Matt. Ferr. (Homoptera, Pseudococcidae). Des glycosides cyanogéniques et trois flavono?des glycosylés majeurs (rutine, kaempferol glycoside-1 et kaempferol glycoside-2) sont transportés par la sève phloémienne du manioc et consommés par la cochenille. Le profil sensiblement différent des composés secondaires du meillat de la cochenille, en particulier l'apparition d'un flavonoide libre et de cyanure, suggère une modification de certains composés lors du transit intestinal. La relation entre les teneurs de ces différentes substances dans le liquide foliaire de 7 variétés de manioc et du ‘faux-caoutchouc’ (hybride deM. esculenta et deM. glaziovii) et l'expression de leur résistance antibiotique vis-à-vis deP. manihoti a également été étudiée. L'infestation par la cochenille se traduit par une importante augmentation des teneurs en rutine et en kaempferol glycoside-2, alors qu'aucune modification des teneurs en cyanure libre n'est enregistrée. La meilleure corrélation entre la résistance par antibiose, exprimée par la capacité intrinsèque d'accroissement r _c , et les teneurs en substances secondaires analysées est observée avec les teneurs en rutine des plants infestés (ρ=−0.73; p=0.05). L'association éventuelle de ce glycoside aux mécanismes biochimiques de défense du manioc à la cochenille farineuse est discutée.

     

Polyamines,indole-3-acetic acid and abscisic acid in rice phloem sap

Putrescine, spermidine, spermine and cadaverine have been identified and quantified in rice phloem sap and shoot extracts by HPLC. It is suggested that diamines, putrescine and cadaverine, easily migrate into the phloem, while movement of a triamine, spermidine, and a tetramine, spermine, tend to be restricted. Spermine especially seems to be the most immobile among polyamines. Thus it is indicated that movement of polyamines into phloem is decreased with increasing number of amino groups. Indole-3-acetic acid and abscisic acid in rice phloem sap were also analyzed by HPLC and it is suggested that indole-3-acetic acid is transported freely into phloem, while abscisic acid is much more actively exuded into phloem.     

Free amino acid content of phloem sap from Brassica plants in relation to performance of Lipaphis erysimi (Hemiptera: Aphididae)

Cultivars of Brassica juncea, B. napus and B. campestris, differing in host-suitability to the mustard aphid (Lipaphis erysimi pseudobrassicae Kalt.), were analysed for their phloem sap content of free amino acids. Sap was collected from excised mustard aphid stylets and analysed by means of high performance liquid chromatography. Aspartic and glutamic acid were, together with glutamine, the dominating amino acids. All plants had very little glycine, γ-aminobutyric acid (GABA), methionine and tryptophan in their sap. This amino acid pattern closely resembled that of cereals. The total concentration and the individual composition of amino acids were not found to be related to the level of aphid resistance previously found in the investigated Brassica species. However, one B. campestris cultivar, yellow sarson YSB-9, on which the aphids were shown to grow more slowly, had less amino acid content than other B. campestris varieties. The slow population growth of aphids on B. juncea, compared to that on B. campestris, is probably due to other than nutritional factors.     

Changes in the amino acid composition and protein modification in phloem sap of rice

Pure phloem sap was collected from insects feeding on rice (Oryza sativa L.) leaves by a laser technique similar to the aphid stylet technique. Rapid circulation of nitrogen in the sieve tubes was demonstrated directly using ¹⁵N as a tracer. Application to the roots of the metabolic inhibitors of amino acids, aminooxyacetate and methioninesulfoximine, changed the amino acid composition in the sieve tubes. Feeding methionine to leaf tips resulted in its bulk transfer into the sieve tubes. In vitro experiments confirmed the existence of protein kinases in the pure rice phloem sap. The phosphorylation status of the sieve tube sap proteins was affected by the light regime. The possibility that changes in chemical composition or protein modification such as phosphorylation in the sieve tubes might affect plant growth are discussed.Analysis of pure phloem sap collected from rice plants by insect laser technique has shown dynamic changes in the chemical composition and the quality of proteins in the sap.     

The nutritional quality of phloem sap utilized by natural aphid populations

Abstract.

• 1 The amino acid content of phloem exudates from leaves and of aphid honeydew were adopted as indices of the nutritional quality of phloem sap for aphids. Four plant species and associated leaf-dwelling aphids were investigated: the sycamore Acer pseudoplanatus and sycamore aphid Drepanosiphum platanoides; Prunus domestica (victoria plum) and the mealy plum aphid Hyalopterus pruni; and the spindle tree Euonymus europaeus and broad-bean Vicia faba, both hosts of the black bean aphid Aphis fabae.
• 2 The concentration of amino acids in the phloem exudates varied with: (a) plant species (greater in the herb Vicia than in the tree species), (b) season (greater in the autumn than summer for Acer and Euonymus), and (c) position (greater in flush leaves than mature leaves of Prunus).
• 3 For Acer and Prunus and their aphids, the concentration of amino acids in phloem exudates was significantly correlated with the amino acid content of the aphid honeydew.
• 4 The amino acids in all exudates and honeydew were dominated by non-essential amino acids (glutamic acid, glutamine, asparagine or serine, varying with season and between plant species). The sole major discrepancy between the amino acid profiles of exudates and honeydew was the production of asparagine-rich honeydew by aphids feeding on leaves, whose exudates were dominated by glutamic acid; this applied to both H.pruni on mature Prunus leaves and Drepanosiphum platanoides on summer-leaves of Acer.
• 5 It is suggested that EDTA-exudation may be a useful technique to study nutritional correlates of aphid life cycles, e.g. the time of migration between primary and secondary plant hosts.

     

Towards the proteome of Brassica napus phloem sap

The soluble proteins in sieve tube exudate from Brassica napus plants were systematically analyzed by 1-DE and high-resolution 2-DE, partial amino acid sequence determination by MS/MS, followed by database searches. 140 proteins could be identified by their high similarity to database sequences (135 from 2-DE, 5 additional from 1-DE). Most analyzed spots led to successful protein identifications, demonstrating that Brassica napus, a close relative of Arabidopsis thaliana, is a highly suitable model plant for phloem research. None of the identified proteins was formerly known to be present in Brassica napus phloem, but several proteins have been described in phloem sap of other species. The data, which is discussed with respect to possible physiological importance of the proteins in the phloem, further confirms and substantially extends earlier findings and uncovers the presence of new protein functions in the vascular system. For example, we found several formerly unknown phloem proteins that are potentially involved in signal generation and transport, e.g., proteins mediating calcium and G-protein signaling, a set of RNA-binding proteins, and FLOWERING LOCUS T (FT) and its twin sister that might be key components for the regulation of flowering time.     

Role of phloem sap quality and exudation characteristics on performance of pea aphid grown on lucerne genotypes

The phloem sap of two lucerne (Medicago sativa L.) genotypes inducing differences in pea aphid,Acyrthosiphon pisum Harris (Homoptera: Aphididae) performances (weight, fecundity, survival), was collected by stylectomy (radio frequency microcautery). Sugars and amino acids were assayed. No significant difference in their concentrations could be established between genotypes. It is thus unlikely that a substantial part of the varietal resistance of these lucerne genotypes to pea aphid could be related to the content in sugars or amino acids. The same conclusion was drawn for the sugar/amino acid ratio. The amino acid profiles were globally similar. They nevertheless allowed a discrimination between the two genotypes on a few amino acids (alanine, leucine, isoleucine, arginine, ornithine) only. Moreover, the degree of resistance can be related neither to GABA concentration, nor to that of sulfur amino acids, because these constituants were either absent, or in trace amounts, whatever the lucerne genotype taken into account. Stylectomy parameters revealed some interesting differences in the sap exudation of the two genotypes. Average volume of exudation by stylet and average duration of exudation were respective 2.8 and 2.3 times higher in the susceptible genotype. These results suggest a reduced flow of phloem sap, after puncturing of the sieve-tubes by the aphid stylets on the resistant genotype; this could result in a greater difficulty in phloem sap ingestion by aphids settled on the resistant genotype and thus in suboptimal performances.     

Immunological analysis of phloem sap of Bacillus thuringiensis corn and of the nontarget herbivore Rhopalosiphum padi (Homoptera: Aphididae) for the presence of Cry1Ab

Phloem sap of transgenic Bacillus thuringiensis (Bt) corn expressing a truncated form of the B. thuringiensis delta-endotoxin Cry1Ab, sap sucking aphids feeding on Bt corn and their honeydew were analysed for presence of Cry1Ab using ELISA. Phloem sap of Bt and non-Bt corn was collected using a newly developed technique with a microcapillary being directly inserted into the phloem tubes. Using this technique, no Cry1Ab was detected in the phloem sap. In contrast, measurable concentrations of Cry1Ab in the range of 1 ppb were detected when phloem sap of pooled leaf samples was extracted using EDTA buffer. This was probably because of Cry1Ab toxin released from damaged cells. When analysing apterous adults of Rhopalosiphum padi L. and their honeydew, no Cry1Ab could be detected. In contrast, Cry1Ab was clearly detected in both larvae of the leaf chewing herbivore Spodoptera littoralis (Boisduval) and their faeces, showing that Cry1Ab is detectable after ingestion and excretion by herbivores. These results suggest that R. padi ingests or contains no or only very low concentrations of Cry1Ab in the range of the detection limit. In consequence it is hypothesized that R. padi as an important prey for beneficial insects in corn is unlikely to cause any harm to its antagonists due to mediating Bt toxin.