不同地理种源杉木根叶功能性状与碳氮磷化学计量分析

为认识地理环境因子对不同种源杉木的影响及其响应和适应特征,对生长在同质园的7个种源3年生杉木王幼树根、叶功能性状及碳氮磷化学计量与环境因子的关系进行了研究。结果表明:不同地理种源杉木功能性状种源间变异系数为7.3%-18.9%,化学计量种源间变异系数为1.6%-18.9%,均小于20%,叶、根C含量变异系数较小,说明C是植物体最稳定的元素。不同地理种源杉木幼树叶厚、比叶面积、叶组织密度、比叶重、叶干物质含量差异显著,表明杉木叶片形态具有较强的可塑性来适应不同种源地地理环境变化。不同地理种源杉木根N含量、根C∶N和根C∶P差异极显著,反映不同种源地气候与土壤养分对根系化学计量特征的潜在影响。江西文公山种源的杉木比叶面积最大,比根长、根干物质含量最小;安徽休宁种源的杉木叶片C含量、C∶P、根C∶N、N∶P最小,细根N、P含量最高;江西安福山种源的杉木细根C∶N最大,叶片N∶P、细根C、N、P含量、C∶N、C∶P最小。不同种源地经度与杉木叶组织密度、叶C∶P呈极显著正相关。随着降水梯度升高,不同种源杉木叶根N∶P、比叶面积、比叶重升高。随海拔升高和气温的降低,叶片厚度增加,叶组织密度密度减小,叶干物质含量减小,比叶面积减小。对不同地理种源杉木功能性状间的权衡关系分析表明,不同种源杉木根比表面积与比叶面积呈显著负相关且与比叶重呈极显著正相关,叶片及细根N含量与P含量均呈极显著正相关,说明杉木叶、根面积存在关联性,根长与根面积生长趋势相同,根叶对N、P元素含量的响应也相同。     

胡杨异形叶结构型性状及其与胸径关系

对胡杨5种类型植株的叶片厚度、叶面积、比叶面积、叶片干重和干物质含量等5个结构型性状指标进行分析,研究叶片的结构型性状与胸径之间及各个结构型性状彼此间相互关系。结果表明:叶面积、叶厚度和叶干重与胸径呈极显著正相关(P0.01),叶片干物质含量与胸径间呈显著正相关(P0.05),比叶面积与胸径之间呈极显著负相关(P0.01),说明随着植株生长发育的进行,叶片的面积、厚度、干重和干物质含量均逐渐增加,而比叶面积逐渐减小。各性状指标间相关性表现为叶面积、叶厚度、叶干物质和叶片干物质含量任意2个指标之间均呈显著或极显著正相关;而比叶面积与这4个指标间均呈显著或极显著负相关。     

大青沟自然保护区主要森林群落优势种的叶性状

植物功能性状是近年来生态学研究的热点。其中叶功能性状与植株生物量和植物对资源的获得、利用及利用效率的关系最为密切。大青沟森林植物群落分布于科尔沁沙地,生境条件非常特殊,在沙沟里存在着一片茂密的森林,与周围浩瀚无垠的沙坨景观形成极为鲜明的对照。从沟底到沟顶,虽然海拔高度仅相差六、七十米,但由于距离沟底水源不同,土壤条件差异大,形成了不同的森林植物群落类型。大果榆群落、蒙古栎群落和水曲柳群落分别分布在大青沟自然保护区的沟顶、沟中和沟底。为了对大青沟自然保护区,不同环境梯度下的森林植物群落叶片功能性状进行研究,以大青沟自然保护区大果榆、蒙古栎、水曲柳3种主要森林群落为研究对象,分别测定不同群落优势种的叶厚度、比叶面积、叶干物质含量、叶大小和叶干重等5项叶功能性状,研究叶功能性状之间的关系,并对不同生长型、不同群落叶功能性状进行比较。相关分析结果表明,叶厚度与比叶面积呈极显著负相关,与叶大小和叶干重呈极显著正相关;比叶面积与叶干物质含量、叶干重呈极显著负相关,与叶大小呈显著正相关;叶干物质含量与叶大小呈极显著负相关,与叶干重呈极显著正相关;叶大小与叶干重呈极显著正相关。不同生长型植物叶片性状的分析表明,草本植物的叶干物质含量比乔木和灌木低,而其比叶面积高于乔木和灌木;对不同群落叶功能性状进行比较发现,大果榆群落和蒙古栎群落乔木、灌木叶厚度与干物质含量显著高于水曲柳群落的叶厚度与干物质含量,二者比叶面积显著低于水曲柳群落。大果榆群落和蒙古栎群落从叶功能性状的角度,它们具有较高的叶干物质含量和较低的比叶面积,体现出适应干旱生境叶片的特征。水曲柳群落呈现出低叶干物质含量、高比叶面积的特征,体现出适应湿润、土壤水分较好生境的特征。不同群落通过调节自身的物种组成,形成不同的功能性状组合来适应环境。     

猫儿山交让木叶性状海拔变异格局及其环境影响因子

分析相近树龄单一树种叶性状、叶性状网络的海拔变异格局及其影响因素，有助于排除遗传因素的影响，准确反映环境因子对叶片的作用规律。本研究对猫儿山海拔梯度上中龄林交让木叶片气孔性状、结构性状、化学性状和叶脉性状进行研究，分析了不同海拔(1100、1500和1900 m)叶性状和叶性状网络的变异格局、生活策略以及影响叶性状变异的主要环境因子。结果表明：交让木叶面积、比叶面积、叶厚、叶干物质含量、叶绿素含量、叶氮含量、叶磷含量、叶片C∶N、叶片C∶P、叶脉密度和叶脉直径在不同海拔间存在显著差异。年均温和总辐射分别解释叶性状42.1%和16.2%的变异，是驱动交让木叶性状海拔变异的关键环境因子。年均温对叶面积影响最大(R²=0.73),总辐射对叶厚影响最大(R²=0.72),均为显著正相关。1100 m低海拔交让木叶氮含量、叶磷含量较低，叶性状网络整体和内部均表现松散，采取资源保守型策略；1500 m中海拔交让木叶氮含量、叶磷含量较高，叶性状网络整体连接紧密局部松散，通过增强叶性状间的依赖性，显著提高磷利用效率，来应对落叶林分的竞争，采取资源获取型策略；...     

云丘山不同海拔梯度橿子栎叶性特征研究

在吕梁山南段云丘山,以橿子栎为主要研究对象,采用野外调查和室内实验相结合的方法,对比叶面积(SLA)、叶面积(LA)、叶干物质含量(LDMC)、叶长宽比(L/W)、叶绿素含量(Chl)以及叶氮含量等有代表性的指标进行分析.结果表明：(1)随着海拔梯度的增加,橿子栎 L/W、Chl、SLA、LA、单位面积的叶氮含量(N area )、单位重量的叶氮含量(N mass )、叶饱和鲜重和叶干重均呈先上升后下降的趋势,在海拔1180 m 处有最大值;LDMC 与海拔呈显著负相关(P ＜0．05);(2)橿子栎 LA 与 SLA 和 N mass 呈极显著正相关关系(P ＜0．01),与 LDMC 呈显著负相关关系(P ＜0．05),与 N area呈极显著负相关关系(P ＜0．01);(3)橿子栎叶片 SLA与 L/W 呈显著正相关(P ＜0．05),与 N mass 呈极显著正相关关系(P ＜0．01),橿子栎叶片 SLA 与 LDMC 和N area呈显著负相关关系(P ＜0．05);(4)橿子栎 LDMC 与 N area 呈显著正相关(P ＜0．05).通过对云丘山景区橿子栎叶片性状随海拔变化规律的研究,可以探索橿子栎叶片性状与海拔梯度变化的关系以及最适生长环境,从而为景区橿子栎林的保护和抚育提供理论依据,为合理开发橿子栎林资源提供基础资料.     

狗娃花叶片生态化学计量特征及功能性状对坡向的响应

坡向对植物叶片生态化学计量特征及功能性状有显著影响。研究不同坡向上植物叶片生态化学计量特征与功能性状有助于了解植物对环境变化的响应及适应。以兰州地区狗娃花叶片为研究对象,分析其生态化学计量及功能性状随坡向的变化特征。结果显示:阳坡叶片C含量(408.12 g·kg~(-1))、C∶P(348.61)与N∶P(29.71)均显著高于阴坡(其值分别为400.75 g·kg~(-1)、196.69和16.60),N含量(34.81 g·kg~(-1))和C∶N(11.72)与阴坡无显著差异(其值分别为34.07 g·kg~(-1)和12.24),而P含量(1.67 g·kg~(-1))显著小于阴坡(2.56 g·kg~(-1));阳坡比叶面积及叶片含水量(10.55 cm2·g~(-1)、59.06%)与阴坡(9.70 cm2·g~(-1)、60.65%)无显著差异,叶干物质含量(0.14 g·g~(-1))和叶面积(0.31 cm2)均显著低于阴坡(0.15 g·g~(-1)、0.41 cm~2);不同坡向狗娃花叶片生长均受P限制;阳坡比叶面积、叶片含水量、叶面积分别是影响叶片C含量、N含量和叶片C∶N的主导因子;阴坡叶干物质含量是影响叶片C含量和P含量的主导因子,而叶片功能性状的4项指标对叶片N含量和C∶N均有影响,且叶面积为主导因子,比叶面积次之。研究表明,坡向对狗娃花叶片生态化学计量特征和功能性状均有影响,将二者结合研究有助于全面了解植物对环境的适应策略。     

引种期同质园翅荚木主要叶功能性状与种源地环境关系

从比叶面积(SLA)、叶片相对含水量(LRWC)、叶组织密度(LTD)、叶片干物质含量(LDMC)、叶片碳(C)氮(N)磷(P)含量及其化学计量比等方面对在引种期同质园的广西桂林市(GGL)、广西靖西市(GJX)、湖南江华县(HJH)、湖南通道市县(HTD)、广东英德市(GYD)等5翅荚木种源叶片主要功能性状进行差异分析及相关性分析,并研究翅荚木叶功能性状与种源地环境因子的关系。结果显示,(1)不同种源间叶片C、N含量及C/N差异显著,5种源间叶片C含量GYD最高、GGL最低,N含量GJX最高、GYD最低,C/N为GYD最高、GJX最低。(2)比叶面积与叶组织密度呈显著负相关;叶片N含量与C/N和C/P呈显著负相关,与N/P呈显著正相关;C/P与P含量呈显著负相关,与C/N呈显著正相关;C/N与N/P呈显著负相关。(3)翅荚木叶功能性状主要受七月均温、年均温和相对湿度等地理环境因子影响。七月均温与C/P、比叶面积呈正相关,与叶片N含量、叶片干物质含量、叶组织密度呈负相关;年均温与C/N呈正相关,与N/P呈负相关;相对湿度与叶片P含量、叶组织密度呈正相关,与C/P、比叶面积呈负相关。研究结果有助于理解翅荚木对引种区的环境适应特征及其在引种期的生长与生存策略,对探究翅荚木的养分利用策略、揭示其对环境变化的响应机制等具有重要意义,并为相关部门选择适宜的翅荚木引种栽培种源及开展其优良种源的遗传改良等提供理论指导与决策参考。     

壳斗科五树种幼苗叶片结构型性状及其相关关系

通过种子的人工采集、低温沙藏和温室培育后,将幼苗移栽到环境条件相同的同质园中自然生长,研究了中亚热带壳斗科丝栗栲(Castanopsis fargesii)、苦槠(C.sclerophylla)、锥栗(Castanea henryi)、板栗(C.mollissima)和麻栎(Quercus acutissima)2年生幼苗生长季节的叶面积、叶厚度、比叶面积、叶干物质含量、叶氮浓度、叶磷浓度和叶氮磷比7种结构型性状的差异及其相关关系。结果表明:(1)在5树种幼苗中,锥栗幼苗的叶面积和比叶面积最大、叶氮浓度最高,其叶干物质含量最低;丝栗栲幼苗的叶面积最小、叶氮浓度和叶磷浓度最低;苦槠幼苗的叶厚度和比叶面积最小,其叶干物质含量最高;板栗幼苗则是叶厚度最小;同时5树种幼苗叶片的氮磷比均无显著差异(P0.1)。(2)Pearson相关和线性回归分析表明,壳斗科5树种幼苗的叶面积与叶氮浓度极显著正相关(P0.01),并与比叶面积显著正相关(P0.05);叶厚度与比叶面积极显著负相关(P0.01),其与叶干物质含量显著正相关(P0.05);比叶面积与叶干物质含量极显著负相关(P0.01),并与叶氮浓度极显著正相关(P0.01),同时与叶磷浓度显著负相关(P0.05);叶氮浓度与叶干物质含量、叶磷浓度分别为显著负相关(P0.05)和显著正相关(P0.05)。该研究结果表明壳斗科5树种幼苗叶片的7种结构型性状表现出了一定的种间差异性,但同一结构型性状的大小顺序和种间差异的显著性不同;同时所有树种幼苗叶片各结构型性状间的相关性及其显著程度不同。     

地下水埋深对灰胡杨叶性状与化学计量学特征的影响

以塔里木荒漠河岸林优势树种灰胡杨为研究对象,分析灰胡杨叶性状与化学计量学特征沿地下水埋深梯度(GWD)的变异格局及其相互关系,以揭示灰胡杨适应极端干旱环境的生态策略。结果表明：(1)不同地下水埋深条件下灰胡杨叶性状和化学计量学特征存在显著差异(P<0.05),叶面积(LA)、叶片含水量(LWC)、叶氮含量(LNC)和叶磷含量(LPC)随GWD增加呈下降趋势,叶干物质含量(LDMC)、叶组织密度(LTD)、叶碳含量(LCC)、碳氮比(C∶N)、碳磷比(C∶P)和氮磷比(N∶P)随GWD增加呈上升趋势;其中C∶P、N∶P、LPC和LA变异系数较大(39.90%、36.09%、30.63%和28.22%),对GWD变化较为敏感。(2)LA与LTD和LDMC, LWC与LTD和LDMC, LNC与C∶N, LPC与LCC、N∶P和C∶P分别呈极显著负相关关系(P<0.01),LA与N∶P和C∶P呈显著负相关(P<0.05);逐步回归分析表明LA、LDMC、LTD、C∶P可作为响应地下水埋深变化的重要指标。(3)C∶N、N∶P、C∶P随GWD增加均呈上升趋势,LCC、LNC和L...     

喀斯特石漠化地区土壤养分对泡核桃功能性状的影响

为了探究喀斯特石漠化地区植物叶片功能性状及影响因素,以及揭示其对石漠化环境的适应机理,该文以中国南方喀斯特高原峡谷地区的泡核桃(Juglans sigillata)为对象,揭示土壤养分对叶片结构和光合性状的影响效应。结果表明：(1)泡核桃叶功能性状随石漠化等级增加,叶面积减小,比叶面积增大,叶干物质含量和叶组织密度先降后升,蒸腾速率、胞间CO₂浓度、气孔导度和光能利用率先下降后升高,其他性状变化趋势不显著。(2)冗余分析表明土壤养分能够解释37.4%的光合性状变异与53.4%的结构性状变异,其中全磷和溶解性有机碳对光合性状影响最大,而对结构性状影响最显著的是碱解氮和速效磷。(3)比叶面积分别与叶干物质含量极显著负相关,与净光合速率极显著正相关,叶厚度与叶组织密度极显著负相关,蒸腾速率与胞间CO₂浓度、气孔导度极显著正相关,水分利用速率与蒸腾速率、胞间CO₂浓度、气孔导度极显著负相关,光能利用率与净光合速率显著正相关。研究结果表明,泡核桃为适应喀斯特石漠化的特殊生境采取增强生长功能性状,同时提高资源获取能力的开拓型生长策略...     

Mechanisms and ecological consequences of plant defence induction and suppression in herbivore communities

Background Plants are hotbeds for parasites such as arthropod herbivores, which acquire nutrients and energy from their hosts in order to grow and reproduce. Hence plants are selected to evolve resistance, which in turn selects for herbivores that can cope with this resistance. To preserve their fitness when attacked by herbivores, plants can employ complex strategies that include reallocation of resources and the production of defensive metabolites and structures. Plant defences can be either prefabricated or be produced only upon attack. Those that are ready-made are referred to as constitutive defences. Some constitutive defences are operational at any time while others require activation. Defences produced only when herbivores are present are referred to as induced defences. These can be established via de novo biosynthesis of defensive substances or via modifications of prefabricated substances and consequently these are active only when needed. Inducibility of defence may serve to save energy and to prevent self-intoxication but also implies that there is a delay in these defences becoming operational. Induced defences can be characterized by alterations in plant morphology and molecular chemistry and are associated with a decrease in herbivore performance. These alterations are set in motion by signals generated by herbivores. Finally, a subset of induced metabolites are released into the air as volatiles and function as a beacon for foraging natural enemies searching for prey, and this is referred to as induced indirect defence.Scope The objective of this review is to evaluate (1) which strategies plants have evolved to cope with herbivores and (2) which traits herbivores have evolved that enable them to counter these defences. The primary focus is on the induction and suppression of plant defences and the review outlines how the palette of traits that determine induction/suppression of, and resistance/susceptibility of herbivores to, plant defences can give rise to exploitative competition and facilitation within ecological communities “inhabiting” a plant.Conclusions Herbivores have evolved diverse strategies, which are not mutually exclusive, to decrease the negative effects of plant defences in order to maximize the conversion of plant material into offspring. Numerous adaptations have been found in herbivores, enabling them to dismantle or bypass defensive barriers, to avoid tissues with relatively high levels of defensive chemicals or to metabolize these chemicals once ingested. In addition, some herbivores interfere with the onset or completion of induced plant defences, resulting in the plant’s resistance being partly or fully suppressed. The ability to suppress induced plant defences appears to occur across plant parasites from different kingdoms, including herbivorous arthropods, and there is remarkable diversity in suppression mechanisms. Suppression may strongly affect the structure of the food web, because the ability to suppress the activation of defences of a communal host may facilitate competitors, whereas the ability of a herbivore to cope with activated plant defences will not. Further characterization of the mechanisms and traits that give rise to suppression of plant defences will enable us to determine their role in shaping direct and indirect interactions in food webs and the extent to which these determine the coexistence and persistence of species.     

Design and Use of Multiplexed Chemostat Arrays

Chemostats are continuous culture systems in which cells are grown in a tightly controlled, chemically constant environment where culture density is constrained by limiting specific nutrients.^1,2 Data from chemostats are highly reproducible for the measurement of quantitative phenotypes as they provide a constant growth rate and environment at steady state. For these reasons, chemostats have become useful tools for fine-scale characterization of physiology through analysis of gene expression^3-6 and other characteristics of cultures at steady-state equilibrium.⁷ Long-term experiments in chemostats can highlight specific trajectories that microbial populations adopt during adaptive evolution in a controlled environment. In fact, chemostats have been used for experimental evolution since their invention.⁸ A common result in evolution experiments is for each biological replicate to acquire a unique repertoire of mutations.^9-13 This diversity suggests that there is much left to be discovered by performing evolution experiments with far greater throughput. We present here the design and operation of a relatively simple, low cost array of miniature chemostats—or ministats—and validate their use in determination of physiology and in evolution experiments with yeast. This approach entails growth of tens of chemostats run off a single multiplexed peristaltic pump. The cultures are maintained at a 20 ml working volume, which is practical for a variety of applications. It is our hope that increasing throughput, decreasing expense, and providing detailed building and operation instructions may also motivate research and industrial application of this design as a general platform for functionally characterizing large numbers of strains, species, and growth parameters, as well as genetic or drug libraries.     

Comparison of Extraction and Shipping Methods for Cysts and Juveniles of Heterodera glycines

Experiments to determine the effects of extraction techniques and the influence of shipping on extraction of Heterodera glycines life stages gave variable results. Shipping did not significantly affect numbers of nematodes extracted. More second-stage juveniles (J2) were extracted with Baermann funnels than with an elutriator, probably because incubation of encysted eggs on the Baermann funnel for 1 week allowed hatching to occur. Sieving was more efficient than elutriation for extracting cysts. Adding air agitation to the water pressure during elutriation increased extraction efficiency of cysts but not J2. Sample sizes of 250 cm³ and 500 cm³ did not influence extraction efficiency of cysts; however, sample size did influence extraction of J2.     

昆虫平衡棒的发育及功能

平衡棒(haltere)是双翅目昆虫后翅特化而成的结构,可在飞行中起重要作用。平衡棒基部的感受器可以检测到飞行中的惯性力,向运动神经元提供反馈,迅速地平衡身体并纠正航向。昆虫的平衡棒由成虫盘发育形成,其特化受HOX基因(Ultrabithorax,Ubx)调控。发育成熟的平衡棒由两层上皮细胞组成,末端球状结构内部充满高度空泡化的细胞,基部具有大量感器。平衡棒的运动由独立的肌肉控制,相对于同侧的翅反向移动,翅与平衡棒的协同运动对于昆虫起飞和维持平衡十分重要。近年来,平衡棒的导航原理越来越多地应用于仿生学研究中,基于果蝇平衡棒的结构和功能,研制出多种飞行器的导航设备。本文结合近年来相关领域的研究成果,就平衡棒的发育、形态结构、功能和仿生应用等方面的研究进展进行综述,为深入理解昆虫平衡棒的发育机制和生物学功能提供参考。     

Unraveling regulation and new components of human P-bodies through a protein interaction framework and experimental validation

The cellular factors involved in mRNA degradation and translation repression can aggregate into cytoplasmic domains known as GW bodies or mRNA processing bodies (P-bodies). However, current understanding of P-bodies, especially the regulatory aspect, remains relatively fragmentary. To provide a framework for studying the mechanisms and regulation of P-body formation, maintenance, and disassembly, we compiled a list of P-body proteins found in various species and further grouped both reported and predicted human P-body proteins according to their functions. By analyzing protein-protein interactions of human P-body components, we found that many P-body proteins form complex interaction networks with each other and with other cellular proteins that are not recognized as P-body components. The observation suggests that these other cellular proteins may play important roles in regulating P-body dynamics and functions. We further used siRNA-mediated gene knockdown and immunofluorescence microscopy to demonstrate the validity of our in silico analyses. Our combined approach identifies new P-body components and suggests that protein ubiquitination and protein phosphorylation involving 14-3-3 proteins may play critical roles for post-translational modifications of P-body components in regulating P-body dynamics. Our analyses provide not only a global view of human P-body components and their physical interactions but also a wealth of hypotheses to help guide future research on the regulation and function of human P-bodies.     

红花水提取液对小鼠系统性硬皮病的防治作用及其机制*

目的:研究红花水提取液对系统性硬皮病(SSc)模型小鼠的防治作用及相关机制研究。方法:60只 BALB /C小鼠随机分为对照组、模型组、强的松组、红花低、中、高剂量组,每组10只。对照组背部注射生理盐水,其余5组均背部皮下注射100 μl浓度为 200 μg /ml的注射用盐酸博来霉素,每天1次,连续注射28 d,制备SSc模型;造模同时对照组和模型组给予生理盐水10 ml/kg灌胃,强的松组给予强的松溶液4.5 mg/kg (10 ml/kg)灌胃,红花低、中、高剂量组分别给予红花1.5、3、6 g/kg (10 ml/kg)灌胃,各组均连续灌胃28 d。给药28 d后,取各组小鼠背部注射博来霉素区皮肤组织切片测量真皮厚度,采用水解法检测皮肤组织羟脯氨酸(HYP)含量;采用ELISA法检测皮肤组织结缔组织生长因子(CTGF)、转化生长因子-β(TGF-β)含量及血清白细胞介素-6(IL-6)、白细胞介素-17(IL-17)水平。结果:与对照组比较,模型组皮肤真皮厚度,皮肤组织CTGF、TGF-β、HYP含量及血清 IL-6、IL-17 水平明显升高(P＜0.05);与模型组比较,强的松组、红花低、中、高剂量组皮肤真皮厚度,皮肤组织 CTGF、TGF-β、HYP含量及血清 IL-6、IL-17水平明显降低(P＜0.05)。结论:红花水提取液可改善SSc小鼠皮肤状况(或真皮厚度),其作用机制可能与减轻免疫炎症反应有关。     

Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues

S-Adenosyl-l-methionine (AdoMet or SAM)-dependent methyltransferases (MTase) catalyze the transfer of the activated methyl group from AdoMet to specific positions in DNA, RNA, proteins and small biomolecules. This natural methylation reaction can be expanded to a wide variety of alkylation reactions using synthetic cofactor analogues. Replacement of the reactive sulfonium center of AdoMet with an aziridine ring leads to cofactors which can be coupled with DNA by various DNA MTases. These aziridine cofactors can be equipped with reporter groups at different positions of the adenine moiety and used for Sequence-specific Methyltransferase-Induced Labeling of DNA (SMILing DNA). As a typical example we give a protocol for biotinylation of pBR322 plasmid DNA at the 5’-ATCGAT-3’ sequence with the DNA MTase M.BseCI and the aziridine cofactor 6BAz in one step. Extension of the activated methyl group with unsaturated alkyl groups results in another class of AdoMet analogues which are used for methyltransferase-directed Transfer of Activated Groups (mTAG). Since the extended side chains are activated by the sulfonium center and the unsaturated bond, these cofactors are called double-activated AdoMet analogues. These analogues not only function as cofactors for DNA MTases, like the aziridine cofactors, but also for RNA, protein and small molecule MTases. They are typically used for enzymatic modification of MTase substrates with unique functional groups which are labeled with reporter groups in a second chemical step. This is exemplified in a protocol for fluorescence labeling of histone H3 protein. A small propargyl group is transferred from the cofactor analogue SeAdoYn to the protein by the histone H3 lysine 4 (H3K4) MTase Set7/9 followed by click labeling of the alkynylated histone H3 with TAMRA azide. MTase-mediated labeling with cofactor analogues is an enabling technology for many exciting applications including identification and functional study of MTase substrates as well as DNA genotyping and methylation detection.     

Physiology of pepper fruit and the metabolism of antioxidants: chloroplasts,mitochondria and peroxisomes

Background and Aims Pepper (Capsicum annuum) contains high levels of antioxidants, such as vitamins A and C and flavonoids. However, information on the role of these beneficial compounds in the physiology of pepper fruit remains scarce. Recent studies have shown that antioxidants in ripe pepper fruit play a key role in responses to temperature changes, and the redox state at the time of harvest affects the nutritional value for human consumption. In this paper, the role of antioxidant metabolism of pepper fruit during ripening and in the response to low temperature is addressed, paying particular attention to ascorbate, NADPH and the superoxide dismutase enzymatic system. The participation of chloroplasts, mitochondria and peroxisomes in the ripening process is also investigated.Scope and Results Important changes occur at a subcellular level during ripening of pepper fruit. Chloroplasts turn into chromoplasts, with drastic conversion of their metabolism, and the role of the ascorbate–glutathione cycle is essential. In mitochondria from red fruits, higher ascorbate peroxidase (APX) and Mn-SOD activities are involved in avoiding the accumulation of reactive oxygen species in these organelles during ripening. Peroxisomes, whose antioxidant capacity at fruit ripening is substantially affected, display an atypical metabolic pattern during this physiological stage. In spite of these differences observed in the antioxidative metabolism of mitochondria and peroxisomes, proteomic analysis of these organelles, carried out by 2-D electrophoresis and MALDI-TOF/TOF and provided here for the first time, reveals no changes between the antioxidant metabolism from immature (green) and ripe (red) fruits.Conclusions Taken together, the results show that investigation of molecular and enzymatic antioxidants from cell compartments, especially chloroplasts, mitochondria and peroxisomes, is a useful tool to study the physiology of pepper fruit, particularly in the context of expanding their shelf-life after harvest and in maintaining their nutritional value.     

Temporal and intraclonal variation of flowering and pseudovivipary in Poa bulbosa

Background and Aims

Versatility in the reproductive development of pseudoviviparous grasses in response to growth conditions is an intriguing reproduction strategy. To better understand this strategy, this study examined variation in flowering and pseudovivipary among populations, co-occurring clones within populations, and among tillers in individual clones of Poa bulbosa, a summer-dormant geophytic grass that reproduces sexually by seed, and asexually by basal tiller bulbs and bulbils formed in proliferated panicles.

Methods

Clones were collected from 17 populations across a rainfall gradient. Patterns of reproduction were monitored for 11 years in a common garden experiment and related to interannual differences in climatic conditions. Intraclonal variation in flowering and pseudovivipary was studied in a phytotron, under daylengths marginal for flowering induction.

Key Results

Clones showed large temporal variability in their reproductive behaviour. They flowered in some years but not in others, produced normal or proliferated panicles in different years, or became dormant without flowering. Proliferating clones did not show a distinct time sequence of flowering and proliferation across years. Populations differed in incidence of flowering and proliferation. The proportion of flowering clones increased with decreasing rainfall at the site of population origin, but no consistent relationship was found between flowering and precipitation in the common garden experiment across years. In contrast, flowering decreased at higher temperatures during early growth stages after bulb sprouting. Pulses of soil fertilization greatly increased the proportion of flowering clones and panicle production. High intraclonal tiller heterogeneity was observed, as shown by the divergent developmental fates of daughter plants arising from bulbs from the same parent clone and grown under similar conditions. Panicle proliferation was enhanced by non-inductive 8 h short days, while marginally inductive 12 h days promoted normal panicles.

Conclusions

Interannual variation in flowering and proliferation in P. bulbosa clones was attributed to differences in the onset of the rainy season, resulting in different daylength and temperature conditions during the early stages of growth, during which induction of flowering and dormancy occurs.