马尾松(Pinus massoniana)、湿地松(Pinus elliottii)挥发性化学物质的昼夜节律释放

用TCT-GC／MS分析了马尾松、湿地松挥发性有机化合物的昼夜节律变化。结果表明：马尾松昼夜节律中检测到的挥发物主要是萜烯类化合物,其中单萜种类最多,且α-蒎烯和β-蒎烯含量约占整个挥发物的80％,其次是含氧化合物等。这些挥发物的释放高峰多数在10：30,少数在1：30;整个变化中有两个低峰期,即13：30和22：30。湿地松中检测到的挥发物组分与马尾松相似,多数释放高峰在12：00-15：00之间;另一些在6：00;α-蒎烯的释放高峰在3：00,而此时其它挥发性化合物的释放量最低。挥发物的释放也受到外界环境的影响,一定范围内随着温度的升高、湿度的减小,其释放量增加。     

外用不同浓度茉莉酸甲酯诱导的茶树挥发物的种类和时序变化

【目的】明确MeJA对茶树挥发物的诱导作用。【方法】采用顶空活体取样法对不同浓度MeJA处理后的茶苗挥发物进行抽提,并利用GC-MS对挥发物进行鉴定。【结果】不同剂量MeJA显著地影响茶树挥发物的种类组成和释放量,50μL MeJA处理可显著诱导茶树释放香叶烯、萜品油烯、罗勒烯等10种单萜类化合物,法呢烯、橙花叔醇和红没药烯等7种倍半萜类化合物,苯甲醇、苯乙腈和吲哚等5种氨基酸衍生物,以及3种未知化合物;而100μL MeJA处理仅能诱导茶树释放7种化合物。不同挥发物对MeJA处理的响应时间不同,但其释放量都具有昼高夜低的趋势。并且,释放量的大小明显受到光照强度的影响。【结论】外用MeJA喷雾处理可诱导茶树挥发物的产生和释放。     

茉莉酸甲酯对水稻幼苗生长的影响(简报)

2．5×10－7mol·L-1茉莉酸甲酯促进水稻幼苗生长,而高于2．5×10-5mol·L－1则抑制。不同水稻品种对茉莉酸甲酯的反应不同。     

内源茉莉酸甲酯的积累改变了大豆叶片与根的发育

茉莉酸甲酯是一种调节植物形态发生、诱导防御相关基因的植物信号转导分子。为了解内源茉莉酸甲酯在植物发育中的作用,将编码茉莉酸甲基转移酶的NTR1基因与CaMV 35S启动子连接并导入大豆植株。PCR及Northern杂交结果表明,NTR1基因稳定整合在大豆基因组并得到过量表达。与野生型植株相比,转基因大豆叶片与根的形态发生了显著的变化。大部分转基因大豆叶片变得细长,初生根生长受到抑制而侧根的生长却受到促进。定量分析结果表明,转基因大豆植株叶片中茉莉酸甲酯的含量比对照高出 2～2.5 倍。这些结果表明,内源茉莉酸甲酯的积累参与了大豆形态发生的调控。     

茉莉酸甲酯喷施和光肩星天牛（Anoplophora glabripennis(Motschulsky)咬食后五角枫释放的挥发物

茉莉酸甲酯喷施和光肩星天牛(Anoplophora glabripennis(Motschulsky)咬食后五角枫(Acer mono Maxim.)植株均诱导产生了乙酸丁酯、2-壬醇、乙酸己酯、3-甲基-2-丁醇、1-辛烯、里那醇、3-蒈烯、1-丁醇、(Z)-2-己烯醛9种正常植株中没有检测到的挥发物。两种处理均诱导或促进了萜烯类、醇类和醛类挥发物的大量释放,且乙酸-3-己烯酯、乙酸乙酯、己醇、反-2-己烯醇、乙基己醇、1-辛烯、石竹烯、法尼烯等挥发物的时序变化趋势也非常相似。因而推测光肩星天牛咬食五角枫后在植物体内诱导产生了茉莉酸信号传导途径。     

茉莉酸甲酯和虫害诱导棉花对棉蚜及其天敌选择行为的影响

以棉蚜及其天敌十一星瓢虫、叶色草蛉为对象,通过Y型嗅觉仪测定了3种昆虫对不同味源物的行为反应。结果表明,健康棉株叶片对棉蚜的吸引作用与虫害棉叶和茉莉酸甲酯(MeJA)处理棉叶未存在显著性差异;十一星瓢虫和叶色草蛉对虫害棉叶的选择性是健康棉叶的2.7和2.2倍(P<0.001),对MeJA处理棉叶的选择性明显高于健康叶片(P<0.001),其中0.02μmol.mL-1处理的棉叶对天敌的吸引力最强,单一的MeJA对十一星瓢虫和叶色草蛉也具有吸引作用。     

茉莉酸甲酯和机械损伤对茶树叶片多酚氧化酶时序表达的影响

以温室中培养的龙井43茶树2年生扦插苗为实验材料,采用分光光度计法测定了正常茶树叶片中多酚氧化酶(PPO)活性变化的日节律,以及茉莉酸甲酯(MeJA)处理和机械损伤诱导不同叶位叶片PPO活性的时序变化.结果表明:正常茶树叶片中PPO活性高峰出现在4:00;MeJA处理茶树30 h后,其叶片PPO活性与对照间呈现显著差异,于处理后174 h达到最高峰;半定量RT-PCR检测结果发现,MeJA诱导茶树叶片PPO基因表达量最高峰出现在处理后126 h,比生化测定结果提前了48 h.机械损伤可诱导处理叶及其下叶位叶片PPO活性迅速升高,表现出一定系统性;机械损伤叶片与系统诱导叶片间PPO活性随时间变化的趋势基本平行,但机械损伤叶片中PPO活性始终高于系统诱导叶片.本研究证明,MeJA和机械损伤均可诱导茶树叶片中PPO活性的显著增强,进一步证实茉莉酸(JA)信号转导途径在茶树的直接防御反应中具有重要作用.     

氨基茚磷酸(AIP)和氨基氧乙酸(AOA)对茉莉酸甲酯诱导的烟草一些酶活性的影响

用茉莉酸甲酯(MJ,1mg ml^-1)处理培养在含0．1mmol／L AIP(水杨酸合成抑制剂)和／或1mmol／L AOA(乙烯合成抑制剂)的MS培养基上的烟草愈伤组织,测定某些酶的活性。结果表明：MJ明显提高过氧化物酶(POD)、β1,3-葡聚糖苷酶和几丁质外切酶的活性,略微促进苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)的活性,抑制几丁内切酶的活性,而AOA和AIP则明显抑制MJ对POD和β1,3-葡聚糖苷酶活性的诱导作用,但对MJ诱导的PAL和PPO的活性影响很小,AOA和AIP可能作为逆境因子促进PAL的活性。AOA能部分解除MJ对几丁内切酶的抑制作用,但对MJ诱导的几丁外切酶的影响较小,而AIP抑制几丁内切酶的活性,也抑制MJ对此酶的诱导作用。因此我们认为：MJ对POD、PPO和几丁内切酶的影响可能是通过乙烯途径,对β1,3-葡聚糖苷酶和几丁外切酶的影响可能是通过水杨酸(SA)途径,而对PAL的影响可能是通过其它途径。     

茉莉酸甲酯(MeJA)对贯叶连翘悬浮细胞生长和贯叶金丝桃素产量的影响

培养基中添加100 μm01·L-1MeJA后,贯叶连翘悬浮细胞生物量增加量和贯叶金丝桃素(HF)的产量分别是未经MeJA处理的1.3倍和1.73倍;培养3 d时,培养基中添加100 μmol·L-1 MeJA能提高HF产量,是未经MeJA处理的2.4倍.     

茉莉酸甲酯对灵芝三萜生物合成的影响及其灵芝应答基因的差异表达

灵芝三萜类化合物是灵芝的主要药用成分之一,灵芝三萜含量的多少是衡量灵芝质量高低的重要指标。经研究发现茉莉酸甲酯能显著提高灵芝三萜含量并上调灵芝三萜生物合成途径中的羟甲基戊二酰辅酶A还原酶、法尼酯焦磷酸合成酶和羊毛甾醇合成酶等基因的表达。通过在灵芝中建立的cDNA-AFLP体系,采用64对引物组合对灵芝的转录组进行分析。64对引物组合共产生3 910个TDFs。茉莉酸甲酯不同诱导条件下约有919个TDFs的表达谱发生了变化,占表达谱的23.5%,其中上调表达的有703个,下调表达的有216个。选择458个TDFs进行回收、TA克隆、测序后,获得390个高质量的TDFs片段。通过蛋白数据库同源性检索,并对蛋白功能进行GO分类,结果表明:390个高质量的TDFs中,BlastX比对后没有显著匹配的序列占77.1%。在有匹配蛋白的序列中,与碳代谢及能量相关的基因为最大一组,占35.2%,转录调控类及信号传导类基因分别占14.3%和11.0%。通过荧光定量PCR技术考察了茉莉酸甲酯对20个基因转录水平的影响,与cDNA-AFLP分析的结果相一致。并通过灵芝基因过量表达技术和基因沉默技术对筛选到的相关基因进行沉默和过量表达,研究筛选到的基因在灵芝三萜生物合成与调控中的作用,获得大量参与灵芝三萜生物合成与调控的候选基因,进一步阐明了茉莉酸甲酯诱导下灵芝基因表达的变化图。     

受害马尾松、湿地松挥发性化学物质的释放

萜烯类化合物是针叶树防御昆虫为害的重要组分。对盆栽马尾松、湿地松进行未受害、接虫咬食为害、人工剪叶处理后,用TCT-GC-M S分析了叶片挥发物的成分及相对含量。结果表明:人工剪叶后马尾松挥发物相对含量除α-蒎烯降低外,其余都有所升高,但挥发性化学物质相对含量出现高峰的时间没有规律性;虫害后,挥发物的相对含量除水芹烯降低外,其余均在1h升高,同时检测到p-薄荷-1.4(δ)-二烯,但其变化较小。湿地松受害后检测到7种挥发性化学物质,除人工剪叶后α-蒎烯、虫害后α-蒎烯、水芹烯相对含量降低外,其它挥发物的相对含量在不同的时间内都有所升高。β-蒎烯在两种松树受害后都有显著的变化,马尾松中的石竹烯没有明显的变化,氨茴酸-3,5-二甲基-3-乙烯基-4-己烯酯、大香叶烯D只在湿地松中检测到,而且其变化显著。     

Allelopathic effects of volatile organic compounds released from Pinus halepensis needles and roots

The Mediterranean region is recognized as a global biodiversity hotspot. However, over the last decades, the cessation of traditional farming in the north part of the Mediterranean basin has given way to strong afforestation leading to occurrence of abandoned agricultural lands colonized by pioneer expansionist species like Pinus halepensis. This pine species is known to synthesize a wide range of secondary metabolites, and previous studies have demonstrated strong allelopathic potentialities of its needle and root leachates. Pinus halepensis is also recognized to release significant amounts of volatile organic compounds (VOC) with potential allelopathic effects that have never been investigated. In this context, the objectives of the present study were to improve our knowledge about the VOC released from P. halepensis needles and roots, determine if these VOC affect the seed germination and root growth of two herbaceous target species (Lactuca sativa and Linum strictum), and evaluate if soil microorganisms modulate the potential allelopathic effects of these VOC. Thirty terpenes were detected from both, needle and root emissions with β‐caryophyllene as the major volatile. Numerous terpenes, such as β‐caryophyllene, δ‐terpinene, or α‐pinene, showed higher headspace concentrations according to the gradient green needles < senescent needles < needle litter. Seed germination and root growth of the two target species were mainly reduced in presence of P. halepensis VOC. In strong contrast with the trend reported with needle leachates in literature, we observed an increasing inhibitory effect of P. halepensis VOC with the progress of needle physiological stages (i.e., green needle < senescent needle < needle litter). Surprisingly, several inhibitory effects observed on filter paper were also found or even amplified when natural soil was used as a substrate, highlighting that soil microorganisms do not necessarily limit the negative effects of VOC released by P. halepensis on herbaceous target species.     

受害马尾松木质素含量及其过氧化物酶活性

木质素是植物防御植食性昆虫危害的重要物质。对林间马尾松针叶进行接虫咬食处理后,测定了不同时间内木质素含量、过氧化物酶活性及其同功酶谱带的变化。结果表明:与对照相比,虫害针叶1h木质素含量就略有升高,而系统针叶木质素含量10h才开始升高,24h两种处理的木质素含量及过氧化物酶活性均显著提高;受害后所取时间点内,其差异逐渐加大,这种差异在于表达量不同。F检验的结果显示,虫害及系统针叶木质素含量、过氧化物酶活性及同工酶谱带变化一致,均在24h达到显著差异(P(0.05)。     

Effects of exogenous methyl jasmonate on the synthesis of endogenous jasmonates and the regulation of photosynthesis in citrus

Methyl jasmonate (MeJA) is an airborne signaling phytohormone that can induce changes in endogenous jasmonates (JAs) and cause photosynthetic responses. However, the response of these two aspects of citrus plants at different MeJA concentrations is still unclear. Four MeJA concentrations were used in two citrus varieties, Huangguogan (C. reticulata × C. sinensis) and Shiranuhi [C. reticulata × (C. reticulata × C. sinensis)], to investigate the effects of MeJA dose on the endogenous JAs pathway and photosynthetic capacity. We observed that MeJA acted in a dose-dependent manner, and its stimulation in citrus leaves showed a bidirectional character at different concentrations. This work demonstrates that MeJA at only a concentration of 2.2 mM or less contributed to the activation of magnesium protoporphyrin IX methyltransferase (ChlM, EC 2.1.1.11) and protochlorophyllide oxidoreductase (POR, EC 1.3.1.11) and the simultaneous accumulation of Chl a and Chl b, which in turn contributed to an improved photosynthetic capacity and PSII photochemistry efficiency of citrus. Meanwhile, the inhibition of endogenous JAs synthesis by exogenous MeJA was observed. This was achieved by reducing the ratio of monogalactosyl diacylglycerol (MGDG) to diagalactosyl diacylglycerol (DGDG) and inhibiting the activities of key enzymes in JAs synthesis, especially 12-oxo-phytodienoic acid reductase (OPR, EC 1.3.1.42). Another noteworthy finding is that there may exist a JA-independent pathway that could regulate 12-oxo-phytodienoic acid (OPDA) synthesis. This study jointly analyzed the internal hormone regulation mechanism and the external physiological response, as well as revealed the effects of exogenous MeJA on promoting the photosynthesis and inhibiting the endogenous JAs synthesis.     

不同温度、光照对虫害紫茎泽兰挥发物释放的影响

用不同温度和光照处理对照和棉蚜侵害的盆栽紫茎泽兰植株后,通过TCT-GC/MS分析了叶片挥发物的成分及相对含量。结果表明:温度与光照均不影响对照和虫害紫茎泽兰挥发物的种类,但影响其挥发物的相对含量。当光照强度达到300400μmol.m-.2s-1时,虫害紫茎泽兰挥发物中,多数单萜的相对含量显著高于对照,而倍半萜含量显著低于对照;绿叶挥发物相对含量与其对照相比差异不显著。当外界温度在15 30℃时,虫害紫茎泽兰挥发物中绿叶气体如己醛、2-己烯醛和多数单萜化合物的相对含量随温度升高而显著增加,而倍半萜含量却低于对照。说明适宜的温度和光照条件影响挥发物化学指纹图的构成,进而可能影响到蚜虫的取食。     

被害马尾松（Pinus massoniana）针叶挥发性物质的提取、鉴定及蚕饰腹寄蝇（Blepharipa zebina）的电生理活性

试验使用动态顶空吸附的方法,提取了被害马尾松(Pinus massoniana)针叶挥发物,并通过气相色谱-质谱联用仪(GC-MS)及标准样品进行鉴定。结果表明,被害马尾松针叶挥发性物质主要包括绿叶挥发物、单萜、含氧单萜及倍半萜等四大类物质;挥发物中以单萜的含量最高,其它3类物质含量较低。通过触角电位图(EAG)技术,测定了23种挥发性物质标准样品的石蜡油溶液对蚕饰腹寄蝇(Blepharipa zebina)雌蝇的电生理活性。结果表明,蚕饰腹寄蝇雌蝇对α-蒎烯、β-蒎烯、长叶烯、莰烯、乙酸冰片酯等的相对触角电位反应值与湿润空气对照相比并无显著差异;雌蝇对其它刺激物的相对触角电位反应值与湿润空气对照相比均存在显著差异。同时测定了雌蝇对8种标样,在5个浓度系列下的剂量-反应,并分析了反应阈值。结果表明,蚕饰腹寄蝇雌蝇对顺-3-己烯-1-醇和里哪醇的反应阈值为0.1μg/μl,对壬醛的反应阈值为1μg/μl,对柠檬烯、α-水芹烯、2,6-二甲基-2,4,6-辛三烯、β-石竹烯4种物质的反应阈值为100μg/μl,而对α-蒎烯,在测试的剂量范围内不存在反应阈值。在确定对蚕饰腹寄蝇有生理活性的物质成分和分析反应阈值的基础上,对寄生蝇类定位寄主的机制进行了讨论。     

Induction of conidiation by endogenous volatile compounds in Trichoderma spp

Light and starvation are two principal environmental stimuli inducing conidiation in the soil micromycete Trichoderma spp. We observed that volatiles produced by conidiating colonies of Trichoderma spp. elicited conidiation in colonies that had not been induced previously by exposure to light. The inducing effect of volatiles was both intra- and interspecific. Chemical profiles of the volatile organic compounds (VOCs) produced by the nonconidiated colonies grown in the dark and by the conidiating colonies were compared using solid-phase microextraction of headspace samples followed by tandem GC-MS. The conidiation was accompanied by increased production of eight-carbon compounds 1-octen-3-ol and its analogs 3-octanol and 3-octanone. When vapors of these compounds were applied individually to dark-grown colonies, they elicited their conidiation already at submicromolar concentrations. It is concluded that the eight-carbon VOCs act as signaling molecules regulating development and mediating intercolony communication in Trichoderma.