Cytoskeleton and mitochondrial morphology of saprotrophs and the pathogen Heterobasidion annosum in the presence of Suillus bovinus metabolites

Ectomycorrhizal fungi are known to synthesize antifungal compounds both in vitro and in symbiosis with the host-plants. Culture filtrates of the ectomycorrhizal fungus Suillus bovinus (at pHs of 2.5–6) showed antifungal activity towards saprotrophs Trichoderma harzianum, and Trichoderma virens and the pathogen Heterobasidion annosum, by significantly suppressing their growth relative to sterile liquid medium at the same pHs. In the presence of the culture filtrates, hyphae of the saprotrophs and the pathogen were characterized by distensions, irregular and frequent branching, tip damage and cytoplasm coagulation. Since hyphal abnormalities may be evoked by disruptions in the cytoskeleton and mitochondria, their structural changes were also examined. Depolymerization of microtubules was confirmed for all of the fungi. Serious damage to mitochondria morphology may cause significant functional impairment. Growth of mycelia was inhibited in the lower pH S. bovinus culture filtrate, and the mitochondrial morphology was altered. This suggests that the activity of antifungal compounds synthesized by ectomycorrhizal fungus is significantly affected by pH.     

The capacity in Heterobasidion annosum s.l. to resist overgrowth by the biocontrol agent Phlebiopsis gigantea is a heritable trait

The necrotrophic pathogen Heterobasidion annosum sensu lato (Fr.) Bref. causes severe root rot on coniferous trees in the boreal and temperate forests. The annual economic losses caused by this fungus in Europe are estimated to at least 790 million €. In managed forests, the major route of infection is via stump surfaces from which the H. annosum s.l. grows through the roots and attacks adjacent healthy trees. A biocontrol method to reduce H. annosum s.l. infection is to apply the wood degrading fungus Phlebiopsis gigantea in a spore solution (Rotstop) directly on the freshly cut stumps immediately after cutting. We investigated the potential risk for a build-up in the capacity of H. annosum s.l. to resist overgrowth by P. gigantea. Wood blocks of Picea abies, precolonized with the two fungal species, were juxtaposed on top of agar and the overgrowth of the P. gigantea strain (Rotstop) on the H. annosum s.l. was measured periodically. We found a natural variation in Heterobasidion parviporum to resist overgrowth by P. gigantea. There was no difference between homo- and heterokaryotic strains. In a mapping population of 91 progenies from a H. annosum hybrid strain we were able to identify one quantitative trait locus (QTL) which controls the examined resistance capacity. We estimated the broad sense heritability to 0.336 for the capacity to resist the P. gigantea overgrowth. We conclude that there exists a theoretical risk for resistance build-up in the H. annosum s.l. population towards its biological control agent P. gigantea.     

水引发对油松种子和幼苗的生理效应及其作用机理

为了解决油松种子在直播造林中发芽率低、幼苗抗旱性差的问题,研究了油松种子水引发的方法及引发对其种子和幼苗的生理效应,并对水引发后幼苗的抗旱机理进行了探索。结果表明:油松种子水引发的最佳条件为10℃,闭光条件下分批给种子加水,加水总量为种子初始质量的30%,引发10 d。水引发可显著(P0.05)降低种子的半数发芽天数和硬实率,显著提高种子的发芽指数、最终发芽率和活力指数;水引发种子的可溶性糖及可溶性蛋白含量相比对照也得到提高。水引发可增加油松幼苗的茎粗、鲜重及根长,降低其相对苗高;并极显著(P0.01)提高了幼苗过氧化氢酶、过氧化物酶和根内脱氢酶活性,降低了游离氨基酸含量。干旱胁迫下,引发种子的幼苗脯氨酸和可溶性糖含量分别高于对照39.6%、118.9%,丙二醛含量低于对照93.4%,均达极显著水平。综合分析认为,水引发提高了种子可溶性糖及可溶性蛋白含量为种子萌发提供了保障,从而提高了种子发芽率,并通过增强幼苗抗氧化酶活性、降低丙二醛含量,进而增强幼苗抗氧化系统的功能,最终明显提高其抗旱性。     

马尾松组培苗对氮素形态的生长响应

马尾松属高氮需求树种,然而在苗木培育中马尾松对氮素,尤其是不同形态氮素的需求尚不明确.该文以马尾松组培苗为试验材料,采用基质培养方法,针对硝态氮、铵态氮两种氮素形态均分别设置了2、4、8、16 mmol·L-14个处理,以不添加氮素为对照,对苗木的高径生长、根构型参数(总根长、总表面积、总体积、平均直径和根尖数)以及生...     

干旱和遮荫对马尾松幼苗生长和光合特性的影响

为探究马尾松对干旱和遮荫胁迫的生理响应规律和适应机制,以2年生马尾松幼苗为对象,设置对照(CK)、模拟干旱(DR)、遮荫(LL)以及干旱与遮荫的交互处理(DRLL)4种环境,研究干旱和遮荫对马尾松幼苗的生长和光合生理特性的影响。结果表明:(1)在干旱、遮荫和二者的交互处理下,马尾松幼苗的基径和株高增长量均显著减小,且二者的交互处理加重了干旱和遮荫单一处理下的减小趋势,二者交互作用的影响符合"相互作用理论"。(2)在干旱处理下,针叶长度和比叶面积减小,净光合速率、蒸腾速率、气孔导度和细胞间隙CO₂浓度显著降低,水分利用效率显著增加,光合色素含量基本不变。(3)在遮荫处理下,针叶长度和比叶面积增大,净光合速率、蒸腾速率、气孔导度显著降低,光合色素含量显著增加,水分利用效率和细胞间隙CO₂浓度基本未发生改变。(4)二者的交互处理下,针叶长度、比叶面积减小,净光合速率、蒸腾速率和气孔导度显著降低,且降低程度大于干旱单一处理,光合色素含量显著增加,但其增加程度小于遮荫单一处理。说明干旱和遮荫均能抑制马尾松植株的生长,但其光合生理特性在干旱和遮荫胁迫下分...     

不同强度净风频繁吹袭对樟子松(Pinus sylvestris var.mongolica)幼苗光合蒸腾特征的影响

为了解不同强度净风频繁吹袭对樟子松幼苗光合蒸腾特性的影响,2013年春季在内蒙古科尔沁沙地研究了0(对照)、6、9、12、15、18 m/s等6个风速处理(分别相当于0、4、5、6、7、8级风)4次吹袭下樟子松幼苗光合速率、蒸腾速率、水分利用效率等指标的变化。结果表明,净风频繁吹袭没有改变樟子松幼苗的光合速率和蒸腾速率的日变化规律,但可使其光合蒸腾的"午休"时间加长、"休眠"程度加深;随着风吹强度的增加,其日均光合能力和蒸腾速率显著降低,其中18 m/s处理较对照分别下降27.6%和22.3%;随着风吹强度增加,气孔导度、胞间CO_2浓度均先下降后回升,除18 m/s处理胞间CO_2浓度显著高于CK外,其他处理均显著低于CK;随着风吹强度增加,水分利用效率和光能利用效率均先增加后下降,其中除18 m/s处理的水分利用效率显著低于CK,6 m/s处理的光能利用效率高于CK外,其他处理的水分利用效率均高于CK,光能利用效率均低于CK;日均光合蒸腾速率的下降主要源于气孔导度的降低,而水分利用效率和光能利用效率的变化均受制于光合速率和蒸腾速率的变化。     

Plant and fungal responses to elevated atmospheric carbon dioxide in mycorrhizal seedlings of Pinus sylvestris

The effects of elevated CO₂ concentration upon the mycorrhizal relationships of Scots pine (Pinus sylvestris) seedlings were investigated. Plants were grown for 4 months with their shoots exposed to ambient (C_AMB=360 μl l⁻¹) or elevated (C_ELEV=700 μl l⁻¹) CO₂ environments while their root systems, either colonised by the mycorrhizal fungi Paxillus involutus or Suillus bovinus, or left in the non-mycorrhizal condition, were maintained in sealed dishes. In one series of these plants the effects of C_ELEV upon the extent of mycorrhizal development and upon their growth and nutrition were determined, while another series were transferred from the dishes after 1 month, to transparent observation chambers before being returned to the two CO₂ environments. In these chambers, the effects of C_ELEV upon development of the external mycelial systems of the two mycorrhizal fungi was determined by measuring the advance of the hyphal fronts of the mycorrhizal fungi across non-sterile peat from the colonised plants. The dry mass and number of mycorrhizal tips were significantly higher in C_ELEV than in the C_AMB condition in plants colonised by both fungi in the dishes. Yields of whole plants and of shoots were higher in the C_ELEV treatment whether or not they were grown in the mycorrhizal condition, but the greater yields were not associated in these sealed systems with enhanced nutrient gain. The dry mass of non-mycorrhizal plants was greater than that of those colonised by mycorrhizal fungi under elevated CO₂. This is thought to be attributable to the energetic cost of production of the larger mycorrhizal systems in this treatment. The extent of development of the mycorrhizal mycelial systems of both fungi was greatly increased in C_ELEV relative to that in C_AMB environments. It is hypothesised that increased allocation of carbon to mycorrhizal root systems and their associated mycelia would provide the potential for enhancement of nutrient acquisition in open systems of greater fertility.     

模拟氮沉降和接种外生菌根真菌对马尾松(Pinus massoniana)幼苗营养元素的影响

量化植物地上部和地下部元素含量对于理解和预测植物养分平衡如何响应大气氮沉降的变化至关重要。通过盆栽试验研究了氮沉降增加背景下外生菌根真菌对马尾松幼苗营养元素的影响。对马尾松幼苗进行了接种两种外生菌根真菌：（彩色豆马勃（Pisolithus tinctorius,Pt）与厚环乳牛肝菌（Suillus grevillei,Sg））以及4种氮素浓度添加：0 kg N hm^-2a^-1（N0）、正常氮沉降30 kg N hm^-2a^-1（N30）、中度氮沉降60 kg N hm^-2a^-1（N60）、重度氮沉降90 kg N hm^-2a^-1（N90）,共12个处理,测定了马尾松地上部和地下部大量元素和微量元素的含量。结果表明：施氮改变了营养元素在马尾松幼苗地上部和地下部的含量,马尾松幼苗磷（P）、钙（Ca）、铁（Fe）、锰（Mn）等元素均在N60时达到临界值,而当输入的量超过了马尾松对氮的需求时,氮沉降会使马尾松营养元素含量较最适浓度时降低,地上部碳（C）随施氮浓度的升高先升高后降低,N随施氮浓度的升高而升高,根系和叶片钾（K）、Ca、镁（Mg）均随施氮浓度的升高而降低,施氮也降低了根系C及微量元素的含量。但在同一施氮浓度下,接种外生菌根真菌（EMF）后能够提高大多数元素的含量,N90时接种厚环乳牛肝菌（Sg）和彩色豆马勃（Pt）的叶片N含量与对照相比分别提高112.6%和138.6%,根系N含量分别提高73.1%、71.6%;N60时接种Sg和Pt的植株叶片P含量比不施氮未接种对照分别提高了166.3%、132.9%,根系P含量分别提高了40.8%、38.5%。EMF能够维持植物养分平衡,从而降低高施氮量对植物的影响效果。这为未来气候变化情景中氮沉降增加下接种EMF可以调节植物元素含量,从而达到更适应环境的元素平衡来促进生长提供理论依据。     

野外模拟酸雨胁迫下接种外生菌根真菌对马尾松幼苗的缓解作用

外生菌根真菌能够提高宿主植物对外界环境胁迫的抵抗力。主要探讨野外条件下外生菌根真菌对酸雨胁迫下马尾松(Pinus massoniana)幼苗生长、养分元素以及表层土壤的影响,以期为酸雨严重区马尾松林恢复提供科学依据。以2年生马尾松幼苗为材料,采用原位试验,共设置6个处理:p H5.6(对照)处理未接种、对照处理接种、p H4.5酸雨处理未接种、p H4.5酸雨处理接种、p H3.5酸雨处理未接种、p H3.5酸雨处理接种。研究表明:(1)酸雨处理与对照处理相比显著降低了非菌根苗总生物量及各部位生物量(根、茎、叶),对株高无显著影响,接种外生菌根真菌可以缓解酸雨对马尾松幼苗生长的不利影响;(2)与对照处理相比,酸雨处理的非菌根苗的针叶中N、P、Ca含量升高,Mg含量降低,根系中N、P、Ca含量降低,Mg含量随p H的降低先升高后降低。接种外生菌根真菌显著提高了p H3.5酸雨处理的马尾松幼苗根系中N、P、Ca、Mg含量,而对针叶中N、P、Ca、Mg含量无显著影响。(3)在非菌根土壤中,p H3.5酸雨处理与对照处理相比显著降低了土壤中有机质、速效磷、速效钾、可溶性碳、可溶性氮、铵态氮、硝态氮含量,而接种外生菌根真菌显著提高了上述指标。酸雨对土壤阳离子交换量无显著影响。总而言之,接种外生菌根真菌促进了酸雨处理的马尾松幼苗生长、缓解了酸雨对马尾松幼苗养分元素和表层土壤的不利影响,由此可见接种外生菌根真菌是减轻酸雨对马尾松危害的一个重要途径。     

Quantitative comparison of chemical, biological and mechanical induction of secondary compounds in Pinus pinaster seedlings

Chemical elicitors and mechanical treatments simulating real insect herbivory have been increasingly used to study induced defensive responses in woody plants. However, simultaneous quantitative comparisons of plant chemical defences elicited by real and simulated herbivory have received little attention. In this paper we compared the effects of real herbivory, simulated herbivory using two chemical elicitors, and mechanical damage treatments on the quantitative secondary chemistry of Pinus pinaster juveniles (namely on non-volatile resin in the stem and total phenolics in the needles). The real herbivory involved phloem wounding by Hylobius abietis and defoliation by Brachyderes lusitanicus (two pine weevils); the chemical elicitors to simulate herbivory induction were 40 mM methyl jasmonate (MJ) and 20 μM benzothiadiazole (BTH); and the mechanical treatments included phloem wounding and needle clipping. We also performed an additional experiment for assessing at what extent insect extracts could increase plant responses over mechanical damage. Chemical induction with MJ, mechanical wounding and real phloem herbivory by H. abietis all produced quantitatively similar results, increasing the concentration of resin in the stem and total phenolics in the needles by equivalent magnitudes. Exogenous application of BTH increased the concentration of phenolic compounds in pine needles, but did not show the same effect on stem resin. Contrastingly, we did not find significant changes in the concentration of resin in the stem or phenolics in the needles after needle clipping and B. lusitanicus feeding. Mechanical damage followed by the application of extracts from the insects B. lusitanicus and H. abietis on the injured tissues did not increase the responses in comparison to mechanical damage alone. The fact that strong induced responses elicited by phloem wounding insects are equally elicited by phloem injuries suggests that defences in pine trees are raised with low specificity regarding biotic enemies. Results from this paper support future methodological approaches using chemical elicitors and mechanical damage as simulated herbivory treatments for the experimental induction of conifer defences.     

New labdane resin acids from Pinus elliottii

Slash pine needles and cortex oleoresin have been found to contain a new major diterpene constituent, imbricataloic acid. The closely related imbricatoloic acid, previously reported only in Araucaria imbricata, was found to be present in small amounts in slash pine needle extract. Spectral data are given for an unidentified diterpene alcohol isolated from the cortex oleoresin.     

Identification of volatiles from felled Pinus radiata and the electroantennograms they elicit from Sirex noctilio

Volatiles emanating from Pinus radiata show only minor changes in composition with time after felling even though attractiveness of the felled timber to the wood wasp Sirex noctilio changes markedly over the first 3 weeks. Monoterpene hydrocarbons account for more than 95% of the volatiles and show little change in relative composition during this time. Oxygenated components (camphor, pinocamphone, isopinocamphone and trans-pinocarveol), which are absent or present in trace amounts immediately after felling, are produced in increasing amounts and account for 1% of the volatiles after 3 weeks. Their presence in the volatile fraction significantly increases the response from the antenna of S. noctilio. The major monoterpene hydrocarbons and oxygenated components were identified by chromatographic and spectral techniques.

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Résumé Les composés volatils émanant de Pinus radiata ne montrent que de faibles changements dans leur composition après l'abattage, bien que l'attractivité des arbres abattus à l'égard de Sirex noctilio change nettement au-delà de 3 semaines. Pendant les premières 3 semaines suivant l'abattage les monoterpènes hydrocarbons représentent plus de 95% des composés volatils et leur composition varie peu durant cette période. Les composants oxygénés (camphor, pinocamphone, isopinocamphone et trans-pinocarveol) qui sont absents ou seulement présents à l'état de traces immédiatement après l'abattage, sont produits en quantités croissantes et représentent 1% des substances volatiles au bout de 3 semaines. Leur présence dans la fraction volatile accroît de facon significative la réponse antennaire de Sirex noctilio. Les principaux monoterpènes hydrocarbonés et les composants oxygénés ont été identifiés par les techniques chromatographiques et spectrographiques.

     

Activity of ligninase peroxidase from Acinetobacter anitratus and the degradation of Pinus pinaster lignin

The ligninase activity of Acinetobacter anitratus cells and enzyme extracted from that bacterium was measured with a model substrate (benzyl alcohol) and two lignin substrates: Pinus pinaster lignin precipitated from kraft black liquor and treated black liquor. The activity of this intracellular enzyme was confirmed on benzyl alcohol, showing a sigmoid Michaelis curve for constant H₂O₂ concentration. Enzymatic activity on treated black liquor leads to the production of low molecular weight compounds and to partial polymerisation of lignin. The evolution of the molecular weight distribution of the original sample with time was followed by high pressure liquid chromatography.