内生真菌对植物抗旱性的影响

内生真菌广泛地存在于植物体内,它们在植物体内的生活不会对植物引起任何感病症状,而且内生真菌侵染对植物生长、生物和非生物胁迫抗性很好的促进作用,理解内生真菌在提高植物干旱胁迫耐受性方面的作用和机理对其在缓解植物干旱胁迫中的应用有重要意义。这篇综述介绍了植物内生真菌的多样性、对植物抗旱性的影响及其作用机理等方面的研究进展。内生真菌对植物抗旱性提高的机理包括:干旱耐受、干旱回避和干旱恢复。文中还对以后的研究进行了展望。     

5种内生真菌对玉米幼苗抗干旱的胁迫作用

为了研究干旱胁迫下内生真菌对玉米幼苗生长的影响和筛选具有提高玉米抗旱潜能的优良菌种,在温室盆栽条件下,将分离自新疆荒漠盐生植物根系的5种内生真菌Curvularia inaequalis、Fusarium avenaceum、Neocamarosporium sp.AS65231、Pleosporales AS29261和Ulocladium sp.AS91282分别接种到玉米幼苗根系,进行干旱胁迫培养6周,测定苗期玉米的株高、生物量、可溶性糖含量、过氧化物酶(POD)活性和丙二醛(MDA)含量。研究结果表明:干旱胁迫下,与不接菌对照相比,接种C.inaequalis、F.avenaceum、Pleosporales AS29261和Ulocladium sp.AS91282可显著提高玉米幼苗地下生物量(P0.05),而接种Neocamarosporium sp.AS65231对其影响不显著;接种F.avenaceum和Ulocladium sp.AS91282可极显著提高玉米根系可溶性糖含量(P0.01);接种C.inaequalis、Pleosporales AS29261和Ulocladium sp.AS91282对根系POD活性影响不显著,而F.avenaceum和Neocamarosporium sp.AS65231显著降低了根系POD活性;接种Neocamarosporium sp.AS65231和Ulocladium sp.AS91282显著降低根系的MDA含量(P0.01)。然而,与不接菌对照相比,接种5种内生真菌对玉米幼苗的株高、地上生物量和总生物量,以及叶片的可溶性糖含量、POD活性和MDA含量等均无显著影响。综合比较来看,干旱胁迫下接种内生真菌Ulocladium sp.AS91282对促进玉米幼苗抗干旱能力优于其他4种菌。     

丛枝菌根真菌对连翘幼苗抗旱性的影响

采用集球囊霉（Glomus fasciculatum Gerd.＆ Trappe emend.Walker ＆ Koske）、缩球囊霉（Glomus constrictum Trappe）单独接种和混合接种于连翘（Forsythia suspensa（Thunb.）Vahl）幼苗,研究丛枝菌根真菌对连翘抗旱性的影响.结果表明：在干旱胁迫条件下,随着菌根侵染率的提高,连翘幼苗叶绿素和脯氨酸含量增加,SOD活性增强,丙二醛含量和膜透性降低,苗木枯死率下降.菌根真菌通过促进苗木快速累积游离脯氨酸,提高SOD酶活性,减缓干旱对细胞膜的破坏,延缓了植物受伤害的速度,提高连翘幼苗的抗旱性;在不同接种处理间,混合接种效果最好,其次为缩球囊霉单独接种.     

不同抗旱性玉米幼苗根系抗氧化系统对水分胁迫的反应

以抗旱性不同的2个玉米品种为材料,研究不同程度水分胁迫下玉米根系活性氧清除系统的变化及膜脂过氧化水平。明确了轻度水分胁迫下玉米根系POD、CAT、APX等保护酶活性明显提高;中、重度胁迫下其活性急剧下降,但几种酶对水分胁迫的敏感程度不同。SOD对水分胁迫表现最不敏感,在中度水分胁迫下仍保持上升趋势;抗氧化剂GSH含量变化趋势与保护酶相似;而AsA含量在不同程度水分胁迫下持续下降;MDA含量随水分胁迫程度加剧而增加。其中抗旱性强的鲁玉14与抗旱性弱的掖单13相比具有较高的保护酶活性和抗氧化剂含量,膜脂过氧化程度较轻,除POD外,品种间抗氧化酶活性（抗氧化剂含量）呈极显著差异,说明抗氧化能力强是抗旱性品种具有较强抗旱性的重要原因之一。     

玉米赤霉烯酮浸种对玉米幼苗抗旱性的影响

玉米赤霉烯酮浸种（２４ｈ）可提高玉米幼苗的抗旱性,在干旱条件下,经玉米赤霉烯酮浸种的玉米幼苗叶片中含水量下降缓慢,相对电导率较低,超氧化物歧化酶活性较高,游离脯氨酸含量升高。０．１ｍｇ·Ｌ＾－１玉米赤霉烯酮浸种的抗逆效果优于０．０１ｍｇ·Ｌ＾－１。     

红豆杉—内生真菌化学成分的研究

从药用植物红豆杉（Taxus chinensis(Pilg.)Rehd.）的内生真菌-粘帚霉属真菌（Gliocladium sp.简称F菌）菌丝体中分离到3个化合物,根据光谱方法确定了它们的结构。其中,（20s,22s)-4a-同-22-羟基-4-氧杂麦角甾-7,24（28）-二烯3-酮为新化合物,化合物4,8,12,16-四甲基-1,5,9,13-四氧杂环十六烷-2,6 ,10,14-四酮为首次从该属真菌中分离到,6,9-环氧麦角甾-7,22-二烯-3-羟基为首次从F菌中分离到。     

钙和钙调素对玉米幼苗抗旱性的调控

用２０ｍｍｏｌ／ＬＣａＣｌ２浸种处理能显著提高玉米种子在干旱胁迫下的萌发率。改善玉米幼苗的生长状况,减轻胚根在干旱胁迫下的膜伤害,提高其活力和玉米幼苗在强干旱迫下的存活率。     

内生真菌对草坪植物病原真菌抑制作用的比较

分别从野生牧草羽茅(Achnatherum sibiricum (L.) Keng)、栽培种高羊茅(Festuca arundinacea Schreb.)(品种Millennium)、栽培种黑麦草(Lolium perenne L.) (品种Justus)中分离出内生真菌Neotyphodium sp.、N. coenophialum和N. lolli,通过体外培养法比较了这3种内生真菌对草坪植物病原真菌的抑制作用.结果表明,从羽茅中分离的内生真菌Neotyphodium sp.在两菌相交前对所有供试的病原真菌都有一定的抑制作用,其中对枝孢霉属(Cladosporium sp.)、弯孢霉属(Curvularia sp.)和拟茎点属(Phomopsis sp.)病原真菌的抑制效果尤为显著,对峙培养3d后的抑菌率分别达70.1%、52.3%和30.9%,营养竞争作用、重寄生作用是其主要的拮抗机制;从高羊茅中分离的内生真菌N. coenophialum对枝孢霉属病原真菌存在一定的抑制作用;而从黑麦草中分离的内生真菌N. lolli与病原真菌对峙培养时,病原真菌菌落慢慢侵占整个营养空间,内生真菌停止生长并逐渐褐变死亡.体外培养结果说明Neotyphodium sp.对供试病原真菌的拮抗效果优于N. coenophialum和N. lolli,由此推测Neotyphodium sp.与宿主植物羽茅的共生可能有利于宿主植物抵抗病原真菌的侵扰.     

药用植物内生真菌研究现状和发展趋势

药用植物内生真菌普遍存在于健康植物组织和器官中,种类繁多,分布广泛。到目前为止,人们已从大量药用植物中分离出不同类型的内生真菌,这些植物广泛分布于除南极洲以外的各种陆地生态系统中。随着现代科学技术的迅速发展,药用植物内生真菌的研究也取得了长足的进步。由于内生真菌对于药用植物的重要性,其相关研究也受到了世界各国学者的高度关注。本文总结现阶段药用植物内生真菌相关研究,并对未来研究的发展趋势做出展望。     

芦荟植物内生真菌的研究 Ⅰ．内生真菌的分离及鉴定

从中华芦荟 [AloeveraL .varChinese(HawBerg) ]、元江芦荟 [A .yuanjiangensisXiong&Zhengsp .nov .(暂拟 ) ]及库拉索芦荟 (A .barbadsisMill.)的根、叶、花柄、花中分离获得内生真菌共 88株 ,经形态观察分类鉴定为 4个目、3个科、2 2个属。且查明 ,芦荟不同种类不同部位微生物的数量、分布、种群及其组成存在有差异。     

干旱胁迫下亚精胺对玉米幼苗抗旱性影响的生理生化机制

为探究外源亚精胺(Spd)在增强玉米干旱胁迫耐受性中的作用,以‘先玉335’(干旱不敏感型)和‘丰禾1’(干旱敏感型)为试验材料,采用营养液水培法,研究了15%聚乙二醇(PEG-6000)模拟干旱胁迫下,外源Spd (0.1 mmol·L^-1)对玉米幼苗生长、光合特性、叶绿素含量、渗透调节物质、活性氧生成、膜质过氧化及根系活力的影响.结果表明:干旱胁迫下,外源Spd处理可显著促进干旱胁迫下玉米幼苗的生长;提高叶绿素含量、净光合速率(P_n)、气孔导度(g_s)、蒸腾速率(T_r)和水分利用效率(WUE),减缓‘丰禾1’叶片中胞间CO₂浓度(C_i)的升高,有效减轻干旱胁迫对玉米幼苗叶片光合作用的气孔限制和非气孔限制;增加脯氨酸和可溶性糖的含量;降低O₂^-·生成速率、H₂O₂和丙二醛(MDA)含量、细胞膜透性,有效缓解了胁迫对膜的伤害;增强幼苗根系活力;其中干旱敏感品种‘丰禾1’变化幅度大于耐旱品种‘先玉335’.表明在干旱胁迫下,外源Spd能促进玉米幼苗对光能的捕获与转换,增强光合作用,促进幼苗的生长,并能够通过减少玉米幼苗体内活性氧的产生,增加渗透调节物质的积累以稳定细胞膜系统,提高根系活力,从而增强幼苗对干旱逆境的适应性,且对干旱敏感品种‘丰禾1’的效果更显著.     

复合外源物质对玉米幼苗抗寒性的影响

该研究采用不同浓度的水杨酸(SA)、脱落酸(ABA)、磷酸二氢钾(KH2PO4)以及抗坏血酸(VC)进行正交组合获得不同浓度组合的复合外源物质,同时以蒸馏水处理作为对照,将其喷施于玉米幼苗后进行低温胁迫处理,在胁迫结束后使幼苗恢复生长,并测定叶片相关生长指标及生理生化指标的变化。结果表明:(1)与对照组相比,9种复合外源物质处理均可显著提高玉米幼苗的相对生长速率、干物质积累速率及脯氨酸含量,降低相对电导率,总体上提高了玉米幼苗的素质。(2)在4℃低温胁迫下,9种复合外源物质处理下幼苗相对生长速率、干物质积累速率、根系活力、脯氨酸含量、可溶性蛋白含量及可溶性糖含量,均显著高于对照,并减缓了丙二醛的积累和相应的膜脂过氧化,降低了相对电导率,提高了玉米幼苗的耐冷性。低温胁迫条件下,水杨酸、脱落酸、磷酸二氢钾和抗坏血酸四种物质组成的复合物可以提高玉米幼苗的耐冷性,其中以SA0.14 g·L~(-1)+ABA 0.015 g·L~(-1)+KH_2PO_4 3.0 g·L~(-1)+V_C 3.0 g·L~(-1)的复合物效果最好。该研究结果为新型复合抗寒剂的应用推广提供了技术支持。     

接种菌根真菌对青冈栎幼苗耐旱性的影响

利用丛枝菌根真菌摩西球囊霉(Glomus mosseae)、根内球囊霉(Glomus intraradices)和外生菌根真菌彩色豆马勃(Pisolithus tinctorius)对石漠化地区造林树种青冈栎(Cyclobalanopsis glauca)幼苗进行接种试验。在大棚盆栽条件下模拟土壤干旱胁迫,研究菌根真菌对青冈栎生长和耐旱性的影响。结果表明:在土壤干旱条件下,接种菌根处理植株生物量显著高于未接种处理(P0.05),菌根依赖性随土壤水分含量降低而升高;未接种处理植株叶绿素含量在土壤干旱条件下显著降低(P0.05),除接种Pisolithus tinctorius处理外,其它接种处理叶绿素含量无显著变化。土壤干旱使植株体内脯氨酸和可溶性糖含量上升,在中度干旱条件下,接种处理可溶性糖含量均显著高于对照处理,接种Glomus intraradices、Pisolithus tinctorius处理脯氨酸含量显著低于对照(P0.05);在重度干旱条件下,接种Glomus mosseae和Glomus intraradices处理可溶性糖含量显著高于对照处理(P0.05),而相应的脯氨酸含量显著低于对照处理。当土壤水分含量在田间持水量55%—65%时,接种处理植株SOD、POD和CAT酶活性显著高于未接种处理(P0.05),在土壤水分含量降至35%—45%时,Glomus mosseae和Glomus intraradices处理SOD酶活性显著高于对照,并且所有接种处理POD酶活性均显著高于对照。此外,在水分干旱条件下,植株全磷和全钾含量也显著高于未接种处理(P0.05)。研究表明,丛枝菌根真菌和外生菌根真菌均能够侵染青冈栎幼苗根系;在干旱胁迫条件下,接种菌根真菌能够提高青冈栎植株生物量、抗氧化酶活性、增加植株可溶性糖含量和促进植株养分吸收,提高植株耐旱性,从而使青冈栎幼苗在岩溶干旱环境下更容易存活。     

Effects of endophytic fungi on some drought tolerance mechanisms of tall fescue in a hydroponics culture

Neotyphodium, a seed-transmissible nonpathogenic fungal endophyte (symbiont) is considered beneficial because endophyte-infected grasses are more drought-tolerant, produce more dry matter, utilize soil nitrogen more efficiently, and deter insects. In this study, the effects of endophytes on physiological mechanisms of drought tolerance in tall fescue (Festuca arundinacea Schreb.) were studied in a greenhouse. Two clonally propagated genotypes of tall fescue (F. arundinacea Schreb.), naturally containing endophyte (EI), and their endophyte-free ramets (EF) were tested at three water stress treatments exerted by PEG 6000 in a hydroponics system. Relative water content (RWC), cell membrane stability (CMS), proline and chlorophyll contents in plant leaves were measured during water stress treatments. After harvest, K⁺, Ca²⁺, and Mg²⁺ contents were measured in plant roots and shoots. After 20 days under stress conditions, plants were transferred to basal hydroponics medium, and their survival after stress relief was evaluated. The results showed that endophyte considerably contributes to host grass water stress tolerance. Both genotypes of EI and EF plants did not differ in RWC, but, regardless of the infection status, genotype 75 had the higher RWC than genotype 83. EI clones of both genotypes maintained slightly higher chlorophyll content and membrane stability than EF clones, although these differences were not significant. The EI plants of genotype 83 concentrated significantly more proline than EF plants, but in the genotype 75, differences between EI and EF clones were not significant. Plant mineral absorption was also influenced by the endophyte presence. EI clones had the higher concentrations of K⁺ in the shoots of both genotypes. The Mg²⁺ and Ca²⁺ contents in EF plants of both genotypes were higher than EI plants in the roots, but in the shoots there were no differences between EI and EF clones. EI clones survived longer after stress removal. These results strongly suggest that Neotyphodium endophytes exert their effects on tall fescue drought tolerance through alteration of various physiological mechanisms involved. Published in Russian in Fiziologiya Rastenii, 2009, Vol. 56, No. 4, pp. 563–570. This test was submitted by the authors in English.     

Effects of ectomycorrhizal and vesicular-arbuscular mycorrhizal fungi on drought tolerance of four leguminous woody seedlings

Seedlings ofAcacia auriculiformis A. Cunn. ex. Benth.,Albizia lebbeck (L.) Benth.,Gliricidia sepium (Jac.) Walp andLeucaena leucocephala (Lam.) de Wit. were inoculated with an ectomycorrhizal (Boletus suillus (l. ex. Fr.) or indigenous vesicular-arbuscular mycorrhizal (VAM) fungi in a low P soil. The plants were subjected to unstressed (well-watered) and drought-stressed (water-stressed) conditions. InGliricidia andLeucaena, both mycorrhizal inoculations stimulated greater plant growth, P and N uptake compared to their non-mycorrhizal (NM) plants under both watering regimes. However, inAcacia andAlbizia, these parameters were only stimulated by either ectomycorrhiza (Acacia) or VA mycorrhiza (Albizia). Growth reduction occurred as a result of inoculation with the other type of mycorrhiza. This was attributed to competition for carbon betweenAcacia and VA mycorrhizas and parasitic association betweenAlbizia and ectomycorrhiza. Drought-stressed mycorrhizal and NMLeucaena, and drought-stressed mycorrhizalAcacia tolerated lower xylem pressure potentials and larger water losses than the drought-stressed mycorrhizal and NMAlbizia andGliricidia. These latter plants avoided drought by maintaining higher xylem pressure potentials and leaf relative water content (RWC). All the four leguminous plants were mycorrhizal dependent. The higher the mycorrhizal dependency (MD), the lower the drought tolerance expressed in terms of drought response index (DRI). The DRI may be a useful determinant of MD, as they are inversely related.     

沼液施用对干旱胁迫下玉米幼苗生长和生理特性的影响*

为了探明干旱胁迫下沼液对玉米幼苗抗旱、光合生理、形态的缓解效应,以中度抗旱玉米杂交种‘先玉335’和较强抗旱杂交种‘中单2号’为材料,采用10%聚乙二醇-6000模拟干旱胁迫,研究50%沼液根部浇灌处理对干旱胁迫下玉米幼苗生长和生理特性的影响。结果表明,沼液根部施用可以显著提高两品种玉米的抗旱性,有效缓解干旱胁迫对玉米幼苗根系和地上部生长的抑制作用,促进两品种玉米幼苗生长和提高根系活力,降低根冠比,且‘先玉335’的变幅更大;同时显著提高两品种叶片SOD、POD和CAT活性以及可溶性糖、脯氨酸、可溶性蛋白含量(‘中单2号’的CAT除外),降低MAD含量,且对‘中单2号’的影响更显著;沼液根施还可以显著提高干旱胁迫下两品种叶片的净光合速率(Pn)、蒸腾速率(Tr),降低气孔导度(Gs)、胞间CO_(2)浓度(Ci),但‘中单2号’的Tr和Gs除外,同时使两品种叶片叶绿素含量和水分利用效率(WUE)均显著增加。可见,沼液根施处理可以有效改善干旱胁迫下玉米幼苗的光合能力,显著提高幼苗抗氧化酶活性和渗透调节物质的含量,减轻膜脂过氧化程度,有效缓解干旱胁迫对2种不同抗旱性玉米幼苗的生长抑制,从而增强玉米耐受干旱胁迫的能力,且对‘中单2号’的缓解效果更明显。     

Sequence of drought response of maize seedlings in drying soil

Leaf elongation in monocotyledonous plants is sensitive to drought. To better understand the sequence of events in plants subjected to soil drying, leaf elongation and transpiration of maize seedlings ( Zea mays L.) of 4 cultivars were monitored continuously and the diurnal courses of the root and leaf water relations were determined. Results from this study indicate the following sequence of drought response: Leaf elongation decreased before changes in the leaf water relations of non‐growing zones of leaf blades were detected and before transpiration decreased. Reductions in leaf elongation preceded changes in the root water potential (ψ_w). Root ψ_w was not a very sensitive indicator of soil dryness, whereas the root osmotic potential (ψ_s) and root turgor (ψ_p) were more sensitive indicators. The earliest events observed in drying soil were a significant increase in the largest root diameter class (1 720 to 1 960 gm) and a decrease in leaf elongation ( P = 0.08) 2 days after withholding water. Significant increases in root length were observed 2 days later. Soil drying increased the number of fine roots with diameters of <240 µm. Slight increases in soil strength did not affect leaf elongation in the drying soil.     

Effects of attack of saprobic fungi on twig litter decomposition by endophytic fungi

Endophytic fungi of healthy twig tissues act as dominant primary colonizers of twig litters and are exposed to the attacks of saprobic secondary colonizers. In the present study, we examined the effect of interspecific interactions on the decay rate of twigs in laboratory experiments using two endophytic ascomycetes (Phomopsis sp. and Xylaria sp.) and two saprobic basidiomycetes (Mycena polygramma and Phanerochaete filamentosa) on twigs of Japanese beech (Fagus crenata) as a model system. Both endophytes were defensive against two saprobes on 2% malt agar, and were not replaced by the saprobes in the twigs during the incubation period. However, inoculation of the saprobes reduced the mass loss of twig components (acid-unhydrolyzable residue (AUR), holocellulose, soluble carbohydrate, and polyphenol). Especially when the twigs previously inoculated with Xylaria sp. or Phomopsis sp. were successively inoculated with M. polygramma the net weight of AUR increased, which was probably due to the synthesized melanin in hyphae of the endophytes. Our findings indicate that interspecific interactions between endophytic and saprobic fungi do affect the total mass of humic substances produced during the litter decay process.     

水分胁迫对不同抗旱类型冬小麦幼苗叶绿素荧光参数的影响

在人工气候室水培条件下,选用3个不同抗旱类型的冬小麦品种,研究了水分胁迫对冬小麦幼苗叶绿素荧光参数的影响。结果表明：水分胁迫下,冬小麦幼苗可变荧光(Fv)、最大荧光(Fm)、可变荧光与最大荧光比(Fv／Fm)、可变荧光与初始荧光比(Fv／Fo)、光化学淬灭系数(qF)均降低;而初始荧光(Fo)与非光化学淬灭系数(qNP)则升高,说明光系统Ⅱ(PSⅡ)受到了伤害,使得PSⅡ原初光能转换效率(Fv／Fm)、PSⅡ潜在活性(Fv／Fo)降低;光合电子传递、光合原初反应过程受到抑制,起光保护作用的热耗散提高。但水分胁迫下品种间各参数变化幅度不同。除qNF外,其余各参数均为抗旱性越强降低幅度越小,而qNF则升高幅度越大。说明水分胁迫对冬小麦幼苗叶绿素荧光参数的影响与其抗旱性密切相关。