秤锤树叶片内生真菌的分离鉴定及其对植株生长的影响

本研究从秤锤树叶片中分离内生菌进行分子鉴定,并接种到无菌组培苗中研究其对植株生长的影响。研究结果显示,从秤锤树叶片中共分离获得7株真菌。扩增出的6个菌株经ITS测序,在NCBI网站上进行基因序列比对,鉴定出其中5株菌株均属于球毛壳菌（Chaetomium globosum）,1株属于子囊菌（As-comycete）。进一步实验表明：分离的菌株多数对组培苗的株高生长影响不大,可初步鉴定其为内生菌,其中的属于球毛壳菌的F和H菌株对无菌苗的株高有明显的促进作用。     

基质未灭菌和灭菌条件下接种AM真菌对香橙砧木生长的影响

在基质灭菌条件下接种丛枝菌根（AM）真菌能促进植物生长发育,但在未灭菌条件下接种AM真菌鲜有报道。因此,在温室盆栽条件下,以香橙砧木（Citrus Junossieb ex Tanaka）为材料,研究了不同处理基质（灭菌和未灭菌）及接种AM真菌对香橙砧木的根系菌根侵染率、株高、茎径、叶片数、生物量及根系性状的影响。探讨在模拟自然条件（基质未灭菌）下接种AM真菌其接种效应是否仍然有效。研究结果表明：在基质灭菌条件下,接种黄雷克囊霉（Redeckera fulvum）(R.f)处理的株高、叶片数、地上干重、生物量和根系性状显著优于未接种菌根处理,而接种凹坑无梗囊霉（Acaulospora excavata）(A.e)处理的株高、茎径、叶片数、生物量低于未接种处理,但无显著差异。而根系性状显著优于对照;在基质未灭菌条件下,除了接种R.f处理的总根长显著高于未接种处理外,其他指标均与未接种处理无显著差异。这表明在盆栽试验中基质灭菌条件下,接种R.f处理能提高香橙砧木生长,接种A.e处理能抑制其生长,但在基质未灭菌条件下,接种两种AM真菌的香橙砧木生长与对照无显著影响,没有表现出在基质灭菌条件下...     

丛枝菌根真菌对镉污染土壤中黑麦草幼苗生长的影响

通过不同浓度的镉污染土壤接种丛枝菌根真菌的黑麦草盆栽试验,研究了丛枝菌根真菌对镉污染条件下黑麦草幼苗生长的影响。结果表明:重度镉污染(Cd2+:180 mg/kg)条件下,Glomus mosseae对黑麦草根系的侵染率仍达到30.23%,对黑麦草的生长有较好的促进作用;丛枝菌根在一定程度上缓解了镉污染对黑麦草株高、根长和生物量积累的抑制;镉污染显著降低黑麦草叶片的叶绿素含量,叶绿素a在重度镉污染时下降幅度最大,不接种丛枝菌根真菌的黑麦草较对照下降37.9%,而接种的黑麦草下降26.7%,接种菌根真菌在中重度镉污染条件下显著提高了黑麦草叶片的叶绿素含量;重度镉污染下接种和不接种的黑麦草根系活力都开始显著下降,但接种植株根系活力下降的幅度小于不接种植株。     

盐胁迫下AM真菌对紫花苜蓿生长及生理特征的影响

利用盆栽试验研究了NaCl胁迫条件下AM真菌对紫花苜蓿(Medicago sativa L.)植株生长及生理特征的影响。结果显示:随土中加入NaCl浓度增高(0%,0.5%,1.0%),与未接菌紫花苜蓿相比,接种AM真菌显著(P<0.05)促进紫花苜蓿生长,并使其可溶性糖、可溶性蛋白及根系脯氨酸累积量增加。1.0%NaCl接种AM真菌处理下根系鲜重(28.1 mg)为0%NaCl未接种下的1.27倍;不同盐处理接种AM真菌植株丙二醛含量均低于对照株;1.0%NaCl浓度下接种AM菌植株叶片及根系脯氨酸含量分别为对照株的0.82,1.41倍。表明AM真菌在胁迫环境下不仅可促进植株生长发育及渗透调节物质的积累,还可维持植物体丙二醛含量较低水平,降低植株遭受盐胁迫,推测叶片与根系脯氨酸对ABA依赖程度不同而导致其脯氨酸含量不同。     

发酵牛粪对苏丹草根系丛枝菌根（AM）真菌侵染及磷吸收效率的影响

通过温室盆栽试验,研究接种苏格兰球囊霉(Glomus caledonium)条件下添加不同比例发酵牛粪(0.33%、0.50%和1.00%)对苏丹草(Sorghum sudanense)根系丛枝菌根(AM)真菌侵染率、土壤孢子密度、植株生物量与根冠比及根系磷(P)吸收效率的影响。结果发现,与对照相比,接种AM真菌处理植株地上部生物量趋于下降、根冠比显著提高(p<0.05),在此基础上添加0.33%或0.50%发酵牛粪处理土壤孢子密度、植株根系生物量和AM真菌侵染率均趋于升高,根冠比没有明显变化,根系P吸收效率显著提高(p<0.05);添加1.00%发酵牛粪显著提高土壤孢子密度、植株生物量和根系AM真菌侵染率(p<0.05),根冠比与仅接种AM真菌处理相同,根系P吸收效率则达到仅接种AM真菌处理的1.83倍。结果表明,添加1.00%发酵牛粪对苏格兰球囊霉扩繁及其宿主植物P吸收均具有突出促进作用。     

丛枝菌根(AM)真菌对土壤中阿特拉津降解的影响

于盆栽高粱(Sorghum,龙杂一号)条件下研究了丛枝菌根(AM)真菌根内球囊霉(Glomus intraradices,GI)和摩西球囊霉(Glomus mosseae,GM)降解土壤中阿特拉津的效用。结果表明,阿特拉津(浓度为50 mg/kg)污染土壤中,供试AM真菌都能够侵染高粱根系形成菌根,而且GM比GI侵染效果好,最高侵染率可达到90.5%,显著提高了植株的生物量。接种AM真菌后土壤中阿特拉津的残留浓度显著低于不接种对照处理,并且接种GM比GI对阿特拉津的降解效果显著。接种GM处理的土壤中阿特拉津最高降解率达到了91.6%,其中菌根效应占22.6%。接种AM真菌的宿主植物根际土壤中微生物数量多于不接种处理,且GM优于GI处理,说明AM真菌能促进根际微生物的繁殖。此外,接种AM真菌后能显著增加土壤中脲酶活性,但对过氧化氢酶活性影响不显著。认为GM是一株比较理想的修复阿特拉津污染土壤的AM真菌。     

丛枝菌根对酸枣实生苗耐盐性的影响

本文研究了在土中加入不同量NaCl条件下 (0、1 5、3 0、4 5gkg-1干土 )接种丛枝菌根真菌 (AMF)Glomusmosseae对盆栽酸枣 (ZizyphusspinosusHu)实生苗生长及耐盐性的影响。结果表明 ,无论接种与否 ,植株的高度、根茎叶的干鲜重均随土壤NaCl浓度的增加而降低 ,而根、茎、叶和整株的Na浓度及Na全量均随土壤NaCl浓度的增加而增大。在土壤盐浓度相同的条件下 ,接种AMF植株的生长量 (株高、鲜重、干重等 )和叶片的叶绿素含量显著高于不接种植株。接种AMF的植株茎、叶中Na浓度低于不接种植株 ,而根中Na浓度、植株Na总量显著高于不接种植株。盐浓度最大的接种处理 ,其植株生长量和叶片叶绿素含量均高于不加盐不接菌处理。播种时进行盐胁迫处理和播种后 4 0d开始进行盐胁迫处理对菌根的侵染率、植株生长的影响差异不显著。上述四种盐浓度播种时进行盐处理的接种AMF植株的总干重比不接种植株分别提高 16 4 %、14 9%、4 8%、35 % ,在播种后 4 0d进行盐处理的接种AMF的植株比不接种植株分别提高 194 %、12 7%、72 %、4 6 %。结果证明 ,酸枣实生苗具有较强的耐盐性 ,其生长对菌根真菌有很强的依赖性 ,接种菌根真菌提高了其耐盐能力。     

丛枝菌根真菌对镉胁迫下黑麦草光合生理的响应

为探究镉(Cd)胁迫下丛枝菌根(AM)真菌对黑麦草(Lolium perenne)光合生理的影响,盆栽条件下以Cd浓度为0、5、15和30 mg·kg-1分别对黑麦草接种AM真菌摩西斗管囊霉(FM)、变形球囊霉(GV)、混合处理(FM+GV)以及不接种对照(NM)共16个处理。结果表明,Cd降低了AM真菌侵染;接种AM真菌则提高了黑麦草株高、叶绿素含量、荧光参数、地上部和根系氮(N)、Cd含量,增强光合作用,显著提高植物体内生理活性。在Cd浓度为30 mg·kg-1水平下,FM+GV处理的菌根侵染率最大,为42.7%,与NM相比,FM+GV处理的黑麦草叶片叶绿素含量、地上部和根系N含量分别提高26.2%、70.6%和85.3%;株高和地上干重分别提高34.1%和18.8%;PSⅡ最大光化学效率和PSⅡ潜在活性分别提高4.9%和19.7%;净光合速率、蒸腾速率和气孔导度分别增加19.8%、28.3%和14.7%;超氧化物歧化酶、过氧化物酶活性、可溶性蛋白含量分别是NM的1.3、1.5和1.8倍,丙二醛含量相较NM下降50.0%。土壤中Cd浓度与植株地上部和根内的Cd含量呈显著正相关,FM+GV处理的黑麦草地上部和根系Cd含量分别达到88.1和606.7 mg·kg-1。总体来看,FM+GV处理增加黑麦草光合生理抗性、吸收固持Cd能力的效果最为显著。     

不同施铜水平下接种AM真菌对海州香薷根际pH的影响

研究表明,丛枝菌根(Arbuscular mycorrhiza,AM)真菌对宿主植物生长和吸收、转运重金属的作用受土壤pH的影响[1-3]。植物可以通过分泌质子或有机酸等改变根际土壤的pH,接种AM真菌可以改变宿主植物的根系分泌物[4-6],从而影响土壤pH。Li等[7]研究发现,接种AM真菌的白三叶在根-土界面、菌丝室及菌丝-土壤界面的pH均降低。Li和Christie[8]发现Zn污染土壤中接种G.mosseae降低了红三叶植物体内Zn浓度和吸收量;菌根处理土壤的pH比对照土壤高,土壤溶液中的Zn浓度低,在施Zn量大时尤为显著。AM真菌菌丝往往在利用NO3-N的同时释放出OH-,导致…     

AM真菌对烟苗生长及某些生理指标的影响

在低浓度营养液条件下,利用漂浮育苗技术培育烟苗,于播种期、小十字期、生根期分别接种不同的AM真菌,研究了它们对烟苗生长、营养和某些生理指标的影响。结果表明,越早接种AM真菌,其侵染率越高;播种期接种,侵染率达到39.2%～59.6%。AM真菌的菌根效应因菌种(株)不同而异,接种球囊霉真菌（BEG-141）后,显著增加烟苗干重、磷含量、氮磷钾吸收量、叶绿素含量,以及根系硝酸还原酶、超氧化物歧化酶和几丁质酶活性。表明在漂浮育苗技术中,播种期接种适宜的AM真菌是培育壮苗的有效措施。     

Responses of directly seeded wetland rice to arbuscular mycorrhizal fungi inoculation

The mycorrhizal enhancement of plant growth is generally attributed to increased nutrients uptake. A greenhouse experiment was conducted to investigate the effect of arbuscular mycorrhizal fungi (AMF) inoculation on the growth and nutrient uptake of directly seeded wetland rice. Seeds were germinated and inoculated with arbuscular mycorrhizal fungi or left uninoculated. The plants were grown at 60% of ‐0.03 MPa to establish the mycorrhizas. After 5 weeks, half of the pots were harvested and the rest were flooded with deionized water to maintain 3–5 cm of standing water until harvesting (122 days after sowing). Mycorrhizal fungal colonization of rice roots was 36.2% at harvest. Mycorrhizal fungi inoculated rice seedlings grew better compared to uninoculated seedlings and had increased grain yield (10%) at the harvesting stage. Shoot and root growth were effectively increased by AMF inoculation at the harvesting stage. The nitrogen (N) and phosphorus (P) acquisition of direct seeding wetland rice were significantly increased by AMF inoculation. The AMF enhanced N and P translocation through the hyphae from soils to roots/shoots to grains effectively.     

Bioguided fractionation and isolation of free radical scavenging components from in vitro propagated chinese medicinal plants Dendrobium tosaense Makino and Dendrobium moniliforme SW

This study was performed to investigate the free radical scavenging active components from in vitro propagated medicinal herbs of the genus Dendrobium, namely, Dendrobium tosaense Makino and Dendrobium moniliforme SW, using a 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical antioxidative assay. Seeds of the capsules derived after 12 weeks of hand-pollination germinated asymbiotically (50-74%) on half-strength Murashige and Skoog's (MS) basal medium with 3% sucrose and solidified with 0.9% Difco agar. Active growth in the germinated seedlings was achieved by reculturing on full-strength MS basal medium supplemented with 8% banana homogenate, 8% potato homogenate, 8% coconut water, 1.5% sucrose, and 0.9% Difco agar. Healthy plantlets transferred to plastic trays containing moss or moss and tree fern successfully acclimatized (84-100%) in the greenhouse. Extracts were prepared from plants grown in the greenhouse for a period of 6 months. Methanolic extracts of D. tosaense and D. moniliforme scavenged DPPH at 95.9 and 83.4%, respectively, at a concentration of 0.4 mg/mL. Therefore, methanolic solubles of D. tosaense and D. moniliforme were subjected to bioguided fractionation and separation by column chromatographic methods individually. After chromatographic separation of these crude extracts, the obtained fractions (Dm 1, Dm 2, Dm 3, Dt 1, Dt 2, and Dt 3) were tested for their activity. Among them, fractions Dm 2 and Dt 1 showed significant antioxidant activity by DPPH radical antioxidative assay. Active fractions were purified further by column chromatography and resulted in identification of the antioxidant components alkyl ferulates from D. moniliforme and quercetin from D. tosaense.     

丛枝菌根真菌(Glomus caledonium)对铜污染土壤生物修复机理初探

利用盆栽试验,研究了丛枝菌根真菌(Glomus.caledonium)在不同程度铜污染土壤上对玉米苗期生长的影响。结果表明,即使在土壤施铜量达150mg/kg时,菌根真菌对玉米仍有近55%的侵染率;接种菌根真菌,能显著促进玉米根系的生长。菌根玉米的根系生物量和根系长度,平均较未接种处理分别提高108.4%和58.8%;接种处理的植株地上部生物量达到每盆(3株)10.58g,显著高于不施铜的非菌根玉米。这些结果表明,丛枝菌根真菌对铜污染具有较好的抗性;并且由于菌根的形成,使宿主植物明显地改善了对磷的吸收和运输,并能通过抑制土壤酸化、降低土壤可溶态铜的浓度等机制,增强宿主植物对铜污染的抗(耐)性。在150mg/kg施铜水平时,与非菌根玉米相比,菌根玉米地上部和根系铜浓度分别降低24.3%和24.1%,吸铜量分别提高了28.2%和60.0%,表明菌根植物对铜污染土壤具有一定的生物修复作用。     

两种丛枝菌根真菌对黄瓜苗期枯萎病的防效及根系抗病相关酶活性的影响

由尖孢镰刀菌引起的黄瓜枯萎病是设施黄瓜生产的主要障碍之一,丛枝菌根真菌(AM真菌)可以和包括黄瓜在内约80%的维管植物的根系形成菌根共生体,适宜的共生体组合对于寄主的生长与抗病性的提高十分有益。为明确Glomus versiforme与Glomus intraradices两种AM真菌对"津绿3号"黄瓜苗期枯萎病的防治效果。试验采用盆钵培养的方法,研究了两种AM真菌对幼苗生长及其根系3种抗病相关酶活性的影响。结果表明:两种AM真菌均可促进黄瓜幼苗的生长,并能减轻病害,但以G.versiforme的促生及生防作用更显著,接种G.versiforme处理的黄瓜幼苗株高、茎粗、叶面积及干重均显著大于对照,该处理的幼苗病情指数较对照降低26.6%。菌根化黄瓜幼苗抗病性的提高一方面与接种病原菌Fusarium oxysporum f.sp.cucumerinum前幼苗生长健壮有关;另一方面与根系抗病相关酶活性的提前诱导有关。接种F.oxysporum f.sp.cucumerinum前,G.versiforme处理的黄瓜幼苗壮苗指数显著高于G.intraradices处理与对照,分别为G.intraradices处理与对照的1.19倍与1.22倍;G.versiforme处理的黄瓜幼苗根系几丁质酶、β-1,3-葡聚糖酶与PAL酶分别比对照提前2 d、7 d、7 d被诱导,且酶活性分别为对照的1.44倍、2.16倍和92.00倍。     

Effect of manganese on endomycorrhizal sugar maple seedlings

Abstract

Manganese (Mn) toxicity may play an important role in the poor survival of seedlings in declining sugar maple (Acer saccharum Marsh.) stands in northern Pennsylvania. To determine the effect of Mn on the growth of sugar maple seedlings, 1‐year‐old seedlings inoculated with vesicular‐arbuscular mycorrhizal (VAM) fungi and growing in sand‐vermiculite‐peat moss medium were irrigated for 7 weeks with nutrient solution (pH 5) containing 0.1 (control), 1, 2, 4, 8, or 16 mg L^?1 Mn. Total seedling dry weight was negatively correlated with Mn, becoming significantly different than the control at 2 mg L^?1 Mn. Stem and root dry weight were reduced by lower Mn levels than leaf dry weight. Manganese had no effect on the root/shoot ratio. The concentration of Mn in roots and leaves increased as the level of Mn in the nutrient solution increased, with the concentration in the leaves 2.2‐ to 3.7‐fold greater than the concentration in the roots. Except for a reduction of P in the roots, Mn had little effect on the concentration of nutrient elements in the roots or leaves. Colonization of the roots by VAM fungi was increased by Mn, with a maximum percentage at 4 mg L^?1 Mn. Manganese toxicity symptoms in the leaves, small discrete chlorotic spots, began to appear at 1 mg L^?1 Mn. The sensitivity of sugar maple seedlings to Mn found in this study supports the hypothesis that Mn may affect regeneration in declining sugar maple stands. However, evaluation of the effects of Mn on seedlings in native soils under field conditions will be necessary before the role of Mn in sugar maple regeneration can be understood.     

Plant growth and nutrition in pine (Pinus thunbergii) seedlings and dehydrogenase and phosphatase activity of ectomycorrhizal root tips inoculated with seven individual ectomycorrhizal fungal species at high and low nitrogen conditions

We investigated the functions and ecological traits of seven individual ectomycorrhizal (ECM) fungal species derived from a Pinus thunbergii-dominated (nitrogen-poor) and a Robinia pseudoacacia-dominated (nitrogen-rich) area. P. thunbergii seedlings were inoculated with seven individual ECM fungal species and cultured under high nitrogen (nitrogen group) and low nitrogen (control group) conditions. Control seedlings were not inoculated with any ECM fungus. Seedlings harvested were examined for the numbers of non-mycorrhizal and ECM root tips, growth, nutrient status (nitrogen, phosphorus and carbon) and the activity of dehydrogenase and phosphomonoesterase in the root tips. The relationships among the growth, nutrient status and the enzymatic activity in the seedlings inoculated with each ECM fungal species were also analyzed by canonical discriminants. As the results, inoculation of ECM fungi made the plant growth significantly higher than those of non-inoculated seedlings. Plant growth significantly correlated with the phosphorus content, N:P ratio and phosphomonoesterase activity, especially in the nitrogen group. Dehydrogenase and phosphomonoesterase activities in the seedlings were higher when the ECM fungi from a nitrogen-rich area were inoculated than when those from a nitrogen-poor area were inoculated. The canonical discriminant analyses separated seedlings by inocula: no fungus, ECM fungi from a nitrogen-poor area, and those from a nitrogen-rich area. These results suggested the possibility that nitrogen deposition caused P shortage and the P absorbing ability was higher in the ECM fungi in nitrogen-rich forests than those in nitrogen-poor forests.     

Protecting Cucumber from Fusarium Wilt with Arbuscular Mycorrhizal Fungi

A pot experiment was conducted to investigate the biocontrol effects of arbuscular mycorrhizal fungi (AMF) Glomus mosseae and Glomus versiforme on Fusarium oxysporum wilt disease of cucumber (Cucumis sativus L.). The results indicated that both AMF improved the growth of cucumber seedlings and reduced disease severity, but G. versiforme was more efficient. Compared with nonmycorrhizal plants infected by F. oxysporum, shoots and roots dry weights increased by 100% and 80% in G. versiforme–inoculated plants, and the qualities of seedlings were significantly improved; meanwhile, nitrogen, phosphorus, and potassium contents in shoots of G. versiforme–inoculated plants were significantly greater than those of G. mosseae–inoculated plants and nonmycorrhizal plants. Moreover, for mycorrhizal plants, soluble sugar and free proline contents in mycorrhizal roots were significantly greater than those of nonmycorrhizal treatment; however, malonaldehyde content in roots and the quantity of fungi in rhizosphere decreased when the plants were attacked by F. oxysporum.     

四种AM真菌接种剂的田间效应及其分子检测研究

采用灭菌土壤生产了 4种AM真菌接种剂。在盆栽条件下测试了接种剂的质量 ,结果显示 ,4种接种剂促进玉米生长效果明显 ,地上部分生物量均显著高于对照 (p <0 .0 1 ) ;以MPN试验检测了接种剂的侵染能力 ,结果表明每克接种剂中真菌的繁殖体数在 95～ 1 4 0 0之间。将AM真菌的预接种技术和农业生产上的营养钵育苗技术相结合 ,进行了玉米的田间试验 ,结果显示 ,玉米根系的AM真菌感染率早期增长较快 ,然后趋于平稳 ;AM真菌接种剂A(Glomusconstrictum)、C (Glomus三种菌混合 )和D (G .intraradices)对玉米籽粒产量有显著的增产效果 (p <0 .0 5 ) ;玉米籽粒的淀粉含量和磷含量也高于对照。运用特异性分子探针和nest ed PCR技术 ,从田间接种AM真菌Glomusintraradices和G .mosseae的玉米根样中粗提DNA进行特异性扩增 ,成功地从感染根段中检测到特定的接种AM真菌。本工作从分子水平为评价高效AM真菌的应用潜力、研究AM真菌之间及其与其他微生物之间的相互关系奠定了基础。     

Promotion of the seed germination and seedling growth of cabbage by root-colonizing fungi isolated from oat and Eucalyptus roots

(Jpn. J. Soil Sci.Plant Nutr., 77, 265–272, 2006)

We isolated fungi colonizing oat (Avena sativa L.) and eucalyptus (Eucalyptus camaldulensis Dehnh) roots to select the strains that promote the seed germination and seedling growth of cabbage. Twenty-nine strains were inoculated into the soil which grows cabbage seedlings. This inoculation test was repeated three times. From the results of the test, we found that MT0011 and EU0013 strains promoted seed germination. In the first inoculation test, seven strains increased the dry weight of cabbage seedling significantly compared with the control. In the second test with the strains that promoted cabbage growth, MT0008 and EU0013 strains significantly increased the dry weight of seedlings. In the third test, only EU0013 significantly increased seedling growth. The ITS region for DNA (ITS1-5·8SrDNA-ITS2) of 28 isolated strains were sequenced to examine the molecular phylogeny. Isolates were classified into two divisions, Zygomycota and Ascomycota. Nine strains showed the highest similarities with same accession number AF504832 (Uncultured fungus clone D50). MT0011 was clustered with fungi belonging to Drechela sp. from the sequence data on the rDNA ITS region. EU0013 was identified as Penicillium sp. from the morphological feature conidiophore and the sequence of the ITS region.