一株溶磷真菌筛选鉴定及其溶磷促生效果

【目的】从高产农田筛选高效溶磷微生物菌株,为溶磷微生物肥料开发提供高效菌种资源。【方法】利用菌株的形态学特性、培养特征和ITS rDNA序列分析方法进行菌株鉴定,结合液体培养和土壤培养方法研究菌株的溶磷能力,进而采用温室盆栽和田间小区试验,明确溶磷菌P83促进作物生长和提高作物产量的作用效果。【结果】溶磷菌株P83鉴定为斜卧青霉菌(Penicillium decumbens)。液体条件下培养10 d,菌株P83表现显著高效的溶磷能力,对Ca3(PO4)2(5g/L)的溶解效果,有效磷达956 mg/L,溶解率为42.68%,对湖南永和磷矿粉的溶液效果,有效磷达到152.8 mg/L;将P83菌株分别接种于施用Ca3(PO4)2、Zn3(PO4)2和磷矿粉(RP)3种磷源的盆栽试验土壤中,结果显示,菌株P83对玉米植株促生效果比对照显著提高,玉米植株鲜重提高9.5%-89.2%、干重增加35%-231%,土壤有效磷提高2.1-40.5 mg/kg。田间小区玉米产量结果显示,溶磷菌P83增产效果最好(P=0.05),玉米子粒产量达9.2t/hm2,比不接种菌剂的对照增加2.4 t/hm2,增产率为35.3%。【结论】获得了一株溶解难溶磷的斜卧青霉菌P83,它能够活化多种难溶磷、显著增加土壤有效磷水平,对玉米生长和增加作物产量具有显著作用,是一株展现良好应用前景的高效溶磷菌种。     

贵州两处茶园溶磷青霉菌的筛选、鉴定及溶磷能力分析

为维持土壤自然完整性、活化利用土壤中难溶性磷,从贵州名茶产地都匀、贵定茶园土壤中筛选高效溶磷真菌,为制备真菌肥料提供菌种资源。利用溶磷指数(SPI)、形态特征和ITS rDNA序列筛选、鉴定菌株,并采用液体摇床培养实验测定鉴定菌株在以磷酸钙、磷酸铁或磷酸铝为唯一磷源的无机磷液体培养基中的溶磷能力。共筛选到7个高效溶磷菌落,经形态观察分属2种菌株,鉴定为微紫青霉(Penicillium janthinellum)和赭绿青霉(Penicillium ochrochloron)。液体培养基接种、摇床培养15 d,微紫青霉菌在以Ca3(PO4)2、Fe3(PO4)2或AlPO4为唯一磷源的上清液中有效磷含量分别为73.47 mg·L^-1、30.93 mg·L^-1和14.00 mg·L^-1,4℃继续保存至30d后对Fe-P和Al-P的溶解量分别达到72.20 mg·L^-1、32.84 mg·L^-1;赭绿青霉菌培养15d的溶磷量分别为30.72 mg·L^-1、4.14 mg·L^-1和1.51 mg·L^-1,30d对Fe-P和Al-P的溶解量分别达到35.19 mg·L^-1和10.98 mg·L^-1。微紫青霉菌溶解无机磷能力明显优于赭绿青霉菌,有望应用于地区缺磷茶园土壤真菌肥料的制备。     

蔗叶堆肥中一株泡盛曲霉溶磷能力的鉴定及其对辣椒的促生效果

[目的]从甘蔗叶堆肥中分离筛选具有高效溶磷及促生功能的菌株,为微生物肥料制备提供一种可利用的菌种资源.[方法]以Ca3(PO4)2和Zn3(PO4)2为磷源,进行平板溶磷筛选实验;采用形态学特征和ITS rDNA序列分析法进行菌种鉴定;采用液体摇瓶培养测定菌株的溶磷能力;将溶磷菌接种至辣椒幼苗根部分析其促生效应.[结果...     

4株溶磷细菌和真菌溶解磷矿粉的特性

溶磷微生物广泛地分布在土壤,根际等生态环境中,了解这些微生物溶解难溶性磷酸盐如磷矿粉的特性,对于开发利用这些微生物,提高磷素利用效率具有重要作用,研究发现;真菌比细菌溶解磷矿粉的能力要强得多,培养基中的铁,镁,锰,钠等成分可以提高真菌的溶磷量,但降低了细菌的溶磷量,培养基中磷矿粉用量越高,溶磷量越低;碳源物质浓度高于3%将显地降低溶磷量,微生物能够破坏磷矿粉的结构。使其中的磷在以后的培养过程中更加容易释放出来,可见利用微生物活化磷矿粉中的磷,具有良好前景。     

一株解磷细菌的筛选、鉴定及其溶磷培养条件的优化

从土壤作物根际筛选分离出的一株解磷能力较强的溶磷菌P0417,对其进行16S r DNA基因水平上的初步鉴定,测定其溶解磷的能力,并对该菌的溶磷培养基条件进行优化。结果表明,经序列分析,确定该菌株P0417为洋葱伯克霍尔德氏菌。且其溶磷能力与培养液p H呈显著相关性,当培养基条件为葡萄糖10 g/L、草酸铵0.5 g/L、Na Cl 1.0 g/L时,菌株P0417对Ca3(PO4)2盐培养基具有较好的解磷能力,其解磷能力可达791.84μg/m L。     

高效溶磷菌的分离、筛选及在土壤中溶磷有效性的研究

从采自全国各地130个土样中.分离出1000余菌株,从中筛选出1株代号为Ap-2号的高效溶磷菌.经鉴定为黑曲霉Aspergillus niger.对该菌株的产酸特性以及接种土壤后的溶磷有效性进行了研究.试验表明,该菌株发酵过程产草酸、柠檬酸等多种有机酸,使土壤速效磷含量增加141.94%.其溶磷作用,与土壤pH含水量有密切关系,对温度要求不严,与接种量是一定正相关.并可与磷酸铵、三料、尿素、硫酸铵等化肥混合使用.     

类芦根际溶磷真菌的筛选、鉴定及其溶磷能力分析

为揭示类芦(Neyraudia reynaudiana)等水土保持植物的耐低磷机制,开发溶磷菌种质资源,提高赤红壤磷素利用率,从类芦根际土壤中筛选到一株溶磷能力较强的真菌FP1,经形态学和ITS序列分析,鉴定为黑曲霉(Aspergillus niger)。3种难溶性磷酸盐液体培养基接种菌株FP1后,其pH值和溶磷量的动态变化显示,培养液的pH值均呈显著下降趋势。溶磷量与培养时间有关,除磷酸三钙外,菌株FP1对其他难溶性磷酸盐的溶磷趋势均为先上升再下降并趋于稳定。菌株FP1对不同磷源的最大溶磷率顺序为:磷酸铝(92.02%)磷酸三钙(41.62%)3种磷酸盐的混合物(35.86%)磷酸铁(19.20%)。FP1对磷酸铝和磷酸铁都具有较强的溶磷能力,表明抗逆性强的水土保持植物类芦根际土壤蕴藏着高效的溶磷微生物资源。     

一株红壤溶磷菌的分离、鉴定及溶磷特性

【目的】为了提高红壤磷素利用率,探讨溶磷菌溶磷机理。【方法】利用难溶性无机盐培养基从花生根际土壤样品中分离到一株溶磷菌C5-A,结合菌落形态特征、生理生化和16S rRNA序列确定该菌株的系统发育地位;通过菌株C5-A在NBRIP液体培养基培养过程中培养液pH变化确定其溶磷能力;利用液体发酵实验测定不同的碳源、氮源对菌株C5-A溶磷的影响;通过高效液相色谱检测C5-A在不同氮源培养液中有机酸的种类和浓度。【结果】菌株C5-A鉴定为洋葱伯克霍尔德氏菌(Burkholderia cepacia),遗传稳定性较好。在FePO4和AlPO4培养液中,菌株C5-A的溶磷量和pH变化呈显著负相关;菌株C5-A对磷酸三钙、磷酸铝、磷酸铁、磷矿粉均有较强的溶解能力,最高溶磷量分别为125.79、227.34、60.02和321.15 mg/L;菌株C5-A对不同浓度的两种磷矿粉有较强的溶解能力;分别以麦芽糖和草酸铵为碳源和氮源时溶磷量最高。高效液相色谱检测出10种有机酸,分别为草酸(葡萄糖酸)、乙酸、苹果酸、琥珀酸和5种未知有机酸,然而,乙酸而非草酸似乎是影响C5-A溶磷的重要有机酸。【结论】从红壤花生根际土壤中筛选到一株对难溶性无机盐具有较强溶解能力溶的菌株C5-A,有望为开发高效红壤微生物磷肥提供种质资源。     

古国槐叶片溶磷内生真菌的筛选及其促生潜力初探

该试验从周公庙古国槐叶片内分离、筛选具有溶磷能力的内生真菌,采用溶磷圈法和钼锑抗比色法测定其溶解无机磷的能力,同时测定其回接后对国槐无菌苗叶绿素、可溶性蛋白质、超氧化物歧化酶(SOD)和过氧化物酶(POD)活性的影响,探究内生真菌的溶磷能力及其促生效应。结果表明:(1)从古国槐叶片中分离出的55株内生真菌中,28株具有溶磷能力,其中溶磷能力较强的活性菌株有12株:ZG-7为拟盘多毛孢(Pestalotiopsis sp.),ZG-9、ZG-23、ZG-32、ZG-36和ZG-53为镰刀菌(Fusariumsp.),ZG-15和ZG-34为曲霉(Aspergillus sp.),ZG-39和ZG-51为链格孢(Alternariasp.),ZG-42为木霉(Trichoderma sp.),ZG-48为附球菌(Epicoccumsp.)。(2)曲霉属真菌ZG-15和ZG-34的溶磷圈直径(d)及其与菌落直径(D)的比值d/D均高于其他菌株,对Ca_3(PO_4)_2的溶解能力最强(1 238.28和941.22mg·L-1),并极显著高于其他菌株(P0.01)。(3)4株内生真菌ZG-7、ZG-9、ZG-15和ZG-48回接至国槐无菌苗10d后,可从苗根组织的皮层细胞中观察到内生菌丝,其中黑曲霉(Aspergillus niger)ZG-15可显著提高国槐无菌苗的叶绿素含量、可溶性蛋白质含量、SOD、POD活性(P0.05),从而维持幼苗的正常生长和提高其抗逆能力,为该试验得到的促生潜力优势内生真菌,可为古国槐的有效保护及林木溶磷生物菌肥的生产提供依据。     

一株伯克霍尔德菌的筛选鉴定及溶磷性能优化

为解决我国大部分耕地土壤中可溶性磷含量不足、植物生长困难的问题,本研究对一株溶磷微生物(PB)进行了筛选鉴定及溶磷性能优化。结果表明: 菌株PB属于伯克霍尔德菌。该菌具有固氮和分泌吲哚-3-乙酸(IAA)等植物促生能力,对大肠杆菌也表现出一定的抑制效果;在pH 8.0～10.0范围内,菌株PB仍然能够保持较高的活性和溶磷能力,具有良好的耐碱性;溶磷性能优化结果表明,在30 ℃、pH 7.0、180 r·min^-1条件下,以葡萄糖为碳源、硫酸铵为氮源、磷酸三钙为磷源和添加50 μmol·L^-1赖氨酸时,菌株PB的溶磷能力达到最优,溶磷量为569.33 mg·L^-1,是优化前的1.9倍。该菌代谢过程中主要分泌柠檬酸、丙二酸和葡萄糖醛酸,添加赖氨酸后其分泌的有机酸种类不变,含量明显增加。盆栽试验表明,施用 PB菌肥能够显著促进大蒜幼苗的生长,而添加赖氨酸后促进效果更明显。与对照相比,PB加赖氨酸处理苗高增长18.6%,苗直径增长16.7%,地上部分鲜重和干重分别增长15.7%和22.1%,地下部分鲜重和干重分别增长22.0%和28.7%。土壤磷含量检测结果表明,PB和PB加赖氨酸处理土壤速效磷含量分别为对照的2.1和2.3倍,表明施用PB菌肥能够提高土壤中可溶性磷含量,而添加赖氨酸可以进一步强化PB菌肥的溶磷性能。     

广陈皮外源真菌的组成及产毒真菌分析

[背景]广陈皮为药食同源中药材,在高温、高湿且贮存不当的条件下容易发霉,从而产生毒素,严重威胁陈皮的质量安全.[目的]分析广陈皮表面外源真菌的组成及其产生毒素的真菌.[方法]采用平板稀释法分离广陈皮表面外源真菌,利用分生孢子形态特征及DNA序列分析进行真菌鉴定,采用高效液相色谱-三重串联四极杆质谱联用技术对青霉属和曲霉...     

Availability of phosphorus in a calcareous soil treated with rock phosphate or superphosphate as affected by phosphate-dissolving fungi

A study was undertaken to isolate some fungi exhibiting phosphate-dissolution ability, and to test whether these fungi are capable of increasing the amount of available P in a calcareous soil treated with rock phosphate (RP) or with triple superphosphate (TSP) and its subsequent uptake by sorghum (Sorghum bicolor L. Moench).Penicillium sp. and twoAspergillus foetidus (Naka) isolates significantly increased the availability of P in soil treated with RP or TSP during the growing season.Penicillium sp. isolate was more effective in increasing available P in the soil treated with RP or TSP than were Aspergillus isolates. However, the dry matter and P uptake responses to inoculation with these fungi were better in the soil treated with RP than in soil treated with TSP. In the TSP treated soil, the fungi achieved their maximum P releasing capacity two weeks earlier than in soil treated with RP. Positive and significant correlation coefficients among available P, P uptake and dry matter production at different periods of the growing season were observed following inoculation. However, none of these variables were found to be significantly correlated with the fungal populations.     

三种松树内生真菌的分离鉴定及其对红脂大小蠹伴生真菌的拮抗作用

【背景】松树的内生真菌会影响红脂大小蠹及其伴生真菌的生长及扩散,从而影响红脂大小蠹的入侵。【目的】掌握赤峰地区红脂大小蠹寄主树种内生真菌的物种多样性,筛选对其伴生菌有拮抗作用的菌株,为红脂大小蠹的生物防治提供资源。【方法】采用组织分离法、形态学鉴定和internal transcribed spacer (ITS)序列分析相结合的方法对红脂大小蠹寄主树种油松、樟子松和潜在寄主落叶松进行内生真菌多样性研究,并用两点对峙法进行拮抗菌株筛选。【结果】松树内生真菌鉴定为2门6纲10目18科19属,其中落叶松韧皮部分离到的内生真菌数量最多,为39株,隶属于9属12种,真菌检出率为43.33%;油松次之,为30株,隶属于7属8种,真菌检出率为33.33%;樟子松最少,为29株,隶属于10属13种,真菌检出率为32.22%。3个树种的内生真菌相似性较低,无共有的菌种,青霉属(Penicillium)和篮状菌属(Talaromyces)是唯二的共有菌属,且青霉属在3个树种中均为优势菌属。平板对峙结果表明90%以上的树木内生真菌均能够与伴生真菌形成稳定的对峙,抑制率在50%-86%之间,且Phialocephala sp.和Pochonia bulbillosa对伴生菌的抑制率能高达93.7%。【结论】松树韧皮部的内生真菌具有较高的生防潜力,Phialocephala sp.和P. bulbillosa对红脂大小蠹伴生菌有较好的抑制效果,可作为红脂大小蠹的生防资源。     

Biodiversity of soil-inhabiting fungi

This survey is concerned only with filamentous fungi living in the soil layer. The observed diversity of soil fungi largely depends on the method used and the numbers of isolates obtained. Particle-plating usually yields higher numbers of taxa than dilution plating. The Centraalbureau voor Schimmelcultures (CBS) preserves a great diversity of soil fungi. The CBS database contained 2,210 species of soil fungi in 2001, an estimated 70% of the known species available in culture. Thus, the current estimate for described culturable soil fungi is approximately 3,150 species, many of which have a cosmopolitan distribution. Adding the ca. 150 spp. of nonculturable Glomerales results in 3,300 species of currently known soil fungi. Molecular studies in such groups as Fusarium, Trichoderma, Penicillium, and Aspergillus are finding a number of more narrowly distributed cryptic species. Thus the number of species of soil fungi is expected to be considerably higher than the 3300 species currently known.     

一株高效解磷菌的筛选鉴定及溶磷性能

【背景】土壤中大部分磷元素是以难溶性磷酸盐的形式存在,不能被农作物有效利用,而传统化学肥料会带来环境污染等问题。【目的】解决土壤磷缺失现状,开发新型、安全、高效的微生物菌肥。【方法】取武汉科技大学图书馆后土壤为试验材料,筛选出一株高效解磷菌。通过个体形态鉴定、生理生化鉴定、16S rRNA基因序列分析鉴定菌株,以NBRIP为基础培养基进行条件优化,借助高效液相色谱进行细菌解磷机理探究。【结果】所筛选的高效解磷菌株为唐菖蒲伯克霍尔德氏菌(Burkholderia gladioli)。在20种氨基酸中,D-蛋氨酸对菌株的生长和溶磷促进作用最好,促进效果达到19.09%和16.16%,甲酸钠对菌株的生长和溶磷有抑制效果,抑制效果达到39.08%和10.66%。该菌株通过分泌葡萄糖醛酸、D-L-苹果酸等有机酸溶解环境中的磷酸盐,将菌株制作成菌肥对辣椒幼苗有明显的促生长作用。【结论】利用唐菖蒲伯克霍尔德氏菌(Burkholderia gladioli)分泌有机酸溶解土壤中的磷酸盐,可为生物肥料的制备和应用提供一定的理论参考。     

Isolation,identification and testing for allergenicity of fungi from air-conditioned indoor environments

Twelve fungi were isolated and identified from air-conditioned rooms. Of the 12, 7 were species of Aspergillus, viz. A. niger, A. oryzae, A. fumigatus, A. terreus, A. nidulans, A. versicolor and A. parasiticus. Other fungi were Penicillium citrinum, Fusarium oxysporum, Trichoderma viride, Neurospora crassa and Alternaria alternata. A. niger was present in 80% of the locations. Some of the fungi isolated in this study could be opportunistic fungal pathogens, like A. fumigatus, A. niger and Penicillium citrinum, and were found to be allergenic. Results of this study indicate that air-conditioned rooms could be reservoirs of fungi and may cause allergic problems or infections in healthy or immunocompromised individuals living in these environments.     

Accumulation of genes for susceptibility to rust fungi for which barley is nearly a nonhost results in two barley lines with extreme multiple susceptibility

Nonhost resistance is the most common type of resistance in plants. Understanding the factors that make plants susceptible or resistant may help to achieve durably effective resistance in crop plants. Screening of 109 barley (Hordeum vulgare L.) accessions in the seedling stage indicated that barley is a complete nonhost to most of the heterologous rust fungi studied, while it showed an intermediate status with respect to Puccinia triticina, P. hordei-murini, P. hordei-secalini, P. graminis f. sp. lolii and P. coronata ff. spp. avenae and holci. Accessions that were susceptible to a heterologous rust in the seedling stage were much more or completely resistant at adult plant stage. Differential interaction between barley accessions and heterologous rust fungi was found, suggesting the existence of rust-species-specific resistance. In particular, many landrace accessions from Ethiopia and Asia, and naked-seeded accessions, tended to be susceptible to several heterologous rusts, suggesting that some resistance genes in barley are effective against more than one heterologous rust fungal species. Some barley accessions had race-specific resistance against P. hordei-murini. We accumulated genes for susceptibility to P. triticina and P. hordei-murini in two genotypes called SusPtrit and SusPmur, respectively. In the seedling stage, these accessions were as susceptible as the host species to the target rusts. They also showed unusual susceptibility to other heterologous rusts. These two lines are a valuable asset to further experimental work on the genetics of resistance to heterologous rust fungi.Electronic Supplementary Material Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s00425-004-1319-1Abbreviations ff. spp Formae speciales - RIL Recombinant inbred line - DC Double cross - DC-S Progeny produced by selfing of double-cross plants     

In vitro evaluation of combination of Trichoderma harzianum and chitosan for the control of sapstain fungi

In vitro assays were undertaken to evaluate the control of two sapstain fungi, Leptographium procerum and Sphaeropsis sapinea by a combination of chitosan or chitosan oligomer and an albino strain of Trichoderma harzianum. Spore germination and hyphal growth of the test fungi were assessed on media amended with chitosan or chitosan oligomer with and without T. harzianum using either simultaneous inoculation with test fungus or inoculation 1, 2, or 3 days after pre-infection with test fungus.There was no mycelial growth of the test fungi regardless of chitosan concentrations used when either L. procerum or S. sapinea was simultaneously inoculated with T. harzianum. However, the dose–response of chitosan or chitosan oligomer on the test fungi was apparent when T. harzianum was not simultaneously inoculated with test fungus but introduced later. There was a greater growth reduction at higher concentrations (0.075–0.1% v/v) of chitosan, and overall chitosan oligomer was more effective than chitosan aqueous solution.Chitosan alone was able to restrict or delay the germination of spores but the combination of chitosan and T. harzianum inhibited spore germination and hence colony formation of test fungi regardless of time delay.     

Synthesis and evaluation of N-alkyl-beta-D-glucosylamines on the growth of two wood fungi, Coriolus versicolor and Poria placenta

Various glucosylamines were synthesized from glucose and different alkyl amine compounds. These amino compounds are beta-d-glucopyranosylamine (GPA), N-ethyl-beta-d-glucopyranosylamine (EtGPA), N-butyl-beta-d-glucopyranosylamine (BuGPA), N-hexyl-beta-d-glucopyranosylamine (HeGPA), N-octyl-beta-d-glucopyranosylamine (OcGPA), N-dodecyl-beta-d-glucopyranosylamine (DoGPA), N-(2-hydroxyethyl)-beta-d-glucopyranosylamine (HEtGPA) and N,N-di(2-hydroxyethyl)-beta-d-glucopyranosylamine (DHEtGPA). They were tested for their antifungal activity against the growth of Coriolus versicolor and Poria placenta. An improvement of the antifungal activity with the increase of alkyl chain length was observed. DoGPA exhibited the best antifungal activity against both strains. It completely inhibited the fungal growth at 0.01x10(-3)molmL(-1) and 0.0075x10(-3)molmL(-1) for C. versicolor and P. placenta, respectively. For other glucosylamines higher concentrations were needed for complete inhibition of fungi.     

A viscometric study of the biodegradation of photographic gelatin by fungi isolated from cinematographic films

The biodegradation of photographic gelatin grade (Bloom 225) material was studied by viscometry in aqueous solution (at 37 °C, 6.67% w/w) using filamentous fungi isolated and identified from cinematographic film stored in different Spanish archives. From viscosity data, different variables such as molecular weight and chain scission were calculated. To ensure initial spore suspension concentration was standardized for all the biodegradation experiments, a correlation between transmittance at 530 nm of fungal spore suspensions and the corresponding cytometric determination of populations was established for all the fungal strains studied in this work. The bioassay experiments were carried out at 25 and 4 °C using an initial concentration of fungi of 4.5×10⁵ conidia/mL except in the case of the genus Alternaria, where the concentration was 10 times lower. The fungal strains were three species of Aspergillus, i.e., A .ustus, A. nidulans var. nidulans, A. versicolor, seven Penicillium chrysogenum strains, and Cladosporium cladosporioides, Alternaria alternata, Mucor racemosus, Phoma glomerata, and Trichoderma longibrachiatum. All were gelatinase positive. Through the viscosity decay profiles with bioassay-time and the corresponding calculated chain scission, the relative quantitative gelatinase efficiency of these fungi has been evaluated.