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1.
苹果树腐烂病拮抗细菌鉴定及其抑菌作用效果测定   总被引:8,自引:0,他引:8  
为了开发一种高效低毒的苹果树腐烂病生防制剂,通过对峙培养法、形态学及分子生物学的方法进行了苹果树腐烂病菌拮抗菌的分离筛选及鉴定,采用离体枝条法测定了拮抗菌对苹果树腐烂病的防效,并采用显微观察和液体培养法分别研究了拮抗菌对苹果树腐烂病菌的抑菌机理和无菌滤液对苹果树腐烂病菌生长的影响。分离筛选结果表明,从甘肃省各苹果产区果园土壤和苹果树枝条上分离得到23株细菌,2株对苹果树腐烂病菌具有较好拮抗作用,分别为LZ-1201和TS-1203,其对苹果树腐烂病菌菌丝生长抑制率分别为79.00%和85.00%。鉴定结果表明,菌株LZ-1201和TS-1203分别为枯草芽孢杆菌(Bacillus subtilis)和解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。离体枝条防效测定表明,拮抗菌无菌滤液对苹果树腐烂病的防效随着稀释倍数增大而降低,原液防效最高,分别为74.43%和77.07%。抑菌作用机理结果表明,两株拮抗菌均可导致苹果树腐烂病菌丝膨大畸形、原生质浓缩、释放及溶解。拮抗菌无菌滤液对腐烂病菌生长的影响测定结果表明,无菌滤液对腐烂病菌分生孢子萌发和菌丝生长量均有显著抑制作用(P0.05),其无菌滤液稀释40倍时对腐烂病菌分生孢子萌发和菌丝生长量的抑制率均高于60%,表明该拮抗菌具有很好的生防潜力。  相似文献   

2.
以生防放线菌长枝木霉T6和放线菌ZZ-9为试材,采用圆盘滤膜法和对口培养法比较T6、ZZ-9所产生的代谢物对苹果树腐烂病病菌、苹果树轮纹病病菌和苹果树早期落叶病病菌的拮抗作用,结果表明:长枝木霉T6的非挥发性物质对这3种病原菌抑制效果明显,抑制率分别达到97.50%、83.69%、68.35%。放线菌ZZ-9的非挥发性物质对苹果树腐烂病菌和苹果树轮纹病菌的抑制率达到了96.25%和72.42%%,对早期落叶病菌的拮抗作用较弱,抑制率小于50%。长枝木霉T6和放线菌ZZ-9的挥发性物质对这3种病原菌的拮抗作用不明显,抑制率均小于50%。长枝木霉T6和放线菌ZZ-9对苹果树三种病害的拮抗作用主要由二者所产生的非挥发性物质起作用。  相似文献   

3.
牛粪发酵沼液对立枯丝核菌的抑制作用   总被引:2,自引:0,他引:2  
为探讨牛粪发酵沼液的抑菌机理,以立枯丝核菌为研究对象,进行了试验研究。采用平皿培养方法:培养基表面分别均匀涂布1mL5种不同处理的沼液,即不作处理沼液、沼液灭菌、离心上清液、无菌膜滤液和菌体溶液,中心接种立枯丝核菌并于生化培养箱中28℃培养96h,每12h测量立枯菌落的生长直径以计算抑制率,同时借助分子生物学手段鉴定了沼液中起抑制作用的微生物。试验结果表明菌体溶液和不作处理沼液对立枯的抑制效果较好,抑制率最高可达74%和53%,离心上清液的抑制效果次之,无菌膜滤液和沼液灭菌处理的抑制作用较弱,一般在20%以下;对沼液中的微生物菌群进行了分离、纯化与筛选,获16株菌种,并与立枯丝核菌进行对峙培养试验,其中nfd-9和nfd-16具有较强抑制作用,其他菌株几乎没有抑制作用。将nfd-9、nfd-16与立枯对峙交界面处进行了扫描电镜观察,结果显示:这2株微生物通过竞争、拮抗和重寄生等作用来抑制立枯丝核菌的生长,从而到达防止病害的目的。经分子生物学技术鉴定,确定了nfd-9和nfd-16分别为米根霉菌(Rhizopus oryzae)和哈茨木霉(Trichoderma harzianum)。研究结果为进一步了解沼液抑制真菌的机理提供一定的理论参考。  相似文献   

4.
为探讨牛粪发酵沼液的抑菌机理,以立枯丝核菌为研究对象,进行了试验研究。采用平皿培养方法:培养基表面分别均匀涂布1 mL 5种不同处理的沼液,即不作处理沼液、沼液灭菌、离心上清液、无菌膜滤液和菌体溶液,中心接种立枯丝核菌并于生化培养箱中28℃培养96 h,每12 h测量立枯菌落的生长直径以计算抑制率,同时借助分子生物学手段鉴定了沼液中起抑制作用的微生物。试验结果表明菌体溶液和不作处理沼液对立枯的抑制效果较好,抑制率最高可达74%和53%,离心上清液的抑制效果次之,无菌膜滤液和沼液灭菌处理的抑制作用较弱,一般在20%以下;对沼液中的微生物菌群进行了分离、纯化与筛选,获16株菌种,并与立枯丝核菌进行对峙培养试验,其中nfd-9和nfd-16具有较强抑制作用,其他菌株几乎没有抑制作用。将nfd-9、nfd-16与立枯对峙交界面处进行了扫描电镜观察,结果显示:这2株微生物通过竞争、拮抗和重寄生等作用来抑制立枯丝核菌的生长,从而到达防止病害的目的。经分子生物学技术鉴定,确定了nfd-9和nfd-16分别为米根霉菌(Rhizopus oryzae)和哈茨木霉(Trichoderma harzianum)。研究结果为进一步了解沼液抑制真菌的机理提供一定的理论参考。  相似文献   

5.
植物内生真菌作为获取生物防治材料的一个潜在且庞大的优质资源库,极具开发应用潜力。采用平板对峙法,对已有的137株小白及内生真菌进行块茎腐烂病菌菌丝生长的拮抗活性筛选,并以6种常见农业病原菌作为靶标菌完成抗菌谱测定,初步获得抗病能力较强的2株内生真菌:菌株GZFJ021和GZLG033。  相似文献   

6.
本研究采用平板对峙法和菌丝块法测定分离自广西北部湾的3株海洋真菌MF-3、MF-6和MF-13的抗荔枝和水稻病原菌抗菌活性,并通过形态学观察结合分子生物学方法鉴定其分类地位。结果表明,当采用平板对峙法时,3株海洋真菌对荔枝霜疫霉、炭疽病菌、水稻纹枯病菌和水稻稻瘟病菌等4种指示菌的菌丝生长均有强的抑制作用;菌丝块法试验结果发现3株真菌的发酵液均对荔枝霜疫霉菌有很好的抑制效果,菌丝相对生长抑制率分别达到94.29%、98.16%和86.37%,对水稻纹枯病病原菌抑制效果次之,其菌丝相对生长抑制率达到80%左右;其中,MF-3菌株对荔枝霜疫霉、炭疽菌、水稻纹枯病菌和水稻稻瘟病菌都有较好的抑制作用,MF-6对霜疫霉、水稻纹枯病菌和水稻稻瘟病菌也有较好的抑菌效果和广谱性。结合形态学观察和ITS rDNA序列比对分析,初步将MF-3鉴定为布雷正青霉(Eupenicillium brefeldianum),MF-6为微紫青霉(Penicillium janthinellum),MF-13为短棒曲霉(Aspergillus clavatonanicus)。耐盐度试验结果表明这3株真菌是适应海洋环境的兼性海洋真菌。该3株海洋真菌表现的抗真菌活性,对应用于荔枝或水稻病害的生物防治和新型农药的研发有潜在的应用价值。  相似文献   

7.
芦笋茎枯病的生物防治及机理研究   总被引:3,自引:0,他引:3  
利用拮抗芽孢杆菌菌株B96-Ⅱ对芦笋茎枯病菌进行了室内抑菌试验和田间防治及机理研究,结果表明:B96-Ⅱ对芦笋茎枯病菌有明显抑制作用.稀释50~1600倍后平皿试验,生长速率抑制率为97.37%~92.98%;离体组织培养对菌斑的抑制率为94.40%;田间防治效果为93.40%.显微观察发现,B96-Ⅱ处理后可导致菌丝和孢子严重破损,使细胞内容物外渗.处理后24h,菌液电导率(μs·cm-1)和总溶解固体(TDS)比不处理的对照明显提高,处理中B96-Ⅱ培养液浓度从1%升到10%时,电导率增加量从47.50%提高到518.33%.总溶解固体增加量从176.10%提高到797.60%.经测定,B96-Ⅱ可抑制病原菌菌丝呼吸,处理中B96-Ⅱ培养液浓度从1%升到10%时呼吸抑制率从25.00%升至196.40%.生物测定表明,B96-Ⅱ的主要抑菌物为拮抗蛋白.  相似文献   

8.
本试验在实验室条件下,研究大中型沼气工程猪沼和牛沼两种沼液原液及其离心上清液对大豆尖孢镰刀菌、大豆菌核病菌、小麦纹枯病菌、小麦根腐病菌、水稻纹枯病菌、玉米大斑病菌和玉米小斑病菌等7种农作物病原真菌的抑制作用。结果表明:猪沼原液和牛沼原液对其中的大豆尖孢镰刀菌、大豆菌核病菌、小麦纹枯病菌、小麦根腐病菌和水稻纹枯病菌5种病原真菌具有较好的抑制作用,其菌丝生长抑制率均在72%以上,但对玉米大斑病菌和玉米小斑病菌的菌丝生长基本没有抑制作用;相比之下,猪沼和牛沼离心上清液对以上5种病菌的菌丝生长抑制作用明显减弱,除猪沼离心上清液对大豆菌核病菌的菌丝生长抑制率大于70%以外,试验中猪沼和牛沼离心上清液对实验病原菌的菌丝生长抑制率基本都在60%以下。试验结果显示猪沼液和牛沼液对农作物病原真菌具有潜在的植物病害防治作用,为养殖场大中型沼气工程沼液应用新技术的开发提供科学依据。  相似文献   

9.
采用自然腐解与接种微生物人工腐解两种方式分别处理棉秆,研究了两种腐解物水浸提液对棉花枯、黄萎病菌的化感效应。结果显示,两种腐解液对枯、黄萎病菌的菌丝生长及孢子萌发均具有较强的抑制作用,抑制强度整体上随着腐解液浓度的升高而增大。棉秆自然腐解液对枯、黄萎病菌菌丝生长的最大抑制率分别为58.17%、48.16%,对孢子萌发的最大抑制率分别为23.29%、16.22%,对枯、黄萎病菌毒力的有效中浓度(EC50)分别为33.20、45.20g·L-1;人工腐解液对枯、黄萎病菌菌丝生长的最高抑制率分别为71.63%、79.04%,对孢子萌发的最高抑制率分别为35.45%、42.74%,EC50分别为24.18、20.75g·L-1。表明棉秆腐解后具有作为植物源抑菌剂的开发潜力,并且采用人工方式腐解棉秆能有效提高腐解液对棉花枯、黄萎病菌的抑制强度。  相似文献   

10.
采用菌丝生长抑制和分生孢子萌发抑制的试验方法,以草莓枯萎病菌为靶标病原菌,研究了江苏省内运行稳定的21个大型沼气工程的沼液及其无菌滤液对草莓枯萎病菌的生长抑制特性。结果表明,所有沼气工程的沼液对草莓枯萎病菌的菌丝生长都有不同程度的抑制作用,不同运行年限的沼气池沼液以及不同发酵原料的沼液对草莓枯萎病菌的抑制效果均无显著差异,但不同贮液阶段的沼液对其抑菌效果影响显著。沼液的无菌滤液对草莓枯萎病菌菌丝的生长没有抑制作用;沼液及其无菌滤液对草莓枯萎病菌分生孢子的萌发均有不同程度的抑制作用,猪粪沼液及其无菌滤液对草莓枯萎病菌分生孢子萌发的抑制作用好于牛粪沼液。沼液对生产上常见的5种植物病原真菌的抑制效果有显著差别。沼液中的拮抗微生物是沼液抑菌防病的主要因子。  相似文献   

11.
Summary -(Isoxazolin-5-on-2-yl)-alanine (IA), a heterocyclic non-protein amino acid from root extracts and root exudates of pea seedlings, acts as a potent growth inhibitor of several eukaryotic organisms, including yeasts, phytopathogenic fungi, unicellular green algae, and higher plants. The antibiotic effect on baker's yeast was reversed by l-methionine, l-cysteine, and l-homocysteine. Phytopathogenic fungi such as Botrytis cinerea, Pythium ultimum, and Rhizoctonia solani grown on agar containing IA were inhibited in the growth of mycelia or in the production of sclerotia. In contrast, no significant inhibition of either Gram-positive or Gram-negative bacteria was observed. Rhizobium leguminosarum, the compatible microsymbiont of Pisum spp., and Rhizobium meliloti were able to tolerate up to 2.9 mM IA (500 ppm) without any effect on the growth rate. Bradyrhizobium japonicum even gave a positive chemotactic response to IA. The ecological significance of IA as a preformed plant protectant during the seedling stage of Pisum spp. and other IA-containing legumes is discussed.  相似文献   

12.
从海产虾壳上分离筛选拮抗放线菌、研究其抑菌活性具有重要的实践意义。藤黄轮丝链霉菌(Streptomyces luteoverticillatus)T0907-107是分离自海产虾壳上并经初筛和鉴定的一株链霉菌,为了进一步开发和利用该菌,本研究采用极性不同的5种溶剂(丙酮、乙酸乙酯、正丁醇、乙醇和甲醇),对该菌株的发酵液菌体活性物质进行了提取,对粗提物的理化性质和抑菌活性进行了初步研究。结果表明,其发酵液活性物质大部分存在于菌体中,采用甲醇和丙酮浸提的效果好,其抑菌圈直径分别为26.20和22.50 mm;对粗提物进行耐热性实验,结果表明,粗提物是一种能够耐60℃高温和一定酸碱的物质;粗提物对蛋白酶K稳定;菌体及粗提物对多种植物病原菌物如烟草炭疽病菌(Colletotrichum nicotianae Sacc.)、烟草赤星病菌(Alternaria alternata(Fries)Keissler)、烟草立枯丝核病菌(Rhizoctonia solani Kühn)、枇杷炭疽菌(C.gloeosporioides(Penz.)Sacc.)和白菜炭疽病菌(C.higginsianum Sacc.)有较强抑制作用。研究结果表明,甲醇和丙酮是较合适的萃取溶剂,该菌发酵液菌体粗提物极性强、稳定性好、有较广的抑菌谱,显示出好的开发前景。本研究为进一步鉴定该活性物质及开发和利用该菌提供了基础资料。  相似文献   

13.
Fungistasis is a widespread phenomenon that can be mediated by soil microorganisms and volatile organic compounds (VOCs). The relationship between soil microorganisms and VOCs is still unclear, however, and many fungistatic compounds remain to be identified. We assessed the effects of soils (soil direct fungistasis) and VOCs produced by natural soils (soil volatile fungistasis) on the spore germination of several fungi. Both strong soil direct fungistasis and soil volatile fungistasis were observed in a wide range of soils. Soil fungistasis and VOC fungistasis were significantly correlated (P<0.001). The volatile fungistatic activity of soils stopped after autoclaving. Some VOCs were identified by using solid-phase microextraction-gas chromatography/mass spectrum. VOC composition and in vitro antagonism of relatively pure commercial compounds also were measured. Some VOCs, trimethylamine, 3-methyl-2-pentanone, dimethyl disulfide, methyl pyrazine, 2,5-dimethyl-pyrazine, benzaldehyde, N,N-dimethyloctylamine and nonadecane, were produced by various fungistatic soils. Moreover, antifungal activity test of above VOCs showed that trimethylamine, benzaldehyde, and N,N-dimethyloctylamine have strong antifungal activity even at low levels (4-12 mg l−1).  相似文献   

14.
Summary The beneficial effect of rhizopseudomonad strains on the growth of the barley cultivar Iban as a function of the storage time under laboratory conditions was evaluated. The beneficial rhizopseudomonad strains were preserved on slants of Modified King Medium B (MKB) at 28 °C and transferred every 2 weeks. After 6 months of storage most strains lost their beneficial effect on plant growth. Cold shock treatment enabled some strains to recover their beneficial effect on plant growth. Evidence is presented that there is a relationship between recovery of the stimulatory effect of certain rhizopseudomonad strains on plant growth and recovery of the capacity to colonize roots and to express antifungal activity.  相似文献   

15.
根霉菌(Rhizopus oryzae)是热带果蔬采后主要病原真菌之一,不仅引起果实腐烂,影响果实风味,还能分泌对人体有毒害作用的麦角类生物碱,对果蔬贮藏具有极大的危害。本研究从腐烂的草莓(Fragaria ananassa L.cv.Zhangji)中分离主要致病菌,经核糖体DNA内转录间隔区(r DNA-ITS)序列分析辅以传统的形态学观察法,鉴定其遗传背景,然后利用碳酸氢钠(SB)、硼酸(BA)、肉桂油(Co)、硝普钠(SNP)、亚磷酸盐(Phi)等5种已知抑菌物质处理病原菌。通过检测处理前后病原菌生理、生化指标的改变以及番茄(Solanum lycopersicum cv.Ailsa Cragi)果实接种实验,确定5种抑菌物质对病原菌发育及致病力的影响。结果表明,分离的病原菌通过回接试验证实为草莓的主要致病菌,致病力强,果实发病部位出现明显的褐化、软化以及水化,病征与匍枝根霉(Rhizopus stolonifer)引起的软腐病表型相似。同时,利用ITS通用引物扩增得到大小约600 bp的产物,经比对后与Rhizopus oryzae的r DNA-ITS序列完全一致,因此判断该病原菌为R.oryzae。此外,本研究选用的5种物质对R.oryzae都有明显的抑制作用,且抑菌效果均与浓度成正比。当SB、BA、SNP、Co以及Phi的浓度分别达到0.2%、0.2%、0.5%、0.02%及5 mmol/L时,培养6 h后对照病原菌的孢子萌发率超过90%,而抑菌物质处理的孢子萌发率均低于10%。因此,这些浓度被确定为5种物质的最低抑菌浓度。在最低抑菌浓度下,尽管抑制效力有所不同,但5种抑菌物质均能明显延缓芽管伸长及菌丝扩展速度,降低病原菌生物量积累,引起病原菌糖吸收障碍,并对R.oryzae诱发的番茄软腐病有明显的防治作用。其中BA、Co和Phi在体外抑菌实验中效果优于SB和SNP;SB、BA以及Phi对番茄R.oryzae软腐病的防治效果优于Co与SNP。本研究为果蔬采后病原菌R.oryzae的防治提供了新的思路,为开发SB、BA、Co、SNP以及Phi等5种抑菌物质的应用潜力提供了理论基础。  相似文献   

16.
In order to identify key adaptive traits which affect productivity in Mediterranean grain and forage legumes and simultaneously determine the agricultural potential of a wide range of Vicia species, germplasm collected from the wild throughout the eastern Mediterranean was grown under semi-arid conditions in Tel Hadya, Syria (313 mm growing season rainfall). These included species currently in use in Mediterranean agriculture, such as V. sativa L., as well as those more widely used in the past-such as V. ervilia L., but also a broad selection from Section Narbonensis (B. Fedtsch. ex Radzhi) Maxted, including V. narbonensis L, V. johannis Tamamsch., V. hyaeniscyamus Mout., V. serratifolia Jacq., and V. kalakhensis Khattab et al. V. faba, a near relative of the taxa in Section Narbonensis, was included as a domesticated control. Where applicable, a representative range of subspecies was used. Accessions were chosen from a wide range of habitats in terms of latitude (31.02–40.72 decimalo), longitude (27.1–43.17 decimalo), altitude (20–1510 m), rainfall (180–1700 mm/yr) and soil depth (5–50 cm) in order to maximise diversity within species. Agricultural potential was determined by measuring seed, hay and biological yield, as well as agronomic traits such as harvest index, standing crop height, and seed size. The comparative influence of phenology and key agronomic traits such as plant habit and seed size on productivity varied tremendously between species, depending on their reproductive strategies. In V. sativa and V. ervilia, the smaller seed species which rely on long vegetative phases and growing seasons to accumulate sufficient biomass to set seed, and in which there was comparatively little agronomic variation, phenology had a large impact on yield. In early emerging taxa such as V. ervilia and V. s. subsp. sativa, with built-in long vegetative phases and growing seasons, seed yield was negatively correlated with flower ing (r = –0.86 to –0.88), whereas the opposite was the case for later emerging taxa such as V. s. subsp. nigra (L.) Ehrh. (r = 0.95). Within V. narbonensis and relatives, the larger seeded Vicia species which rely on more conservative reproductive strategies where high seedling vigour associated with large seeds enables the species to enter reproductive phases relatively early, phenology had a much smaller impact on yield than did variation of key traits such as seed weight, plant habit and pod shattering. Among the undomesticated germplasm harvest indices ranged from 0.09–0.31, hay yields from 0.1–3.4 t/ha, seed yield from 0–2.0 t/ha, and dry matter at maturity from 1.6–6.5 t/ha. Sub-specific taxonomy was crucial in determining agronomic potential. V. narbonensis var. aegyptiaca Kornhuber ex Asch. et Schweinf. showed the most potential, combining an upright habit, large seeds (212 mg) and tendency to retain intact pods after maturity, with the highest yield, harvest index and crop height of all the wild Vicia species. V. sativa subsp. sativa, V. ervilia and V. narbonensis var. narbonensis were less productive, but still showed agricultural potential. The smaller seeded V. narbonensis, var. affinis, var. jordanica H. Schäf. and var. salmonea (Mout.) H. Schäf., and their close relatives V. johannis, V. hyaeniscyamus, V. serratifolia and V. kalakhensis have little to offer Mediterranean agriculture on the basis of poor agronomy.  相似文献   

17.
Plant growth promoting rhizobacteria (PGPR) enhance the plant growth directly by assisting in nutrient acquisition and modulating plant hormone levels, or indirectly by decreasing the inhibitory effects of various pathogens. The aim of this study was to select effective PGPR from a series of indigenous bacterial isolates by plant growth promotion and antifungal activity assays. This study confirmed that most of the isolates from maize rhizosphere were positive for PGPR properties by in vitro tests. Azotobacter and Bacillus isolates were better phosphate solubilizers and producers of lytic enzymes, hydrocyanic acid (HCN), and siderophores than Pseudomonas. Production of indole-3-acetic acid (IAA) and antifungal activity were the highest in Azotobacter, followed by Bacillus and Pseudomonas. The most effective Azotobacter isolates (Azt3, Azt6, Azt12) and Bacillus isolates (Bac10, Bac16) could be used as PGPR agents for improving maize productivity. Further selection of isolates will be necessary to determine their efficiency in different soils.  相似文献   

18.
species were isolated from the rhizosphere of green gram [Vigna radiata (L.) Wilczek] and some of the rhizobacterial isolates were found to have a wide range of antifungal activity inhibiting growth of the phytopathogenic fungi Aspergillus sp., Curvularia sp., Fusarium oxysporum and Rhizoctonia solani in culture. These isolates also showed slight inhibition of the growth of a Bradyrhizobium strain (Vigna) in a spot test which was mainly a result of nutrient competition as culture supernatants of the Pseudomonas isolates did not inhibit the growth of bradyrhizobia but inhibited the growth of fungi. The rhizobacterial isolates produced siderophores in Fe-deficient succinate medium. However, the inhibition of fungal growth by different Pseudomonas isolates in Luria Bertani and King's medium B which were not limiting in Fe3+ ions suggested that, besides siderophores, other antifungal compounds (antibiotics) produced by these rhizobacteria were involved in antagonism. On coinoculation of green gram with Pseudomonas strains MRS13 and MRS16 and Bradyrhizobium sp. (Vigna) strain S24, there was a significant increase in nodule weight, plant dry weight and total plant N as compared to inoculation with Bradyrhizobium strain S24 alone, suggesting that the nodule-promoting effects of Pseudomonas sp. lead to an increase in symbiotic N fixation and plant growth. Received: 27 October 1997  相似文献   

19.
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