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1.
郑磊  张静  麻文建  彭燕  朱天辉 《广西植物》2016,36(6):651-657
为分析腐皮镰刀菌(Fusarium solani)引起的核桃根腐病对寄主抗性生理指标的影响,该研究以铁核桃、绵阳早熟、新疆2号3个核桃品种为材料,研究各品种受腐皮镰刀菌侵染后,不同时期的发病情况以及丙二醛(MDA)、可溶性糖含量以及过氧化物酶(POD)、多酚氧化酶(PPO)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)的动态变化。结果表明:接种病原菌50 d 后,铁核桃100%发病,属高感品种,绵阳早熟和新疆2号发病率分别为23.33%和16.67%,属抗病品种;在50 d 监测范围内,各品种对照组各项叶部生理指标仅出现上下波动且差异不显著;然而,各品种核桃接入腐皮镰刀菌后,可溶性糖含量随时间迅速下降,MDA 含量和 POD 活性总体表现为先上升趋势,SOD、PPO 活性总体则表现为先上升后下降的变化趋势,并且5个指标品种间差异显著,而 CAT 活性变化无规律。相关性分析表明,可溶性糖、MDA 含量、SOD、POD、PPO 活性可以作为核桃品种抗性评价指标。该研究结果为深入研究由腐皮镰刀菌引起的核桃根腐病发病机制以及通过诱导等措施提高其抗性的研究提供了理论基础。  相似文献   

2.
【背景】腐皮镰刀菌(Fusariumsolani)是一种分布较为广泛的致病性真菌,可引起多种植物的土传病害,是枸杞根腐病的主要致病菌之一。马铃薯糖苷生物碱(potatoglycosidealkaloids,PGA)为一类植物源提取物,其原材料种植广泛、成本低廉,对腐皮镰刀菌具有较强的抑菌活性。【目的】探究PGA对腐皮镰刀菌呼吸作用及活性氧(reactive oxygen species, ROS)代谢的影响,从能量代谢角度揭示其可能的抑菌机理。【方法】以马铃薯芽为原材料,采用乙酸-氨水沉淀法提取PGA,以腐皮镰刀菌为供试病原菌,通过PDA和PDB培养体系考察PGA对腐皮镰刀菌菌丝生长的抑制作用,并确定半最大效应浓度(EC50);采用氧电极仪检测PGA对腐皮镰刀菌呼吸作用的影响;并通过PDB液态培养试验体系,研究PGA对腐皮镰刀菌抗氧化酶系统、ROS及其代谢产物丙二醛(malondialdehyde, MDA)的影响。【结果】PGA处理下菌丝体呼吸速率明显下降,且随着PGA处理时间的延长,表现出一定的时间浓度效应。PGA处理使胞内过氧化氢(H2O2)和超氧阴离子(O2-)含量显著增高(P...  相似文献   

3.
本文对镰刀菌3个种的47个菌株用豌豆发芽抑制和鸽子呕吐试验筛选雪腐镰刀菌烯醇(Nivalenol)的产生菌株。化学分析的结果表明雪腐镰刀菌M-24和禾谷镰刀菌M-46能够产生雪腐镰刀菌烯醇。菌株M-24适宜的产毒温度为25-30℃;25℃条件下,M-24的产毒高峰出现在接种后的第三、四周,雪腐镰刀菌烯醇的含量分别达到447.9mg/kg和538.0mg/kg;五种培养基上产毒量也有所不同,在大麦和大米上产毒量最高(毒素含量分别为819.7mg/kg和780.5mg/kg),玉米和小麦上次之(521.9mg/kg,402.4mg/kg),绿豆上产毒量最低(167.6mg/kg)。菌株M-46在上述条件下产毒量均低于5mg/kg。  相似文献   

4.
腐皮镰刀菌SYBR Green实时荧光定量PCR快速检测方法的建立   总被引:1,自引:0,他引:1  
目的建立一种能够快速、灵敏、特异的鉴定腐皮镰刀菌的SYBR Green实时荧光定量PCR。方法运用SYBR Green实时荧光定量PCR反应体系检测腐皮镰刀菌,并对此方法的特异性、灵敏度和稳定性进行评价。结果通过对45例样品的检测,结果显示SYBR Green实时荧光定量PCR特异性好,其检出率高于普通PCR;灵敏度高,对重组质粒标准品的检测灵敏度为1.0×10~2copies/μL;稳定性好,对质粒为1.0×10~7copies/μL、1.0×10~5copies/μL、1.0×10~3copies/μL的标准品重复检测10次,结果显示扩增反应Ct值的变异系数为0.96%~1.68%。结论SYBR Green实时荧光定量PCR检测腐皮镰刀菌,不仅特异性好,灵敏度高,稳定性好,而且简便、快速、易操作。  相似文献   

5.
脱氧雪腐镰刀菌烯醇(DON)是镰刀菌属产生的一种真菌毒素,具有细胞毒性,可导致胚胎发育畸形,严重威胁人畜健康.为了获得大量的高纯度DON用于毒理学及脱毒研究,本研究开发了一种操作简单、经济高效的DON分离纯化方法:抽提:采用84%的乙腈水溶液从禾谷镰刀菌PH-1的大米培养物中提取DON获得DON粗毒素;净化:使用石油醚...  相似文献   

6.
抗赤霉病小麦品种苏麦3号、繁9能转化镰刀菌毒素脱氧雪腐镰刀菌烯醇(DON)成产物X,而感病小麦品种宁麦6号、徐州21无转化能力。产物X对小麦黄化芽鞘的伸长生长无抑制作用,而对禾谷镰刀菌分生孢子的萌发有明显抑制。说明抗性小麦品种对赤霉病菌毒素的脱毒是小麦重要的抗赤霉病机制。  相似文献   

7.
脱氧雪腐镰刀菌烯醇对胃癌细胞增殖及细胞周期的影响   总被引:2,自引:0,他引:2  
探讨脱氧雪腐镰刀菌烯醇(DON)对体外培养的两株胃癌细胞(SGC-7901和BGC-823)增殖及细胞周期的影响。采用细胞培养、噻唑蓝(MTF)比色法、流式细胞定量检测(FCM)、蛋白质免疫印迹(Western印迹)以及免疫细胞化学染色(ICH)等方法,研究不同浓度DON处理72h对体外培养胃癌细胞的增殖、细胞周期及细胞周期相关蛋白—细胞周期蛋白依赖性激酶抑制蛋白(CKIs)P21WAF/CIP1和细胞周期蛋白E(cyclin E)表达的影响。MTT法检测结果显示DON可明显抑制两株胃癌细胞的增殖,SGC.7901和BGC.823细胞100、500、1000ug/L DON处理组的增殖抑制率分别为4.28%、36.20%、45.35%和14.89%、32.30%、51.61%。FCM检测结果显示,给予1000ug/LDON处理72h可使两株细胞周期阻滞在GffM期。在100~1000u扎浓度范围内,两株细胞P21WAF/CIP1表达量均高于对照组,P21WAF/CIP1的表达与DON浓度呈显著正相关关系(SGC-7901细胞:r=0.886,P〈0.01;BGC-823细胞:r=0.943,P〈0.01);两株细胞的细胞周期蛋白E表达量均低于对照组,与DON浓度有明显剂量依赖关系(SGC.7901细胞:r=-0.923,P〈0.01;BGC-823细胞:r=-0.854,P〈0.01)。Western印迹及免疫细胞化学检测进一步证实了DON处理对蛋白质表达的影响。综合结果表明,DON可抑制体外培养胃癌细胞的增殖活性,G2/M期阻滞、P21WAF/CIP1表达增高及细胞周期蛋白E表达下降可能是DON抑制胃癌细胞增殖的可能机制,DON对分化程度不同的胃癌细胞的影响没有明显差别。  相似文献   

8.
抗病小麦对脱氧雪腐镰刀菌烯醇的脱毒及产物的生物活性   总被引:3,自引:0,他引:3  
抗赤霉病小麦品种苏麦3号、繁9能转化镰刀菌毒素脱氧雪腐镰刀菌烯醇(DON)成产物X,而感病小麦品种宁麦6号、徐州21无转化能力。产物X对小麦黄花芽鞘的伸长生长无抑制作用,而对禾谷镰刀菌分生孢子的萌发有明显抑制。说明抗性小麦品种对赤霉病菌毒素的脱毒是小麦重要的抗赤霉病机制。  相似文献   

9.
雪腐镰刀菌烯醇产生菌株的筛选   总被引:3,自引:0,他引:3  
陆仕华  张树荣 《真菌学报》1989,8(2):148-153
  相似文献   

10.
饲料中脱氧雪腐镰刀菌烯醇的薄层层析测定法   总被引:2,自引:0,他引:2       下载免费PDF全文
确立了一个适于测定饲料中脱氧雪腐镰刀菌烯醇(Deoxynivslenol,简称DON)的薄层层析法。该方法可排除样品中成分复杂、干扰物质多的不利条件,提取和净化效率高,操作较简便,具有较好的实用性和可靠性。标准点最低检出量为10ng,样品的最低检出限是40ppb。该方法对100、300和500ppb水平的回收率分别是85.9、88.9和79.2%。  相似文献   

11.
Fusarium species are common soil saprophytes and plant pathogens that have been frequently reported as etiologic agents of opportunistic infections in humans. We report eight cases of onychomycosis caused by Fusarium solani (4) and Fusarium oxysporum (4) in São Paulo, Brazil. These species were isolated from toenails in all cases. The infections were initially considered to be caused by dermatophytes. The clinical appearance of the affected toenails was leukonychia or distal subungual hyperkeratosis with yellowish brown coloration. The eight cases reported here suggest that Fusarium spp. should be taken into consideration in the differential diagnosis of tinea unguium.  相似文献   

12.
Eumycetoma is a mycotic disease caused by saprophytic soil fungi that are usually inoculated through minor injuries. A case of mycetoma in a Brazilian farmer aged71 years is reported. This patient presented erythema and edema on the dorsal surface of the left hand with multiple crusted and cicatricial lesions. No macroscopic grains were observed. The histopathological findings showed grains consisted of numerous hyphae which stained well with Gomori-Grocott method. This material obtained by cutaneous biopsy was submitted to culture on Sabouraud’s medium and the colonies were identified as Fusarium solani. The radiological studies revealed bone osteolytic lesions and the ultrasound showed pseudocysts and fistulae at the site of this infection. The patient was treated with oral ketoconazole with a good clinical response. This revised version was published online in June 2006 with corrections to the Cover Date.  相似文献   

13.
Fusarium root rot (Fusarium spp.) is one of the most important seedling diseases of coneflower (Echinacea spp.) in Alberta greenhouses. Effects of microbial antagonists (Trichoderma spp.) and fungicides, including difenoconazole, fludioxonil, and a mixture of fludioxonil, metalaxyl and difenoconazole, on the management of this disease, were investigated in Alberta. Twenty Trichoderma isolates demonstrated antagonistic activity to Fusarium in agar plate bioassays, with inhibition rates ranging from 44 to 65%. Some Trichoderma isolates significantly ( p < 0.05) reduced disease incidence and severity on seedlings in greenhouse experiments. An in vitro bioassay indicated that difenoconazole and the mixture equally inhibited the growth of both Fusarium and Trichoderma, but, while fludioxonil strongly inhibited the growth of Fusarium, it had little effect on Trichoderma, according to the dose--response models developed ( p < 0.01, R2= 0.902-0.998). Two Trichoderma isolates, T1 and T13 were applied singly or in combination with a low rate of fludioxonil in greenhouse evaluations. The results suggested that fludioxonil and Trichoderma could be integrated into a disease management program for fusarium root rot in coneflower.  相似文献   

14.
A fungus was isolated from the stem cuttings of Taxus celebica, which produced paclitaxel in liquid-grown cultures. The fungus was identified as Fusarium solani based on colony characteristics, morphology of conidia and the 26S rDNA sequence. Paclitaxel was identified by chromatographic and spectroscopic comparison with authentic paclitaxel and its cytotoxic activity towards Jurkat cells in vitro.  相似文献   

15.
聚丁二酸丁二酯是一种可替代传统塑料的生物降解塑料,有助于解决白色污染问题,但其在自然界中往往降解缓慢,获取高效降解微生物或解聚酶是使其快速有效降解的关键。通过构建重组毕赤酵母表达体系以实现腐皮镰孢聚丁二酸丁二酯解聚酶(FSC)的高效表达。根据毕赤酵母偏好密码子优化FSC的基因密码子,并导入毕赤酵母X33中实现其重组表达。进一步通过单因素实验优化了菌株的产酶条件。结果显示,优化后的FSC基因序列中的157个碱基发生改变,G+C含量由59.6%降低到48.3%,序列同源性为77.34%;构建的重组表达载体p PICZα-FSC转入毕赤酵母X33,结合抗性平板初筛、SDS-PAGE和Western bolt验证,以及摇瓶发酵酶活力测定获得了1株具有较高产酶能力的重组菌株L1;进一步确定其摇瓶发酵培养条件:培养基起始p H 6.0,摇床转速220 r/min,甲醇补加量1%,接种量8%,培养时间72 h,培养温度30℃,此优化条件下菌株发酵液酶活力可达110 U/mL。  相似文献   

16.
A component ofFusarium solani (F. solani), identified as the major allergen,Fus sI3596 * was purified to homogeneity from culture filtrate (CF) by means of anion-exchange column chromatography, gel filtration and FPLC. The homogeneity ofFus sI3596 * was assessed by IEF, PAGE, SDS-PAGE (non-reducing), immunoblot and HPLC.Fus sI3596 * was isolated as a glycoprotein of MW 65 kd and pI 3.6. The IgE ELISA-inhibition assay after periodate treatment of the fraction showed a lower IgE binding capacity suggesting involvement of carbohydrate moiety in IgE binding reactions of the allergen. Peptide fragments ofFus sI3596 * obtained after CNBr and trypsin treatment were analysed by immunoblotting for their allergenicity. This study indicated that there could be at least 3 allergenic determinants in the major allergen,Fus sI3596 * ofF. solani CF.Abbreviations DEAE diethyl aminoethyl - HPLC high performance liquid chromatography - SDS sodium dodecyl sulphate - PBS-Tween phosphate buffer saline containing 0.1% tween - DTT dithiothreitol - TFA triflouroacetate - FPLC fast protein liquid chromatography  相似文献   

17.
High-level extracellular production of Fusarium solani cutinase was achieved using a Pichia pastoris expression system. The cutinase-encoding gene was cloned into pPICZαA with the Saccharomyces cerevisiae α-factor signal sequence and methanol-inducible alcohol oxidase promoter by two different ways. The additional sequences of the c-myc epitope and (His)6-tag of the vector were fused to the C-terminus of cutinase, while the other expression vector was constructed without any additional sequence. P. pastoris expressing the non-tagged cutinase exhibited about two- and threefold higher values of protein amount and cutinase activity in the culture supernatant, respectively. After simple purification by diafiltration process, both cutinases were much the same in the specific activity and the biochemical properties such as the substrate specificity and the effects of temperature and pH. In conclusion, the high-level secretion of F. solani cutinase in P. pastoris was demonstrated for the first time and would be a promising alternative to many expression systems previously used for the large-scale production of F. solani cutinase in Saccharomyces cerevisiae as well as Escherichia coli.  相似文献   

18.
Hartung C  Lugo MR 《Mycopathologia》1996,135(3):183-185
A strain ofFusarium solani sensu Snyder & Hansen invaded the eggs of the insectPanstrongylus geniculatus in a vivarium. None of the invaded eggs hatched. To establish experimentally the pathogenicity of thisFusarium species against the eggs ofP. geniculatus, the fungus and the eggs were incubated together under different relative humidities and temperatures. At 64% relative humidity and 26 °C, the fungus grew well colonizing and penetrating all of the chorions.Three embryos died and were also colonized byF. solani. Only 4 nymphs hatched and survived to day 20. It is concluded that the isolate ofF. solani was capable of colonizing and invading the chorion of the eggs under certain humidity and temperature conditions and cause the death of the embryos.  相似文献   

19.
内切-β-1,4-葡聚糖酶(endo-1,4-β-glucanases,EGases)广泛参与植物细胞壁的编辑,在组织伸长、果实成熟和脱落等过程中起重要作用。该研究采用RT-PCR方法,从三七(Panax notoginseng)中克隆了1个EGases基因(PnCel1),并对其进行表达和功能分析。结果显示:(1)外源茉莉酸甲酯、水杨酸、赤霉素、脱落酸和乙烯利处理显著诱导PnCel1的表达,而根腐病菌茄腐镰刀菌(Fusarium solani)、尖孢镰刀菌(Fusarium oxysporum)以及交链格孢(Alternaria alternata)、木贼镰刀菌(Fusarium equiseti)侵染三七后显著抑制PnCel1的表达。(2)亚细胞定位分析表明,PnCel1-GFP融合蛋白定位于洋葱表皮细胞的细胞壁中。(3)采用染色体步移技术克隆出PnCel1的启动子序列[(-1)~(-828)bp],进而成功构建PnCel1启动子驱动的β-葡萄糖醛酸酶植物表达载体(pBI121-PPnCel1-GUS)并转入烟草(Nicotiana tabacum L.),经PCR筛选鉴定获得阳性转基因烟草7株。(4)GUS活性检测结果表明,5种植物激素能诱导PnCel1的启动子活性,但茄腐镰刀菌等4种病原菌侵染明显降低了PnCel1启动子的转录活性,且PnCel1启动子受三七WRKY转录因子PnWRKY5/9/12/15/27的负调控。(5)PnCel1过表达转基因烟草与野生型烟草相比,对茄腐镰刀菌的易感性增加,木质素含量降低,表明PnCel1可能参与改变细胞壁的结构。研究表明,植物激素能上调三七根中PnCel1的表达,而病原菌侵染降低了PnCel1的表达水平,并抑制PPnCel1的活性,推测三七PnCel1可能通过改变细胞壁结构而增加三七对根腐病菌的易感性。  相似文献   

20.
Computerized equipment to control soil temperature and soil water matric potential, at high soil hydric conditions was developed and evaluated. A series of experiments demonstrated the accuracy and reproducibility of the equipment's performance and its adequacy for the assessment of the inoculum potential of soil-borne pathogens in soils with different characteristics. Control of soil water potential is achieved by variation in the height of the water table in a medium with high water conductivity supporting the soil. The equipment consists of double-walled tanks, permitting the adjustment of soil temperature. It is provided with sensors, control software and valves for automatic operation. in a growth chamber at 24°C. with RH 70% and irradiation of 90 W.m–2, was maintained in dynamic equilibrium for pF values ranging from 1 (–1 kPa) to 2 (–10 kPa) in various arable soil samples during the four to five weeks period of the bioassays. During the fourth week of pea or iris growth at pF=2, the system controlled within an amplitude of 0.4 pF-units. Between replications, variance was approximately 0.1 at F=2, decreasing with increasing. Soil temperature in the system could be maintained at a constant level with a variance below 0.1, within an amplitude of 0.3°C.  相似文献   

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