申嗪霉素高产技术研究

本论文研究了在培养基中添加不同种类以及不同量的植物油对申嗪霉素产量的影响,结果表明大豆油最有利于提高申嗪霉素的产量。通过将大豆油与培养基其他几个主要成份的正交实验,确定了培养基的最佳成份,摇瓶申嗪霉素产量达到1 700mg/L。通过在发酵罐中进行扩大实验,申嗪霉素产量达到1 587mg/L,达到工厂化生产要求。     

新型微生物源农药申嗪霉素对水稻纹枯病菌的毒力测定及田间药效

在室内离体条件下,申嗪霉素对水稻纹枯病菌的毒力回归方程为y=0.200 9x-2.090 4(r2=0.968 1),其EC50为0.08μg/mL。田间试验结果表明,在水稻植株喷施申嗪霉素有效成分剂量6.45μg/mL后第1、8、15天接种水稻纹枯病菌,其对水稻纹枯病的防治效果分别为100.00%、99.37%和81.23%;喷施申嗪霉素3.23μg/mL后第1、8、15天接种水稻纹枯病菌,其对水稻纹枯病的防治效果分别为100.00%,60.54%和64.58%,且与喷施井冈霉素有效成分剂量50μg/mL的防治效果94.43%、64.17%和30.49%差异均不显著。     

申嗪霉素对广东水稻稻瘟病的防治效果

2012和2013两年田间药效试验结果表明,申嗪霉素1%悬浮剂对水稻稻瘟病(叶瘟和穗颈瘟)均有较好的防治效果,施用13.5g(a.i.)/hm2处理对叶瘟的防效分别为88.92%和86.83%,对穗颈瘟的防效分别为90.72%和88.58%,保产率均在17%以上。该药剂是防治水稻稻瘟病较为理想的药剂,有较好的保产效果,在推荐用量下使用对水稻安全。     

新农药介绍

中文通用名称:申嗪霉素英文通用名称:Phenazine-1-Carboxylic农药登记名称和商品名:1%申嗪霉素悬浮剂(绿群)理化性质:申嗪霉素为我国自主开发的抗生素类杀菌剂,是由荧光假单胞菌菌株M18经发酵、提取而成的。申嗪霉素原药的质量分数≥95.0%;外观为黄绿色或金黄色针状结晶;熔点:241～242℃;溶解性:溶于醇、醚、氯仿、苯,微溶于水;稳定性:在偏酸性及中性条件下稳定。     

新型微生物源农药中嗪霉素对水稻纹枯病菌的毒力测定及田间药效

在室内离体条件下,申嗪霉素对水稻纹枯病菌的毒力回归方程为y=0.200 9χ-2.0904(r2=0.9681),其EC50为0.08μg/mL.田间试验结果表明,在水稻植株喷施申嗪霉素有效成分剂量6.45μg/mL后第1、8、15天接种水稻纹枯病菌,其对水稻纹枯病的防治效果分别为100.00%、99.37%和81.23%;喷施申嗪霉素3.23μg/mL后第1、8、15天接种水稻纹枯病菌,其对水稻纹枯病的防治效果分别为100.00%,60.54%和64.58%,且与喷施井冈霉素有效成分剂量50μg/mL的防治效果94.43%、64.17%和30.49%差异均不显著.     

申嗪霉素在水稻中的高效液相色谱残留分析方法

建立了一种用高效液相色谱仪分析稻苗、稻壳及糙米中的申嗪霉素残留量方法。样品以丙酮和乙酸混合溶液提取,稻苗样品过SPE小柱净化,稻壳和糙米直接浓缩、定容,最后用液相色谱DAD检测器测定。该方法条件下,申嗪霉素的最小检出量为0.2ng,在稻苗、稻壳、糙米中最低检测浓度分别为0.01、0.01、0.004mg/kg,申嗪霉素在样品中的回收率为76.6%～106.5%,变异系数为2.6%～11.5%,满足农药残留分析要求。     

申嗪霉素在辣椒和土壤中的液相色谱残留分析方法

建立了一种用高效液相色谱仪分析辣椒和土壤中的申嗪霉素残留量方法。样品以丙酮和冰醋酸混合溶液提取,辣椒样品过SPE小柱净化,土壤直接浓缩、定容,最后用液相色谱DAD检测器测定。该方法条件下,申嗪霉素的最小检出量为:0.2ng,在辣椒和土壤中最低检测浓度分别为0.001mg/kg,申嗪霉素在样品中的回收率为78.9～101.8%,变异系数为0.99～8.29%,满足农药残留分析要求。     

高效液相色谱法快速测定黄瓜和土壤中申嗪霉素的残留

建立了一种用高效液相色谱法分析黄瓜和土壤中申嗪霉素残留量的方法。样品用乙腈(1%乙酸)混合溶剂提取,吸取上清液,过有机滤膜,最后选择DAD检测器用250nm波长测定。该方法条件下,申嗪霉素的最小检出量为:0.2ng,在黄瓜和土壤中最低检测浓度为0.01mg/kg,申嗪霉素在黄瓜和土壤样品中的添加回收率为79.8%~100.5%,变异系数为1.5%~4.8%,满足农药残留分析要求。     

枯草芽胞杆菌YB-05与申嗪霉素复配防治小麦茎基腐病

由假禾谷镰刀菌Fusarium pseudograminearum引起的小麦茎基腐病是一种重要的土传病害,生产上亟需一种安全有效的防控方法。本研究利用枯草芽胞杆菌YB-05与申嗪霉素复配,并评价该复配制剂对小麦茎基腐病的防治效果。结果显示当申嗪霉素浓度低于500 μg/mL时,对YB-05的菌落生长无显著影响;室内盆栽及田间试验表明枯草芽胞杆菌YB-05菌液和1%申嗪霉素悬浮剂按照19：1（vt/vt）复配生物制剂20 mL/kg拌种处理发病最轻,均超过了高剂量的单剂处理（枯草芽胞杆菌YB-05 30 mL/kg拌种处理和1%申嗪霉素2 mL/kg拌种处理）和4.8%适麦丹水悬浮剂2 mL/kg拌种处理,并且具有一定的促生和增产作用。其中,复配制剂室内防治效果为69.8%,在两次的田间调查中防治效果分别为57.8%和45.7%。由此可见,枯草芽胞杆菌与申嗪霉素复配具有协同增效的作用,可以减少化学农药的使用,具有较好的开发利用潜力。     

申嗪霉素1％悬浮剂在水稻及稻田环境中的残留动态研究

采用高效液相色谱法测定了申嗪霉素在水稻及稻田环境中的残留动态。结果表明中嗪霉素l％悬浮剂在水稻田土壤、田水、稻杆、糙米和稻壳中的添加回收率为77．5％～108％。申嗪霉素在3地的稻秆和田水中的消解半衰期分别为1．96～2．53d、2．06～3．61d。14d稻杆、稻壳最终残留量〈0．01mg／kg（最低检出浓度）,稻米、土壤最终残留量〈0．004mg／kg（最低检出浓度）。     

Phenazines are Involved in Biocontrol of Pythium myriotylum on Cocoyam by Pseudomonas aeruginosa PNA1

Root rot of cocoyam (Xanthosoma sagittifolium) caused by Pythium myriotylum is the most devastating disease of this important tropical tuber crop with yield reductions of up to 90%. Bioassays were conducted in vitro and in sterile volcanic soil artificially infested with Pythium myriotylum, isolate CRPm, to test whether Pseudomonas aeruginosa PNA1 can control the cocoyam root rot disease. P. aeruginosa PNA1 (wild type) produces phenazine-1-carboxylic acid and phenazine-1-carboxamide (oxychlororaphin), while its tryptophan auxotrophic mutant FM13 is phenazine negative and secretes anthranilate in vitro. PNA1 and FM13 have previously been shown to control Pythium debaryanum and Pythium splendens on lettuce and bean. PNA1 and FM13 significantly inhibited growth of P. myriotylum in dual cultures, while their supernatants highly reduced mycelial dry weight in potato dextrose broth. However, in the presence of tissue culture derived cocoyam plantlets, only strain PNA1 strongly reduced root rot disease severity. Soil experiments involving strain PNA1 in comparison to phenazine-deficient mutants suggested that the biocontrol activity of PNA1 against P. myriotylum may involve phenazines. Phenazine involvement was further strengthened by the fact that FM13 fed with exogenous tryptophan (so that phenazine production is restored) significantly reduced disease severity on cocoyam. The efficiency of PNA1 to control P. myriotylum on cocoyam was significantly improved when the strain and the pathogen were allowed to interact for 24 h prior to transplanting cocoyam plantlets, while doubling the inoculum density of the pathogen negatively affected its efficiency.     

吩嗪-1-羧酸及其类似物的合成研究进展

天然产物吩嗪-1-羧酸（PCA）作为重要的微生物代谢产物,在假单胞菌属（Pseudomonads）和链霉菌属（Streptomycetes）等微生物分泌物中广泛存在,具有医用抗肺癌活性及抗水稻纹枯病、西瓜枯萎病、辣椒疫病、小麦全蚀病、西瓜炭疽病和油菜菌核病等病原菌的广谱性农用抗菌活性,对人畜和环境无害,并具有独特的化学结构,是研发绿色农药的理想化合物。文章综述了吩嗪-1-羧酸及其类似物在微生物发酵及化学合成方面的研究进展,分析了各合成路线中的关键反应,讨论了各合成方法的优缺点。     

嗜线虫致病杆菌SN313的次生代谢产物及其抑制植物病原真菌活性研究

通过化学与生物活性筛选从土壤中分离得到一株菌株,利用16S rDNA方法将其鉴定为嗜线虫致病杆菌Xenorhabdus nematophila并命名为SN313。采用微生物发酵、液相萃取、柱层析及半制备高效液相色谱等技术,对SN313的发酵液进行分离纯化,得到3个化合物。利用质谱和核磁共振等波谱技术并依据文献数据确定了这3个化合物分别是N-(2-羟基苯基乙酰) 色胺 ( 1 )、吩嗪-1-羧酸 ( 2 ) 和环(脯氨酸-色氨酸) ( 3 ),其中化合物 1 是一个新的β-吲哚基乙胺类衍生物。通过微量稀释法测定了3个化合物对4种植物病原真菌的抑制活性。结果表明:化合物 1 对辣椒疫霉Phytophthoa capsici和番茄灰霉病菌Botrytis cinerea具有明显的抑制作用 (IC₅₀值分别为11.20 μg/mL和28.94 μg/mL);化合物 2 对番茄灰霉病菌、辣椒疫霉、水稻纹枯病菌Thanatephorus cucumeris和小麦根腐病菌Bipolaris sorokiniana有明显的抑制作用 (IC₅₀ < 40 μg/mL);化合物 3 对番茄灰霉病菌具有较好的抑制效果 (IC₅₀ = 41.58 μg/mL)。从土壤微生物中获取化合物 2 ,为生物农药申嗪霉素有效成分的天然获取提供了一种新的途径。     

利用新鲜蓝藻藻泥发酵制备苏云金芽胞杆菌杀虫剂

富营养化湖泊中蓝藻爆发已成突出的环境问题。以云南昆明滇池蓝藻新鲜藻泥为培养基,通过接种Bt菌种,测定发酵过程中Bt的活菌数及芽胞数,观察其生长过程中形态变化,同时测定杀虫晶体蛋白产量及杀虫毒力效价,并与常规LB培养基比较,探究了利用蓝藻藻泥培养基接种苏云金芽胞杆菌(Bt GIM1.32)制备生物源杀虫剂的可行性。结果表明:Bt GIM1.32菌种在新鲜蓝藻培养基中能正常生长并完成芽胞和伴胞晶体的合成;当蓝藻含固率为3%、pH 7、接种活化10 h处于对数生长期的菌种、接种量为2%、培养温度为30℃时,灭菌后直接接种Bt GIM1.32菌株,发酵48 h左右其活菌数可达到相对稳定值(6.79×108CFU/mL),54 h左右芽胞数达到相对稳定值(7.01×108CFU/mL);其130 ku晶体蛋白的质量浓度为4.81 mg/mL,对3龄小菜蛾的致死中浓度(LC50)为0.180μL/mL,毒力效价为1 600.00 IU/μL;发酵性能较常规LB培养基优良。研究表明,蓝藻作为发酵生产生物源农药Bt的培养基具有很好的开发潜力和应用前景。     

Efficacy of bacterial antagonists and different commercial products against Fusarium wilt on rocket

Seven experimental trials were carried out to evaluate the efficacy of the bacterial strains Achromobacter xylosoxydans AM1 and Serratia sp. DM1 obtained from suppressive soils and from soilless used rockwool substrates (Pseudomonas putida FC6B, Pseudomonas sp. FC7B, Pseudomonas putida FC8B, Pseudomonas sp. FC9B and Pseudomonas sp. FC24B) against Fusarium wilt on rocket caused by Fusarium oxysporum ff. spp. raphani and conglutinans. Along with these strains, two commercial bioproducts (RootShield—Trichoderma harzianum T22; Cedomon—Pseudomonas chlororaphis MA342) were also tested. Different application strategies such as soil treatment (trials I to VI; 10⁷ and 10⁸ CFU ml⁻¹) and root dipping (trial VII; 10⁸ and 10⁹ CFU ml⁻¹) were performed in a glasshouse in order to test the efficacy of the bacterial strains against Fusarium oxysporum ff. spp. raphani and conglutinans. The lowest disease incidence (16.7%) was observed with the application of Achromobacter sp. AM1, Serratia sp. DM1 at 10⁸ CFU ml⁻¹ and Pseudomonas sp. FC9B at 10⁷ CFU ml⁻¹ against F. oxysporum f. sp. conglutinans (experiment I). Maximum plant biomass (5.0 g/plant) was registered in Serratia sp. DM1 at 10⁸ CFU ml⁻¹ treated plants in trial IV. The trials against F. oxysporum f. sp. raphani (experiment II) showed that the application of Pseudomonas sp. FC7B, P. putida FC8B at 10⁸ CFU ml⁻¹ and P. chlororaphis MA342 at 7.5 × 10⁶ CFU ml⁻¹ significantly reduced disease incidence to values ranging between 87% and 92%. The highest plant biomass was recorded with the application of Achromobacter sp. AM1 and P. putida FC6B at 10⁷ CFU ml⁻¹ (3.9 to 4.2 g) carried out 7 days before the artificial inoculation of the pathogens (trial IV). The present study showed the potential biocontrol activity of the bacterial strains Achromobacter sp. AM1, Serratia sp. DM1 and Pseudomonas sp. FC9B against F. oxysporum f. sp. conglutinans and of Pseudomonas sp. FC7B, P. putida FC8B, P. chlororaphis MA342, Achromobacter sp. AM1 and P. putida FC6B against F. oxysporum f. sp. raphani. Growth-promoting activity of biocontrol bacteria used during the trials was observed.     

模拟酸雨对小麦条锈病流行学组分的影响

为明确酸雨对小麦条锈病菌孢子萌发及病害流行学组分的影响,采用不同酸度的模拟酸雨(p H值分别为2.5、3.5、4.5和5.6)处理小麦条锈病菌Puccinia striiformis f.sp.tritici生理小种CYR32和CYR33的夏孢子,并将不同酸度模拟酸雨处理后的夏孢子接种小麦感病品种铭贤169,采用组分分析法研究酸雨对病害发展过程的影响。结果表明,随着模拟酸雨酸度的增强,生理小种CYR32和CYR33的夏孢子萌发率均显著降低,萌发24 h后对照组生理小种CYR32和CYR33的夏孢子萌发率分别为93.7%和79.8%,在p H 3.5模拟酸雨处理后萌发率仅为13.1%和8.6%。温室内接种试验结果显示,不同酸度模拟酸雨对生理小种CYR32和CYR33的侵染概率、潜育期和病情指数均有显著影响,对生理小种CYR32和CYR33病斑扩展率影响不显著;与对照相比,p H 3.5模拟酸雨处理后生理小种CYR32和CYR33的侵染概率分别下降79.9%和79.8%。结果表明强酸雨(p H 3.5)显著延长小麦条锈菌潜育期,减少病斑产孢量,降低病情指数和病害进展曲线下面积。     

Relationship between the incidence of latent infections caused by <Emphasis Type="Italic">Monilinia</Emphasis> spp<Emphasis Type="Italic">.</Emphasis> and the incidence of brown rot of peach fruit: factors affecting latent infection

Five field experiments were performed in commercial orchards located in Lleida (Spain) over three growing seasons, 2000–2002, in order to estimate the relationship between the incidence of latent infection caused by Monilinia spp. in peaches and the incidence of post-harvest brown rot. No latent infection was recorded at popcorn and the maximum incidence occurred pre-harvest; in some orchards a second peak was detected during the pit hardening period. Monilinia laxa is the most prevalent species isolated from peaches with brown rot. There was a positive correlation between the incidence of latent infection and that of post-harvest brown rot. The average incidence of latent infection during the crop season explained 55% of the total variation in the incidence of post-harvest brown rot. The effect of temperature (T) and duration of wetness (W) on the incidence of latent infection in peach and nectarine orchards was analysed using multiple regression. The regression analysis indicated that T and W jointly explained 83% of the total variation in the incidence of latent infection. The model predicts no latent infections when T < 8°C, and >22 h wetness are required when T = 8°C but only 5 h at 25°C are necessary for latent infection to occur. The incidence of brown rot and latent infection of peaches caused by M. laxa under controlled experimental conditions were also affected by T and W, as well as by fruit maturity and inoculum concentration. Latent infections were produced in fruit when T was not suitable for the development of brown rot symptoms. In these experiments more than 4–5 h of daily wetness were required after embryo growth in fruit sprayed to run-off with an inoculum concentration higher than 10⁴ conidia ml⁻¹ of M. laxa for brown rot and latent infections to develop. The fitted model obtained from the field data was able to predict the observed data obtained under controlled environmental conditions.     

Germination and appressorium formation of wheat leaf rust on susceptible,partially resistant and resistant wheat seedlings and on seedlings of other Gramineae

Germination and appressorium formation of wheat leaf rust urediospores were studied in two experiments. No consistent differences could be detected between 11 genotypes of wheat (Triticum aestivum), two of barley, one ofTriticum dicoccum, one ofT. dicoccoides, one ofT. boeoticum and one ofAegilops squarrosa. Host genotypes and non-hosts gave similar results, suggesting that the phases before penetration of the host leaf are not affected by the resistance mechanisms operating in hosts and non-host genotypes.Samenvatting Kieming en de vorming van appressoria is bestudeerd in twee experimenten. Er zijn geen systematische verschillen waargenomen tussen 11 genotypen van tarwe, twee van gerst, een vanTriticum dicoccum, een vanT. dicoccoides, een vanT. boeoticum en een vanAegilops squarrosa. Waard en niet-waard genotypen gaven overeenkomstige resultaten, dit suggereert dat de fasen voor penetratie van het gastheerblad niet beïnvloed worden door de resistentiemechanismen werkzaam in waard en niet-waard.