木霉复合菌剂与拮抗性腐熟堆肥对水稻生长及稻瘟病的防控研究

稻瘟病是水稻三大病害之一,在水稻整个生长发育期间都有发生。研究了2种木霉复合菌剂(堆肥腐熟菌剂和水稻拮抗菌剂)与2种有机肥(普通有机肥和拮抗有机肥)对稻瘟病原茵的抑制作用、水稻生长的影响以及对叶瘟病和穗瘟病的防治作用。结果表明,添加木霉腐熟复合菌剂堆肥制备而成的有机肥即拮抗有机肥,拮抗堆肥60 d的0.2%无菌浸提液对稻瘟病病原菌ACCC 37361具有显著抑制作用,平板抑菌率为9.92%,显著高于普通有机肥(1.19%,P<0.05)。水稻品种“沪早香软1号”移栽第2、4、6周时,拮抗有机肥和拮抗生物有机肥处理的水稻株高显著高于普通有机肥处理,拮抗生物有机肥比普通有机肥处理的株高平均分别高出6.99、5.62和6.80 cm。上海崇明岛北湖农场的田间试验结果表明,与普通有机肥相比,拮抗有机肥和水稻拮抗菌剂共同施用显著增加成熟期间的水稻株数,并提高水稻产量(P<0.05);拮抗有机肥加施2次水稻拮抗菌剂处理对稻瘟病防治效果最好,扬花授粉期的叶瘟病相对防效为42.09%,灌浆期和成熟期的穗瘟病相对防效分别为65.76%和61.80%。研究表明拮抗有机肥与水稻拮抗菌剂同时施用,具有较好的促生抑菌效果,可广泛应用于绿色水稻的生产。     

拮抗菌防治西瓜枯萎病的试验

从西瓜、黄瓜、柑桔等作物根际中分离到的B11等芽孢杆菌的哈茨木霉T31、粘帚霉G1等拮抗真菌，对防治西瓜枯萎病菌有一定抑制作用。经田间防治试验，T31、G1和B11单个菌的田间防效在35％－50％左右；施用T31、G1、B11的混合拮抗菌和采取的一些措施（有机肥作基肥，拮抗菌与杀菌霸混施）对西瓜枯萎病有更好的防治作用，防效达60％－85％。     

复合生物制剂防治西瓜连作病害的研究

T42木霉与枯草芽孢杆菌Bs-6对尖孢镰刀菌FUS3有明显的拮抗作用。田间试验表明，由T42木霉与枯草芽孢杆菌Bs-6组成的复合生物制剂能显著增加连作西瓜的茎长和茎粗，经济产量、生物产量分别增加50．9％、66．7％，对连作病害的防效可达75％。     

木霉Fg菌株防治韭菜白绢病研究

通过筛选生防菌和高效杀菌剂来防治韭菜白绢病,并通过盆栽试验、小区试验、田间试验,测得木霉(T richod erm a sp.)Fg菌株与丙环唑对韭菜白绢病的防治效果分别为6 6.7 5%~9 6.5 4%与77.29%~89.65%。     

利用玉米秸秆制备生物杀菌剂的研究

通过拮抗性筛选分离出对蔬菜枯萎病原菌具有很强抑制作用的木霉菌株“T41”,并利用玉米秸秆粉为原料,采用液固两相发酵工艺制成木霉菌剂。该菌剂对多种蔬菜枯萎病防治效果显著且稳定,并高于化学杀菌剂防治效果     

9种杀菌剂对番茄灰霉病的田间防治效果

通过田间药效试验的方法,评价9种常用杀菌剂对番茄灰霉病的防治效果,根据结果筛选出防治番茄灰霉病的有效药剂。结果表明,按推荐剂量施药3次,400 g/L嘧霉胺悬浮剂,250g/L嘧菌酯悬浮剂和50%腐霉利可湿性粉剂的防效分别为86.4%、82.3%和80.5%,高于其他供试药剂的防治效果。因此在生产上推荐轮换使用上述3种防效较高的杀菌剂,在灰霉病发生前期或初期开始喷药,每7～14 d施药1次。     

三种虫生菌协同毒性及与有机肥耦合效应研究

为揭示白僵菌、绿僵菌、淡紫紫孢菌对蚜虫、地蚕、根结线虫的毒性大小,并探索生防菌与有机肥耦合效应,采用PDA对白僵菌、绿僵菌、淡紫紫孢菌对其进行活化培养,并稀释不同浓度菌剂作用于蚜虫,观察单独菌种下毒性大小,然后利用对峙法研究菌种间拮抗性,寻求最优协同杀虫组合,最后利用响应面法设计不同有机肥与菌剂配比,分析混合配方对防效...     

复合菌剂防控香蕉枯萎病的效果及其微生物学机制

由病原菌尖孢镰刀菌古巴专化型(Fusarium oxysporum f. sp. cubense(Foc))侵染引起的香蕉枯萎病对全世界香蕉产业带来了毁灭性的影响,且目前尚无广泛采用的防治方法。研究复合生防真菌制剂对香蕉枯萎病的防治效果,以期为大田香蕉枯萎病的防治提供依据。设置3组不同的菌剂处理,分别为对照组CK、复合菌剂NFP、复合菌剂NFPT,通过两季的盆栽试验,研究复合菌剂对香蕉枯萎病的防治效果及其对土壤微生物多样性的影响;利用IlluminaMiseq高通量测序平台对细菌16S rRNA基因和真菌ITS区域进行测序分析,采用实时荧光qPCR定量分析各处理病原菌的数量。结果表明:(1)复合菌剂处理(NFP和NFPT)对香蕉枯萎病有较好的防治效果,其防效分别为43%和48%。(2)施用复合菌剂增加了细菌和真菌群落丰富度和多样性。基于Bray-curtis距离矩阵的主坐标分析(PCoA)结果表明NFP和NFPT改变了细菌和真菌群落结构。NFP和NFPT处理增加了潜在有益微生物中与香蕉枯萎病病情指数呈显著负相关的大理石雕菌属Marmoricola、类诺卡氏菌属Nocardioides、野野村式菌属Nonomuraea、norank_c__Acidobacteria、DS-100属和norank_f__Blastocatellaceae__Subgroup_4属的相对丰度,显著减少了病原菌尖孢镰刀菌的数量,重塑了土壤微生物结构和功能,增强其抗病性。     

绿色木霉复配有机肥的筛选及对黄瓜枯萎病的防治作用

单独应用绿色木霉控制蔬菜土传病害效果不理想,探索筛选适合的有机肥与绿色木霉复配,达到提高生防菌防病作用的目的。从供试的7种有机肥中,筛选出菇渣和牛粪适合与木霉菌复配。菇渣和牛粪的提取液对黄瓜枯萎病菌菌丝生长的抑制率分别为74.91%和69.38%,对绿色木霉菌丝生长抑制率分别为-7.90%(促进作用)和4.94%,使黄瓜种子胚根伸长增长率分别为36.75%和28.21%。菇渣、牛粪分别与绿色木霉混合后对黄瓜枯萎病的防效分别为64.98%和63.32%。菇渣和牛粪按1:1混合后再与绿色木霉复配制成菇渣、牛粪木霉有机肥的防效高达85.31%,黄瓜地上部分鲜重和地下部分鲜重分别增加了71.50%和79.55%。菇渣和牛粪与绿色木霉复配可显著提高木霉对黄瓜枯萎病的防病效果和对黄瓜生长的促进作用。     

2种生防菌对苹果树3种病害病原菌拮抗作用比较

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。     

牛场沼液对几种蔬菜病原菌抑制作用的研究

实验室条件下,用不同贮存时间的牛场沼液原液和滤液对7种蔬菜病原菌（番茄叶霉病菌、番茄灰霉病菌、番茄早疫病菌、辣椒疫病病菌、辣椒绵腐病菌、黄瓜炭疽病菌、茄子灰霉病菌）的抑菌效果进行试验研究,为沼液应用于蔬菜病害农田防治的进一步研究提供基础数据。结果表明,新鲜沼液原液对番茄灰霉病菌、番茄早疫病菌、辣椒疫病病菌、辣椒绵腐病菌、黄瓜炭疽病菌和茄子灰霉病菌均有较强的抑制作用;随着贮存时间的增加,沼液原液对番茄灰霉病菌、番茄早疫病菌、黄瓜炭疽病菌和茄子灰霉病菌的抑菌率没有显著变化,对辣椒疫病病菌和辣椒绵腐病菌的抑菌率显著下降;新鲜沼液滤液对番茄灰霉病菌和茄子灰霉病菌有较好的抑制作用,但对其他5种病原菌的抑制效果不明显;随着贮存时间的增加,滤液对番茄灰霉病菌的抑菌率没有显著变化,对茄子灰霉病的抑制率表现出先显著下降而后升高的趋势。     

尿素配施木霉菌剂提高甜瓜产量、品质及土壤微生物功能多样性

【目的】明确尿素配施木霉菌剂对甜瓜产量、品质及土壤微生物功能多样性的影响,为生产中甜瓜的减肥增效提供理论依据。【方法】试验于2017年3月至7月在沈阳农业大学园艺科研基地的日光温室内进行,栽培土壤组成为土∶草炭∶牛粪 = 4∶2∶1,供试材料为薄皮甜瓜(Cucumis melo var. makuwa Makino)‘玉美人’。通过测土配方施肥测算,供氮水平100%为尿素(普通尿素,N≥46.4%) 25.9 g/株。以此为基础,设9个氮肥处理:50%普通尿素(T1)、50%控释尿素(T2)、50%普通尿素 + 木霉菌剂(T3)、50%控释尿素 + 木霉菌剂(T4)、30%控释尿素 + 20%普通尿素 + 木霉菌剂(T5)、100%普通尿素(T6)、100%控释尿素(T7)、150%普通尿素(T8),以不施氮肥为对照(CK)。测定了不同处理对薄皮甜瓜果实产量、品质及土壤微生物功能多样性的影响。【结果】与100%施氮处理(T6、T7)相比,减量施氮50%在一定程度上提高了成熟甜瓜果实的品质,其中50%控释尿素配施木霉菌剂(T4)处理不仅降低了甜瓜果实的硝酸盐含量,提高了果实的可溶性糖、Vc含量及香气物质含量,单株产量也有所增加。此外,随着施氮量的增加,氮肥表观利用率和农学利用率均明显降低,其中T1～T4处理与100%和150%施氮处理(T6～T8)间的差异均达显著水平,且T3、T4处理的氮肥利用率均较高。在土壤微生物多样性方面,与施用普通尿素相比,控释尿素处理(T2、T4、T5、T7)的AWCD值均高于其他处理,其中以T4处理的AWCD值最高。【结论】在供试栽培土壤条件下,减少氮肥施用量50%,无论是否采用控释尿素或配施木霉菌剂,都有助于促进甜瓜植株的生长,提高成熟甜瓜果实品质,增加甜瓜的单株产量,但只有配施木霉菌剂后,其产量、品质与施氮100%处理的差异达到了显著水平。微生物多样性只在施用了理论施氮量的150%后显著下降。因此,在有机质丰富的土壤上栽培甜瓜,建议采用施氮量减少50%、配施木霉菌剂的高产优质可持续的施肥技术。     

电子束辐照对灰葡萄孢菌分生孢子萌发活性及致病力的影响

以灰葡萄孢病菌(Botrytis cinerea)分生孢子为研究对象,采用0.5、1.0、2.0和3.0kGy剂量电子 束辐照灰葡萄孢分生孢子,分别测定5℃和25℃培养条件下辐照对灰葡萄孢分生孢子的萌发活性及致病力的影响.结果表明:25℃和5℃培养条件下,2.0kGy处理分生孢子完全萌发时间比对照分别延迟了5和9d;2...     

Antifungal activity and biotransformation of diisophorone by Botrytis cinerea

Diisophorone (1) was tested against two strains of the necrotrophic plant pathogen Botrytis cinerea.Fungal sensitivity varied according to the strain. B. cinera 2100 was more sensitive than B. cinereaUCA992: its mycelial growth was significantly inhibited at concentrations of 50 ppm and above. Although diisophorone (1) showed an effective control of B. cinerea, a detoxification mechanism was present. The detoxification of racemic diisophorone (1) by B. cinerea was investigated. Incubation with two strains of B. cinerea gave one and four biotransformation products (2-5), respectively. Their structures were established as the known 8beta-hydroxydiisophorone (2), 6alpha-hydroxydiisophorone (3), 6beta-hydroxydiisophorone (4) and 8beta,14beta-dihydroxydiisophorone (5) on the basis of their spectroscopic data, including two-dimensional NMR analysis [heteronuclear multiple quantum coherence (HMQC), heteronuclear multiple bond correlation (HMBC), and nuclear Overhauser enhancement spectroscopy (NOESY)] and an X-ray crystallographic study.     

水稻白叶枯病菌蛋白质激发子HarpinXoo诱导植物的防卫反应

摘要：含质粒pHRF4的表达菌株BLHR4在培养基中经IPTG诱导，16 h Harpin表达接近最高量。用不同浓度的HarpinXoo喷雾处理烟草(Nicotiana tabacum L.)、油菜(Brassica campestris L.)和番茄(Lycopersicon esculentum Mill.)，对烟草花叶病毒(TMV)、油菜菌核病(Sclerotinia sclerotiorum (lib.) de Bary)和番茄灰霉病(Botrytis cinerea Pers.)均表现较好的诱导抗病效果。以浓度为100μg/mL的HarpinXoo处理烟草后，分别测定叶片中与防卫反应相关的一些生理指标的变化，结果表明，处理叶片中苯丙氨酸解氨酶（PAL）、过氧化物酶（PO）和多酚氧化酶（PPO）活性都有不同程度的增强。三种酶活性高峰分别在处理后24、12和3 h出现，分别比清水对照增加110.1%、211.2%和273.0%；同时，两个病程相关蛋白（pathogenesis-related protein，PR）基因PR-1b和PR-1a在HarpinXoo处理24 h后被诱导显著表达，36 h时达到最高表达水平。这表明HarpinXoo与HarpinEa一样能够诱导烟草不同抗病信号通路的启动，从而使植株获得抗病性。     

Induction of resistance to gray mold with benzothiadiazole modifies amino acid profile and increases proanthocyanidins in grape: primary versus secondary metabolism

Field treatments of grapevine (cv. Merlot) with the plant activator benzothiadiazole (BTH, 0.3 mM) induced resistance against gray mold caused by Botrytis cinerea. Both incidence and severity of the disease were reduced. The resistance was associated with an increase of total polyphenols in berry skins, in particular, the proanthocyanidin fraction, that increased up to 36%. The amino acid profile of leaves was also modified by treatments, particularly lysine, that augmented 4-fold. Other amino acids involved in resistance mechanisms to either biotic or abiotic stress increased as well. These results indicate that BTH treatments can be used to control gray mold, thereby limiting an excessive use of fungicides, and could be exploited to increase the content of micronutrients of high nutritional value, arising from both primary and secondary metabolisms.     

Preharvest L-arginine treatment induced postharvest disease resistance to Botrysis cinerea in tomato fruits

L-arginine is the precursor of nitric oxide (NO). In order to examine the influence of L-arginine on tomato fruit resistance, preharvest green mature tomato fruits (Solanum lycopersicum cv. No. 4 Zhongshu) were treated with 0.5, 1, and 5 mM L-arginine. The reduced lesion size (in diameter) on fruit caused by Botrytis cinerea, as well as activities of phenylalanine ammonia-lyase (PAL), Chitinase (CHI), β-1,3-glucanase (GLU), and polyphenoloxidase (PPO), was compared between L-arginine treated fruits and untreated fruits. We found that induced resistance increased and reached the highest level at 3-6 days after treatment. Endogenous NO concentrations were positively correlated with PAL, PPO, CHI, and GLU activities after treatment with Pearson coefficients of 0.71, 0.94, 0.97, and 0.87, respectively. These results indicate that arginine induces disease resistance via its effects on NO biosynthesis and defensive enzyme activity.     

Physicochemical properties and bioactivity of fungal chitin and chitosan

Chitinous material was extracted from mycelia of Aspergillus niger and Mucor rouxii grown in yeast peptone dextrose broth for 15 and 21 days, respectively. The extracted material was characterized for purity, degree of acetylation, and crystallinity and tested for antibacterial and eliciting properties. The maximum glucosamine level determined in the mycelium of A. niger was 11.10% dw and in the mycelium of M. rouxii was 20.13% dw. On the basis of the stepwise extraction of freeze-dried mycelia, it appeared that M. rouxii mycelia contained both chitin and chitosan, whereas A. niger contained only chitin. The yields of crude chitin from A. niger and M. rouxii were 24.01 and 13.25%, respectively, and the yield of chitosan from M. rouxii was 12.49%. Significant amounts (7.42-39.81%) of glucan were associated with chitinous compounds from both species and could not be eliminated by the extraction method used. The degrees of acetylation were determined to be 76.53 and 50.07% for chitin from A. niger and M. rouxii, respectively, and 19.5% for M. rouxii chitosan. The crystallinity of fungal chitin and chitosan was estimated to be less intense than in corresponding materials from shrimp shells. The extracted chitin and chitosan in a concentration of 0.1% reduced Salmonella Typhimurium DT104 2576 counts by 0.5-1.5 logs during a 4 day incubation in tryptic soy broth at 25 degrees C. Furthermore, all tested chitinous materials from fungal sources significantly reduced lesions caused by Botrytis cinerea and Penicillium expansum in harvested apples.     

The antifungal activity of widdrol and its biotransformation by Colletotrichum gloeosporioides (penz.) Penz. & Sacc. and Botrytis cinerea Pers.: Fr

Widdrol (1) was tested against the necrotrophic plant pathogens Botrytis cinerea and Colletotrichum gloeosporioides. While 1 was found to be inactive against C. gloeosporioides, it showed a selective and effective control of B. cinerea, significantly inhibiting the mycelial growth of the fungus at concentrations of 100 ppm and above. In addition, the biotransformation of 1 by both fungi was studied. Incubation with C. gloeosporioides and B. cinerea afforded four and one biotransformation products (2-6), respectively. Biotransformation with C. gloeosporioides was highly regioselective, yielding for the most part oxidation products at C-10: 10-oxowiddrol (2), 10beta-hydroxywiddrol (3), 10alpha-hydroxywiddrol (4), and 14alpha-hydroxywiddrol (5). The structures of all products were determined on the basis of their spectroscopic data, including coupling constants, two-dimensional NMR analysis (heteronuclear multiple quantum coherence, heteronuclear multiple bond correlation, and nuclear Overhauser enhancement spectroscopy), and nuclear Overhauser effect. The biotransformation products were then tested against B. cinerea and found to be inactive. These results shed further light on the structural modifications, which may be necessary to develop selective fungal control agents against B. cinerea.     

Isolation, antifungal activity, and structure elucidation of the glutarimide antibiotic, streptimidone, produced by Micromonospora coerulea.

The antibiotic Ao58A,which showed strong antifungal activity against some plant pathogenic fungi, was purified from the culture broth and mycelial mats of Micromonospora coerulea strain Ao58 using various chromatographic procedures. The molecular formula of the antibiotic Ao58A was deduced to be C(16)H(23)NO(4) (M + H, m/z 294.1707) by high-resolution FAB mass spectroscopy. Analyses of (1)H NMR, (13)C NMR, and 2D NMR spectral data revealed that the antibiotic Ao58A is the glutarimide antibiotic streptimidone, 4-(2-hydroxy-5, 7-dimethyl-4-oxo-6,8-nonadienyl)-2,6-piperidinedione. The antibiotic Ao58A was very effective in inhibiting growth of Phytophthora capsici,Didymella bryoniae, Magnaporthe grisea, and Botrytis cinerea in the range approximately 3-10 microg mL(-)(1) of MICs. In vivo evaluation of the antibiotic Ao58A under greenhouse condition showed strong control efficacies against the development of P. capsici, B. cinerea, and M. grisea on pepper, cucumber, and rice plants, respectively. The antibiotic Ao58A was equally as effective as metalaxyl, vinclozolin, and tricyclazole in the control of these plant diseases. However, it did not show any phytotoxicity on the plants even when treated with 500 microg mL(-)(1).