黄芪种衣剂对黄芪根腐病菌(Fusarium oxysporum)抑制作用的研究

目的：以不同浓度的黄芪种衣剂和ND牌种衣剂测定其对尖孢镰刀菌（F．oxySpomm）的抑制效果。方法：以5mL灭菌水代替种衣剂作为对照,把从菌落边缘打出的菌丝块移到带药的PSA培养基中,观察菌落直径。结果：同等有效浓度下,黄芪种衣剂的抑制率高,与ND大豆种衣剂相比有较好的抑菌效果。结论：黄芪种衣剂对黄芪根腐病菌-尖孢镰刀菌具有明显的抑制效果,抑菌效果好于ND大豆种衣剂。     

18种中草药提取物对芒果蒂腐病菌的抑制作用研究

采用Duncan测验法研究探讨了18种中草药提取液对芒果蒂腐病菌的抑制作用。结果表明：乌药提取液对芒果蒂腐病菌具有很强的抑制作用,而瓜蒌、旋覆花、五味子等五种中草药提取液对蒂腐病菌却具有促进作用;通过紫外-可见吸收光谱分析得知,乌药提取物有2个明显的最大吸收峰。     

大豆疫霉根腐病菌的rDNA ITS序列分析

采用真菌核糖体基因转录间隔区(ITS)通用引物,PCR扩增了大豆疫霉根腐病菌具有差异的17个菌株的ITSI与ITS2,经过与DL2000的标准分子量DNA进行比较,得到了大约800~1000bp左右的片段,并对PCR产物进行了序列测定。以USA为外类群利用最大简约法构建了大豆疫霉根腐病菌的系统发生树,并分析了菌株之间的遗传进化关系。结果表明:不同菌株ITS1和ITS2在碱基构成上有很大差异,17个菌株大致分为4个谱系中,且来自于同一地区的菌株大都分布在同一谱系中,显示出地理上的差异。     

贝莱斯芽孢杆菌GDND-2对烟草镰孢根腐病菌生长发育的影响

【背景】尖孢镰孢(Fusariumoxysporum)引起的烟草根腐病在世界烟区普遍发生,严重影响烟草产量和质量,化学农药无法有效防治病害,利用生防菌防治该病成为研究热点。【目的】明确贝莱斯芽孢杆菌(Bacillusvelenzensis)GDND-2对尖孢镰孢生长发育的抑制作用。【方法】制备含10%和20%GDND-2发酵滤液的PDA培养基,涂在玻片上,接种尖孢镰孢分生孢子,观察滤液对尖孢镰孢孢子萌发、菌丝生长、孢子形成和色素产生的影响,应用扫描和透射电镜观察菌体超微结构的变化。【结果】贝莱斯芽孢杆菌GDND-2发酵滤液延迟孢子萌发2h以上,造成芽管膨大畸形,促使菌丝提早分枝,抑制菌丝延伸,使菌丝产生畸形球状结构,10%和20%发酵滤液对菌丝生长抑制率达53.41%和61.58%。滤液延迟病菌产孢,显著抑制产孢量,影响孢子形态,刺激病菌产生色素。10%和20%发酵滤液延迟产孢20h和28h,对产孢量的抑制率为52.11%和78.85%,滤液促使病菌形成小型分生孢子。观察尖孢镰孢的超微结构,部分菌丝膨大、畸形,细胞壁变薄,细胞膜消失,细胞质渗出,胞内呈空腔结构;部分菌丝严重皱缩、扭曲、...     

中国烟草根黑腐病菌根串珠霉菌群及rDNA-ITS序列分析

从国内不同烟区分离获得烟草根黑腐病菌根串珠霉45个野生型和11个白色变异菌株,参照Punja & Sun的分群鉴定方法,对这56个菌株进行了分群鉴定。结果表明：中国根串珠霉分离株存在明显形态变异,共划分出6个群,其中群Ⅰ、Ⅱ、Ⅲ为野生型,群Ⅳ、Ⅴ为白色变异型;另有白色变异型HLL-1￠不同于其他各群,将其建为新群—群Ⅵ。统计分析发现,群Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ及Ⅵ的分离频率分别占42.9%、33.9%、3.6%、16.0%、1.8%和1.8%。群Ⅰ主要分布重庆和云南,群Ⅱ在5个调查省市均有分布,群Ⅲ、群Ⅴ仅分布     

金属离子对十字花科黑腐病菌不同致病系统表达的影响

十字花科黑腐病菌(Xanthomonas campestris pv.campestris,Xcc)是引起十字花科植物黑腐病的病原细菌,其通过不同的分泌系统将效应物蛋白或细胞毒素转运进真核宿主细胞产生病害,对农业生产造成巨大的经济损失。本研究利用GUS融合报告系统,考察了常见的几种不同金属离子(Zn2+,Mn2+,Ca2+,Fe2+)对Xcc 8004不同致病系统装置基因表达的影响。结果表明,在1滋mol/L~1 mmol/L浓度区间,Zn2+和Mn2+同时对Xcc域型分泌系统(T2SS)和芋型分泌系统(T3SS)有抑制作用;在大于0.01 mmol/L浓度时,Ca2+特异性抑制T3SS;Fe2+同时抑制T2SS、T3SS和郁型分泌系统(T4SS)装置基因的表达。金属离子Zn2+、Mn2+、Ca2+和Fe2+在不同浓度下对Xcc的致病系统均有抑制作用。本研究为分析病原细菌不同分泌系统响应金属离子的模式奠定了基础。     

广东花生主要品种对黑腐病菌的抗性水平鉴定

【背景】寄生帚梗柱孢霉是花生黑腐病的病原菌,被我国列为重要的进境植物检疫性有害生物。该病菌2009年已入侵我国广东,造成花生植株基部腐烂而死亡,严重威胁花生生产安全。筛选与种植抗病品种是防控该病害的重要措施。【方法】收集广东推广种植的15个主要花生品种,通过人工接种方法,鉴定这些品种对花生黑腐病菌的抗性水平。【结果】15个供试花生品种中,湛红2号、湛油62等2个品种表现为抗病;湛油75、湛油82、粤油390、粤油410、仲恺花44、仲恺花99、汕油诱1号等7个品种表现为中抗;花育33号、汕油523、汕油辐1号、粤油18、湛油53等5个品种表现为感病;仲恺花332表现为高感。【结论与意义】目前广东生产上推广种植的花生品种多数对黑腐病菌表现为抗病或中抗水平,部分品种表现为感病或高感。该结果可为我省花生品种的推广与布局提供依据。     

腐婢根化学成分研究

从马鞭草科腐婢属植物腐婢(Premna microphylla)的根中分离鉴定出7种化合物,其结构分别为β-谷甾醇(1).棕榈酸(2).反式-对甲氧基肉桂酸(3).丁香色酮(4).木犀草素(5).木犀草素-7-O-β-D-葡萄糖甙(6)和胡麻素(7).这些成分均为首次从腐婢根中分离得到.     

三七重茬根际土壤中化感物质的测定及其对三七根腐菌的生长作用

目的 研究三七重茬根际土壤中化感物质的组成及其对病原微生物的化感作用。方法 采用乙酸乙酯提取三七重茬根际土壤中的化感物质,利用GC-MS对萃取液进行物质组成分析,并研究检测中出现的特征性化感物质对羟基苯甲酸、邻苯二甲酸二异丁酯对三七根腐病菌（F.oxysporum Schlecht）的化感作用。结果 采集的三七重茬土壤中共检测到29种化感物质,其中2种特征性化感物质对羟基苯甲酸、邻苯二甲酸二异丁酯对三七病原菌的生长呈现低促高抑的现象,且当对羟基苯甲酸浓度为5 mmol/L时能促进病菌小孢子的产生。结论 检测到的化感物质组分为苯甲酸及其衍生物类、酚类、酯类、饱和烃类等物质。这些物质在低浓度时有利于病原菌的生长,对病害的发生具有促进作用。     

桃褐腐病菌拮抗性内生细菌的筛选及其抑病效果

【目的】从桃树根部组织中分离、筛选能够防治桃褐腐病害的内生拮抗菌,从细胞学水平探究其抑制机理。【方法】采用平板对峙法筛选对桃褐腐病菌(Monilinia fructicola)有拮抗作用的内生细菌,调查内生拮抗细菌的形态学及生理生化特性,通过细菌16S r RNA基因测序分析菌株的系统发育学特征。通过果实离体实验检测内生拮抗细菌的抗菌效果,使用电子显微镜观察受抑制的桃褐腐病菌菌丝、孢子的形态以及细胞内部结构变化。【结果】从桃树根部组织中复筛分离得到的3株内生细菌对桃褐腐病防效高且稳定,经鉴定3株细菌均为枯草芽孢杆菌。果实离体试验表明,3株内生细菌均明显抑制了桃褐腐病的发生。通过电子显微镜观察,发现受拮抗细菌抑制的桃褐腐病菌的菌丝明显变细、杂乱且打结缠绕成团,孢子普遍干瘪变大、破裂,细胞质外渗。【结论】从桃树根部组织中得到的3株内生细菌对桃褐腐病菌的生长均有明显的抑制作用,可以作为防治桃褐腐病的新选择,具有进一步研究价值。     

阿勒泰黄芪提取物对蛋白酪氨酸磷酸酯酶1B的抑制作用

本文探讨了阿勒泰黄芪不同提取物对蛋白酪氨酸磷酸酯酶1B(PTP1B)的抑制作用.采用分光光度法测定了提取物中的黄酮和皂苷含量;通过体外酶促动力学方法检测了不同提取物对PTP1B的影响,并确定了抑制类型;并采用氧化酶法检测了阿勒泰黄芪提取物对细胞利用葡萄糖能力的作用.结果表明,阿勒泰黄芪8种提取物(E1 ～8)中黄酮含量分别为5.09、10.46、3.58、3.23、53.91、21.77、5.76和7.49 mg/mL,其中E1、E2、E6、E7、E8皂苷含量分别为16.53、27.45、21.90、10.21和8.96 mg/mL;各提取物对PTP1B活性均表现出抑制作用,其中E1、E2、E7、E8的IC50分别为34.8、4.7、7.35和7.15 μg/mL,E1、E7和E8是竞争性抑制,E2是混合型竞争性抑制.E1、E2、E5、E7和E8较明显的提高了CHO-K1细胞对葡萄糖的利用.提示皂苷可能是阿勒泰黄芪抑制PTP1B活性的主要物质,通过PTP1B途径有效了提高细胞利用葡萄糖的能力.本研究为阿勒泰黄芪开发为防治糖尿病及改善胰岛素抵抗的药物或保健品提供实验依据.     

大蒜秸秆水浸液的抑菌作用和抑茵成分初步分析

以不同浓度的大蒜秸秆水浸液对西瓜枯萎病菌、黄瓜枯萎病菌、辣椒疫霉病菌进行抑菌圈试验,并对3种不同的大蒜秸秆萃取液中的可挥发抑菌物质进行鉴定,以了解大蒜秸秆水浸液的抑菌作用。结果表明,大蒜秸秆水浸液对3种靶标菌均有抑菌活性,其最有效的抑菌浓度为100 g.L-1,对辣椒疫霉病菌的抑菌活性最强。大蒜秸秆水浸液的石油醚、三氯甲烷和乙酸乙酯3种有机溶剂萃取相对3种靶标菌均有抑菌活性,以乙酸乙酯相抑菌活性最强,在50 g.L-1浓度下对辣椒疫霉菌抑菌率达100%,在100 g.L-1浓度下对西瓜枯萎病菌和黄瓜枯萎病菌的抑菌率也达100%;三氯甲烷相的抑菌活性稍高于石油醚相;抑菌活性基本随浓度增加而增大。经GC-MS分析,初步确定大蒜秸秆水浸液中的可挥发抑菌物质主要有2-甲氧基-1,6-己二酸甲酯、邻苯二甲酸二丁酯等有机酯类,3-甲氧基-4-羟基-苯甲酸、对羟基苯甲酸等有机酸,以及对甲氧基苯酚等酚类化合物。     

丹参根腐病生防芽孢杆菌2-1海藻菌剂的研制

以海藻渣为载体,菌种为丹参根腐病生防菌:枯草芽孢杆菌Bacillus subtilis2-1(已保存CGMCC No.9256),研制成的海藻菌剂兼具营养及生物防治功能,同时菌剂的基本指标符合农用微生物菌剂产品的技术指标。用物理方法选择菌种发酵液与海藻渣混合的最佳配比,结合农用微生物菌剂产品质量检测方法对菌剂中有效活菌数、含水量、p H值及有效期进行测定,并进行大田试验,测试生防效果。菌剂有效活菌数为2.32×109/g,含水量为6.75%,p H值为6.7,常温下有效期约为11.6个月,均符合颗粒菌剂的技术指标,大田试验中根腐病的发病率比空白对照组降低37.6%,生防效果较为显著。     

当归根腐病主要病原菌生物学特性及室内毒力测定

为探究当归(Angelica sinensis)根腐病的病原菌种类、生物学特性并筛选其防治药剂。采用常规组织法分离培养病原菌,利用柯赫氏法则验证其致病性,通过形态学和分子生物学相结合的方法进行病原菌的鉴定、生物学特性分析;采用菌丝生长速率法对病原菌进行室内毒力测定。结果表明,当归根腐病菌菌落毛毡状,在三种培养基上大型分生孢子呈镰刀形,大小为(27.5～47.5)μm×(3.8～7.5)μm(n=50),隔膜数1～5;小型分生孢子在PDA和SNA培养基上呈椭圆形、肾型、纺锤形,在CLA培养基上肾型,(3.8～15.0)μm×(2.5～7.5)μm(n=50),隔膜数0～3;厚垣孢子在CLA培养基上呈串生,PDA培养基上呈不规则形,SNA培养基上厚垣孢子着生方式多样,串生、圆形、不规则形,大小为(2.5～3.8)μm×(2.5～5.0)μm(n=50)。EF-1α、RPB1和RPB2联合序列系统发育树表明,该菌为锐顶镰刀菌(Fusarium acuminatum),其最适生长温度为25℃,最适生长初始pH为9,光照对其生长无显著影响。室内毒力测定显示10%苯醚甲环唑(WG)对其抑制效果最好...     

不同加工方法丹参花茶的质量评价

目的：研究丹参非传统药用部位花的制茶工艺,以及不同加工条件下丹参花茶的主要化学成分分析和感官评价。 方法：于盛花期采摘紫花丹参花蕾和开放初花,采用正交试验设计不同制茶工艺,采用紫外分光光度计测量制得花茶中叶绿素、类胡萝卜素、可溶性糖、总多酚、氨基酸以及总黄酮含量,进一步采用HPLC法检测丹参花茶中迷迭香酸、丹酚酸B、丹参素的含量,并进行了相应的方法学考察。结果：丹参花茶中含有较高的总多酚,具体成分以迷迭香酸为主,含有少量的游离氨基酸、黄酮、多糖以及微量的叶绿素和类胡萝卜素。结论：采摘丹参花蕾利用微波杀青、微波干燥,可以保留较多活性成分,同时具有较好的外观和口感。     

基于SSR标记分析大豆疫霉根腐病抗源的遗传多样性

丰富的遗传多样性可为大豆育种提供宽阔的遗传基础,本研究基于35对SSR标记,对60份东北地区大豆疫霉根腐病抗性品种进行了遗传多样性分析,共检测到189个等位基因,平均每个位点等位变异数5.4个,多态性信息含量指数(PIC)为0.1550~0.8195,平均为0.6636;遗传相似系数的变异范围为0.31~0.74。利用5对高多态性SSR引物构建了60份抗性材料的指纹图谱,这5对SSR引物构建的指纹图谱可以将60份疫霉根腐病抗性材料逐一区分开。采用NTSYS2.10基于遗传距离的聚类分析,将60份抗性材料分为7个类群,其中78.33%的抗性品种(系)的遗传相似系数在0.45~0.74间,表明遗传差异相对较窄,品种间遗传多样性水平较低。聚类分析与群体遗传结构分析结果有部分重合,均反映出不同地区的抗性材料间存在一定的渗透和交流。     

Evaluation of Inhibitory Effect of Organic and Inorganic Salts Against Ilyonectria liriodendri,The Causal Agent of Root Rot Disease of Kiwifruit

This study evaluated efficacy of 42 organic and inorganic salts as possible alternatives to synthetic fungicides for the control of Ilyonectria root rot of kiwifruit. Preliminary in vitro tests showed that ammonium bicarbonate, ammonium carbonate, potassium benzoate, potassium sorbate, sodium benzoate and sodium metabisulphite at 2% completely inhibited mycelial growth of the fungus. No significant differences were observed among these salts and disodium EDTA (P ≤ 0.05). However, the ED₅₀, minimum inhibition concentration (MIC), and minimum fungicidal concentration (MFC) values indicated that sodium metabisulphite was more toxic to Ilyonectria liriodendri than these other six salts. Soil bioassays showed that sodium metabisulphite, sodium benzoate and potassium sorbate at 0.25% completely inhibited mycelial growth of the fungus, whereas potassium benzoate reduced the mycelial growth of fungus by 90.30%; however, the differences in inhibitory effects were statistically insignificant (P ≤ 0.05). Moreover, there was no significant difference between 0.1% sodium metabisulphite and 0.5% ammonium carbonate, 0.75% ammonium bicarbonate and 1.5–2.0% disodium EDTA (P ≤ 0.05). Unlike disodium EDTA, complete inhibitory was observed with ammonium carbonate and ammonium bicarbonate at higher concentrations. However, in root bioassays, applications of 2% ammonium bicarbonate, 1.5% ammonium carbonate and 2% disodium EDTA were phytotoxic to kiwifruit seedlings, but 0.25% four other salts were neither phytotoxic to kiwifruit seedlings nor did it adversely affect root length, root fresh weight and root dry weight of seedling. This study also showed I. liriodendri to be capable of growth in both acidic and basic environments. However, while the fungus showed uninhibited growth at pH values of 5–11, growth decreased significantly at both higher and lower pH values (P ≤ 0.05) and was completely inhibited at pH 12.     

Study on Rosellinia necatrix Isolates Causing White Root Rot Disease in Taiwan

White root rot is a serious soil‐borne disease of several woods and crops. Recently, white root rot of tea shrubs and ornamental trees has increasingly been observed in Taiwan. Thirty‐six isolates of white root rot pathogen, showing pear‐shape swellings adjacent to the hyphal septa, had been isolated from samples of white root rot collected from Taiwan for about 4 years. The pathogen isolates produced Dematophora anamorph. Conidia of the pathogen were one‐celled, hyaline, subglobal, with truncate base, 2.9–5.8 × 1.9–3.5 μm . Ascospore dimensions were in the range of 37.0–55.0 × 5.4–7.9 μm with a short, longitudinal and straight germ slit, which complied with Rosellinia necatrix. Based on molecular studies, the pathogen isolates collected from Taiwan except R701 were identified as R. nectarix. Isolate R701, which was relatively polymorphic in internal transcribed spacer DNA sequence than other isolates, was temporarily considered as R. necatrix‐related pathogenic Rosellinia spp. All the tea cuttings (Camellia sinensis) inoculated with isolates developed typical white root rot symptoms. Pathogenicity tests demonstrated the presence of variation in virulence among the Rosellina isolates. Most of the R. necatrix isolates originating from Acer morrisonense were less virulent than those that originated from other hosts. The pathogenic Rosellinia spp., isolate R701, was also highly virulent to both cultivars of tea cuttings.     

Evaluation of Durum Wheat Genotypes for Resistance against Root Rot Disease Caused by Moroccan Fusarium culmorum Isolates

Fusarium culmorum is one of the most important causal agents of root rot of wheat. In this study, 10 F. culmorum isolates were collected from farms located in five agro-ecological regions of Morocco. These were used to challenge 20 durum wheat genotypes via artificial inoculation of plant roots under controlled conditions. The isolate virulence was determined by three traits (roots browning index, stem browning index, and severity of root rot). An alpha-lattice design with three replicates was used, and the resulting ANOVA revealed a significant (P < 0.01) effect of isolate (I), genotype (G), and G × I interaction. A total of four response types were observed (R, MR, MS, and S) revealing that different genes in both the pathogen and the host were activated in 53% of interactions. Most genotypes were susceptible to eight or more isolates, while the Moroccan cultivar Marouan was reported resistant to three isolates and moderately resistant to three others. Similarly, the Australian breeding line SSD1479-117 was reported resistant to two isolates and moderately resistant to four others. The ICARDA elites Icaverve, Berghisyr, Berghisyr2, Amina, and Icaverve2 were identified as moderately resistant. Principal component analysis based on the genotypes responses defined two major clusters and two sub-clusters for the 10 F. culmorum isolates. Isolate Fc9 collected in Khemis Zemamra was the most virulent while isolate Fc3 collected in Haj-Kaddour was the least virulent. This work provides initial results for the discovery of differential reactions between the durum lines and isolates and the identification of novel sources of resistance.