紫茎泽兰提取物对番茄青枯菌的抑菌作用及其机理

用紫茎泽兰70%的乙醇提取物及其氯仿萃取物,对番茄青枯菌生理、生化、细胞形态等方面进行抑菌试验。乙酸乙酯、正丁醇、乙醚、氯仿萃取物具有较好的热稳定性,在pH9时抑菌圈最大;通过透射电镜观察,菌体经紫茎泽兰提取物作用后,鞭毛消失,中央出现空腔。SDS-PAGE显示紫茎泽兰氯仿萃取物处理番茄青枯菌后,菌体内蛋白质减少,随着作用时间的延长,完全抑制或破坏了小分子蛋白质的合成,对大分子蛋白质也有一定的抑制作用。POD、CAT、SDH酶活测定显示,随着加入的紫茎泽兰氯仿萃取液浓度的增大,酶活显著降低。以上试验结果表明,紫茎泽兰氯仿萃取液对番茄青枯菌有明显的抑制作用,对于番茄青枯病防治的应用有一定的指导意义。     

一种有趣的泽兰尾孢菌

紫茎泽兰Eupatorium adenophorum Spreng是近年蔓延于云南省的一种杂草,今年在该草上除出现一种天敌泽菌实蝇Procecidochares utilts Stone并形成虫瘿外,近来又发现在该草上感染了一种寄生真菌,能形成叶片死斑,迄今尚未发现其他寄主,可望对草势蔓延有所抑止。现简报如下:     

链格孢菌毒素对莱茵藻光合电子传递的抑制及其作用位点

以野生型莱茵藻和其抗除草剂突变体为材料,检测链格孢菌毒素在光合电子传递链上的作用位点和模式的结果表明,此种毒素抑制Q_A至Q_B的电子传递,是光系统Ⅱ抑制剂,其作用模式类似于苯酚类型的除草剂。     

链格孢菌毒素对莱茵藻光系统Ⅱ的多重影响

链格孢菌毒素是从杂草紫茎泽兰中分离的天然致病真菌产生的植物毒素。以莱茵藻为实验材料,采用氧电极法和快速叶绿素荧光诱导动力学方法,研究链格孢菌毒素的作用位点。结果表明,该毒素对莱茵藻光系统Ⅱ有多重影响:(1)阻断受体侧QA到QB的电子传递;(2)使反应中心失活;(3)降低光能转化效率;(4)破坏(或迁移)反应中心的部分天线色素;(5)降低活性与非活性反应中心的天线色素之间能量传递的协调性。研究证实链格孢菌毒素在莱茵藻光系统Ⅱ中存在多个作用位点。     

链格孢菌毒素合成相关基因研究进展

链格孢属真菌(Alternaria Nees)是世界上分布最广泛的真菌之一,可产生9种寄主选择性毒素(host-selective toxins,HST)。根据寄主不同,将链格孢属的7个生理小种称为链格孢菌(Alternaria alternata)的7种致病型。每种致病型产生的HST都是低分子质量的次级代谢产物,只对敏感品种有毒,对抗性品种无毒。参与HST生物合成的基因都是多拷贝的,这些共同表达的基因组成基因簇。各致病型的基因簇位于一条小于2.0Mb的conditionally dispensable(CD)染色体上,CD染色体的不稳定性不影响菌丝的生长和孢子的萌发,但与菌株的致病力有关。从链格孢菌6种致病型得到毒素合成基因,日本梨致病型、草莓致病型、橘致病型有共同的EDA结构域,这3种致病型有很多EDA生物合成需要的同源基因。HST的分子遗传学研究为链格孢菌不同致病型的演化提供了新的见解。     

链格孢菌粗毒素对菊花‘神马’幼苗生长及生理代谢的影响

由链格孢菌引起的菊花黑斑病严重降低了菊花的品质和产量.链格孢菌在代谢过程中分泌的粗毒素是菊花黑斑病发生的主要致病因子之一.本文从菊花黑斑病发病叶片中分离筛选出致病真菌链格孢菌1株,研究其粗毒素对菊花幼苗‘神马’生长的影响以及测定盆栽幼苗叶片细胞膜相对透性、抗性物质含量、诱导酶活性及代谢物质含量变化.结果表明:链格孢菌粗毒素对菊花‘神马’幼苗的株高、茎粗、根长均有抑制作用,毒素浓度与抑制效果呈正相关,且粗毒素原液处理14 d后,菊花幼苗株高、茎粗、根长受到显著抑制,分别比对照减少了28.9%、21.4%和23.3%;链格孢菌粗毒素处理菊花‘神马’幼苗后,根系组织细胞膜透性随着毒素浓度的增加而增加,在同一毒素浓度处理下,菊花幼苗叶片细胞膜透性随着处理时间的增加呈先增大后降低的趋势;毒素原液处理菊花幼苗后,菊花幼苗叶片中抗性物质可溶性蛋白、丙二醛(MDA)以及脯氨酸含量均显著提高.链格孢菌10倍稀释液处理对叶片苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)和多酚氧化酶(PPO)活性的提高最为显著.链格孢菌粗毒素对切花菊‘神马’幼苗的致病作用主要通过抑制菊花幼苗根、茎的正常生长,增加菊花幼苗叶片细胞膜透性,影响切花菊幼苗叶片中抗性物质代谢以及提高叶片保护酶活性而影响植株正常生理代谢.     

链格孢菌粗毒素对菊花‘神马’幼苗生长及生理代谢的影响

由链格孢菌引起的菊花黑斑病严重降低了菊花的品质和产量.链格孢菌在代谢过程中分泌的粗毒素是菊花黑斑病发生的主要致病因子之一.本文从菊花黑斑病发病叶片中分离筛选出致病真菌链格孢菌1株,研究其粗毒素对菊花幼苗‘神马’生长的影响以及测定盆栽幼苗叶片细胞膜相对透性、抗性物质含量、诱导酶活性及代谢物质含量变化.结果表明: 链格孢菌粗毒素对菊花‘神马’幼苗的株高、茎粗、根长均有抑制作用,毒素浓度与抑制效果呈正相关,且粗毒素原液处理14 d后,菊花幼苗株高、茎粗、根长受到显著抑制,分别比对照减少了28.9%、21.4%和23.3%;链格孢菌粗毒素处理菊花‘神马’幼苗后,根系组织细胞膜透性随着毒素浓度的增加而增加,在同一毒素浓度处理下,菊花幼苗叶片细胞膜透性随着处理时间的增加呈先增大后降低的趋势;毒素原液处理菊花幼苗后,菊花幼苗叶片中抗性物质可溶性蛋白、丙二醛(MDA)以及脯氨酸含量均显著提高.链格孢菌10倍稀释液处理对叶片苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)和多酚氧化酶(PPO)活性的提高最为显著.链格孢菌粗毒素对切花菊‘神马’幼苗的致病作用主要通过抑制菊花幼苗根、茎的正常生长,增加菊花幼苗叶片细胞膜透性,影响切花菊幼苗叶片中抗性物质代谢以及提高叶片保护酶活性而影响植株正常生理代谢.     

百日草链格孢菌毒素对加拿大一枝黄花叶片伤害的生理生化研究

采用植物抗性生理和光合生理的分析测定技术,研究了分离自罹病苍耳植株上的百日草链格孢菌产生的毒素对加拿大一枝黄花叶组织细胞膜透性、丙二醛(M DA)含量、光合能力以及过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)和过氧化氢酶(CAT)活性的影响.结果表明,百日草链格孢菌毒素使加拿大一枝黄花叶组织细胞膜透性上升,膜脂过氧化加强,M DA含量上升;叶片的POD、APX和CAT的活性均较对照降低,其光合作用明显地受到抑制.说明百日草链格孢菌毒素具有防除加拿大一枝黄花的潜力.     

芸薹链格孢致病毒素分析Ⅰ.腐败菌素B的分离提纯和结构鉴定

芸薹链格孢（Alternariabrassicae）在PD液体培养基中黑暗培养20d,过滤获得培养滤液,经乙酸乙酯萃取、45℃减压浓缩得AB-粗毒素。AB-粗毒素经HPLC分析,在λ＝206nm波长下具有Ⅰ、Ⅱ、Ⅲ和Ⅳ4个明显的吸收峰。组分Ⅲ的红外吸收光谱、FAB／MASS和1H-NMR与已报道的腐败菌素B基本一致,由此确定组分Ⅲ为腐败菌素B。试验发现,芸薹链格孢的培养滤液、AB-粗毒素和毒素组分Ⅲ均能造成白菜叶片黄化、萎蔫和坏死。     

Mycelium of Alternaria alternata as a potential biological control agent for Eupatorium adenophorum

The fungus, Alternaria alternata (Fr.) Keissler Strain 501, has been evaluated as a bioherbicide for control of Eupatorium adenophorum Spreng., but the biology of the pathogen–host interaction and the optimal environmental conditions for disease development and effective weed control are unknown. Disease development of A. alternata Strain 501 mycelia on E. adenophorum was assessed under several factors including pathogen inoculum concentration, plant age, dew period duration, post-dew temperature, storage temperature and duration. The minimum inoculum concentration required to kill E. adenophorum seedlings was 3.2×10⁶ mycelial fragment mL^?1. E. adenophorum seedlings at the four-leaf-pair stage were more susceptible than the older plants, especially those at the older than seven-leaf-pair stage. With a dew period of at least 14 h, 100% mortality occurred. The optimal post-dew temperature for disease development was 18–25°C. Storage at <4°C maintained the infectivity of A. alternata strain 501 mycelia on E. adenophorum longer. Using light and scanning electron microscopy to examine the infection process of A. alternata Strain 501 mycelia, it was shown that the time from initiation to completion of infection with mycelia was much shorter (14 h) than with conidia (72 h). It was further shown that mycelial infection occurred predominately through direct penetration at intercellular junctions, while conidial infection occurred predominately through stomatal penetration. This suggests that mycelia are more suitable as infection propagules for A. alternata strain 501 in a bioherbicide for the control of E. adenophorum.     

不同地理种源紫茎泽兰的生态适应性比较

采用交互移植法,对移栽在6种不同生境中的5个不同种源紫茎泽兰幼苗的存活率、株高、分枝数、生物量、单株花序数、产种量和种子萌发率进行了为期1年的对比研究.结果表明：各种源紫茎泽兰的幼苗生长和繁殖特性对样地环境条件变化均表现出很强的可塑性.试验样地因素对幼苗株高、分枝数、生物量、单株花序数和产种量的影响均达到极显著水平(P＜0.001).随着样地纬度和海拔的升高,各种源的幼苗株高、分枝数量、单株生物量、每株花序数量和单株产种量均呈下降趋势,且各样地间的差异均达到显著水平(P＜0.05).但种源因素对幼苗株高、分枝数、生物量、单株花序数和产种量的影响均不显著（P＞0.05）.除单株产种量外,种源与试验样地的交互作用对上述各指标的影响均不显著.在各样地内,当地种源幼苗的存活率、生长能力和繁殖能力均未表现出显著的优势.说明紫茎泽兰在我国西南地区入侵成功主要依靠其较高的表型可塑性,而局域适应的作用相对较小.     

紫茎泽兰叶片化感作用对10种草本植物种子萌发和幼苗生长的影响

化感作用在生物入侵过程中具有重要的意义,紫茎泽兰具有化感物质,它能抑制豌豆等一些对化感物质敏感的植物种子萌发,但它对入侵早期直接与之竞争的植物的影响以及化感作用与其入侵性的关系还知之甚少。以紫茎泽兰入侵早期直接与之竞争的草本植物和为替代控制紫茎泽兰引进的牧草为材料,利用培养皿滤纸法研究紫茎泽兰叶片提取液对种子萌发和幼苗生长的影响,探讨化感作用与其入侵性的关系,并为其替代控制提供依据。紫茎泽兰叶片提取液对10种受体植物种子萌发和幼苗生长均有化感作用,不同浓度的提取液对植物的化感作用强度不同,低浓度时较弱,高浓度时较强。高浓度的提取液能降低种子发芽率、发芽速率、胚轴和胚根长度,增加幼苗丙二醛含量,其中发芽速率更敏感,可能是衡量化感作用的最敏感指标。不同植物对紫茎泽兰化感作用的敏感程度不同,无芒虎尾草、白三叶、细叶苦荬和莎草砖子苗对紫茎泽兰化感作用较敏感,紫花苜蓿最不敏感。     

外来植物紫茎泽兰18个种群的茎叶形态结构比较研究

利用石蜡切片法结合光学显微镜观察了紫茎泽兰18个种群的茎叶形态结构。结果表明：各种群间茎和叶的形态结构均表现出一定的变化,其中茎的维管束束数、叶表皮的部分特征变化较明显。应用SPSS统计软件对叶表皮的特征分析后,发现种群间的气孔器密度、气孔器指数、气孔器长度、气孔器宽度、上下表皮细胞数目均随地理条件的变化而表现出明显差异。相关分析表明气孔器密度、气孔器指数与海拔高度呈正相关。但紫茎泽兰各种群间的叶表皮细胞形状无明显变化,均为无规则型,垂周壁式样均为浅波状深波状;气孔器类型均为无规则型。     

Fungitoxic activity of fruit extracts of Syzygium cumini (L.) Skeels against plant pathogenic fungi Alternaria alternata and Fusarium oxysporum

This study was aimed to evaluate the effectiveness of the aqueous and ethanolic extracts of fruits of Syzygium cumini, against the mycelial growth of Alternaria alternata and Fusarium oxysporum. The results showed that ethanolic extract at the concentrations of 7.5 and 9?mg/ml completely inhibited the mycelial growth of A. alternata and F. oxysporum, respectively. While the aqueous extract at a highest tested concentration (37.5?mg/ml) exhibited only 27.86 and 37.23% inhibition of mycelial growth of A. alternata and F. oxysporum, respectively. The spore germination assay also showed the complete inhibition of spore germination of A. alternata and F. oxysporum by ethanolic extract at 50 and 60?mg/ml concentrations, respectively. Minimum inhibitory concentration was recorded as 0.039 and 0.156?mg/ml in ethanolic extract and 20 and 6.25?mg/ml in aqueous extract against A. alternata and F. oxysporum, respectively. Phytochemical analysis also showed the presence of high amount of phenolics, tannins, flavonoids, alkaloids and saponins.     

Utilization of Caffeic Acid Phenethyl Ester to Control Alternaria alternata Rot in Tomato (Lycopersicon esculentum Mill.) Fruit

Chemical fungicides that are related with resistant strains develop negative effects on human health and environment. Propolis is a resinous substance collected by bees with positive effects on human health and inhibitory activity against Alternaria alternata. Caffeic acid phenethyl ester (CAPE) is a component of the propolis. The objective of this experiment was to test the effect of CAPE on fungi infecting tomato fruit using as a model the pathosystem A. alternata‐tomato. CAPE was chemically synthesized in our laboratory and analysed with nuclear magnetic resonance spectroscopy. Different concentrations (0, 16, 32, 48, 64, 80, 90 and 100 μm ) of CAPE were tested on A. alternata growing in vitro. For the in vivo experiment, red ripe tomato fruit was inoculated with A. alternata and untreated or treated with 1, 50 and 100 μm of CAPE. After that, the fruit was stored at 25°C for up to 20 days. Colony size (CS) was recorded in vitro. In tomato fruits, the severity of infection (SI), respiration rate (RR), ethylene production (EP), pH, total soluble solids (TSS), weight loss (WL) and titratable acidity (TA) were evaluated during the storage time. CAPE melting point and spectral data probed to be the right molecule. In vitro, 64 and 100 μm of CAPE reduced CS by 30%. In vivo, 50 and 100 μm of CAPE reduced SI higher than the fungicide Captan^® with no effects on RR, EP, WL, pH, TSS and TA. It was concluded that CAPE controls A. alternata infection better than a commercial fungicide without negative effects on tomato fruit ripening and fruit quality.     

深层培养云芝菌丝体蛋白多糖的提取及性质

采用水、３％草酸和０．２ｍｏｌ／Ｌ氢氧化钠为溶剂分别提取深层培养的云芝〔Ｐｏｌｙｓｔｉｃｔｕｓｖｅｒｓｉｃｏｌｏｒ（Ｌ．）Ｆｒ．〕菌丝体蛋白多糖,其得率分别为菌丝体干重的２０％、４９％和１９％。提取的粗多糖经ＳｅｐｈａｄｅｘＧ１００柱层析进一步分离、纯化,纯化的多糖经聚丙烯酰胺凝胶电泳分离后呈现一个斑点,在ＳｅｐｈａｄｅｘＧ１００柱层析中有一个对称峰,紫外及红外光谱分析和蛋白质含量测定表明该多糖为蛋白多糖,其蛋白质和多糖含量在水提多糖中分别为１３％和６２％、在酸提多糖中分别为５％和９１％。在碱提多糖中分别为３％和８１％。组成该多糖的主要单糖是葡萄糖和木糖。     

Mechanisms of the Anticancer Action of Ganoderma lucidum (Leyss. ex Fr.) Karst.: A New Understanding

Ganoderma lucidum (Leyss. ex Fr.) Karst., a medicinal fungus called "Lingzhi" in China, has been used in traditional Chinese medicine in China for the prevention and treatment of various types of diseases, such as cancer, hepatopathy, arthritis, hypertension, neurasthenia, and chronic hepatitis. It is clear that the anticancer activity of G. lucidum is mainly due to polysaccharides and/or triterpenoids of the fungus. However, until now, the mechanism of the anticancer action of G. lucidum has not been well understood and, previously, the activation of the immune response of the host was widely considered to be the only mechanism by which G. lucidum prevented and/or treated cancer. However, recent studies reviewed in the present paper have shown that the potential mechanisms of anticancer action include not only the activation of the immune response of the host, but also the induction of cell differentiation, the induction of Phase Ⅱ-metabolizing enzymes, the inhibition of angiogenesis, and the inhibition of the expression of the urokinase-type plasminogen activator (uPA) and the uPA receptor in cancer cells. To further elucidate the mechanisms of action of G. lucidum, more in vivo tests and randomized controlled clinical trials should be carried out, and the molecular mechanisms should be studied intensively. Additionally, whether the anticancer compounds in G. lucidum act synergistically or independently should be further studied.     

中华蚊母树染色体制片及核型分析

以中华蚊母树根尖为材料,采用常规压片法制片,比较材料的不同采集时间、预处理方法、固定剂、解离方法、解离时间及染色方法对中华蚊母树根尖染色体制片的影响.结果表明最佳的制片技术为:取材时间为上午9:00~11:00或下午13:00~15:00,以饱和对二氯苯预处理3 h,用1 mol?L-1盐酸60℃下解离8 min,卡诺固定剂固定,以改良石碳酸品红染色10 min.以该最佳制片方案对中华蚊母树进行体细胞染色体核型分析,首次揭示了中国特有植物———中华蚊母树体细胞染色体数目为2n=2x=24,染色体基数x=12,染色体核型公式为2n=2x=24=12m(2SAT)+10sm+2st,主要由中部和近中部着丝点染色体组成;染色体相对长度组成为2n=24=2L+8M2+12M1+2S,核型不对称指数为64.29%,属于2A型.结果显示中华蚊母树核型对称程度较高,在进化中属于比较原始的类型.     

金针菇(Flammulina Velutipes(Curt.ex Fr.)Sing.)子实体多糖PA_(5)DE的提取及性质研究

 从金针菇Flammulina velutipes(Curt.ex Fr.)Sing.子实体中提取水溶性多糖,经乙醇分级,DEAE-Sephadex A-25纯化,得PA_5DE。以聚丙烯酰胺凝胶电泳和凝胶柱Sepharose 4B层析证明是化学均一性多糖,分子量是47.1万。用GLC、I.R、~13C-N.M.R.分析表明含有D-葡萄糖、D-甘露糖、D-岩藻糖。PA_5DE分子可能具有分支结构,含β-型糖苷键,存在β(1→3)和β(1→6)型糖苷键连接,并有抑制肿瘤S-180的活性。