芳樟醇对灰葡萄孢生长的影响及对番茄灰霉病的防控效果

通过平板抑菌试验和孢子萌发试验研究了芳樟醇对灰葡萄孢的生长抑制作用,并通过盆栽试验进一步验证了芳樟醇对番茄灰霉病的防控效果。结果表明: 芳樟醇能够显著抑制灰葡萄孢菌丝的生长,半最大效应浓度(EC₅₀)值为0.581 mL·L^-1。孢子萌发试验中,芳樟醇能够有效抑制灰葡萄孢孢子的萌发,并表现出浓度依赖性。芳樟醇处理提高了灰葡萄孢菌菌丝体的相对电导率和丙二醛(MDA)含量,说明芳樟醇可引起氧化损伤效应导致灰葡萄孢菌的膜系统被破坏;芳樟醇处理后灰葡萄孢菌中的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)活性较对照组分别下降了27.4%、68.9%和26.0%,说明芳樟醇抑制了灰葡萄孢菌体内的抗氧化系统。盆栽试验结果显示,芳樟醇处理的病斑直径较对照组显著降低;番茄叶片中的SOD、CAT、POD、多酚氧化酶和苯丙氨酸解氨酶活性均显著高于对照组;而MDA含量下降了41.5%,说明芳樟醇可减轻灰葡萄孢菌对番茄植株造成的氧化损伤以提高植物抗病性。综上,芳樟醇对灰葡萄孢的生长有显著抑制作用并对番茄灰霉病有较好的防治效果,研究结果可为开发新型植物源抑菌剂防控番茄灰霉病提供理论依据。     

灰葡萄孢产孢新基因的功能分析

灰葡萄孢是一种重要的植物病原真菌,其寄主范围广泛,能危害世界上230多种双子叶植物,常给农业生产造成重大的经济损失[1-3].由灰葡萄孢引起的灰霉病是目前我国温室蔬菜生产中最主要的病害之一,一般造成全年减产20％-25％,严重时达到40％以上[4].因此,研究该病菌的致病机理对该病防治具有重要意义,并且随着灰葡萄孢基因组测序的完成,灰葡萄孢已成为发育生物学、分子植物病理学研究的模式生物之一.     

灰葡萄孢交配型基因的分析与检测

通过生物信息对灰葡萄孢的MAT1‐1‐1与MAT1‐2‐1氨基酸序列进行了系统进化与结构域保守氨基酸分析,表明灰葡萄孢的交配型蛋白与核盘菌的亲源关系最近,结构域氨基酸比对结果表明该基因具有保守氨基酸的一致性与部分氨基酸的相似性。应用PCR技术检测灰葡萄孢交配型基因MAT1‐1‐1与MAT1‐2‐1,结果表明各种植区交配型菌株所占比例有较大的差异,多数种植区灰葡萄孢同时存在MAT1‐1与MAT1‐2两种交配类型,快速检测灰葡萄孢的交配型等位基因对于灰葡萄孢种群结构分析非常有意义。     

两种葡萄孢属真菌对不同品种百合花瓣和叶片的侵染能力比较

为明确两种葡萄孢属真菌对不同百合品种叶片和花瓣的侵染能力,采用离体叶片接种法测定灰葡萄孢Botrytis cinerea和椭圆葡萄孢Botrytis elliptica对4个百合品种叶片和花瓣的侵染时间和病斑扩展速度。结果表明,供试百合花瓣接种灰葡萄孢病斑出现时间明显早于叶片,而不同品种花瓣接种椭圆葡萄孢病斑出现时间差异显著。此外,百合品种‘木门’叶片接种椭圆葡萄孢96 h后仍没有病斑出现,而花瓣接种后48 h病斑出现,说明‘木门’叶片对椭圆葡萄孢抗性较强,而花瓣较易感病。     

灰葡萄孢霉菌固体培养产生的（＋）ABA及其生物学效应

目前已筛选出的Botrysits、Cercospord等属菌株能合成次生代谢物ABA。这已成为化学家和生物学研究者感兴趣的课题,然而尚存在一些亟待解决的问题。由于天然ABA纯度高(大于97％),且生理活性较强,是开发ABA的新途径。本文应用草莓灰葡萄孢菌株,通过试探性的实验,用简便的固体发酵方式培养,经吸     

葡萄孢盘菌属一新种——蚕豆葡萄孢的有性阶段

本文报道了葡萄孢盘菌属的一个新种:蚕豆葡萄孢盘菌(Botryotinia fabae Lu et T.H.Wu sp.nov.)即蚕豆葡萄孢的有性阶段;并报道了新种子囊盘形成过程;新种有汉文和拉丁文描述。     

抗灰葡萄孢霉菌乳酸菌的筛选

从80株乳酸菌中筛选出45株具有抗灰葡萄孢霉菌活性的乳酸菌菌株,其中10株具有较强抗灰葡萄孢霉菌活性。对这10株乳酸菌菌株的抗植物致病真菌谱进行了研究,这10株乳酸菌对番茄早疫病菌,甜瓜疫霉菌,甜瓜枯萎病菌,苹果炭疽病菌的生长均有较强的抑制作用。其中1株具有广谱抗植物致病真菌活性的乳酸菌菌株BX6-4为植物乳杆菌。经番茄离体叶片接种试验发现,植物乳杆菌BX6-4的发酵液能够在体外强烈地抑制灰葡萄孢霉菌的生长。     

灰葡萄孢侵染垫缺失突变体的筛选及其相关生物学特性的研究

【目的】从农杆菌介导获得的灰葡萄孢RoseBC-3的突变体库中筛选侵染垫缺失突变体菌株,并明确其相关生物学特性。【方法】将菌株接种于洋葱表皮,利用棉兰染色观察侵染垫形成情况,筛选得到一个侵染垫缺失突变体(AT19)。采用形态学方法、离体叶片接种法、钌红染色法、小麦种子幼芽生长抑制法分别对该菌株的菌落培养性状、侵染垫产生情况、致病力、产果胶酶能力以及产植物毒性代谢产物能力进行测定。【结果】筛选灰葡萄孢突变体168株,根据侵染垫形成可分为三类：快速形成侵染垫型(158株)、缓慢形成侵染垫型(9株)和侵染垫形成缺陷型(1株,AT19)。AT19在接种洋葱120 h后依然无法形成成熟侵染垫。该菌株生长较为缓慢,菌落扩展均匀,可以产生分生孢子,对烟草、草莓、蚕豆和豌豆叶片均不能致病,可以产生果胶酶和植物代谢毒性物质。【结论】突变体菌株AT19可以产生果胶酶和植物代谢毒性物质,其致病力缺失与侵染垫产生缺陷相关。研究结果为了解灰葡萄孢侵染垫形成分子机制提供基础材料。     

灰葡萄孢BC7-3菌株除草活性组分的纯化与结构鉴定

[目的]植物病原真菌毒素是一类重要的微生物源除草剂,本研究旨在找到一个新的具有除草活性的化合物结构.[方法]在前期薄层层析法、柱层析法和高效液相色谱法分析的基础上对灰葡萄孢诱变菌株BC7-3的代谢产物中具有除草活性的5个不同组分分别进行了液相色谱制备.[结果]本研究得到了一个纯度达99.38%对单子叶杂草马唐具有较强杀除活性的纯组分,通过对纯组分的物理性状测定并结合紫外-可见吸收光谱、红外光谱以及核磁共振波谱等分析方法鉴定化学结构为10-顺-二氢化灰霉二醛.[结论]研究的结果为微生物除草剂的创新和开发奠定了理论基础.     

Induction of Volatile Organic Compounds of Lycopersicon esculentum Mill. and Its Resistance to Botrytis cinerea Pers. by Burdock Oligosaccharide

In the present study, we investigated the induction of volatile organic compounds （VOCs） of Lycoperslcon esculentum Mill. and its resistance to Botrytis cinerea Pers. by burdock oligosaccharlde. The disease severity of L. esculentum was evaluated 48 h after treatment with 0.6% burdock oligosaccharlde, followed by inoculation with a spore suspension of B. cinerea. The formation of O2＇, the activity of lipoxygenases （LOX）, peroxidase （POD）, cataiase （CAT）, and superoxide dismutase （SOD）, and the quantity and quallty of changes In VOCs were determined a period of time after treatment with 0.6% burdock ollgosaccharide. The results demonstrated that the disease index in treated plants was decreased by 42.5% compared with control 96 h after Inoculation. The production of O2＇ reached a maximum 6 h after treatment （1.36-fold compared with control）. There was an increase in LOX, POD, CAT and SOD activity in response to burdock oligosaccharide treatment and the enzymes showed different trends in the time-course of induction. At 120 h after treatment, （E）-2-hexenal was increased by 92% compared with control, whereas methyl salicylate showed a gradual Increase with induction period. Previous results had demonstrated that chitosan elicitor enhanced the production VOCs of L. esculentum and decreased plant susceptibility towards B. clnerea. Together, these findings suggest that increasing the production of VOCs in response to burrdock oligosaccharide may be an important mechanism for L. esculentumin its defense against pathogens, in addition, burrdock oligosaccharlde may act as a potent elicitor of resistance to disease in L. esculentum.     

不同方法收集番茄叶挥发性物质的GC/MS指纹图谱比较

以番茄（Lycopersicon esculentum）叶为材料．分别采用溶媒萃取、水蒸气蒸馏、气体吸附法收集挥发性物质;气相色谱-质谱联用技术分离鉴定挥发组分。结果表明,3种收集方式共获得19种挥发性组分．其中的14种组分相同．且多为主要组分。水蒸气蒸馏法获得的挥发性物质总含量最高,为27．13μg FW;其次是气体吸附法．为23．38μg FW,溶媒萃取法获得的最少,仅为12．40μg FW。溶媒萃取法共获得15种挥发组分．含量最高的组分为β-水芹烯。水蒸气蒸馏法获得18种组分．含量最高的组分为（E）-2-己烯醛。气体吸附法获得17种组分．含量最高的组分为（E）-2-己烯醛。不同方法所获各组分的相对含量和绝对含量均不同．可根据研究目的综合分析运用。     

Effects of crop development on the emission of volatiles in leaves of Lycopersicon esculentum and its inhibitory activity to Botrytis cinerea and Fusarium oxysporum

Volatiles emitted from the leaves of Lycopersicon esculentum at the two-, ten-leaf and anthesis periods were collected by a gas absorbing method and analyzed by gas chromatography （GC）-mass spectrometry. In total, 33 compounds of volaUles emitted from three developmental stage plants were separated and identified, and quantitatively analyzed by the internal standard addition method. All of the samples of volatile were found to be rich in monoterpenes and sesquiterpenes.β-phellandrene and caryophyllene predominated in the volatiles of the leaves of plants at the two- and ten-leaf stages. Furthermore, （E）-2-hexenal were the dominant components in the volatiles emitted from anthesis plants. The results of volatiles analyzed show that the compositions varied depending on the developmental stages. The volatiles emitted from crushed tomato leaves of plants at the anthesis stage had the most strongly inhibitory activity against the spore germination and hyphal growth of Botrytis cinerea and Fusarium oxysporum, followed by ten- and two-leaf plants. However, the activity of volatiles, emitted from the leaves of plants at the two-leaf stage, in inhibiting F. oxysporum was greater than B. cinerea.     

Nitrogen availability and susceptibility of tomato leaves to Botrytis cinerea

The aim of this study was to investigate the effect of nitrogen availability on susceptibility of tomato leaves to the fungal pathogen Botrytis cinerea. Plants with varying nitrogen availability were grown by adding N daily in exponentially increasing amounts to a nutrient solution at different rates. Leaves of plants grown at low nitrogen availability had a high leaf C/N ratio (21 g g^-1) and were about 2.5 times more susceptible to primary lesion formation by B. cinerea compared to plant grown at high nitrogen availability, which had a low leaf C/N ratio (11 g g^-1). Leaf C/N ratio accounted for 95% of variation in susceptibility. This relationship between C/N ratio and susceptibility persisted when plants were grown with exponential P addition and optimal N supply, and was thus independent of plant growth rate or related factors. We could not explain the effect of nitrogen availability by variation in the most obvious N-based resistance compound α-tomatine because more susceptible leaves with a high C/N ratio contained more α-tomatine. These leaves also contained more soluble carbohydrates. The level of soluble carbohydrates correlated positively with susceptibility, independent of the growth method. We therefore suggest that the effect of N availability on susceptibility must be explained by variation in levels of soluble carbohydrates and speculate about the role of these carbohydrates in the infection process. This revised version was published online in June 2006 with corrections to the Cover Date.     

Organic hydroponics induces systemic resistance against the air-borne pathogen,Botrytis cinerea (gray mould)

Here, we propose that organic hydroponics trigger induced systemic resistance (ISR) in lettuce against air-borne Botrytis cinerea, which causes gray mold. We compared effects of organic and chemical hydroponics, assessed presence of ISR elicitors in the hydroponic nutrient solution, and investigated molecular mechanism of ISR. Organic hydroponics significantly reduced gray mold lesions in lettuce (cultivated hydroponically) and cucumber (cultivated in soil and foliar sprayed with nutrient solution). The 1-aminocyclopropane-1-carboxylic acid synthase gene in lettuce and lipoxygenase and ethylene receptor-related gene in cucumber showed heightened expression, suggesting that the jasmonic acid/ethylene (JA/ET)-signaling pathway was involved in ISR for both crops. Low salicylic acid β-glucoside levels confirmed role of the ISR signaling pathway. ISR in both lettuce and cucumbers indicated that elicitors in organic hydroponics were nonhost-specific and that the JA/ET pathway was activated without microbe–root interaction. Thus, organic hydroponics can be an effective method for both soil-borne and air-borne disease control.     

枯草芽孢杆菌对几种灰霉病菌的抑制效果研究

分别研究了枯草芽孢杆菌(BacillussubtilisCohn)培养液、过滤液和灭活液对葡萄灰霉病菌(GB)、草莓灰霉病菌(SB)、辣椒灰霉病菌(PB)和番茄灰霉病菌(TB)菌丝生长的抑制作用。结果表明:枯草芽孢杆菌培养液对GB、SB、PB和TB都有很好的抑制作用。在菌液浓度达到10 5CFU/mL时,对4种灰霉病菌的抑制率均达到了10 0 % ;当浓度降低为10 4CFU/mL时,抑制率明显降抵。而菌液浓度为10 8CFU/mL时的过滤液,对GB、PB和TB的抑制率也均在5 0 %以上。灭活液对灰霉菌的抑制作用明显减弱,菌液浓度为10 8CFU/mL时,对PB、GB、TB和SB的抑制率分别为73.6 %、39.5 %、5 0 %和2 5 %。     

Differential Behaviour of Mycelial Growth of Several Botrytis cinerea Strains on either Patchoulol- or Globulol-amended Media

Botrydial and dihidrobotrydial are two characteristic metabolites of the phytopathogenic fungus Botrytis cinerea , which are involved in the development of necrotic lesions on grapevine and tobacco. Patchoulol and globulol, two natural products which are analogues to precursors of botrydial and dihidrobotrydial, were tested on 10 B. cinerea strains which were isolated from different hosts and varied in aggressiveness on grapevine leaves. Mycelial growth of all strains was prevented when they were grown on either patchoulol- or globulol-amended malt agar media (200  μ g/ μ l). Each strain displayed a specific response pattern to those products, according to the high variability previously described for this species. Furthermore, strains were different from one another with regard to their level of aggressiveness against leaves detached from sherry grapevine vineyards.     

拟南芥灰霉病抗性相关转录因子

病原菌的侵染激发植物大量防御响应基因的表达, 其中转录因子在协调庞大的抗病防御网络中发挥重要作用。灰葡萄孢菌(Botrytis cinerea)是最具破坏力的死体营养型病原真菌之一, 在农业生产上造成严重的经济损失。文章综述了ERF(Ethylene response factors)、WRKY、MYB等家族中参与灰霉病防御反应的转录因子的功能研究进展。转录因子通过复杂的mRNA或蛋白水平的互作方式构成了精细的调控网络, 以激活下游防卫基因的表达, 从而诱导抗病反应。一部分转录因子是协调不同激素信号通路交叉响应的重要节点和调节器, 将植物抵御不同类型病原菌的分子机制联系起来。对这类转录因子的研究将为研究植物其他病原菌防御机制提供线索, 另外深入理解抗病机制将有助于研究者在作物改良和保护中更高效地利用抗病基因。     

灰葡萄孢菌（Botrytis cinerea）基因组中T-DNA整合模式分析

摘要：【目的】研究灰葡萄孢菌（Botrytis cinerea）基因组中T-DNA插入位点的整合模式特征。【方法】利用农杆菌（Agrobactirium tumfacience）介导法构建灰葡萄孢菌T-DNA插入突变体库。利用热不对称交错PCR（TAIL-PCR）技术对转化子中T-DNA的旁侧序列进行扩增和克隆,对获得的旁侧序列进行比对分析。【结果】T-DNA插入在灰葡萄孢菌基因组非编码区的占69%,插入在外显子的占30%。T-DNA在插入的过程中发生了碱基缺失、增加等重组现象,其中左边界（left border,LB）整合到基因组碱基缺失较少,有的保持完整,而右边界（right border,RB）及其近邻的T-DNA区域缺失碱基较多。T-DNA的插入位点还发现有额外的序列插入。【结论】对灰葡萄孢菌中插入T-DNA的整合模式的分析为开展该菌的功能基因组学奠定了基础。