核盘菌致病性分化研究

为了解核盘菌的致病性分化,本研究用活体定位穿刺接种法,以油菜为供试寄主,对采自四川省10个地区23个县、9种寄主的108个菌株进行了致病性测定,结果表明,所有菌株对供试油菜品种均能致病,但各菌株所致病斑长度差异很大(2.7～82.0 mm),说明核盘菌种群内存在明显的致病性分化,这种分化与地理来源和寄主来源没有明显的关系。     

十三株球孢白僵菌酯酶同工酶的初步研究

对13株球孢白僵菌的酯酶同工酶进行比较分析,结果表明,从不同寄主分离的白僵菌具有不同的酯酶图形,同一寄主分离的菌株,其酯酶同工酶也表现一定的异形性。球孢白僵菌对寄主的毒力有较大的专一性,并与酯酶同工酶有一定的相关性。     

稻纹枯菌酯酶同工酶、可溶性蛋白及致病力的研究初报

 对稻纹枯菌25个菌株分别进行了致病力测定、酯酶同工酶和可溶性蛋白分析.结果表明:不同菌株存在明显的致病力分化,而酯酶同工酶图谱却有较强的一致性,它们的主酶带基本相同,但在副酶带上反映出一定的同工酶表型异质性,表现为带的数目及着色深浅的不同;可溶性蛋白图谱比酯酶同工酶图谱表现出更多的多样性,大多数致病力较强的菌株都有l~2条特征谱带,而且它们的谱带数目比弱毒力菌株的多出3~5条.     

核盘菌对油菜、向日葵和大豆的侵染及其致病性分化研究

 通过对从陕西大荔采集的油菜及其后茬向日葵上的核盘菌和从新疆阿勒泰的向日葵上采集的核盘菌样品进行单菌核分离、培养和纯化,比较其菌丝生长速度、菌丝干重、菌落形态、致病力强弱及菌株的草酸累积等,将两地的核盘菌分成A、B、C三种类型,其中A类来源于陕西大荔的油菜和向日葵,B、C类来源于新疆阿勒泰的向日葵。不同类型核盘菌对于不同寄主的致病力存在着分化现象,A、B类菌株生长正常、菌落均匀旺盛,B类对油菜、向日葵和大豆的致病力较强,草酸产量较高;而A类仅对油菜和向日葵的致病力较强对大豆的致病力很弱,但草酸产量最高。C类菌株生长异常,菌落稀疏不均匀,对3种作物的致病力均弱,草酸产量较低。2001年田问调查亦表明:A类菌株可导致油菜、向日葵菌核病的发生,但未见其使大豆致病,由此提出油菜茬后,不宜种植向日葵,二者应与小麦、玉米等实行较长周期的轮作。本文也同时对各菌株进行了RAPD分析和菌丝体亲和性研究,结果表明,菌株间的遗传多样性表现丰富但未发现与其致病性分化相关。     

来源于佳木斯茄子上的核盘菌菌株多样性的研究

 从我国黑龙江省佳木斯市同一茄子田采集的菌核样品中分离得到21个核盘菌(Sclerotinia sclerotiorum)单菌核分离物。通过比较其菌丝生长速度、菌落形态、菌核产量及菌核在PDA平板上的萌发特性等将其分成3种类型,记为A、B、C,各包含5、7、9个菌株。A型和B型菌株为正常菌株,而C型菌株为异常菌株,生长慢且呈扇形扩展。对代表性菌株转代培养物及菌核后代的培养特性比较结果说明它们的分化特性是稳定的。致病性的测定结果表明参比菌株Cor-6的致病性最强,A型菌株Ep-1PB和B型菌株Ep-1次之,C型菌株Ep-1PD最弱。3类菌株菌核的可溶性蛋白和酯酶同工酶电泳谱带没有明显的差异。     

稻瘟病菌生理小种酯酶同工酶特性的研究

 运用聚丙烯酰胺凝胶等电聚焦法对来自江、浙两省经全国统一鉴别寄主鉴定的稻瘟病菌7群12个生理小种的16个菌株菌体酯酶同工酶进行了分析比较,结果表明:稻瘟菌菌体酯酶同工酶带有13-18条,等电点分布范围为3.00-7.20。在等电点4.15附近有各生理小种共同的酶带。同一生理小种群内不同生理小种间酯酶同工酶的相似系数一般大于0.50,不同生理小种群的生理小种间酶谱的相似系数一般为0.30-0.40,也存在少数不是同一生理小种群的小种间酶谱相似系数大于0.50的情况。供试各生理小种间酶谱均存在不同程度的差异,表现为酶带数目的多少和等电点分布的不同。来自不同省份经鉴别寄主鉴定为同一生理小种的菌株间酶谱的相似系数亦仅0.05-0.60。研究结果初步反映出供试大多数稻瘟菌菌株间菌体酯酶同工酶的相似性与致病性有关,但也受菌株来源地生态环境影响。本文还讨论了稻瘟病菌致病性容易变异但在许多情况下又具有相对稳定性的可能机制,探讨在一定范围内聚丙烯酰胺凝胶等电聚焦技术用于稻瘟菌生理分化和致病性变异监测的可能性及尚待进一步研究解决的穗题等。     

莴苣上发现一种新的核盘菌菌核病

 从湖北省神农架莴苣上分离到一种产菌核病原真菌。这种真菌在培养特性上同核盘菌、三叶草核盘菌和小核盘菌既存在着明显差异,又存在着相似之处。其菌核易萌发成子囊柄,但在一般散射光下柄顶端难以发育成子囊盘。对偶得的1枚子囊盘观察表明这种真菌符合核盘菌属真菌的特征,并且同核盘菌属3个近缘种的菌株不同。可溶性蛋白质和多种酶同功酶电泳分析结果表明这种真菌不同于核盘菌属的3个常见种,但同它们亲缘关系较近。这种新菌核病的初侵染来源是菌核萌发产生的菌丝,通过病健接触构成再侵染,在病组织上形成菌核越冬越夏。离体和活体致病性测定结果表明这种真菌只能侵染莴苣,不能侵染油菜、小白菜、萝卜和胡萝卜。     

利用SRAP分析油菜品种对核盘菌遗传分化的影响

 本文采用茎秆牙签接种法将单一核盘菌菌株接种至不同油菜品种茎秆,再从46个油菜品种茎秆内收集接种后形成的菌核进行分离纯化和培养,并采用SRAP技术对46株核盘菌菌株进行了遗传分化分析。从12对检测引物中共获得357个位点,其中多态性位点273个,占76.47%;UPGMA聚类分析显示,在相似系数为0.77时,46株核盘菌菌株能够分为7组。当以寄主的抗(耐)病程度、寄主品种类型和品种选育地来源为标准将菌株分为不同群体时,AMOVA(analysis of molecular variance)结果表明核盘菌菌株在各群体内变异率分别为98.50%、105.16%和95.36%,均达到极显著水平(P<0.001),而寄主品种选育地群体间的遗传变异达到极显著,变异率为4.64%。结果表明:核盘菌菌株接种不同油菜品种后,菌株间存在明显的遗传分化,这种分化与油菜品种的选育地来源有密切关系。     

稻瘟菌不同小种间可溶性蛋白及酯酶同工酶的多态性

采用聚丙烯酰胺凝胶泳对来自太湖稻区的１０个水稻稻瘟病菌株和２个禾本科杂草瘟菌的可溶性蛋白及酯酶同工酶进行了分析比较。结果表明：供试菌株菌体可溶性蛋白可呈现１６～２５条电泳谱带，其中在Ｒｆ值为０．２３，０．３３，０．４４的３条谱带为１２２个菌株所共有，没有发现生理小种特异性谱带。对１２个菌株α－酯酶同工酶的分析表明，在所有菌株同工酶谱带中可分辩出具有不同迁移率的１２条谱带，据此可将菌株分为８个不同的同工酶谱带类型。同时表明，α－酯酶同工酶在太湖稻区稻瘟病菌中存在丰富的多态性，但其谱带类型与病菌生理小种无相关性。     

核盘菌转录因子SsMCM1原核表达与亚细胞定位

 核盘菌是一类寄主范围广泛的植物病原真菌。MADS-box蛋白家族基因广泛存在于生物体中,参与调控细胞识别、新陈代谢、细胞周期等。核盘菌转录因子SsMCM1调控其子实体形成与致病性。为进一步揭示SsMCM1的作用机制,本研究以核盘菌野生型菌株uf-70的cDNA为模板,扩增得到SsMCM1基因,并与原核表达载体pET-28a(+)连接,构建成重组质粒转化到大肠杆菌BL21(pLysS)表达菌株中,通过IPTG的诱导和亲和层析,纯化得到SsMCM1蛋白,并以此为抗原获得抗血清,完成效价测定。利用特异性抗血清,与核盘菌不同组织蛋白进行特异性的免疫结合,发现其与核盘菌总蛋白、核盘菌细胞质总蛋白均特异性结合,而核盘菌细胞壁蛋白未发现特异性反应,表明SsMCM1不定位于核盘菌细胞壁上。此外,构建其亚细胞定位载体pCG-1301::SsMCM1,并将重组质粒转化到农杆菌中,通过农杆菌的侵染作用进行瞬时表达,发现SsMCM1定位于细胞核中,作为转录因子调控核盘菌的子实体发育与致病性。     

温度和光照对中黑盲蝽滞育解除的影响

本文研究了中黑盲蝽滞育卵经过4℃低温、35℃高温和全日照处理不同天数后的滞育解除情况。结果表明:4℃低温可以有效地解除中黑盲蝽卵的滞育,随着低温处理时间的延长,中黑盲蝽滞育卵的孵化率逐渐上升,处理60d后孵化率可达80.00%以上。全日照也能够有效地解除中黑盲蝽卵的滞育,滞育卵解除滞育后的孵化率也随处理天数增加而增加,处理40d,孵化率可达到40.00%以上。35℃高温不能解除中黑盲蝽卵的滞育。相对于非滞育卵,经4℃低温和全日照处理后,解除滞育的卵初孵时间有延迟现象,整个孵化历期延长。     

The use of isozyme patterns to distinguish sweetpotato whitefly (Bemisia tabaci) biotypes

Recent collections ofBemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) from California desert regions represent a mixture of biotypes. One biotype was identical to a culture originally obtained in 1981 and since maintained in the laboratory. The other and most prevalent biotype could not be distinguished morphologically but could be distinguished by esterase isozyme banding patterns. The banding patterns of the biotypes were not affected by culturing the whiteflies on different plant species. Different developmental stages, and adults of both sexes, had the same isozyme patterns.     

捕食线虫真菌酯酶同工酶谱和可溶性蛋白质谱分析

 本文对5个属14个种共33株捕食线虫真菌采用聚丙烯酰胺凝胶电泳技术进行了酯酶同工酶谱和可溶性蛋白质谱分析。结果表明,酯酶同工酶谱带少,但各属的特征谱带是明显的,在同属不同种之间,多数种都有自已的特征谱带与其它种相区别,同种不同菌株间没有明显差异;蛋白质谱带较多,在每个属中均有1~2条特征谱带,属间的谱带差异明显,同属不同种和同种不同菌株间蛋白质谱带都表现出多型性。     

用酯酶同工酶技术检测球孢白僵菌株型

对40个不同来源的球孢白僵菌分离株的酯酶同工酶图谱进行了测定,根据图谱对林间放菌前和放菌后发生的白僵菌菌株类型进行分析,进而评价了冬前放菌防治马尾松毛虫的效果。结果表明,酯酶同工酶图谱可以用来鉴别白僵菌菌株的株型。根据图谱将所试菌株归为10个类型。菌株类型与其采集地点、寄主具有相关性。采自同一试验区不同小生境的18个分离株属于同一酯酶型。冬前所放菌株在次年春季4月底前未能检出,导致春季马尾松毛虫种群白僵病的仍是当地菌源。     

Isoenzyme diversity in Xanthomonas campestris pv. mangiferaeindicae

A collection of 26 strains of Xanthomonas campestris pv. mangiferaeindicae isolated from three different host species in eight countries was investigated for variation in isozyme patterns. Three enzyme systems were analysed: esterase (EST), phosphoglucomutase (PGM) and superoxide dismutase (SOD). Four groups of strains were identified: nonpigmented strains isolated from mango and pepper-tree in Australia, Comores, India, Reunion Island, South Africa, and Taiwan; nonpigmented Brazilian strains from mango; nonpigmented strains from ambarella isolated in the French West Indies; heterogeneous yellow pigmented strains from mango (Brazil and Reunion Island). The value of isozyme profiling as markers of the pathogenicity groups in X. c . pv. mangiferaeindicae is discussed.     

Isozyme and RAPD-PCR analyses of Fusarium avenaceum strains from Finland

Differences in isozyme and RAPD-PCR polymorphisms amongst 33 isolates of Fusarium avenaceum were compared using native polyacrylamide gel electrophoresis and agarose gel electrophoresis. The isolates were collected from different regions of Finland. Amongst eight enzymes analysed clear isozyme polymorphism was detected in five enzymes which could be grouped into 20 different electrophoretic phenotypes and three main groups at the similarity level of 70% in unweighted pair group method with arithmetic average (UPGMA) analysis. RAPD-PCR analysis differentiated all F. avenaceum strains from each other. The phenotypes resulting from RAPD-PCR analysis were grouped into five main groups by UPGMA analysis at the similarity level of 55%. These main groups had several similarities with the main groups from isozyme analysis. RAPD-PCR patterns of 16 isolates of Fusarium graminearum F. culmorum F. equiseti F. oxysporum and F. redolens were also studied and strains from each Fusarium species formed individual groups in UPGMA and principal components analyses. Thus, the extent of isozyme and RAPD-PCR polymorphisms found in Fusarium strains potentially provides a method for identifying the fungi both at strain and species level.     

Pyrethroid synergists suppress esterase-mediated resistance in Indian strains of the cotton bollworm, Helicoverpa armigera (Hübner)

The role of esterase in pyrethroid resistance was studied in the final larval instar of different strains of the cotton bollworm, Helicoverpa armigera. The resistant strains viz., Nagpur strain and the Delhi strain were found to have elevated midgut esterase activity in comparison to the susceptible strain. Nagpur strain and Delhi strain have 2.24 and 1.73-fold higher esterase activity, respectively, than that of the susceptible strain. The Native PAGE displayed important differences in the midgut esterase isozyme pattern between the susceptible and the pyrethroid-resistant strains. Out of the 10 esterase isozyme observed, susceptible strain lacked three bands, E2, E6 and E10 that were found in the resistant strains. The potency of the synergists piperonyl butoxide (PBO) and dihydrodillapiole (DDA) as esterase inhibitor were also studied both in vitro and in vivo. The in vitro results clearly show that both PBO and DDA inhibited esterase activity in the two resistant strains, while there was almost no esterase inhibition in the homogenate of the susceptible strain. The in vivo inhibition studies (topical application of PBO and DDA followed by biochemical analysis) illustrated that PBO- and DDA-esterase binding is rather slow and non permanent process. Esterase inhibition did not occur immediately after the synergist treatment but at 4 and 8 h post treatment in case of PBO and DDA, respectively. Native PAGE revealed that the in vivo esterase inhibition caused by both PBO and DDA was due to the binding of the synergist with the E6 isozyme which was not present in the susceptible strain.     

我国棉花枯萎病菌酯酶同工酶测定初报

 利用聚丙烯酰胺薄层凝胶电泳法测定了我国主产棉区棉花枯萎病菌的酯酶同工酶。三年试验结果表明,供试42个菌系的酯酶同工酶有三种类型主酶带,根据主酶带可以将供试的菌系区分为三个类型。类型一,有E₄、E₅、E₆、E₈四条主酶带,包括13省的菌系31个,类型二,有E₄、E₆、E₈三条主酶带,包括6省的菌系10个;类型三,有E₆、E₈两条主酶带,包括新疆吐鲁蕃的菌系一个。     

桃树黄化病对不同桃树品种防御相关酶活性的影响

本文对比4个不同品种的健康桃树叶片和黄化桃树的黄叶及绿叶过氧化物酶和多酚氧化酶的活性,并采用聚丙烯酰胺凝胶电泳方法观察过氧化物同工酶的酶谱变化。结果表明:桃树感染黄化植原体后POD活性先升高后降低,而PPO活性呈持续降低趋势;通过聚丙烯酰胺凝胶电泳发现过氧化物同工酶的酶带减少,颜色变浅,酶活性下降。