恶疫霉致病力和对甲霜灵敏感性的遗传与变异

以分离自黑龙江腐烂苹果的恶疫霉Phytophthora cactorum Schroeter野生型菌株Ap14为亲本,采用菌丝块创伤接种法测定了恶疫霉对苹果的致病力在无性繁殖和有性生殖后代的遗传,结果是连续2代单游动孢子后代在苹果上所致病斑半径分别为22.4-24.1mm和21.8-23.4mm,与亲本所致病斑半径22.5mm无显著差异;而其20个自交后个体所致病斑半径为21.4-25.8mm,与亲本有显著差异。其中2株致病力显著强于亲本,其余与亲本相似,表明恶疫霉对苹果的致病力在无性后代可稳定遗传,而在自交有性后代发生分离,同时在含甲霜灵0.05μg/ml的LBA平板上测定了恶疫霉菌丝生长对甲霜灵的敏感性在无性和有性后代的遗传。结果是恶疫霉对甲霜灵的敏感性在无性单孢后代无显著变异,而在50个自交有性后代中,上述浓度对其菌丝生长的抑制率分布范围为67.3-97.1%,与亲本78.3%有极显著差异。其中3株高于亲本,3株低于亲本,44株与亲本相似,表明恶疫霉对甲霜灵的敏感性同样在无性后代稳定遗传而在有性后代发生变异,上述结果提示,供试恶恶疫霉菌株中上述性状可能由细胞核杂合基因控制。     

掘氏疫霉遗传学研究I．自交后代(S1)生物学性状的遗传与变异

测定了掘氏疫霉Phytophthora drechsleri Tucker生长速率、菌落形态、菌体形态、产孢能力、耐35℃高温生长、致病力等生物学性状在自交S1代的遗传与变异。2个掘氏疫霉野生型菌株(A1、A2各1株)经聚碳膜间隔配对诱导自交产生的卵孢子(保存110天左右)在S+L培养基上萌发,分别获得30和35个单卵孢株(萌发率10％左右)。测定结果表明,上述生物学性状在2个亲本自交后代均发生变异,部分单卵孢株生长速度减慢、菌落形态明显改变、孢子囊和菌丝畸形、产孢能力增强、在35℃中弱生长或不生长,或致病力减弱直至无致病力。在亲本的单游动孢子无性系后代未观察到上述变异,说明上述生物学性状变异主要因亲本遗传因子的杂合性所致。从遗传学角度探讨了在自然条件下掘氏疫霉生物学性状的变异性以及某些分类依据不稳定的原因,指出有性生殖过程中的基因重组可能是引起掘氏疫霉生物学性状变异的主要原因之一。     

掘氏疫霉遗传学研究 Ⅰ.自交后代(S_1)生物学性状的遗传与变异

测定了掘氏疫霉Phytophthora drechsleri Tucker生长速率、菌落形态、菌体形态、产孢能力、耐35℃高温生长、致病力等生物学性状在自交S_1代的遗传与变异。2个掘氏疫霉野生型菌株(A_1、A_2各1株)经聚碳膜间隔配对诱导自交产生的卵孢子(保存110天左右)在S+L培养基上萌发,分别获得30和35个单卵孢株(萌发率10%左右)。测定结果表明,上述生物学性状在2个亲本自交后代均发生变异,部分单卵孢株生长速度减慢、菌落形态明显改变、孢子囊和菌丝畸形、产孢能力增强、在35℃中弱生长或不生长,或致病力减弱直至无致病力。在亲本的单游动孢子无性系后代未观察到上述变异,说明上述生物学性状变异主要因亲本遗传因子的杂合性所致。从遗传学角度探讨了在自然条件下掘氏疫霉生物学性状的变异性以及某些分类依据不稳定的原因,指出有性生殖过程中的基因重组可能是引起掘氏疫霉生物学性状变异的主要原因之一。     

苎麻疫霉有性生殖导致菌落形态和生长速率在无性后代持续变异的研究

研究了苎麻疫霉菌株ＪＳ－５自交Ｓ１代单卵孢株的菌落形态和生长速率在其单游动孢子无性系后代的遗传与变异。结果表明,亲本菌株ＪＳ－５的菌落形态和生长速率在连续４代单游动孢子无性系后代稳定遗传,而该菌株的自交Ｓ１代的约２／３单卵孢株的上述性状在其单孢无性后代中发生分离,且上述性状的分离在无性单孢后代中至少可连续保持３￣４代。本研究结果提示,苎麻疫霉有性生殖导致上述性状的变异,除有性生殖过程中的基因重组外     

苎麻疫霉有性生殖导致菌落形态和生长速率在无性后代持续变异的研究

研究了苎麻疫霉（PhytophthoraboehmeriaeSawada）菌株JS-5自交S1代单卵孢株的菌落形态和生长速率在其单游动孢子无性系后代的遗传与变异。结果表明,亲本菌株JS-5的菌落形态和生长速率在连续4代单游动孢子无性系后代稳定遗传,而该菌株的自交S1代的约2／3单卵孢株的上述性状在其单拘无性后代中发生分离,且上述性状的分离在无性单孢后代中至少可连续保持3～4代。本研究结果提示,苎麻疫霉有性生殖导致上述性状的变异,除有性生殖过程中的基因重组外,有性生殖还诱导产生了一种新的机制参与上述性状遗传与变异的调控。本研究还观察到供试菌株及其后代所产生的游动孢子绝大多数细胞内只含有1个细胞核,表明上述机制与异核现象无关。有性生殖导致生物学性状在随后的无性后代中发生持续变异可能是苎麻疫霉生物学性状遗传多样性的重要机制之一。     

苎麻疫霉有性生殖导致菌落形态和生长速率在无性后代持续变异的研究*

研究了苎麻疫霉（PhytophthoraboehmeriaeSawada）菌株JS-5自交S1代单卵孢株的菌落形态和生长速率在其单游动孢子无性系后代的遗传与变异。结果表明,亲本菌株JS-5的菌落形态和生长速率在连续4代单游动孢子无性系后代稳定遗传,而该菌株的自交S1代的约2／3单卵孢株的上述性状在其单拘无性后代中发生分离,且上述性状的分离在无性单孢后代中至少可连续保持3～4代。本研究结果提示,苎麻疫霉有性生殖导致上述性状的变异,除有性生殖过程中的基因重组外,有性生殖还诱导产生了一种新的机制参与上述性状遗传与变异的调控。本研究还观察到供试菌株及其后代所产生的游动孢子绝大多数细胞内只含有1个细胞核,表明上述机制与异核现象无关。有性生殖导致生物学性状在随后的无性后代中发生持续变异可能是苎麻疫霉生物学性状遗传多样性的重要机制之一。     

紫外光诱导苎麻疫霉抗甲霜灵突变及对苎麻疫霉生物学特性的影响

作者研究了紫外光对苎麻疫霉的抗甲霜灵诱变效应及对苎麻疫霉生物学特性的影响。结果表明,供试6个苎麻疫霉野生型菌株经菌丝块紫外光药剂诱变和菌丝块药剂驯化3周后,均获得抗甲霜灵突变体,且紫外光药剂诱变处理角变区出现频率明显高于药剂驯化处理,说明紫外光对苎麻疫霉抗甲霜灵突变有一定促进效应。紫外光显著地抑制苎麻疫霉游动孢子的萌发和菌丝的生长,在一定范围内,处理时间愈长,抑制率愈高。苎麻疫霉耐紫外光菌株(经亚致死剂量的紫外光照射处理后存活的游动孢子所形成的菌株)与野生型亲本相比,对温度和pH的敏感性大致相同,但菌落形态有一定变异,菌丝生长速率、卵孢子产生量均显著下降,表明紫外光对苎麻疫霉的菌丝生长和卵孢子产生量有明显的抑制作用。苎麻疫霉耐紫外光菌株EC50值比野生型亲本菌株EC50值提高了23.21%-56.70%,即耐紫外光菌株对甲霜灵敏感性比野生亲本菌株显著下降,这与紫外光诱变试验结果是相一致的。     

大豆疫霉菌抗甲霜灵特性研究

大豆疫霉Phytophthorasojae易对甲霜灵产生抗性,从大豆疫霉野生型菌株可诱变筛选到对甲霜灵有抗性的菌株Mtr。Mtr抗性菌株的抗性水平可达野生型单游动孢子菌株的870倍以上。Mtr性状在无性后代稳定遗传,在游动孢子后代连续三代未发生抗药性分离。大豆疫霉Mtr性状的保持对甲霜灵没有表现依赖性。Mtr单游动孢子菌株在不含甲霜灵的胡萝卜培养基(CA)平板上培养30d后对甲霜灵的抗性没有下降,其单游动孢子后代也未出现抗药性分离。     

苎麻疫霉生物学性状遗传与变异研究*

在实验室条件下研究了不同地区和不同寄主来源的苎麻疫霉（PhytophthoraboehmeriaeSawada）的菌落形态、菌丝线性生长速率（以下简称为生长速率）及同宗配合性状的遗传与变异,结果指出苎麻疫霉菌落形态和生长速率的遗传存在3种类型：（1）在单游动孢子后代和自交后代均可稳定遗传;（2）在单游动孢子后代稳定遗传而在自交后代发生变异;（3）在单游动孢子后代和自交后代均发生变异。结果表明,该菌菌落形态和生长速率的遗传具有多样性,上述两性状可以由细胞核杂合基因控制,也可以由细胞核纯合基因控制,还可能由细胞质因子控制。试验结果还指出,苎麻疫霉的同宗配合性状在单游动孢子和单卵孢后代均可稳定遗传,表明在供试的苎麻疫霉菌株中控制该性状的遗传因子是纯合的。     

苎麻疫霉生物学性状遗传与变异研究

在实验室条件下研究了不同地区和不同寄主来源的苎麻疫霉（PhytophthoraboehmeriaeSawada）的菌落形态、菌丝线性生长速率（以下简称为生长速率）及同宗配合性状的遗传与变异,结果指出苎麻疫霉菌落形态和生长速率的遗传存在3种类型：（1）在单游动孢子后代和自交后代均可稳定遗传;（2）在单游动孢子后代稳定遗传而在自交后代发生变异;（3）在单游动孢子后代和自交后代均发生变异。结果表明,该菌菌落形态和生长速率的遗传具有多样性,上述两性状可以由细胞核杂合基因控制,也可以由细胞核纯合基因控制,还可能由细胞质因子控制。试验结果还指出,苎麻疫霉的同宗配合性状在单游动孢子和单卵孢后代均可稳定遗传,表明在供试的苎麻疫霉菌株中控制该性状的遗传因子是纯合的。     

Aberrant genome size and instability of Phytophthora ramorum oospore progenies

The functionality of the sexual cycle in the heterothallic pathogen Phytophthora ramorum, causal agent of Sudden Oak Death, has recently been demonstrated. Sexual reproduction could create genotypic variation and increase the pathogen's ability to adapt to other host plants or changing environments. Genetic characterization using co-dominant microsatellite markers and flow cytometry of single-oospore progeny of crosses between a European A1 isolate and North American or European A2 isolates revealed a considerable number of non-Mendelian inheritance events. This includes inheritance of more than two alleles at a locus and non-inheritance of alleles from one parent at another locus. The progenies were mitotically unstable: zoospore and hyphal tip derivatives of the progenies showed genotypic rearrangements and phenotypic variation. Flow cytometry confirmed variation and instability in DNA content of the single-oospore progenies. This indicates that single-oospore progenies not only display aberrant genomic and phenotypic variation due to meiotic irregularities, but also extra variation as a result of post-meiotic genomic rearrangements.     

恶疫霉有性杂交后代的竞争能力研究*

５株杂交个体（携带对甲霜灵和地茂散的抗性标记）的游动孢子分别与其野生型亲本ＡＰ １４和ＰＫ９（对甲霜灵和地茂散均敏感）按１：１、１：９和１：４９比例混合涂布在１０％Ｖ６培养基平板上或接种在苹果片上生长４～５ｄ,并诱导产生孢子囊和释放游动孢子,经单孢分离建立单孢无性系,在含甲霜灵或地茂散的培养基上检测杂交个体的单孢后代所占的百分率。结果是,在Ｖ６培养基平板上混合生长的３０个组合中,仅３个组合杂交个体的单孢后代在其单孢无性系群体中所占的比例显著低于野生型亲本,其余组合均符合其各自的期望值;在苹果片上混合接种的３０个组合,杂交个体与野生型亲本的单孢后代所占比例均符合其各自的期望值。进一步测定２个杂交个体（ＡＰＫ－９和ＡＰＫ－２４）分别与其野生型亲本以１：１的比例混合后在Ｖ６培养基平板上和苹果片上连续３个产孢循环过程中两者比例的变化,结果是,在杂交个体与野生型亲本ＡＰ１４的组合中,杂交个体的后代所占的百分率由第一个产孢循环的４９．１％～５４．２％至第三个产孢循环下降为１．８％～６．２％：而在与另一野生型亲本ＰＫ９的组合中,杂交个体的后代所占的百分率从第一个产孢循环的４４．４％～５４．２％至第３个产孢循环上升为７２．１     

In vitro Selection of Strains of Phytophthora cactorum Resistant to Metalaxyl

Isolates of Phytophthora cactorum resistant to the systemic fungicide metalaxyl were obtained by exposing them to sequentially increased concentrations of metalaxyl. A linear relationship was observed between the concentrations of metalaxyl and percentage inhibition of mycelial growth of P. cactorum. The stability of metalaxyl-resistant isolates 150R and 250R was confirmed after six serial transfers on corn meal agar without fungicide. The in vitro metalaxyl-resistant isolate (Ph10) was less aggressive on apple rootstocks compared with the Ph07 isolated from metalaxyl-treated trees and the Ph03 isolated from untreated trees. Metalaxyl-resistant and sensitive isolates remained sensitive to the chemically unrelated fungicide fosetyl-Al at high concentration (600 μg/ml), to mancozeb, and to a mixture of metalaxyl + mancozeb. Significant differences in resistance to metalaxyl existed among P. cactorum field isolates.     

Effects of Metalaxyl, Fosetyl-Al, Dimethomorph and Cymoxanil on Phytophthora cactorum of Peach Tree

The activity of metalaxyl, fosetyl-Al, dimethomorph, and cymoxanil against Phytophthora crown rot of peach trees was examined. Application of fosetyl-Al or metalaxyl by painting the trunk (150 g/l) of 3-year-old PR204 trees was inhibitory to growth of the fungus but neither dimethomorph nor cymoxanil were effective. Application of metalaxyl as a soil drench suppressed canker development when the treated trees were subsequently inoculated with Phytophthora cactorum . Fosetyl-Al significantly reduced the growth of fungus compared with cymoxanil, dimethomorph and control but it was not as effective as metalaxyl. Again, dimethomorph and cymoxanil did not influence canker development. Both metalaxyl and fosetyl-Al were active for at least 21 days after applications. Strips of trunk bark were removed from trees, that had been drenched with the tested chemicals 20 days before, and inoculated on the cambium side with P. cactorum . Metalaxyl was the most effective fungicide and fosetyl-Al significantly reduced the development of fungus compared with dimethomorph, cymoxanil and the untreated strips. Colonization of strips treated with dimethomorph was significantly less than untreated strips. In contrast, cymoxanil did not inhibit the growth of fungus. Application of metalaxyl or fosetyl-Al as trunk paint or a soil drench appear to be effective procedures for preventing Phytophthora rot of peach trees.     

Seasonal variations in the activity in soil of Phytophthora cactorum, P. syringae and P. citricola in relation to collar rot disease of apple

Phytophthora species were isolated from infested orchard soils using apple fruit as bait at intervals during the periods July 1956 to August 1957, January 1957 to June 1958, and January 1969 to June 1970. When baited soils were kept out of doors P. cactorum and P. citricola were isolated only from April or May to October, when mean temperatures exceeded 8 and 10 °C respectively; P. syringae was isolated in all months except June, July and August.
The results did not suggest that the incidence of these species was particularly associated with apple as a host plant, but the periods of activity of P. cactorum and P. syringae in soils coincided closely with the periods when apple trees were susceptible to infection by either pathogen. With collar rot disease caused by P. cactorum it was considered that, the time of commencement of activity of the pathogen in the soil, together with the availability of water, might be critical in determining the severity of disease outbreaks.     

不同寄主来源的串珠镰孢生物学性状及其在无性后代的遗传与变异

自安徽、山东、湖北、江苏等地多点采集棉花红腐病、水稻恶苗病、玉米穗腐病的病组织,经分离、鉴定和纯化,获得107个串珠镰孢（Fusarium monilifoFine）菌株。对上述来源于棉花、水稻、玉米的串珠镰孢的菌落形态、生长速率和产孢量等生物学性状及其在无性后代的遗传与变异进行了研究。结果表明,不同寄主来源的串珠镰孢菌株的菌丝生长温度范围和最适温度大致相同,但在菌落形态特别是色素方面存在明显差异,生长速率和产孢量也存在显著差异。棉花菌株的平均生长速率最大,玉米菌株生长速率最小,水稻菌株生长速率居中,相同群体的不同菌株间生长速率有极显著差异;玉米菌株产孢量最大,棉花菌株产孢量最小,水稻菌株产孢量居中。方差分析显示,不同寄主菌株群体间产孢量存在显著差异,而同一寄主群体的不同菌株间产孢量均无显著差异,说明菌株产孢量大小主要与其寄主种类有关,而与地区来源关系不大。遗传测定结果表明,分离自棉花、玉米和水稻的串珠镰孢的菌落形态和生长速率在单分生孢子后代均可稳定遗传;产孢量性状遗传有两种情况：分离自棉花和水稻的串珠镰孢菌株Fm1和Fm31的产孢量性状在单分生孢子后代均可稳定遗传,而分离白玉米的串珠镰孢菌株Fm19的产孢量性状在单分生孢子第一代（CG1）发生变异。