栗疫病菌不同毒力菌株生物学特性的研究

 对不同毒力、含或不含dsRNA的菌株的生物学特性进行了研究。含dsRNA的弱毒力菌株的分生孢子后代,可分化出白色和桔黄色两种菌落类型,菌落白色的单孢菌株,可继续分化出白色和桔黄色两种类型的后代,桔黄色的单孢菌株保持稳定,不再发生分化。不含dsRNA的菌株,未见培养性状发生分化。对含dsRNA的弱毒力菌株的第2代两种类型的单孢菌株进行了毒力测定和dsRNA检测,桔黄色的单孢菌株的毒力水平达到或接近强毒力菌株的水平,不含dsRNA;而白色的单孢菌株的毒力很弱,且都含有dsRNA。对不同毒力菌株分生孢子的萌发率进行了比较,结果表明,孵育12h后,即有部分菌株的分生孢子开始萌发,强毒力菌株孢子的萌发最早,含dsRNA的弱毒力菌株的孢子萌发较迟;孵育24h后,各菌株孢子的萌发率均达到90%以上,差异已不太明显,但芽管的分枝程度有较大差异,强毒力菌株的芽管(菌丝)大量分枝,初步形成菌丝丛,而弱毒力菌株,不论是否含有dsRNA,芽管的分枝程度均较低。     

核盘菌中dsRNA种类及其与致病力的关系

 对源于我国黑龙江省佳木斯市同一块茄子田的14个核盘菌(Sclerotinia sclerotiorum)菌株及其中1个菌株Ep-1PNA5的2个衍生菌株的致病力和菌丝中的双链RNA(dsRNA)进行了分析。在马铃薯琼脂培养基(PDA)上对5个弱致病型核盘菌菌株(Ep-1PD、Ep-1PI、Ep-1PL、Ep-1PM、Ep-1PN)异常性状(菌丝生长慢,且异常分枝)的传染特性进行了测定。结果表明:在离体油菜叶片上,16个供试核盘菌菌株中,7个属于强致病类型,7个属于弱致病类型,2个属于中等致病类型。从10个核盘菌菌株的菌丝中检测到dsRNA因子,并可分成3类dsRNA电泳谱型。第一类dsRNA谱型只含有7.4kb大小的dsRNA分子,3个强致病型核盘菌菌株属于这种谱型;第二类dsRNA谱型含有2种dsRNA分子,大小分别为6.4kb和7.4kb。6个弱致病型菌株属于这种谱型;第三类dsRNA谱型只含有6.4kb大小的dsRNA分子。1个弱致病类型菌株属于这种谱型。从另外6个核盘菌菌株的菌丝中没有检测到任何dsRNA因子,其中4个菌株属于强致病型菌株,2个菌株属于中等致病型。可见,6.4kb大小的dsRNA因子与核盘菌弱致病性状密切相关。5个弱致病型核盘菌菌株(Ep-1PD、Ep-1PI、Ep-1PL、Ep-1PM、Ep-1PN)的异常性状可以通过菌丝接触传染给一些强致病型核盘菌菌株,使其菌丝生长变慢及分枝异常。结果还表明:这些弱致病型核盘菌异常性状的传染具有菌株特异性。     

小麦纹枯病菌中的dsRNA及其与菌株生物学特性的相关性

对采集自我国山东、河南、江苏、安徽4省的21株小麦纹枯病菌菌丝中的dsRNA进行了检测,研究了dsRNA条带与菌株菌落形态、生长速度、对小麦的致病力等生物学特性之间的相关性。结果表明,从16个菌株中检测到dsRNA条带,dsRNA的大小和数量存在丰富的多样性。致病力测定结果表明这21个小麦纹枯病菌菌株的致病力差异显著。目前暂未发现小麦纹枯病菌中dsRNA的类型和数量与菌株的菌落形态、生长速度及致病力等性状存在明显的相关性,但是致病力较弱的菌株中dsRNA条带的多样性更丰富。本研究为今后发掘小麦纹枯病菌中的低毒真菌病毒作了初步的探索。     

我国棉花黄萎病菌致病力分化及ISSR指纹分析

 来自我国12个省84个县（市）的棉花黄萎病菌,在PDA培养基上存在5种不同的培养类型,其中,产生微菌核较多的B型菌株为优势类群,占72.9%。长江流域的菌株培养性状变异最大,新疆棉区的变异最小。致病力测定结果和ISSR指纹图谱均将167个单孢菌株划分为强、弱、中3个致病力类型,供试菌株的ISSR指纹图谱与菌株的致病力存在明显的相关性。中等致病力类型菌株在我国占主导地位;强致病力类型的菌株主要分布在河北、河南、湖北等省;弱致病力类型菌株主要分布在新疆和江苏。     

大丽轮枝菌培养性状的变异

 对棉花,茄子、辣椒等植物上分离到的大丽轮枝菌(Verticillium dahliae kleb)的36个菌株进行了培养性状变异的研究。试验以分生孢子稀释培养的方法,检查单孢菌落的变异率.从野生型(A型)菌株分离到的92,947个单孢菌落,形成绒毛型(B型)菌落的变异率为5.85%,形成菌膜型(C型)菌落的变异率为0.07%。在绒毛型菌株的1,773个单孢菌落中,形成菌膜型菌落的变异率为0.19%。大丽轮枝菌野生型菌株培养性状的变异率,因试验菌株、单孢个体、营养条件、菌种保存温度和保存时间不同而异.绒毛型变异体的培养性状比较稳定.根据试验结果,探讨了菌种保存的方法.     

2013-2014年北方5省(区)致病疫霉抗药性监测及交配型分析

为明确致病疫霉对甲霜灵的敏感性及其交配型的动态变化,采用菌落生长速率法监测了2013-2014年采自我国北方5省(自治区)的马铃薯致病疫霉对甲霜灵的抗性,并利用标准菌株与待测菌株对峙培养测定了交配型.2013年和2014年检测的致病疫霉均以中抗菌株占主导,分别占当年检测菌株总量的82.7％和75.6％,但高抗菌株从3.3％增加到16.3％.测定甲霜灵对部分敏感、中抗和高抗菌株的EC50,发现该药剂对高抗菌株的EC50分别是中抗菌株和敏感菌株的5倍和400倍.2013年采自内蒙古、黑龙江、辽宁3省(自治区)6个县(市)的92个菌株中,A1交配型占主导地位,还发现了A2和A1A2;2014年采自内蒙古、黑龙江2省(自治区)4个县(市)的52个菌株仍以A1为主,A2所占比例较低,但未检测到A1A2.2013-2014年北方5省(自治区)致病疫霉群体结构呈现复杂化的趋势.     

梨果生刺盘孢子囊孢子单孢菌系的生物学特性及致病性研究

 梨果生刺盘孢的2种致病型菌株FJ-85(产生黑点症状)和菌株FJ-11-2(产生坏死斑症状)的子囊孢子单孢培养均可产生正、负2种类型的单孢菌系。正型单孢菌系菌落浅白色,其上形成的子囊孢子再培养,可形成正、负2种类型的单孢菌系;负型单孢菌系菌落深灰色,其上形成的子囊孢子再培养,只形成负型一种单孢菌系。2个菌株的单孢菌系中,负型的比例明显多于正型。2种类型的单孢菌系对峙培养,在菌落内及菌落交界处均可形成子囊壳,而同种类型的单孢菌系对峙培养仅在菌落内产生子囊壳。结果表明同种类型的单孢菌系可同宗配合,而正、负2种类型的单孢菌系还可异宗配合。正型单孢菌系的致病力与原始菌株相近,而负型单孢菌系较原始菌株弱。2个菌株的单孢菌系在翠冠梨上引起的症状差异与其原始菌株相同。研究结果为解析梨果生刺盘孢的有性生殖与致病的关系提供了新的有用信息。     

单克隆抗体检测栗疫菌dsRNA的研究初报

 本文利用dsRNA特异性单克隆抗体,采用双抗体夹芯ELISA法,检测了栗疫菌中的dsRNA.结果表明,该法不仅能从栗疫菌低毒力菌株的总核酸粗提液中检测到dsRNA而不受其它核酸的干扰,而且比电泳法简便迅速,在大量筛选含dsRNA的栗疫菌低毒力菌株中具有很大的实用价值。     

我国棉花黄萎病菌类型分化及培养特性研究

作者选用5个代表性的棉花黄萎菌系与美国落叶型黄萎病菌 T_9,进行比较试验。结果表明,落叶型菌系 V_B 与T_9的致病力强于非落叶型菌系。根据寄主反应型,划分出致病力强(V_B、T_9、V_6)、中(V_3、V_5)、弱(V_(10))三个类型;各菌系在33℃不能生长,在低温9℃和高温32℃均生长缓慢,表现适宜温度为20—30℃,最佳生长温度25℃,其中落叶型菌系比非落叶型菌系耐高温,在30℃下、菌落生长仍迅速;在培养性状上差异也很明显,落叶型菌系 V_B 与 T_9的菌落为白色,菌丝致密绒毛状,微菌核不明显,有成束状膨胀菌丝团;而非落叶型菌系 V_3、V_5、V_6、V_(10)菌落为黑色和灰黑色,菌丝絮状,有的稀松成束,微菌核呈串状葡萄形或似瘤状根须。鉴于落叶型黄萎菌系 V_B 与 T_9在三大棉种的6个品种上致病程度和对不同温度反应及培养性状等方面特性均表现基本一致的结果,进而肯定我国江苏常熟棉花黄萎菌系 V_B 是落叶型的致病菌。     

高毒力白僵菌的筛选及对花生甜菜夜蛾的致病力

为了筛选高毒力的生防白僵菌菌株,本研究对分离获得的4株白僵菌F1、F5、F6、F8的生物学特性进行分析,并在室内检测对甜菜夜蛾的致病力,最后通过大田试验验证了高毒力菌株对花生甜菜夜蛾的防治效果。结果表明,上述4株白僵菌的菌落形态、生长速率、产孢量等生物学性状及致病力存在显著差异。生长最快以及产孢量最高的是菌株F1,菌落呈粉状。菌株F6的分生孢子较其他菌株的分生孢子大。致病力检测结果表明,菌株F1对甜菜夜蛾的致病力最强,LC₅₀和LT₅₀分别是2.9383×10⁶ cfu/mL和5.79 d;其次是菌株F8,菌株F5和F6的毒力相对较弱。大田防效试验表明,菌株F1对花生甜菜夜蛾的防治效果可达75%以上,并在花生叶片具有较强的定殖能力。     

陕西苹果树腐烂病菌(Cytospora spp.)不同分离株的生物学特性与致病性研究

 本研究从陕西分离得到61株苹果树腐烂病菌(Cytospora spp.),选取在PDA培养基上菌落特征差异明显的7个分离株,对其生物学特性及致病性进行研究。结果表明:各分离株在PDA、PSA、PMA和PEMA培养基上菌落生长较好,但在PDA上的菌落颜色和产孢情况差异很大。各分离株均能在5~32℃生长,但对35℃以上高温的适应性差异较大;所有分离株均能在pH4~9的条件下生长,最适pH为5~6;光照对各分离株的菌落生长有明显的促进作用。各分离株均可在Cza-pek培养基及供试的其它碳、氮源培养基上生长,其中以葡萄糖、麦芽糖和酵母膏为最佳碳、氮源。采用烫伤接种的方法,分别以菌饼和分生孢子悬浮液接种苹果离体枝条,两者均可侵染发病,但各分离株致病力差异显著,其中菌落颜色为黄褐色的菌株致病性强。因此,根据菌落颜色、致病性和生物学特性可将这7个分离株划分为3个类群:Ⅰ型为黄褐色强致病类群,但各分离株的产孢量有差异;Ⅱ型为乳白色不产孢的弱致病类群;Ⅲ型为灰褐色易产孢弱致病类群。其余54株分离物均属于Ⅰ型,这些结果说明陕西省苹果树腐烂病菌具有多样性和致病性分化现象。     

甘薯地土壤线虫群体分布规律及取样方法研究

在5年连作甘薯地,于甘薯移栽前和移栽后的不同时间,调查甘薯穴中和穴间、株间和行间以及各样点不同土层线虫的群体数量。分析明确了植物寄生线虫和非植物寄生线虫群体的水平分布和垂直分布规律,提出了研究和了解甘薯地土壤线虫发生动态的调查取样方法。     

Components of spore production in apple powdery mildew (Podosphaera leucotricha)

The morphology of powdery mildew ( Podosphaera leucotricha) colonies of known age was studied on six apple cultivars inoculated in a glasshouse. Colonies were observed first on the older leaves. Mean disease incidence ranged from about 20% for cv. Laxton's Superb to about 45% for cv. Crispin. Conidiophore density (number of conidiophores/mm² colony) was higher on younger than on older leaves, higher on the upper than on the lower leaf surface, and decreased with the age of the colony. The number of hyphal interceptions on a line transect was higher on the upper surface but the ratio of conidiophore density to hyphal interceptions was greater on the lower surface and did not differ with colony age or cultivar. The number of conidia/conidiophore was higher on the younger leaves and on the upper surfaces and was at a maximum 7-12 days after the first sign of the colony. The number of conidia/mm² colony was higher on lower surfaces, especially of the younger leaves. There was generally an associated variation between cultivars in components of spore production.     

Evaluation of a DNA probe for the detection of Erwinia herbicola strains pathogenic on Gypsophila paniculata

A 7.5-kb DNA fragment carrying genes for indole acetic acid biosynthesis in Erwinia herbicola pv. gypsophilae was used as a DNA probe to detect gall-forming E. herbicola strains. A quick colony hybridization procedure was developed to detect E. herbicola pv. gypsophilae in cuttings of gypsophila, and was compared with ELISA and pathogenicity tests. The sensitivity of the colony hybridization procedure was sufficient to detect less than 100 colony-forming units after enrichment culture. In mixed cultures, with saprophytes associated with the gypsophila plant, about 10⁴ CFU/ml of the pathogen were detectable. The colony hybridization is specific to gall-forming E. herbicola strains, and is relatively easy to perform. The advantages of using the colony hybridization procedure for practical purposes, compared with ELISA and pathogenicity tests, are discussed.     

Histological Characterization of Resistance to Uromyces viciae-fabae in Faba Bean

ABSTRACT Components of resistance to the faba bean rust (Uromyces viciae-fabae) were studied at the histological level in seedlings and adult plants of nine faba bean (Vicia faba) lines differing in their level of resistance. Resistance of these lines was previously shown to be characterized macroscopically by an increased latent period, a decreased colony size, and a relatively decreased infection frequency. In some lines, the resistance also was associated with macroscopically visible necrosis. Histological investigations revealed few differences in spore germination and appressorium formation. Significant levels of aborted stomatal penetration by the rust fungus were found on all resistant lines. However, differences among lines were more evident once the stomata were penetrated by the infection structures. Resistance was mainly due to a restriction of haustorium formation with varying levels of early abortion of the colonies, a reduction in the number of haustoria per colony, and smaller colony size. In addition, necrosis of the host cells associated with infection hyphae was detectable in some lines from the beginning of colony development. This microscopically visible necrosis became stronger from 4 days after inoculation, resulting in a reduced growth of the colony. Differences in resistance levels were more marked in adult plants than in seedlings.     

广西防城港首次截获苜蓿黄萎病菌

从一批进境的美国苜蓿草样品中保湿分离得到了3株与苜蓿黄萎病菌(Verticillium albo-atrum)相似的目标菌VA1、VA2、VA3.该目标菌在PDA上生长迅速,菌丝生长0.8～1.0 mm/d,2～3 d即可生成肉眼可见的菌落斑点;菌落为白色圆形,边缘规则呈圆形,菌丝较致密,气生菌丝较少,菌落中心乳白色奶冻状;培养4～5 d即后产生气生典型轮枝状分生孢子梗,7～8 d后菌落中央表面因产生休眠菌丝开始变成黑褐色至黑色,20 d后菌落的表面和背面大部分均变黑色,仍不产生微菌核和厚垣孢子.该目标菌的纯培养用V.albo-atrum特异引物Vaa1/Vaa2进行检测,PCR 扩增后得到预期330 bp的产物片段,产物序列与V.albo-atrum相应序列的相似性为100％.从而鉴定确认该目标菌为苜蓿黄萎病菌V.albo-atrum.     

三种杀真菌剂对金龟子绿僵菌产孢及生长的影响

三种杀真菌剂在一定浓度范围为（氯硝胺在１００×１０￣（－６），孟加拉红在５００×１０￣（－６）和柯赛得在２０００×１０￣（－６）内）对金龟子绿僵菌的菌落形成没有明显影响，而柯赛得（含５０％氢氧化铜）和孟加拉红分别在１００×１０￣（－６）和５００×１０￣（－６）浓度时显著地增加金龟子绿僵菌的产孢量，柯赛得还具有缩短产孢时间的作用。氯硝胺在１～１００×１０￣（－６）浓度范围内，随着浓度增加，对产孢能力和菌落大小的抑制作用加强。柯赛得和孟加拉红随浓度的增加对菌落的抑制作用也有所加强，但没有像氯硝胺的抑制作用那么强。     

Incidence and density relationships of powdery mildew on apple

ABSTRACT The relationships between disease incidence and colony density and between leaf and shoot disease incidences for apple powdery mildew were investigated over four seasons in order to derive a simple relationship for predicting density using incidence. The Neyman type A distribution generally provided a good fit to the observed number of colonies per leaf and shoot, and provided a significantly better fit than the Poisson distribution, indicating a degree of aggregation of mildew colonies. In general, Taylor's power-law satisfactorily described the observed variance-mean relationship for colony density; however, Taylor's power-law broke down at very high levels of mean density. Incidence of leaf infection could be determined based on average number of colonies per leaf assuming a fixed variance-mean relationship and a Neyman type A distribution for colony density. Regression models using the complemen- tary log-log transformation of incidence also provided accurate predictions of leaf (or shoot) disease incidence from colonies per leaf (or per shoot). Similar accuracies of these incidence-density models suggested that variance-mean ratio of colony density was more or less constant over time. Unlike the case with colony density, the number of mildewed leaves per shoot generally had a random pattern, as indicated by the good fit of the binomial distribution. Thus, it was possible to estimate the leaf incidence of the youngest unrolled leaves on a shoot using the shoot incidence. It is argued that the leaf incidence-density relationships developed in the present study may be used in making practical disease management decisions.     

Measuring the sterol content of barley leaves infected with powdery mildew as a means of assessing partial resistance to Erysiphe graminis f.sp. hordei

Barley leaf segments of several partially resistant cultivars were inoculated with different strains and combinations of strains of Erysiphe graminis. Infection level was assessed by counting colony number, by measuring sporulating area using an image analyser, and by extracting and quantifying sterols. Good correlations were obtained between data from all three methods. One cultivar contained less sterol than expected from the number of colonies present but some of the resistance in that cultivar was known to be due to reduced colony size. Image analysis was unable to discern the reduced colony size component of resistance. Scanning electron microscopy revealed that single colonies may originate from more than one conidial infection. Sterol analysis was the most representative method of comparing the reaction of diverse host genotypes to mildew infection.     

保护性耕作方式对土壤微生物生理类群和酶活性的影响

通过设置在甘肃省定西市安定区李家堡乡的田间定位试验,研究了不同耕作方式下（NTS、TS、NT、T）土壤微生物生理类群数量和酶活性及其相关性。结果表明：与传统耕作（T）相比,保护性耕作方式（NTS、TS、NT）能显著增加土壤微生物生理类群数量,增强酶活性。不同的保护性耕作方式下,土壤微生物生理类群数量和酶活性存在显著差异...