运用随机扩增多态性DNA标记检测中国东北大豆灰斑病菌株遗传变异

 运用致病力和DNA多态性检测中国东北地区的35个大豆灰斑病菌分离物的遗传变异、根据菌株在9个品种(系)上的致病力反应可将其分为7个组。利用13个随机引物扩增供试菌株共计产生105个RAPD标记,其中78.1%具有多态性。通过聚类分析计算了各菌株间的遗传距离,并产生树状图,发现同一地区内及不同地区间的病菌表现遗传变异、致病性和DNA多态性间具有一定相关性。     

稻瘟菌致病力变异初探

 从保存的4个单孢分离菌后代再分离若干单胞菌株,接种子国内7个鉴别品种上,又可以分成不同的小种。从6个单孢分离菌再分离的后代单孢菌株,分别对13个抗病品种作致病力测定,各后代单孢菌株仅对少数品种的致病力是一致的。从27个品种的小病斑或个别大病斑上分离的病菌,回接原品种后,有11个分离菌对原品种不致病。这些结果说明大多数分离菌对许多品种的致病力是易变的,只对少数品种的致病力保持相对稳定性。作者认为进行抗性遗传和抗性机制研究需要的致病力稳定分离菌,只能根据其对供试品种致病力的稳定性来选择,而不能按一组鉴别品种来选择。     

辽宁省稻曲病菌遗传多样性和致病力分析

为有效防治辽宁省稻曲病菌Ustilaginoidea virens,利用重复序列PCR(repetitive elementbased PCR,rep-PCR)分子指纹技术,对2017年自辽宁省8个市8个主产稻区采集的51株稻曲病菌菌株进行遗传多样性和致病力分析。结果显示,在3对引物中,以BOX1/BOX2和ERIC1/ERIC2为引物扩增的DNA指纹图谱的遗传多样性值分别为0.764、0.707,均大于0.7,故选择这2种引物扩增的DNA指纹图谱进行遗传多样性分析;当DNA指纹相似系数为0.78时,以BOX1/BOX2为引物和以ERIC1/ERIC2为引物扩增的DNA指纹图谱分别将供试菌株划分为12个和10个遗传类群;供试菌株致病力可划分为弱致病型、中等致病型和强致病型3个致病型,所占比例分别为33.33%、58.82%和7.85%,强致病型菌株仅在沈阳市、鞍山市和大连市出现;所有优势类群均包含3种致病型菌株。表明辽宁省稻曲病菌遗传结构复杂,不同地理来源的稻曲病菌菌株致病力存在一定差异,相同致病型的稻曲病菌菌株分属于不同的遗传类群,同一遗传类群中包含不同的致病型菌株。     

水花生病原菌——蕉斑镰刀菌菌株的筛选及其致病性测定

蕉斑镰刀菌 Fusarium stoveri是从自然界水花生植株上分离的、可致水花生表现萎蔫、地上茎腐烂、叶片上呈现褐色斑或黄化的生防真菌.室内研究表明,蕉斑镰刀菌菌丝生长和产孢的适宜温度范围在25～28℃,多数菌株的最适温度为28℃;菌株间的产孢量有较大差异.在供试菌株中,32-6菌株的产孢量最大;供试的9个蕉斑镰刀菌菌株对水花生均有较强的致病力,但菌株间的致病力有所差异.根据菌株的生长速率、产孢量和致病力等因素,菌株32-6被选为水花生生防菌的优选菌株.采用生产上常见农作物、蔬菜、经济作物及物候上与水花生一致的杂草等42种植物对菌株32-6进行寄主专化性测定,结果表明,蕉斑镰刀菌菌株32-6具有高度的寄主专化性,只对水花生表现出较强的致病性,对其他植物均不致病.     

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。     

黑龙江省马铃薯干腐病菌种类鉴定及致病性

本研究将采自黑龙江省不同地区的马铃薯干腐病病样进行分离和病原菌纯化,得到27个镰刀菌菌株,通过致病性鉴定,其中的18个菌株具有致病性。运用培养性状和形态特征综合分析的方法,对上述18个菌株进行鉴定,结果显示为6种镰刀菌,分别为拟枝孢镰孢(Fusarium sporotrioides)、茄镰孢(F.solani)、接骨木镰孢(F.sam-bucinum)、拟丝孢镰孢(F.trichothecioides)、燕麦镰孢(F.avenaceum)和茄病镰孢蓝色变种(F.solanivar.coerule-um)。同时对上述6种镰刀菌进行致病性测定,结果表明不同种类镰刀菌致病性不同,以接骨木镰孢、燕麦镰孢和拟丝孢镰孢致病力最强,拟枝孢镰孢致病力最弱。     

洛阳市牡丹灰霉病病原菌的鉴定

为明确洛阳市牡丹灰霉病的病原菌种类,于2015年从该地区的4个牡丹Paeonia suffruticosa Andr.种植区采集灰霉病样品,采用组织分离法对病原菌进行分离和纯化,结合形态学和基因序列分析对分离物进行鉴定,采用针刺接种法测定不同分离物对牡丹离体叶片的致病性。结果表明,从灰霉病样品中分离获得40株分离物,这些分离物分别属于Ⅰ、Ⅱ、Ⅲ型菌,I型菌不易产孢但产菌核;Ⅱ型菌易产孢,后期产生少量菌核;Ⅲ型菌易产孢,且产生大量菌核。产孢分离物均形成直立状分生孢子梗,孢子梗分枝顶端聚生葡萄穗状分生孢子,分生孢子卵圆形或长卵圆形。供试菌株的ITS序列和G3PDH基因序列与灰葡萄孢Botrytis cinerea Pers.的同源性达到99%;致病性测定结果表明,各菌群代表菌株对牡丹均有致病性,但不同菌群间致病力有差异。研究表明,引起洛阳市牡丹灰霉病的病原菌为灰葡萄孢,且菌群类型多样。     

东北地区烟草靶斑病菌(Rhizoctonia solani)融合群、致病力分化及品种抗病性研究

2013-2014年吉林省、黑龙江省烟区首次大面积发生烟草靶斑病,损失严重。采集吉林省柳河县、黑龙江省林口县和宾县的烟草靶斑病病叶进行分离纯化,得到了12个菌株。将所获菌株分别与立枯丝核菌标准融合群菌株AG-2、AG-3、AG-4进行对峙培养,结果表明:此12个菌株均与立枯丝核菌AG-3标准菌株发生融合反应,不与其他融合群发生融合;随机选取3个县各1个代表性菌株,提取菌丝基因组DNA并对其ITS序列进行分析与比对,菌株HLJ-2、JL-1、HLJ-7的ITS序列与辽宁省烟草靶斑病菌菌株LF-2、LJT-8(AG-3融合群)的同源性达99%~100%,因此推断,吉林省、黑龙江省烟草靶斑病菌与辽宁省烟草靶斑病菌应属于同一个融合群,即立枯丝核菌AG-3融合群(Rhizoctonia solani AG-3)。根据各菌株在鉴别寄主上表现的抗性不同,可将吉林省、黑龙江省所有菌株分为3个致病类型,即致病类型Ⅰ、致病类型Ⅱ和致病类型Ⅲ。同一省份不同菌株间致病力有差异,黑龙江省菌株3种致病类型均有,以致病类型Ⅱ为主;吉林省菌株只有致病类型Ⅰ和致病类型Ⅱ。分别选取3个省的强致病力菌株,用离体叶片接种法鉴定了我国21个主栽烟草品种对靶斑病的室内抗病性,结果表明:没有免疫或抗病品种,‘龙江981’、‘NC297’对供试的3个强致病力菌株均表现中抗,‘中烟202’、‘云烟97’、‘NC55’、‘KRK26’和‘G28’分别对不同菌株表现中抗。     

Rep-PCR技术对中国水稻条斑病菌的遗传多样性初析

 采用Rep-PCR技术,对30个水稻细菌性条斑病菌株(Xanthomonas oryzae pv.oryzicola)进行遗传多样性分析,同时对李氏禾条斑病菌等其它10个参试菌株也进行了比较。Rep-PCR是利用一些基于细菌的短的重复序列引物(ERIC和BOX)的DNA扩增特性,2种引物组合的电泳图谱结合并分析,以水稻细菌性条斑病菌各自的指纹谱型在相似率80%时可分为6簇,初步表明我国水稻细菌性条斑病菌群体的遗传分化明显;发现自然界存在的弱或无毒性菌株与毒性菌株的Rep-PCR指纹图谱差异很大;毒性菌株的遗传分簇与其致病性具有一定的相关性。用ERIC扩增水稻条斑病菌基因组DNA的指纹比BOX更为多样,两者对菌株的分辨率不同。因此,Rep-PCR技术可有效地用于监测水稻细菌性条斑病菌的遗传变异,还可应用于菌株的鉴定和分类学研究。     

甘肃省辣椒疫霉菌的交配型分布及其致病力差异

采用单孢菌落直接配对法和游动孢子悬浮液土壤接种法对2003—2006年采自甘肃省辣椒主产区的辣椒疫霉菌菌株Phytophthora capsici进行交配型和致病力测定。结果表明,甘肃省辣椒疫霉菌存在A1、A2和A0三种交配型,以A1和A0占优势,在各产区均有分布,发生频率分别达57.9%和31.6%;A2交配型仅2株,分别出现在酒泉和庆阳。不同来源菌株对5个辣椒品种的致病力存在显著差异,平均病情指数为2.2～56.4,强、中、弱致病性菌株分别占10.6%、52.6%和36.8%,不同地区的菌株致病力有强弱之分,以中、弱致病性菌株为主,同一地区中不同菌株的致病力也存在一定的差异性。     

抗稻瘟病单基因系对籼稻稻瘟病菌小种鉴别力分析

 利用30个抗稻瘟病单基因系对146个来自广东的稻瘟病菌单孢分离菌株进行了致病性测定,其中17个单基因系对测试菌株的致病性有较好的鉴别力。基于主成分因子分析法,对17个单基因系和146个菌株组成的互作变量矩阵进行了因子提取,结果表明其中9个单基因系为代表的抽提因子可以解释变量总方差的80.437%,初步组建了这9个单基因系构成的稻瘟病菌生理小种单基因系鉴别寄主。按其对变量方差贡献大小排序,它们分别是:IRBLkp-K 60(Pi-k^p)、IRBLi-F5(Pi-i)、F128-1(Pi-ta²)、IRBL9-W (Pi-9(t))、IRBLsh-S (Pi-sh(1))、IRBLz-Fu (Pi-z)、IRBLkh-K3(Pi-k^h)、IRBL1-CL (Pi-1)、IRBLz5-CA (Pi-z⁵)。用这9个单基因系可将146个菌株划分为83个小种,将来自广西、福建、湖南、湖北、浙江、四川等籼稻区的54个菌株划分为26个小种。筛选的9个单基因系对我国籼稻区稻瘟病菌致病性有较强的鉴别力。在抗病性鉴定中,基于单基因系的代表菌株选择,可考虑应用聚类和变量分析法进行。     

Host Specificity of Phytophthora infestans on Tomato and Potato in Ecuador

ABSTRACT Sixty Ecuadorian isolates of Phytophthora infestans from potato and 60 isolates from tomato were compared for dilocus allozyme genotype, mitochondrial DNA haplotype, mating type, and specific virulence on 11 potato R-gene differential plants and four tomato cultivars, two of which contained different Ph genes. Restriction fragment length polymorphism (RFLP) fingerprints of subsamples of isolates from each host were compared by using RG57 as the probe. All potato isolates had the allozyme genotype, haplotype, and mating type of the clonal lineage EC-1, which had been previously described in Ecuador. With the same markers, only one isolate from tomato was classified as EC-1; all others belonged to the globally distributed US-1 clonal lineage. RFLP fingerprints of isolate subsets corroborated this clonal lineage classification. Specific virulence on potato differentials was broadest among potato isolates, while specific virulence on tomato cultivars was broadest among tomato isolates. Some tomato isolates infected all tomato differentials but no potato differentials, indicating that specific virulence for the two hosts is probably controlled by different avirulence genes in P. infestans. In two separate experiments, the diameters of lesions caused by nine isolates from potato and 10 from tomato were compared on three tomato and three potato cultivars. All isolates produced larger lesions on the host from which they were isolated. No isolates were found that were highly aggressive on both tomato and potato. We conclude that there are two different populations of P. infestans in Ecuador and that they are separated by host.     

粳籼89穗颈瘟分离菌株的致病力及遗传宗谱研究

通过对粳籼89的穗颈瘟分离菌株的致病力及遗传宗谱研究,结果表明:(1)粳籼89分离菌株的小种类型复杂,既有籼型小种如ZA1、ZB1、ZB5、ZB13、ZC13等,又有粳型小种如ZD5、ZG1等;(2)该类菌株的致病力存在不稳定性,同一菌株不同时间接种,鉴别寄主上表现的小种类型不同,对其他主栽水稻品种或抗源的致病力也表现不同,但这些菌株无论何时接种到粳籼89上均能使其表现感病,病级在4级以上;(3)该类菌株接种到粳籼89衍生品种或其他粳籼杂交后代上,一般能侵染这类品种;(4)利用RFLP技术,采用探针MGR586与限制性内切酶EcoRI组合对病菌进行DAN指纹分析,结果16个粳籼89分离菌株被分在2个相邻的遗传宗谱里,即宗谱1(9个菌株)和宗谱2(7个菌株),这两个宗谱恰好是广东的优势宗谱。     

华南籼稻稻瘟病菌致病型单基因鉴别寄主筛选

采用35个单基因鉴别寄主、12个日本清泽鉴别寄主共47个具有已知基因的鉴别寄主,对147个籼稻稻瘟病菌株进行致病型鉴别研究.147个菌株被划分为28个不同的致病类型.优势致病型为I-04、I-02和I-01,这3个致病型的菌株数占参试菌株的74.8%,而且与其他致病型比较,它们的致病谱较广.研究结果表明,F80-1(Pi-k)、IRBL7(Pi-kp)、IRBL16(Pi-sh(1))、IRBL21(Pi-7(t))、IRBL3(Pi-i)、IRBL9(Pi-z)、IRBL19(Pi-3)、IRBL10(Pi-z5)、IRBL11(Pi-zt)、IRBL18(Pi-1)、IRBL10(Pi-2)、F128-1(Pi-ta2)、F145-2(Pi-b)和IRBL22(Pi-9(t))等14个单基因鉴别寄主对以广东为主的华南籼稻区稻瘟病菌具有较好的鉴别能力,其中尤以F80-1(Pi-k)、IRBL7(Pi-kp)、IRBL16(Pi-sh(1))、IRBL21(Pi-7(t))、IRBL3(Pi-i)、IRBL9(Pi-z)、IRBL19(Pi-3)和IRBL10(Pi-z5)等8个单基因鉴别寄主的鉴别能力最强,IRBL11(Pi-zt)、IRBL18(Pi-1)、IRBL10(Pi-2)、F128-1(Pi-ta2)、F145-2(Pi-b)和IRBL22(Pi-9(t))等6个单基因鉴别寄主可鉴别抗性反应表现型较相似的致病型,并可用于细分致病型.3个优势致病型I-04、I-02和I-01对F80-1(Pi-k)等上述8个单基因鉴别寄主的抗性反应表现型依次为RRSRSSSMR、SSSSSSSS和SSRSSSSR.     

Genetic Diversity in South American Colletotrichum gloeosporioides Isolates from Stylosanthes guianensis, a Tropical Forage Legume

The degree of genetic diversity of 127 Colletotrichum gloeosporioides isolates from Stylosanthes guianensis genotypes in South America was measured at the molecular level by random amplified polymorphic DNA (RAPD) with nine arbitrary primers of 10 bases, and by restriction fragment length polymorphism (RFLP) with a non-LTR (long terminal repeats) retrotransposon DNA sequence. The RAPD products revealed scorable polymorphism among the isolates, and a total of 80 band positions were scored. Sixty-three of the 127 isolates were clustered into 13 distinct lineages usually correlating with geographic origin. Where isolates from various regions were clustered together, most had identical host genotype origin. The pathogen population sampled from Carimagua, Colombia, a long-time Stylosanthes breeding and selection site, with a savanna ecosystem, was highly diverse. A set of 12 S. guianensis genotype differentials was used to characterize pathogenic variability of 104 isolates and their virulence patterns were grouped into 57 pathotypes. However, when they were tested on four Australian differentials, they grouped into 11 pathotypes. As shown in previous studies, no strict correlations existed between genetic diversity measured by RAPD or RFLP, and pathotype defined by pathogenicity pattern on the differentials. Southern blot analysis of the 127 isolates revealed 23 hybridizing fragments, resulting in 41 fingerprint patterns among the 127 isolates. Relationships between RFLP and RAPD variables were examined using Spearman's Rank Correlation Coefficient, which showed that the two measures of genotypic variation are in agreement.     

利用PCR标记对水稻稻瘟病菌适合度的测定

 本文利用2个对水稻抗病基因Pi-1(t)有致病力的突变菌株和它们的各自原始菌株共同接种高度感病的品种CO39。第一轮接种后,采集病叶并收集分生孢子用于下一轮接种,同时分离单孢菌株。经过3或4轮接种后,共分离905个单孢菌株。然后,利用稻瘟病菌的一段倒位重复序列Pot2所设计的一对引物,Pot2-1和Pot2-2,直接进行DNA聚合酶链式反应(PCR),检测各菌株在群体中所占比例,结果表明,2个突变体均比各自的原始菌株有更高的适合度,突变体RC1(4+5)141与原始菌株9248-6组合中,前者由第1轮接种循环的71%上升到第4轮的100%;突变体MC1(3+4)57-9与原始菌株101-7-2-1-1组合中,前者在各代中均比原始菌多一倍。2个突变体适合度也相差很大,经过3代接种循环后,MC1(3+4)57-9则逐渐消失,RC1(4+5)141完全占优势,达到100%。突变体RC1(4+5)141的产孢能力是其它菌株的2~3倍,这表明,稻瘟病菌菌株在自然条件下成为优势群体,除了致病力发生突变外,还要在产孢能力上有明显的优势以适应新的抗病基因     

Characterisation ofFusarium oxysporum isolated from carnation in Australia based on pathogenicity,vegetative compatibility and random amplified polymorphic DNA (RAPD) assay

Isolates ofF. oxysporum collected from symptomless carnation cuttings from Australian carnation growers properties, together with isolates from national collections, were screened for pathogenicity and grouped according to vegetative compatibility and random amplified polymorphic DNA (RAPD) patterns. The collection of 82 Australian isolates sorted into 23 different vegetative compatibility groups (VCGs). Of 69 isolates tested for pathogenicity, 24 were pathogenic to carnations, while the remaining 45 were non-pathogenic. All pathogenic isolates were within two VCGs, one of which was also compatible with an isolate obtained from an international culture collection, and which is known to represent VCG 0021 and race 2. Race status of the two pathogenic VCGs remains unknown. The RAPD assay revealed distinct DNA banding patterns which could distinguish pathogenic from non-pathogenic isolates as well as differentiate between isolates from the two pathogenic VCGs.     

Selection for Pathogenicity to Strawberry in Populations of Colletotrichum gloeosporioides from Native Plants

ABSTRACT Colletotrichum gloeosporioides causes a serious crown rot of strawberry and some isolates from native plants are pathogenic to strawberry. C. gloeosporioides from lesions on wild grape and oak were sampled at two sites adjacent to commercial strawberry fields in Florida and two distant sites. Random amplified polymorphic DNA (RAPD) marker data and restriction enzyme digests of amplified rDNA were used to determine whether isolates were from the same C. gloeosporioides subgroup that infects strawberry. There were 17 to 24 native host isolates from each site that clustered with a group of strawberry crown isolates based on RAPD markers. Among strawberry isolates, there were two rDNA genotypes identified by restriction enzyme analysis. Both genotypes were present among native host isolates sampled from all four sites. There was some evidence that the different rDNA genotypes differentiated two closely related subpopulations, although the proportion of pathogenic isolates from native hosts among the two different genotypes was not different. The incidence of isolates pathogenic to strawberry was greater at sites close to strawberry fields relative to sites distant from strawberry fields for isolates with a BstUI(-)/MspI(+) rDNA genotype (44 versus 13%), a BstUI(+)/MspI(-) genotype (57 versus 16%), or when both genotypes were analyzed together (46 versus 15%). Based on these results, it appears that the C. gloeosporioides subgroup that causes crown rot on strawberry is widely distributed in Florida and that selection for pathogenicity on strawberry occurs in the area where this host is grown in abundance.