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1.
由致病疫霉(Phytophthora infestans)引起的马铃薯和番茄的晚疫病是一种世界范围内的重要病害。在致病疫霉与寄主的互作过程中,效应因子对于病原物能否成功入侵与定殖起到了关键作用。本研究检测了来自中国马铃薯主产区的37株致病疫霉菌中编码Avr3a, Avr4和IPI-O三种效应因子的基因组成,研究发现来自国内的所有供试菌株中只有1株分离自马铃薯的和2株分离自番茄的致病疫霉含有无毒基因Avr3a(编码Avr3aKI),其余菌株均含有avr3a(编码Avr3aEM);所有菌株中都仅含有编码截断蛋白的avr4;从37株国内菌株中共检测到了25种ipiO变异型,其中20种为新变异型,聚类分析的结果表明这些变异型仍属于已报道的三类ipiO,而且大部分ipiO变异型为无毒型,可以被抗性基因Rpi-blb1识别。该研究结果表明avr3a和avr4普遍存在于中国致病疫霉菌株中,推测大部分菌株可以克服R3a和R4抗性基因;而ipiO的无毒变异型仍然普遍存在于国内致病疫霉群体中,可以被马铃薯含有的抗性基因Rpi-blb1识别。 相似文献
2.
由致病疫霉引起的马铃薯晚疫病是马铃薯生产中最具危害性的病害,明确现有马铃薯品种(品系)中抗病基因组成情况对于指导抗病品种合理布局及抗病育种具有重要意义。本研究利用农杆菌介导的无毒基因瞬时表达技术分析了致病疫霉8个无毒基因在29个马铃薯品种(品系)中的分布情况。结果表明,品种间抗病基因组成各异,平均每个品种含有4个抗病基因。同时,不同抗病基因在品种中的分布频率也不同,抗病基因R1(76%)、Rpi-blb1(66%)和Rpi-blb2(66%)在品种中分布较普遍;而含抗病基因R3b(45%)、R4(38%)和Rpi-vnt1(34%)的品种较少。此外,品种中所含抗性基因的数量和品种室内抗性程度呈显著正相关(r=0.915 6,P=0.029 1)。 相似文献
3.
为明确宁夏回族自治区马铃薯主产区晚疫病菌的交配型种类以及生理小种的类型、组成和分布,为宁夏有针对性地选育马铃薯抗晚疫病品种提供科学依据。利用A1、A2交配型标准菌株和含有11个主效抗性基因的鉴别寄主,对2018年-2019年从原州区、泾源县、彭阳县、隆德县、西吉县、盐池县、海原县采集得到的130个马铃薯晚疫病菌菌株进行交配型和生理小种鉴定。结果表明,宁夏马铃薯晚疫病菌群体的交配型和生理小种存在多样性,采集地交配型有A1、A2、SF(自育型)3类,分别占被测菌株的30%、63.8%、6.2%;生理小种有8种类型,其中生理小种1.2.3.4.5.6.7.8.9.10.11发生频率最高,占被测菌株总数的44.62%,是宁夏马铃薯主栽区的晚疫病菌优势小种,在各测试地均有分布;次优势小种为3.4.10和1.2.3.5.6.7.8.11,发生频率分别为12.31%和10.77%。宁夏不同种植区域马铃薯晚疫病菌交配型组成复杂,采集地组成差异较大,但西吉县、原州区交配型年度间的变化基本趋于一致。宁夏马铃薯晚疫病菌群体组成日趋复杂,生产中要合理布局已有抗病品种,挖掘培育抗病水平高的新品种,综合防控马铃薯... 相似文献
4.
马铃薯晚疫病的病原是致病疫霉菌(Phytophthora infestans),该病原菌在侵染马铃薯过程中分泌大量RxLR型效应子,但目前绝大多数RxLR效应子的功能和作用机制尚不明确。本研究成功克隆了致病疫霉菌的一个RxLR效应子PITG_16427.2,在本氏烟中瞬时表达PITG_16427.2,发现该效应子能够抑制6种激发子(INF1、PsojNIP、BAX、SIF2、Avh238、Avh241)激发的植物免疫反应。进一步研究发现,效应子PITG_16427.2在晚疫病菌侵染马铃薯早期上调表达。在15个致病疫霉菌株和3个同属菌株中克隆该基因,克隆到氨基酸序列一致性超过93%的同源基因,这些同源基因均能在本氏烟中抑制INF1和Avh241引起的HR,揭示了该效应子在病原卵菌中序列和功能的高度保守性。在本氏烟和马铃薯感病品种Désirée中瞬时表达PITG_16427.2,发现该效应子能够显著促进晚疫病菌的侵染。通过qRT-PCR方法检测发现,乙烯信号的相关基因ERF1显著上调,而水杨酸信号相关基因PR1b显著下调,表明PITG_16427.2在晚疫病菌侵染过程中可抑制寄主的SA信号途径,促进晚疫病菌侵染。因此,RxLR型效应子PITG_16427.2是致病疫霉菌中一个重要的侵染致病因子。 相似文献
5.
为明确云南省元阳县籼/粳型稻瘟病菌中无毒基因的多样性及差异,采用接种鉴别品种方法和PCR技术对9株粳型菌株和21株籼型菌株中无毒基因Avr-Pia、Avr-Pita1和Avr-Pii的多样性进行分析。结果显示,30株稻瘟病菌菌株中无毒基因Avr-Pia、Avr-Pita1和Avr-Pii的平均出现频率分别为33.3%、76.7%和0,Avr-Pia、Avr-Pita1在籼型菌株和粳型菌株中的出现频率分别为4.8%和100.0%、66.7%和100.0%。Avr-Pia主要为存在/缺失的多样性,籼/粳型菌株对水稻品种Achiasahi(Pia)的致病率分别为100.0%和11.1%;Avr-Pita1可划分为3个类群,除菌株CH1195属于PO类群外,籼型菌株均属于J1类群,粳型菌株则属于J2/J3类群;籼/粳型菌株对水稻品种K1(Pita)的致病率分别为100.0%和0,且籼/粳型菌株Avr-Pita1蛋白序列在第83位和第192位氨基酸差异明显。Avr-Pii在30株稻瘟病菌菌株中均未检出,籼/粳型菌株对水稻品种Fujisaka No. 5(Pii)的致病率分别为100.0%和0。表明Avr-Pia和Avr-Pita1在粳型菌株中的出现频率高于籼型菌株且变异程度低,粳型菌株中可能存在其它无毒基因使其不能侵染Fujisaka No. 5。 相似文献
6.
植物病原细菌通过III型分泌系统将大量的效应蛋白分泌到宿主细胞内,从而抑制宿主的先天免疫。效应蛋白Avr Xcc E1广泛存在于黄单胞菌中,然而关于Avr Xcc E1具体的作用机理尚不清楚。本研究利用野油菜黄单胞菌野油菜致病变种(Xanthomonas campestris pv.campestris,Xcc)和拟南芥互作模式系统,发现来自Xcc8004菌株的效应蛋白Avr Xcc E1对于Xcc8004在拟南芥上的全毒性是必须的,并且Avr Xcc E1可以抑制病原细菌鞭毛蛋白Flg22诱导的FRK1(Flg22-induced receptor-like kinase)基因表达,但并不抑制Flg22诱导的M AP激酶活性。通过Avr Xcc E1与GFP蛋白的融合表达,证实效应蛋白Avr Xcc E1定位在细胞膜上,并且细胞膜定位对于Avr Xcc E1在拟南芥上发挥毒性功能和在大白菜品种"中白83"上发挥无毒功能是必须的。 相似文献
7.
稻瘟菌对水稻品种梅雨明的无毒性的遗传分析和分子标记 总被引：18，自引：3，他引：15
将交配型为a、并对水稻品种梅雨明不致病的稻瘟菌菌株S1522与对该品种致病的、交配型为A的稻瘟菌菌株P131和S159杂交,分别获得了24和56个单子囊孢子后代。致病性测定结果表明:这些后代菌株在梅雨明上的致病个体与不致病个体的比例近等于1:1。由此推测:稻瘟菌对水稻品种梅雨明的无毒性是由1个基因控制的。进一步通过RAPD比较了致病后代与不致病后代的DNA多态型,获得了1个与该无毒基因连锁的、长度为1000 bp的RAPD标记,两者间的遗传距离为3.7cM。 相似文献
8.
利用来自国际马铃薯中心含有单显性抗性基因的11个鉴别寄主,采用离体叶片测定方法对2007—2008年采自甘肃和内蒙古地区的共91个致病疫霉马铃薯分离物的致病型进行了测定。结果显示：来自甘肃的65株菌中共有28个致病型,有3株可以克服11个抗性基因;总共有超过一半的菌株对抗性基因R7、R9、R11有毒性,而能克服抗性基因R1、R2、R8的菌株数最少。在来自内蒙古的26株菌中,共检测到24个致病型,有2株菌能克服11个抗性基因,共有84.6%的菌株可以对含有抗性基因R7的植株表现出毒性,而R1、R2、R8被克服比例最低。因此,研究结果表明,在这两个地区可优先考虑在菌株采集地种植和选育含有R1,R2和R8这3个抗性基因的马铃薯品种。 相似文献
9.
黑龙江省马铃薯晚疫病菌交配型及对甲霜灵敏感性测定 总被引：1，自引：0，他引：1
马铃薯晚疫病(late blight)是由致病疫霉(Phytophthora infestans)引起的马铃薯生产中最具毁灭性的病害,明确晚疫病菌的交配型及对甲霜灵敏感性状况对制定马铃薯晚疫病防治策略具有重要意义。本研究对2011—2013年采自黑龙江省黑河、齐齐哈尔、绥化等马铃薯主产区的晚疫病菌菌株进行交配型及对甲霜灵敏感性测定,采用对峙培养法对153株晚疫病菌进行交配型检测,结果表明,A1交配型79株,占51.63%,A2交配型74株,占48.37%;采用菌落直径法对143株晚疫病菌进行甲霜灵敏感性测定,结果表明,83.92%表现高抗,8.39%表现中抗,7.69%表现敏感。 相似文献
10.
基因对基因假说阐明了病原菌无毒基因(avirulence gene,Avr gene)与寄主植物抗性基因(resistance gene,Rgene)相互识别和互作的关系。禾谷类白粉菌无毒基因产物作为重要的激发子,能够与其寄主R基因产物发生特异性互作,诱导植物细胞防卫反应。为了更加深入地了解这些无毒基因的作用,作者总结了最近关于AVRa1、AVRa10、AVRa13、AVRk1、AvrPm2和AvrPm3a2/f2等已克隆无毒基因的研究进展,讨论了它们作为效应蛋白(effector)或激发子的双重功能,在毒性菌株中的变异规律,与其对应R基因之间的互作模型,以及与转座子等重复序列之间的关联等。本文还对无毒基因研究方法的改进和未来的研究方向提出了建议。 相似文献
11.
双重PCR检测马铃薯晚疫病菌和青枯病菌方法的建立及应用 总被引：3，自引：0，他引：3
利用真菌通用引物ITS1和ITS4扩增马铃薯晚疫病菌转录间隔区并进行序列测定,通过序列比较,设计了1对马铃薯晚疫病菌的特异引物INF1/INF2,并对15种不同真菌、细菌和7种疫霉属和腐霉属卵菌基因组DNA进行PCR扩增,结果只有不同来源的马铃薯晚疫病菌株可获得324 bp的特异带。将引物INF1/INF2与卵菌通用引物进行巢式PCR扩增后,其检测灵敏度在DNA水平上可达30 fg。运用设计的引物与马铃薯青枯病菌特异引物结合建立了双重PCR体系,能从马铃薯晚疫病菌和马铃薯青枯病菌总基因组DNA以及人工接种和自然发病的马铃薯植株中分别或同时扩增到324 bp和281 bp的特异片段。实现了同时对马铃薯晚疫病菌和马铃薯青枯病菌的快速可靠检测。 相似文献
12.
ABSTRACT Sensitive and specific primer sets for polymerase chain reaction (PCR) for Phytophthora infestans, the oomycete that causes late blight of potato and tomato, were developed based on families of highly repeated DNA. The performance of these primers was compared to those developed previously for the internal transcribed spacer (ITS) of ribosomal DNA. The detection limit using the new primers is 10 fg of P. infestans DNA, or 0.02 nuclei. This is about 100 times more sensitive than ITS-directed primers. Nested polymerase chain reaction (PCR) allows the measurement of down to 0.1 fg of DNA using the new primers. To enhance the reliability of diagnostic assays, an internal positive control was developed using an amplification mimic. The mimic also served as a competitor for quantitative PCR, which was used to assess the growth of P. infestans in resistant and susceptible tomato. A key dimension of this study was that two laboratories independently checked the specificity and sensitivity of each primer set; differences were noted that should be considered when PCR is adopted for diagnostic applications in any system. 相似文献
13.
A specific and sensitive PCR assay for the detection of Phytophthora infestans , the cause of late blight of potato, in soil and plant tissues was developed. A P. infestans -specific primer pair (INF FW2 and INF REV) was designed by comparing the aligned sequences of rDNA internal transcribed spacer regions of most of the known Phytophthora species. PCR amplification of P. infestans DNA with primers INF FW2 and INF REV generated a 613 bp product, and species specificity was demonstrated against DNA from nine other Phytophthora species and seven potato-blemish pathogens. In a single-round PCR assay, 0·5 pg pure P. infestans DNA was detectable. Sensitivity was increased to 5 fg DNA in a nested PCR assay using Peronsporales-specific-primers in the first round. As few as two sporangia or four zoospores of P. infestans could be detected using the nested assay. Procedures are described for detection of P. infestans in leaves, stem and seed potato tubers before expression of symptoms. A soil assay in which 10 oospores per 0·5 g soil were detectable was developed and validated using samples of field soil. The PCR assay was used to examine the long-term survival of sexual (oospores) and asexual (sporangia and mycelium) inoculum of P. infestans in leaf material buried in a replicated experiment under natural field conditions. Oospores were consistently detected using the PCR assay up to 24 months (total length of the study) after burial in soil, whereas the sporangial inoculum was detected for only 12 months after burial. Sporangial inoculum was shown to be nonviable using a baiting assay, whereas leaf material containing oospores remained viable up to 24 months after burial. 相似文献
14.
以我国11个省(市、区)和6种不同寄主植物的43个青枯菌Ralstonia solanacearum代表性菌株和4个国外青枯菌菌株为试材,采用15条随机引物进行了RAPD分析,筛选到了1条青枯菌所特有的DNA片段,根据其碱基序列,设计了特异PCR引物,对不同青枯菌DNA进行扩增,并以马铃薯的其它病原细菌为对照,只有青枯菌可获得773bp的DNA扩增产物.经过对PCR反应模板制备程序的简化和优化,利用本研究设计的特异引物,直接对马铃薯病块茎的DNA粗提液进行扩增,获得了773bp片段.此技术可望用于马铃薯种薯青枯病菌的检测,大大缩短检测时间,提高检测效率. 相似文献
15.
Genetic variation among asexual progeny of Phytophthora infestans detected with RAPD and AFLP markers 总被引：6，自引：1，他引：6
Genotypic variation among 32 single-zoospore isolates (SZI) of Phytophthora infestans , derived asexually from two hyphal-tip parental isolates (PI-105 and PI-1) of the US-8 genotype, was assessed with 80 random amplified polymorphic DNA (RAPD) primers and 18 amplified fragment length polymorphic DNA (AFLP) primer pairs. In previous investigations, the SZIs from parental isolate PI-105 showed high levels of virulence variability and were differentiated into 14 races, whereas the SZIs from PI-1 showed identical virulence to the parent. The purpose of this investigation was to determine if phenotypic variation observed among SZIs of P. infestans could be detected at the DNA level in these isolates. Polymorphism was detected with 51 RAPD primers and with all 18 AFLP primer pairs in PI-105 SZIs. In SZIs from PI-1, polymorphism was also detected with 25 RAPD primers and 17 AFLP primer pairs. Cluster analysis using the unweighted pair-group method with arithmetic averages (UPGMA) separated the SZIs from parent PI-105 into six virulence groups, 11 RAPD groups and three AFLP groups. Cluster analysis of PI-1 SZIs, which all belong to the same virulence group, differentiated them into four RAPD groups and six AFLP groups. No close correlation among RAPD, AFLP and virulence groups could be established within the two progenies of SZIs. Results of this study suggest that there is a considerable level of inherent genetic variability among SZIs derived asexually from the same parental isolate. The possible mechanisms and implications of this genetic variation are discussed. 相似文献
16.
采用实时荧光PCR技术建立了瓜炭疽病菌(Colletotrichum orbiculare)的检测方法。根据瓜炭疽病菌甘油醛-3-磷酸脱氢酶(GAPDH)基因和谷氨酰胺合成酶(GS)基因序列,设计了该病菌特异性引物和TaqMan探针,并对所设计的引物和探针的反应条件进行了优化。采用本试验建立的实时荧光PCR方法对瓜上的其他菌株及近似菌株进行检测,可将瓜炭疽病菌与其他病原菌区分开。灵敏度试验表明,25μL体系中只要有39.6pg的核酸量就可以被检测到,检测灵敏度达到1.584pg/μL,比普通PCR检测灵敏度高100倍。同时对田间采集的病株和未知样品进行的检测证明了引物和TaqMan探针的特异性。 相似文献
17.
Pcr Assay for specific detection of European stone fruit yellows phytoplasmas and its use for epidemiological studies in France 总被引：2，自引：0，他引：2
W. Jarausch M. Lansac C. Saillard J.M. Broquaire F. Dosba 《European journal of plant pathology / European Foundation for Plant Pathology》1998,104(1):17-27
DNA amplification by polymerase chain reaction was used to specifically detect phytoplasmas associated with severe decline diseases of European stone fruits. PCR primers were designed according to the partial sequence of a nonribosomal genomic fragment of European stone fruit yellows phytoplasmas obtained by direct sequencing of a specific PCR product. A PCR assay was developed which resulted in specific amplification of a 237 bp-DNA fragment from total DNA extracts derived from over 300 stone fruit samples. No PCR product was obtained with DNA from healthy controls or plants diseased with various other phytoplasmas, e.g. the closely related apple proliferation and pear decline phytoplasmas. Phytoplasma infection was checked in all samples by PCR amplification with universal ribosomal primers. Detection rate with specific and universal primers was correlated by 97%. European stone fruit yellows phytoplasmas were detected in samples of 114 out of 139 examined orchards which represent the major stone fruit growing regions of France. Typical symptoms like chlorotic leaf roll in summer and off-season growth in winter were correlated by 95% to the presence of phytoplasmas. However, phytoplasmas were also detected in 51% of samples derived from trees showing non-specific symptoms. A comparison study including 201 samples showed that 81% of the PCR-positive samples were also tested positive using fluorescence microscopy with DAPI staining. 相似文献
18.
本研究根据番茄斑萎病毒(TSWV)S RNA上的核衣壳蛋白(N)基因保守序列设计特异性引物,比较了4种检测方法的灵敏度。结果表明,特异性引物可扩增出397bp的片段,序列和已发表的TSWV核苷酸序列同源性高达99%,可用于常规PCR和荧光定量RT-PCR(qRT-PCR)检测。qRT-PCR的灵敏度比快速检测试纸条、双抗体夹心酶联免疫吸附法(DAS-ELISA)和常规PCR分别高出15 625倍、3 125倍和125倍,且能够准确定量;常规PCR灵敏度较高,但不能准确定量;DAS-ELISA方法适用于批量定性测定,但检测时间较长;试纸条法检测速度最快,但灵敏度最低,使用时可根据症状程度和试验条件选择适宜的检测方法。 相似文献
19.
DNA amplification by polymerase chain reaction (PCR) was used specifically to detect the mycoplasma-like organism (MLO) associated with lethal yellowing disease of palms in Florida. For PCR, a pair of oligonucleotide primers was synthesized according to partial sequences of a cloned 1·3 kbp fragment of lethal yellowing MLO-specific genomic DNA isolated from a diseased windmill palm ( Trachycarpus fortunei ). A DNA product of about 1 kbp was specifically amplified by PCR in reaction mixtures containing template DNA derived from either heart, inflorescence or leaf tissues of lethal yellowing-affected palms. PCR performed for 35 cycles with as little as 5 pg of DNA template, in some instances, was sufficient consistently to amplify the same lethal yellowing MLO DNA product from hearts of 11 species comprising 30 symptomatic palms. Similar reliable and reproducible detection of the lethal yellowing MLO in palm inflorescence spikelets was also achieved after 35 cycles of PCR. When template DNA for PCR was derived from tissues of the the most immature emerging leaf, a 40-cycle reaction was sufficient for consistent foliar detection of the pathogen in all coconut palms including palms with earliest visible symptoms of disease. 相似文献
20.
Deahl KL Cooke LR Black LL Wang TC Perez FM Moravec BC Quinn M Jones RW 《Pest management science》2002,58(9):951-958
In recent years, late blight, caused by Phytophthora infestans (Mont) De Bary, has increased in severity in many parts of the world, and this has been associated with migrations which have introduced new, arguably more aggressive, populations of the pathogen. In Taiwan, late blight has been endemic on outdoor tomato crops grown in the highlands since the early 1900s, but recent epidemics have been more damaging. To ascertain the present status of the Taiwanese population of P infestans, 139 isolates of the pathogen collected and maintained by the Asian Vegetable Research and Development Center (AVRDC) were characterized using mating type, metalaxyl sensitivity, allozyme genotype, mitochondrial haplotype and RFLP fingerprinting. Up to 1997, all isolates were found to belong to the old clonal lineage of P infestans (US-1 and variants), but in isolates from 1998 a new genotype appeared, and by 2000 this had apparently completely displaced the old population. This new genotype was an A1 mating type and has the dilocus allozyme genotype 100/100/111, 100/100 for the loci coding for glucose-6-phosphate isomerase and peptidase, respectively. These characters, together with RG57 fingerprinting, indicated that these isolates belonged to the US-11 clonal lineage, a minority (11%) being a previously unreported variant of US-11. Whereas metalaxyl-resistant isolates were not detected in the old population, 96% of the new genotypes proved resistant, with the remainder being intermediate in sensitivity. It may be inferred from this sudden, marked change in the characteristics of the Taiwanese P infestans that a new population of the pathogen was introduced around 1997-98 and that this may well have already been metalaxyl-resistant when it arrived, although a role for in situ selection cannot be excluded. 相似文献