Assessment of Genetic Diversity in Thai Isolates of Pyricularia grisea by Random Amplification of Polymorphic DNA

One hundred and seventy‐four isolates of Pyricularia grisea were collected from various hosts such as barley, rice, weed and wild rice in Thailand. Seven arbitrary decamer primers from the set of University of British Columbia were employed and nine lineages were classified. Lineages B, C and H were predominant, contributing up to 70% of total pathogens in this study. Analysis showed that the distribution of each lineage differs from the predominant lineages across Thailand in such that other lineages were restricted in particular area. For instance, lineage A was limited only in southern Thailand, whereas wide distribution of lineages B and C reflected an influence of both biological and physical effects on pathogen variation. Principal component analysis resulted in a total of four groups of blast pathogen with small distinctions between barley‐, rice‐, weed‐ and wild rice‐infected blast. Bridging relationships occurred among border isolates of weed and rice blast suggesting a chance of migrations between hosts. Higher diversity was observed in northern, north‐eastern and central Thailand while eastern and southern parts were rather low. Genetic diversity indices elucidated an abundance of pathogen lineages existing in northern Thailand suggesting that it should be the centre of diversity.     

云南稻瘟病菌系谱与致病型的关系

为探究稻瘟病菌无性世代DNA水平的变异,寻找云南稻瘟病菌谱系(genetic lineage,G)和致病型之间的对应关系,根据稻瘟病菌散布的重复序列Pot2(Pyricularia oryzac transposon),对云南水稻主产区稻瘟病菌菌株DNA进行了rep-PCR(repetitive polymerase chain reaction)扩增,获得rep-PCR指纹。聚类分析将134个稻瘟病菌代表菌株划分为G1～G8等8个谱系,揭示云南水稻主产区稻瘟病菌无性系丰富的遗传多样性。进一步接种分析了8个谱系的29个稻瘟病菌菌株对33个云南主产区水稻品种的亲和性,依其毒性谱,采用STATISTICAL5．0软件的UP-GMA程序进行聚类分析,将其划分为P1～P6等6个致病型群(pathotype group)。结果表明同一谱系的稻瘟病菌菌株多数对应2～3个致病型群,少数1个或4个致病型群;但G1～G8等8个谱系中的部分菌株都可对应致病型群P2。因此,云南水稻主产区稻瘟病菌谱系和致病型群之间属于复杂关系类型。此外,33个水稻品种中的合系16和京国92抗全部29个稻瘟病菌株,云粳20和合系30对全部供试菌株表现感病,这对云南水稻主产区品种布局提供了稻瘟病抗性方面的依据。因此,从育种应用和生产实际需要出发,水稻品种抗瘟谱测定仍然必要。     

Differential responses in vitro of rice cultivars to Italian lineages of the blast pathogen Pyricularia grisea (Cooke) Sacc. 1. Oxidative burst

Suspension cultured cells of six rice cultivars differing in their sensitivity to blast were treated with mycelial wall hydrolysates prepared from seven isolates belonging to different Pyricularia grisea lineages. Soon after elicitor addition, rice cells produced significant amounts of superoxide anion, which was rapidly converted into diffusible peroxide. Maximal effects were achieved at 50 mg L-1 elicitor. In all cases, a 7 to 13-fold increase in the basal rate of reactive oxygen species production was found. Neither differential effects among strains nor clear relationships between lineage and the resulting oxidative burst were evident. Interestingly, a good correlation was found between basal (and elicited) levels of peroxide generation and the overall tolerance of rice cultivars to the pathogen. About two days after elicitation, cell death occurred proportional to the amount of hydrogen peroxide released. Peroxide was required to trigger loss of cell viability, but the latter was not due to a direct toxic effect, suggesting the induction of programmed cell death. Results represent the first data aimed to develop in vitro tests for pathogenicity prediction of Italian blast lineages toward rice cultivars.     

Pathogénie Comparée et Signe Sexuel des Isolats Marocains de Pyricularia grisea (Magnaporthe grisea) Originaires de Riz et de Stenotaphrum secundatum

Moroccan isolates of Pyricularia grisea (Magnaporthe grisea) from Oryza sativa and Stenotaphrum secundatum were crossed with standard fertile isolates (mating type Mat 1.1 and Mat 1.2) by using the three‐points culture method on oatmeal agar. Only Mat 1.1 isolates from rice were identified and considered to have only the ability to function as males, whereas no isolate from S. secundatum mated with Mat 1.1 or Mat 1.2. Cross‐inoculation studies using Pyricularia grisea (M. grisea) isolates from Oryza sativa and S. secundatum show that rice varieties cultivated in Morocco were susceptible to isolates originating from S. secundatum. Similarly, S. secundatum was susceptible to some isolates from O. sativa.     

Azoxystrobin induces lignification-related enzymes and phenolics in rice (Oryza sativa L.) against blast pathogen (Pyricularia grisea)

Abstract

This study was conducted to analyse the induction of lignification-related enzymes and phenolic content in rice to blast disease caused by Pyricularia grisea using azoxystrobin. The severity of rice blast was reduced (70% over control) through treatment by azoxystrobin. This reduction in disease severity was mainly associated with induction of host defense mechanisms by azoxystrobin. Increased production of secondary metabolite – phenolic and lignification – related enzymes, namely, peroxidase (POD), polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL) were observed in rice plants treated with azoxystrobin.     

Genetic Diversity in Ralstonia solanacearum Strains from Mauritius using Restriction Fragment Length Polymorphisms

Race 1, biovar III of Ralstonia (synonym Pseudomonas ) solanacearum , causal organism of bacterial wilt, has been reported in Mauritius on several crops and plant species. The genetic relationship among 38 strains isolated from potato, tomato, bean and anthurium was determined by restriction fragment length polymorphisms (RFLPs). After hybridization with probe 5a67, five RFLP patterns could be distinguished. Types V and I were most commonly encountered. A common band of approximately 6.5 kb was found in 35 strains. Type I pattern consisted of only this band and was observed in 12 out of 16 anthurium strains tested. Type V was associated with 12 out of 16 potato strains and consisted of a band of approximately 3.3 kb in addition to the one observed in type I. RFLP patterns II, III and IV were less frequently encountered. The RFLP analysis showed that genetic diversity was present in race 1, biovar III strains. The relationship between the host and RFLP pattern is discussed.     

羚牛的遗传多样性及其种群遗传结构分析

羚牛是亚洲大陆一种特有的大型珍稀动物,目前正面临着栖息地丧失、片段化和人类活动的严重威胁。为了有效地保护这种濒危动物,全面了解羚牛的种群结构、进化历史和整个分布区内遗传多样性的分布是至关重要的。本研究以mtDNA D-loop330bp基因片段为分子标记,比较分析了来自陕西秦岭、甘肃南部、四川岷山、邛崃山和云南贡山的40个样品的序列差异,根据分布特点将所采集到的羚牛分为3个地理单元,即秦岭、四川和云南。结果表明,在3个地理单元中存在4种单倍型,且地理单元间不存在共享单倍型,相互单倍型之间的平均序列差异为1.66％。进一步分析表明,3个地理单元间的基因流较低,存在着显的遗传分化 ,说明羚牛具有明显的系统地理分布格局。同时提出应将分布于秦岭山区、唐家河青川地区、天全以及云南贡山地区作为独立的管理单元分别加以保护。     

Population Structure and Genetic Diversity within Peach latent mosaic viroid Field Isolates from Peach Showing three Symptoms

To gain insight into the molecular basis of field isolates inducing the symptoms of leaf yellowing, discolouration along leaf sides and leaf mosaic, six isolates from peach showing the three different symptoms in the field were studied by single-strand conformation polymorphism analysis and nucleotide sequence analysis. Results revealed that each Peach latent mosaic viroid (PLMVd) isolate is composed of a population of genetically related variants (haplotypes), one being predominant with frequencies from 32% to 57%, and most of others having a low frequency of 4–5%. Each predominant haplotype was sequenced, as well as some non-predominant haplotypes selected randomly for comparative purposes. In each isolate, sequence alignment among the predominant and non-predominant haplotypes demonstrated that the predominant haplotype had the least variation with others among them, and its sequence was identical to the consensus sequence, which inflected that the predominant haplotype displayed a wide representative of sequence for others in a PLMVd isolate. The similarities and genetic distance between the predominant sequences from peach showing the same symptoms were higher and smaller, respectively, than that with different symptoms; they were more than 98.8% and <1%, respectively, between the predominant sequences with same symptomatic source, and were <98.5% and more than 1%, respectively, between the predominant sequences with different symptomatic source. Some particular variations were indicated for these isolates, and it revealed that the isolates with the symptom of discolouration along leaf sides on their source peach trees had a G or U in position 169 nt, and the isolates with the symptom of leaf yellowing had U and C in 115 and 116 nt, respectively, and the isolates with the symptom of leaf mosaic showed diversity at (3 nt: delete C; 5 nt: A and 54 nt: U).     

Low Levels of Genetic Diversity within Populations of Hosta clausa (Liliaceae)

Abstract Genetic diversity of Korean populations in Hosta clausa was investigated using starch gel electrophoresis. Hosta clausa is widespread, grows only along streamsides, and has both sexual and asexual reproduction. Populations of the species are small and isolated. Thirty-two percent of the loci examined were polymorphic, and mean genetic diversity within populations (He_p=0.082) was lower than mean estimates for species with very similar life history characteristics (0.131), particularly for its congener H. yingeri (0.250). The mean number of multilocus genotypes per population was 8.7, and genotypic diversity index (D_G) was 0.84. Significant differences in allele frequencies among populations were found in all seven polymorphic loci (P < 0.001). About one-fifth of the total allozyme variation was among populations (G_ST=0.192). Indirect estimate of the number of migrants per generation (Nm=0.48, calculated from mean G_ST) and nine private alleles found indicate that gene movement among populations was low. The low levels of genetic diversity within populations and the relatively high levels of genetic diversity among populations suggest that strong moist habitat preferences, clonal reproduction, low level of gene flow among populations, genetic drift, and historical events may have played roles in the genetic structuring of the species.     

茄子种质遗传多样性及群体结构的SRAP分析

采用SRAP标记法对183份茄子(Solanum melongena)种质资源的遗传关系和群体结构进行分析。结果表明, 33对多态性SRAP引物组合共扩增出215条清晰稳定的多态性条带, 平均每对引物组合产生7条多态性条带。183份茄子种质的遗传相似系数介于0.276-0.813之间, 平均值为0.623, 表明茄子种质资源间遗传背景存在一定的差异。在遗传距离为0.345 6处可将183份茄子种质分为4组。通过群体结构分析可将种质划分为4个群体, 不同群体间的界限十分明显, 且群体间的基因渗透较高。     

基于线粒体控制区的序列变异分析青海东部甘肃鼢鼠遗传多样性

甘肃鼢鼠(Myospalax cansus)是一种终年营地下独居生活的小型掘土类动物。本文通过测定mt DNA的控制区部分序列(530 bp)变异,分析青海东部地区8个甘肃鼢鼠地理种群遗传多样性与遗传结构。158个样本共发现26个变异位点,定义了39种单倍型,整体的平均单倍型多样性高(h=0.953 2)、核苷酸多样性低(π=0.006 36)。歧点分布和中性检验均说明青海东部甘肃鼢鼠种群在历史上存在着快速扩张的事件。基于邻接法构建的网络关系图中,单倍型呈星状分布,没有按地理位置形成对应类群。基因流(Nm)数据显示多数地理种群间基因交流贫乏,AMOVA结果显示种群内与种群间遗传变异分别为48.82%和51.18%,遗传分化明显。IBD分析表明,甘肃鼢鼠的遗传分化与地理距离呈正相关,说明距离隔离对甘肃鼢鼠种群分化具有重要作用。甘肃鼢鼠的这种遗传多样性与种群遗传结构特点,可能是地下生活方式靠挖掘迁移带来的较小扩散能力的结果。     

在韩国境内Potentilla fragarioides var .sprengeliana的遗传多样性和种群结构(英文)

根据 2 2个等位酶位点遗传变异 ,探讨了韩国境内委陵菜 (PotentillafragarioidesL .var.sprengeliana)的遗传多样性和种群结构。酶位点的多态位点百分比为 5 9 1%。种和种群水平上的遗传多样性比较高 ,分别为Hes=0 .2 10 ,Hep=0 .199;而种群的分化水平则相对较低 (GST=0 .0 74)。 19个种群中随机交配的偏差为FIS=0 .331。每代迁移数的间接估计 (Nm=3.15 )表明该种在韩国的种群间基因流较高。另外 ,固定指数分析显示在一些种群和位点有轻微的杂合子缺乏。种群间平均遗传一致度为 0 985。这些韩国委陵菜种群存在于较为均一的生境 ,这很有可能是造成其种群遗传一致性较高的原因。     

Phylogenetic distribution and genetic mapping of a (GGC)n microsatellite from rice (Oryza sativa L.)

DNA microsatellites are ubiquitously present in eukaryotic genomes [30] and represent a vast source of highly informative markers [30, 33, 34, 2]. We describe in this article a (GGC)_n microsatellite which is widely distributed in eukaryotic genomes. Using polymerase chain reaction (PCR) techniques and DNA sequencing, we demonstrated for the first time in plant species that a (GGC)_n microsatellite locus is moderately polymorphic. Six alleles are present at this locus in rice and length polymorphisms are caused by variation in the number of tandem GGC repeats. By scoring a backcross mapping population, we were able to demonstrate that this locus is stably inherited and does not link to any known RFLP markers on the rice RFLP map. Our results suggest that DNA microsatellites should be useful in plants for construction of genetic linkage maps, extension of the existing genetic linkage maps, linkage analysis of disease and pest resistance genes, and the study of population genetics.     

Patterns of Genetic Diversity and Population Structure of the Clonal Herb,Potentilla fragarioides var. sprengeliana (Rosaceae) in Korea

The genetic diversity and population structure of Potentilla fragarioides var. sprengeliana(Rosaceae) in Korea were investigated using genetic variation at 22 allozyme loci. The percent of polymorphic loci within the enzymes was 59.1%. The genetic diversity at the species level and at the population level was high (Hes=0.210; Hep=0.199, respectively), whereas the extent of the population divergence was relatively low (GST=0.074). FIS, a measure of the deviation from random mating within the 19 populations, was 0.331. An indirect estimate of the number of migrants per generation (Nm=3.15) indicates that gene flow is high among Korean populations of the species. In addition, analysis of fixation indices revealed a slight heterozygote deficiency in some populations and at some loci. The mean genetic identity between populations was 0.985. It is highly probable that the trend of genetic uniformity in a relatively homogeneous habitat is thought to be operated among Korean populations of P. fragarioides var.sprengeliana.     

濒危物种山红树居群遗传结构的RAPD分析

利用RAPD分子标记检测了云南省3个山红树居群的遗传多样性和居群遗传结构。10个引物共检测到54个位点, 其中多态位点11个, 占20 37%。与其他的濒危物种相比, 山红树居群内的遗传多样性很低, 居群间的遗传分化很大(78 65%)。大量经济植物的种植加上人为的破坏, 使山红树的生境遭到严重破坏, 数量大为减少, 可能导致了山红树遗传多样性的丧失、居群间较高的遗传分化。基于以上结果, 探讨了山红树进一步的迁地保护措施。     

Characterization of Trichoderma Isolates from Philippine Rice Fields by UP-PCR and rDNA-ITS1 Analysis: Identification of UP-PCR Markers

Forty-two isolates of Trichoderma from rice fields in four provinces in the Philippines were characterized using rDNA-ITS1 analysis and universally primed polymerase chain reaction (UP-PCR). Two groups were clearly distinguishable on the basis of length and restriction pattern of the internal transcribed spacer (ITS) region of the ribosomal DNA and UP-PCR banding profiles using UP primer, L45. The 40 isolates comprising the largest group were very similar with respect to their UP-PCR banding profiles and were assigned to Trichoderma harzianum Rifai following morphological identification of four of the isolates. The two isolates belonging to the second group were identified as Trichodermaviride Pers. ex. Gray on the basis of their morphology, rDNA-ITS1 analysis and distinct UP-PCR banding profiles. One of the T. harzianum isolates with good cellulolytic and competitive saprophytic abilities was analysed using single and pair-wise combinations of UP primers in order to distinguish it from the remaining 41 isolates. A suitable diagnostic marker was identified and this marker will be valuable for monitoring the isolate in field tests.     

Sixteen polymorphic microsatellite markers from Zizania latifolia Turcz. (Poaceae)

Sixteen polymorphic microsatellite markers were isolated and identified in Zizania latifolia Turcz. (Poaceae), a perennial aquatic plant widespread in Eastern Asia. The microsatellite-enriched library was constructed using the fast isolation by AFLP of sequences containing repeats method. These markers revealed two to 14 alleles, with an average of 5.6 alleles per locus. The observed and expected heterozygosities varied from 0.071 to 0.690 and from 0.174 to 0.812, respectively. These markers will be useful for studying of gene flow and evaluating the genetic diversity of the Zizania latifolia population.     

普遍野生稻和亚洲栽培稻遗传多样性的研究

用 ４４个 ＲＦＬＰ标记对来自中国、印度、泰国等亚洲 １０个国家的普通野生稻（简称普野,下同）和来自多个国家的７５个栽培稻品种,从多态位点的比率、等位基因数、基因型数、平均杂合度及平均基因多样性等多个方面,比较了不同国家和不同地区的普通野生稻、栽培稻籼粳亚种及栽培稻与普野之间遗传多样性的差异。结果表明：中国普野的遗传多样性最大;其次是印度普野;南亚普野的平均基因多样性大于东南亚普野,而多态位点的比率、等位基因数及基因型数等却低于东南亚普野;栽培稻的遗传多样性明显小于普通野生稻。在所检测的４４个位点中,栽培稻的多态位点数仅为野生稻的３／４,等位基因数约为野生稻的６０％,基因型种类约为野生稻的１／２。栽培稻中籼稻的遗传多样性高于粳稻。在平均每个位点的实际杂合度上,以中国普野杂合度最高,普通野生稻是栽培稻的２倍。说明从野生稻演化成栽培稻的过程中,经过自然选择和人工选择,杂合度降低,等位基因减少,基因多样性下降。