5种松树的遗传多样性和遗传分化研究

利用cpSSR和AFLP标记,比较了巴山松、黄山松、油松、马尾松以及云南松的遗传多样性与遗传分化结果.发现5种松树都表现出较高水平的遗传多样性与种间分化,但cpSSR比AFLP标记提供了更多的种内种间分化,而展示的遗传多样性相对要少.在对不同方法指标的对比研究后,认为将cpSSR的单倍体关系考虑在内,能够更有效地分析植物居群遗传分化造成的地理结构.对于显性标记AFLP,传统的平方根法会低估遗传参数,而Lynch和Milligen的方法因样本量的限制也可能高估遗传多样性,Bayesian法被认为是最精确的方法,提供了与分子方差分析(AMOVA)近似的遗传分化值.     

梵净山尖叶拟船叶藓的遗传多样性分析

采用RAPD技术,选取10个引物对梵净山的尖叶拟船叶藓(Dolichomitriopsis diversiformis)自然分布区的南、北坡14个居群153个个体的总DNA进行了扩增,共得到196个位点.统计分析表明:(1)梵净山尖叶拟船叶藓在物种水平有较高的遗传多样性,居群水平的遗传多样性相对较低.(2)该藓种遗传多样性高低与海拔高度无关,但南坡的多样性水平略高于北坡.(3)遗传多样性的63.29%发生在居群间,只有36.71%发生在居群内.研究结果提示尖叶拟船叶藓的遗传多样性受小生境的影响较大,遗传漂变和环境适应可能是影响居群分化的主要原因.     

APM 和Liped两程序在β-地中海贫血特征连锁分析上的比较研究

Liped程序已广泛地应用在遗传方式明确的遗传病（或性状）家系的连锁分析上，具有精确、简便、快速和多效（如还可考虑突变的影响，以及依赖年龄的外显率等分析上）诸优点；但显然不能用于遗传方式尚未搞清的遗传病家系的连锁分析。晚近由Weeks编制的APM (Affected-Pedigree-Member,暂译为家系患者连锁分析）程序突破了Liped程序的上述局限性，具有更广泛的应用，日有更强的检测力；但该法只能估计连锁的显著性，还不能揭示连锁强度，因而还难以用于人类整个基因组的序列分析研究上。 本文以一个β-地巾海贫血特征的大家系为实例，对上述两程序作了具体的比较研究。结果表明了上述两程序分别具有的优缺点，因此在连锁分析上可相互补充。     

共显性分子标记基因型数据转换为二元型数据的处理软件及其在研究种质资源遗传多样性中的意义

遗传多样性研究是植物种质资源有效利用和保护的重要基础.遗传多样性研究所采用的分子标记工具主要有显性和共显性两种,两种不同类型的分子标记将产生不同的数据类型.显性分子标记产生二元型数据,共显性分子标记产生基因型数据.数据形式不同给遗传多样性的分析和研究带来多方面的困难,不同类型数据之间的互相比较也需要将基因型数据进行二元型转换.本研究基于Excel平台,设计开发了将基因型数据转换成二元型数据的处理软件.该软件按照基因型数据向二元型数据转换的原理,可以将庞大的分子标记基因型数据矩阵,迅速、高效、准确地转换成二元型(0、1)数据矩阵.利用显性分子标记ISSR和共显性分子标记SSR对野生大豆居群遗传多样性的案例分析表明,将共显性分子标记的基因型数据转换为二元型数据,有利于种质资源遗传多样性研究中不同分子标记获取的遗传多样性结果之间的比较和综合分析.     

遗传多样性概述

遗传多样性作为生物多样性的重要组成部分,是物种多样性、生态系统多样性和景观多样性的基础。随着研究方法和实验技术的发展,遗传多样性研究从形态学水平、细胞学（染色体）水平、生理生化水平逐渐发展到分子水平。形态标记、细胞学标记、等位酶分析、DNA多态性分析等方法,为我们研究遗传多样性提供了有效的工具。特别是DNA多态性分析是一种更为直接而有效的方法。     

基于cyt b基因序列的我国西部地区马麝遗传多样性

为了解我国西部地区马麝Moschus chrysogaster的遗传多样性及其系统发育关系，对来自四川省、甘肃省、西藏自治区及重庆市的88份组织样品的cyt b基因序列进行分析。研究发现，马麝cyt b基因序列长1 032 bp，共定义了54个单倍型。A+T的平均含量为59.9%，存在明显的A、T偏倚；平均单倍型多样性和平均核苷酸多样性水平都较高，分别为0.974 1和0.024 88。基于单倍型和遗传距离分别构建的邻接树结果一致，各支系的聚类不存在明显的地理界限，各种群均无扩张现象。我国西部地区马麝的遗传多样性水平总体较高，但张掖市和阿坝县种群的遗传多样性水平略低。本文为马麝的遗传多样性研究与种质资源保护提供了分子基础资料。     

棉立枯丝核菌(Rhizoctonia solani)中的一个dsDNA病毒

自从1962年Hollings在栽培蘑菇(Agaricus bisporus)中发现第一例真菌病毒以来,迄今已在100多种真菌中发现了病毒,多数含双链RNA基因组。1972年Bozarth报道在R.solani中发现病毒,但未报道该病毒的理化性质。1975年Finlker在R.solani的一个强致病力菌株中分离到双链RNA病毒,它含3个组分dsRNA。     

四川省东、南部稻区水稻纹枯病菌系研究

将来自川东、南稻区代表性的38个县(市)的水稻纹枯病标样296份,按不同的品种、海拔、土质、前作和症状归粪,选代表性的标样分离得到108个丝核菌菌株。按HCL—Giemsa染色程序和菌丝融合测定法,将108个菌株分为3个菌系：Rhizoctonia solani AG-1和AG-4,以及双核丝核菌的AG—Bb。其比例分别为97％、1％和2％。经致病性测定表明,该各菌系对水稻的致病性有显著差异：R. solani AG-1的大多数菌株最强,双核丝核菌AG—Bb最弱,R. solani AG一4居中。 对上述各菌系的培养性状、非特异性酯酶和过氧化物酶同工酶谱进行比较研究发现,不同菌系间在上述诸方面均存在明显差异,而同菌系不同菌株间却具一致性。由此说明,按菌丝融合与否区分丝核菌种群较之现行的其它分类法更能反映其遗传本质和亲缘关系。     

川金丝猴遗传多样性研究进展

川金丝猴目前处于生境破碎、种群隔离的濒危境地,对其进行遗传多样性研究是有效保护这一物种的必要前提.结合文献,比较了川金丝猴遗传研究中常用样品采集方法、标记方法等,总结了其遗传多样性的研究现状,并建议通过建立遗传谱系来避免或减少近交,建立生态走廊促进群间基因交流,以提高其遗传变异能力.     

几种常用分子标记遗传多样性参数的统计分析

对235篇文献中314种野生种子植物的遗传多样性参数进行了统计分析.结果表明,目前常用的五种分子标记中,ISSR、等位酶和SSR的参数值间差异显著,彼此不宜直接比较,且与RAPD和AFLP的参数也不宜直接比较;显性标记RAPD和AFLP的参数之间可以直接比较.基于Hardy-Weinberg平衡的遗传分化指数GR值明显低于基于AMOVA分析的ΦR值,两者亦不宜直接比较.对基于RAPD和AFLP标记的179种植物的遗传多样性参数进行联合分析,结果表明:在种群水平上,裸子植物的遗传多样性比双子叶植物和单子叶植物都要高,而其遗传分化值较低;乔木的遗传多样性比草本和灌木高,而分化值更低;克隆植物具有比有性生殖更高的遗传多样性,在有性生殖植物中,异交植物最高,而自交植物最低;广布种的遗传多样性明显高于濒危和狭域分布种.     

Genetic diversity of Rhizoctonia solani associated with potato tubers in France

The soilborne fungus Rhizoctonia solani is a pathogen of many plants and causes severe damage in crops around the world. Strains of R. solani from the anastomosis group (AG) 3 attack potatoes, leading to great yield losses and to the downgrading of production. The study of the genetic diversity of the strains of R. solani in France allows the structure of the populations to be determined and adapted control strategies against this pathogen to be established. The diversity of 73 French strains isolated from tubers grown in the main potato seed production areas and 31 strains isolated in nine other countries was assessed by phylogenetic analyses of (i) the internal transcribed spacer sequences (ITS1 and ITS2) of ribosomal RNA (rRNA), (ii) a part of the gene tef-1α and (iii) the total DNA fingerprints of each strain established by amplified fragment length polymorphism (AFLP). The determination of the AGs of R. solani based on the sequencing of the ITS region showed three different AGs among our collection (60 AG 3 PT, 8 AG 2-1 and 5 AG 5). Grouping of the strains belonging to the same AG was confirmed by sequencing of the gene tef-1α used for the first time to study the genetic diversity of R. solani. About 42% of ITS sequences and 72% of tef-1α sequences contained polymorphic sites, suggesting that the cells of R. solani strains contain several copies of ITS and the tef-1α gene within the same nucleus or between different nuclei. Phylogenetic trees showed a greater genetic diversity within AGs in tef-1α sequences than in ITS sequences. The AFLP analyses showed an even greater diversity among the strains demonstrating that the French strains of R. solani isolated from potatoes were not a clonal population. Moreover there was no relationship between the geographical origins of the strains or the variety from which they were isolated and their genetic diversity.     

Genetic diversity of Rhizoctonia solani AG-3 from potato and tobacco in North Carolina

Anastomosis group 3 (AG-3) of Rhizoctonia solani (teleomorph = Thanatephorus cucumeris) is frequently associated with diseases of potato (AG-3 PT) and tobacco (AG-3 TB). Although isolates of R. solani AG-3 from these two Solanaceous hosts are somatically related based on anastomosis reaction and taxonomically related based on fatty acid, isozyme and DNA characters, considerable differences are evident in their biology, ecology, and epidemiology. However, genetic diversity among field populations of R. solani AG-3 PT and TB has not been documented. In this study, the genetic diversity of field populations of R. solani AG-3 PT and AG-3 TB in North Carolina was examined using somatic compatibility and amplified fragment length polymorphism (AFLP) criteria. A sample of 32 isolates from potato and 36 isolates from tobacco were paired in all possible combinations on PDA plus activated charcoal and examined for their resulting somatic interactions. Twenty-eight and eight distinct somatic compatibility groups (SCG) were identified in the AG-3 PT and AG-3 TB samples, respectively. AFLP analyses indicated that each of the 32 AG-3 PT isolates had a distinct AFLP phenotype, whereas 28 AFLP phenotypes were found among the 36 isolates of AG-3 TB. None of the AG-3 PT isolates were somatically compatible or shared a common AFLP phenotype with any AG-3 TB isolate. Clones (i.e., cases where two or more isolates were somatically compatible and shared the same AFLP phenotype) were identified only in the AG-3 TB population. Four clones from tobacco represented 22% of the total population. All eight SCG from tobacco were associated with more than one AFLP phenotype. Compatible somatic interactions between AG-3 PT isolates occurred only between certain isolates from the same field (two isolates in each of four different fields), and when this occurred AFLP phenotypes were similar but not identical.     

Soil suppressiveness to Rhizoctonia solani and microbial diversity

Rhizoctonia solani anastomosis group 2-2IIIB causes damping-off, black root rot and crown rot in sugar beet (Beta vulgaris). Based on experiences of growers and field experiments, soils can become suppressive to R. solani. The fungus may be present in the soil, but the plant does not show symptoms. Understanding the mechanisms causing soil suppressiveness to R. solani is essential for the development of environmentally friendly control strategies of rhizoctonia root rot in sugar beet. A bioassay that discriminates soils in their level of disease suppressiveness was developed. Results of bioassays were in accordance with field observations. Preliminary results indicate an active role of microbial communities. Our research is focused on the disentanglement of biological mechanisms causing soil suppressiveness to R. solani in sugar beet. Therefore, we are handling a multidisciplinary approach through experimental fields, bioassays, several in vitro techniques and molecular techniques (PCR-DGGE).     

Highly polymorphic in silico-derived microsatellite loci in the potato-infecting fungal pathogen Rhizoctonia solani anastomosis group 3 from the Colombian Andes

Fourteen polymorphic microsatellite DNA markers derived from the draft genome sequence of Rhizoctonia solani anastomosis group 3 (AG-3), strain Rhs 1AP, were designed and characterized from the potato-infecting soil fungus R. solani AG-3. All loci were polymorphic in two field populations collected from Solanum tuberosum and S. phureja in the Colombian Andes. The total number of alleles per locus ranged from two to seven, while gene diversity (expected heterozygosity) varied from 0.11 to 0.81. Considering the variable levels of genetic diversity observed, these markers should be useful for population genetic analyses of this important dikaryotic fungal pathogen on a global scale.     

Detection of phlD gene in some fluorescent pseudomonads isolated from Iran and its relative with antifungal activities

Fluorescent pseudomonads that produce antibiotic 2,4-diacetylphloroglocinol (2,4-DAPG) are important group of PGRP that inhibit a broad spectrum of plant pathogenic fungi. Studying on genetic diversity of 2,4-diacetylphloroglucinol-producing fluorescent pseudomonads has been shown with special importance. The first step to investigate the genetic diversity of these bacteria is detecting of the genes required for the biosynthesis of this antibiotic. The objectives of the current study were detection of phlD gene within fluorescent pseudomonads by a PCR-based assay, and comparison of phenotypic and genotypic characteristics of fluorescent pseudomonads with proven biocontrol potential against some soil-borne phytopathogenic fungi. We used a collection of 47 fluorescent Pseudomonas spp. some with known biological control activity against Macrophomina phaseolina, Rhizoctonia solani, Phytophthora nicotianae var. parasitica, Pythium sp. and Fusarium sp. in vitro and the potential to produce known secondary metabolites such as, siderophore, HCN and protease. The results indicated that 66, 40.42, 63.82,48.94 and 27.65% of strains revealed antagonistic activity against R. solani, M. phaseolina, Pythium sp., P. nicotianae and Fusarium sp., respectively. Rhizoctonia solani recognized as the most vulnerable fungus. Among 47 strains, 76.59, 97.87 and 17% of strains produced protease, siderophore and HCN, respectively. We could detect phlD gene in strains P-5, P-32, P-47. Strain CHA0 was used as positive control for the detection this gene. Overall, there was no obvious link between the existence of phlD gene and inhibition of fungal growth or production of the antifungal metabolites in vitro. But in some strains such as CHA0 and P-5, we saw a link between the existence of phlD and antifungal activities. Studying on detection and diversity of phlD provides a fundamental knowledge for developing a rapid genetic screening system to identify a potential biocontrol strains.     

Comparative analysis of genetic diversity in pea assessed by RFLP- and PCR-based methods

DNA-based molecular-marker techniques have been proven powerful in genetic diversity estimations. Among them, RFLP was the first and is still the most commonly used in the estimation of genetic diversity of eukaryotic species. The recently developed PCR-based multiple-loci marker techniques, which include RAPD, AFLP, Microsatellite-AFLP and inter-SSR PCR, are playing increasingly important roles in this type of research. Despite the wide application of these techniques, no direct comparison of these methods in the estimation of genetic diversity has been carried out. Here we report a direct comparison of DNA-based RFLP with various PCR-based techniques regarding their informativeness and applicability for genetic diversity analysis. Among ten pea genotypes studied, all the PCR-based methods were much more informative than cDNA-RFLP. Genetic diversity trees were derived from each marker technique, and compared using Mantel's test. By this criterion, all trees derived from the various molecular marker techniques, except for the tree derived from inter-SSR PCR, were significantly correlated, suggesting that these PCR-based techniques could replace RFLP in the estimation of genetic diversity. On the basis of this result, AFLP analysis was applied to assess the genetic diversity of a sample of accessions representing the various species and subspecies within the genus Pisum.     

Development of a specific PCR assay for the detection of Rhizoctonia solani AG 1-IB using SCAR primers

AIMS: The aim of this study was to develop a specific and sensitive identification method for Rhizoctonia solani AG 1-IB isolates based on phylogenetic relationships of R. solani AG-1 subgroups using rDNA-internal transcribed spacer (rDNA-ITS) sequence analysis. METHODS AND RESULTS: A neighbour-joining tree analysis of 40 rDNA-ITS sequences demonstrated that R. solani AG-1 isolates cluster separately in six subgroups IA, IB, IC, ID, IE and IF. A molecular marker was generated from a random amplified polymorphic DNA fragment (RAPD). After conversion into a sequence-characterized amplified region (SCAR), a specific primer set for identification of subgroup AG 1-IB was designed for use in a polymerase chain reaction (PCR). The primer pair amplified a single DNA product of 324 bp. CONCLUSIONS: R. solani AG-1 subgroups were discriminated by sequence analysis of the ITS region. The designed SCAR primer pair allowed an unequivocal and rapid detection of R. solani AG 1-IB in plant and soil samples. SIGNIFICANCE AND IMPACT OF THE STUDY: Sequence analysis of the rDNA-ITS region can be used for differentiation of subgroups within AG-1. The use of the developed SCAR primer set allowed a reliable and fast identification of R. solani AG 1-IB and provides a powerful tool for disease diagnosis.     

Changes in fungi with age. Chemical composition of Rhizoctonia solani and Sclerotium bataticola

Gottlieb, David (University of Illinois, Urbana), and James L. Van Etten. Changes in fungi with age. I. Chemical composition of Rhizoctonia solani and Sclerotium bataticola. J. Bacteriol. 91:161-168. 1966.-The chemical composition of the mycelium of Rhizoctonia solani and Sclerotium bataticola was determined in cells of various ages. The percentage, per unit of dry weight, of soluble amino nitrogen, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), ergosterol, and protein decreased with age in both fungi. Total lipids and fatty acids increased with age in S. bataticola but remained constant in R. solani. Total carbohydrate increased with age in R. solani and decreased in S. bataticola. Fewer changes with age were observed when the results were calculated in ratio to DNA. There was no change in the ratios of protein, RNA, and soluble amino nitrogen to DNA with age in either fungus, but the ergosterol-DNA ratio decreased. The total lipid-DNA ratio and the total fatty acid-DNA ratio increased with age in both fungi, whereas the total carbohydrate-DNA ratio increased in R. solani but remained constant in S. bataticola. Both fungi contained myristic, palmitic, palmitoleic, stearic, oleic, and linoleic acids. In addition, R. solani contained pentadecanoic acid, and S. bataticola had myristoleic, linolenic, and arachidic acids. No marked change in the fatty acid pattern of S. bataticola was observed with age, whereas in R. solani the percentage of linoleic acid per total fatty acids decreased slightly when oleic acid increased.     

Genetic Diversity of Thanatephorus cucumeris Infecting Tomato in Iran

The necrotrophic fungus Thanatephorus cucumeris (anamorph Rhizoctonia solani) is among the most important soil‐borne pathogens which causes tomato foot and root rot worldwide. We investigated virulence and genetic relationships among and within different taxonomic groups of R. solani from the tomato‐growing regions in the north‐east of Iran. Characterization of R. solani taxonomic groups revealed that, of 56 isolates, four were AG‐2‐1, 16 were AG‐3 PT, 21 were AG‐4 HG‐I and 15 were AG‐4 HG‐II. Because interprimer binding site (iPBS), which is based on amplification of retrotransposons, is known as novel and powerful DNA fingerprinting technology, we selected four iPBS primers, which can detect polymorphisms of tomato foot root and root rot pathogen, for investigating genotypic variability of the isolates. The iPBS analyses separated various taxonomic groups of R. solani and showed great diversity among the isolates, demonstrating that the R. solani isolates obtained from tomato were not a clonal population. Crop rotation strategies and geographic location seem to be important factors affecting genetic structure of the isolates. Pathogenicity tests on tomato cultivar ‘Mobil’ showed significant differences in the virulence of various isolates. The overall results indicated that isolates of AG‐3 and AG‐4 were more virulent than AG‐2‐1. There was no significant correlation between genetic diversity and virulence of the isolates. This is the first report of R. solani AG‐4 HG‐II, causing tomato foot and root rot. Also, our research is the first in assessment of genetic diversity in fungal populations using iPBS molecular markers.     

Typing of anastomosis groups of Rhizoctonia solani by restriction analysis of ribosomal DNA

A method based on restriction analysis of polymerase chain reaction (PCR)-amplified ribosomal DNA was developed for the rapid characterization of large populations of Rhizoctonia solani at the anastomosis group (AG) level. The restriction maps of the internal transcribed spacers (ITS) sequences were compared for 219 isolates of R. solani belonging to AG-1 to AG-12 and AG-BI, representing diverse geographic and host range origins. Four discriminant restriction enzymes (MseI, AvaII, HincII, and MunI) resolved 40 restriction fragment length polymorphism (RFLP) types among the 219 ITS sequences of R. solani. Each RFLP type could be assigned to a single AG except for two RFLP types, which were common to two AG. A fifth enzyme allowed the discrimination of AG-6 and AG-12. In addition, the combination of four enzymes allowed the discrimination of subsets within AG-1, AG-2, AG-3, and AG-4. The efficiency of the typing method was confirmed by analyzing PCR-amplified ITS sequences of 30 reference strains. Furthermore, the PCR-RFLP method was used to characterize at the AG level 307 isolates of R. solani originating from ten sugar beet fields exhibiting patches of diseased plants in France. The PCR-based procedure described in this paper provides a rapid method for AG typing in R. solani.