松球壳孢菌菌落生长速度对比研究

利用79个辽宁松球壳孢菌菌株与4个A型和B型标准菌株及4个美国菌株进行菌落生长速度对比研究发现,除个别菌株外,辽宁菌株的菌落生长速度显著慢于供试的A型标准菌株和美国菌株;美国菌株与A型标准菌株的菌落生长速度相近,显著快于B型标准菌株;辽宁菌株之间也有显著差别。同时发现,光照可加快菌落离体生长速度。     

松树枯梢病发生的立地条件及其主要诱因分析

对福建、江西等地227块松树枯梢病的林分病情标准地进行调查.结果表明:病害的发生与林分及立地因子关系密切.由上杭、连城、永新和安福4县有代表性的49块样地推导出了松树枯梢病的病情指数与林龄、造林密度、郁闭度、土壤质地、土壤湿度、坡位、坡向的生态数学模型:y=41.38-16.56x11 5.08x21 18.62x31-3.78x41 19.94x51 7.32x61 4.27x62-4.06x71-10.56x72-13.88x73,其偏相关系数分别为0.606 9、0.612 5、0.268 9、0.1789、0.681 1、0.279 0、0.473 7.该模型模内判别准确率为79.59%,其中病情指数与土壤湿度、郁闭度、林龄和坡位间关系紧密,在0.01水平上显著,它们是松树枯梢病在我国南方发生的主要诱导因子.提出以营林措施为主的生态控制技术.     

DNA fingerprinting of Populus trichocarpa clones using RAPD markers

Nine trees from a single, natural population of black cottonwood (Populus trichocarpa Torr. et Gray) in Alaska were screened for randomly amplified polymorphic DNA (RAPD) markers with ten different 10-base random oligonucleotide primers in order to evaluate the use of RAPD analysis for distinguishing black cottonwood clones. Nine primers amplified the genomic DNA targets; two primers were able to differentiate all clones. Eight clones were distinguished among the nine tree samples assayed. Two trees showed identical banding patterns with all primers used; therefore it is suggested that these trees are from the same clone. The RAPD fingerprinting method is simple and powerful-one primer can distinguish different clones, while the use of multiple primers reduces fingerprint similarity and resolves discrepancies.     

家蚕来源肠球菌RAPD反应体系的优化

通过调整 Mg2 、DNA模板、引物、Taq DNA聚合酶和dNTPs的浓度及PCR反应循环次数,建立并优化了家蚕来源肠球菌的RAPD-PCR反应体系及条件.结果表明,在25 μl反应体系下,当Mg2 的浓度为3.5 mmol/L、DNA模板加入量为50 ng、引物浓度为15 pmol/L、Taq DNA聚合酶加入量为3U、 PCR反应循环次数为45时,扩增结果最好.在本实验室,为家蚕来源肠球菌的遗传多样性研究建立了一种最佳RAPD分析模型.     

Persistence of some pine pathogens in coarse woody debris and cones in a Pinus pinea forest

The persistence of Sphaeropsis sapinea, Leptographium serpens and Heterobasidion annosum s.s. in artificially inoculated pine branch pieces (S. sapinea and L. serpens) and wood blocks (L. serpens and H. annosum s.s.) was investigated in order to discuss the alternative of leaving coarse woody debris in stands of Italian stone pine (Pinus pinea). Also, natural colonization by S. sapinea of pine cones of different ages was assessed. Methods used for inoculating branch pieces and wood blocks were highly effective for all fungi. Type of a forest stand in which branch pieces and wood blocks have been incubated did not affect the persistence of the pathogens in the inoculated samples. For branch pieces, the success of re-isolation of L. serpens dropped as the sample incubation time increased, while S. sapinea was always successfully (100%) re-isolated (even 12 months after the inoculation). L. serpens and H. annosum s.s. were re-isolated from most of the buried wood blocks (from more than 95% samples) up to 3 months following the inoculation. Of the observed P. pinea cones (in most cases, more than 2 years old), 74% were naturally infected byS. sapinea. All three investigated pathogens were able to survive in dead plant tissues for long periods of time (at least for several (3–12) months). The persistence of these pine-pathogenic species in dead plant material questions the feasibility of leaving coarse woody debris in managed Italian stone pine forests meant for landscape conservation and leisure activities.     

Genetic diversity of Quercus glandulifera var. brevipetiolata populations in three forest communities with different succession stages

In order to understand the relationship between population succession and its genetic behavior, random amplified polymorphic DNA (RAPD) technique was used to analyze the genetic diversity of Quercu glandulifera var. brevipetiolata populations in three forest communities with different succession stages (coniferous forest, coniferous and broad-leaved mixed forest, evergreen broad-leaved forest). The results showed that 145 repetitive loci were produced in 60 individuals of Q. glandulifera using 11 primers, among which 120 loci were polymorphic, and the total percentage of polymorphic loci was 82.76% with an average of 64.14%. Estimated by the Shannon information index, the total genetic diversity of the three populations was 0.4747, with an average of 0.3642, while it was 0.3234, with an average of 0.2484, judged from the Nei index. Judged from percentage of polymorphic loci, Shannon inform at ion index and Nei index, the genetic diversity followed a decreasing order: coniferous forest > broad-leaved mixed forest > evergreen broad-leaved forest. Analysis of molecular variance (AMOVA) showed that 69.73% of the genetic variance existed within populations and 30.27% of the genetic variance existed among populations. The coefficient of gene differentiation (GST) was 0.2319 and the gene flow (N _m) was 1.6539. The mean of genetic identity among populations of Q. glandulifera was 0.8501 and the mean of genetic distance was 0.1626. The genetic identity between the Q. glandulifera population in the coniferous forest and that in the coniferous and broadleaved mixed forest was the highest. UPGMA cluster analysis based on Nei’s genetic distance showed that the population in the coniferous forest gathered with that in the coniferous and broad-leaved mixed forest firstly, then with that in the evergreen broad-leaved forest. The genetic structure of Q. glandulifera was not only characteristic of the biological characteristics of this species, but was also influenced by the microenvironment in different communities. __________ Translated from Journal of Northwest Forestry University, 2008, 23(1): 18–22 [译自: 西北林学院学报]     

Sapstain fungi on <Emphasis Type="Italic">Pinus radiata</Emphasis> logs – from New Zealand Forest to export in Japan

Supply of clear, unstained logs from an export country is important economically as well as in reducing the biosecurity risk to an importing country such as Japan. Although conditions found within the holds of ships containing logs are thought to be ideal for rapid colonization of sapstain fungi, no research has been conducted. Research-focussed log export trials were designed to determine the extent of sapstain colonisation at specific points in the processing of logs from harvesting to arrival at the export destination. Two trials were established, in the austral summer and winter, in which mature Pinus radiata logs were harvested in New Zealand and shipped to export ports in Japan. Data loggers placed above and below deck of the ships recorded microclimatic conditions. Nine species of sapstain fungi were isolated from logs during the summer trial; the most common species isolated were Ophiostoma floccosum, Ophiostoma querci, and Ophiostoma setosum. In contrast, ten sapstain species were detected during the winter trial, with Sphaeropsis sapinea, O. querci, O. floccosum, and O. setosum most commonly isolated. This research was the first successful attempt at measuring visual sapstain development and isolating sapstain fungi from the time of harvesting to arrival at an export destination.     

Monitoring genetic stability inQuercus serrata Thunb. somatic embryogenesis using RAPD markers

Genetic stability of propagules regeneratedvia somatic embryogenesis is of paramount importance for its application to clonal forestry. Random amplified polymorphic DNA (RAPD) markers were used to determine the genetic stability in somatic embryogenesis ofQuercus serrata Thunb. (Japanese white oak). Forty samples from an embryogenic line, consisting of regenerated plantlets, somatic embryos, and embryogenic calli, were examined using 54 decanucleotide primers. A total of 6520 clear reproducible bands obtained from these studies exhibited no aberration in RAPD banding pattern among the tested samples. Our results show that somaclonal variation is absent in our plant propagation system. The genetic stability is discussed in terms of the origin of somatic embryos.     

Site-related influences on cone-borne inoculum and asymptomatic persistence of Diplodia shoot blight fungi on or in mature red pines

The shoot blight and canker pathogens Diplodia pinea and D. scrobiculata sporulate abundantly on cones of many pine hosts. Variation in incidence and abundance of potential inoculum from cones and frequency of asymptomatic persistence on or in shoots was examined for mature red pines in sites differing in dominant presettlement vegetation and soil type in Bayfield and Douglas counties in northern Wisconsin. Collections were made in each county from 6 plantations, 3 each in areas historically vegetated with jack pine and soils mapped as sands and three in areas historically vegetated with red pine with soils mapped as loamy sands. At each site, 5 cones were collected from each of 5 red pines and 10 shoots were collected from up to 5 red pines. Conidia from cones were quantified with a water wash and filtration technique. Diplodia species were cultured from surface-disinfested asymptomatic shoots. A species-specific PCR assay was used to identify the Diplodia species from cones and shoots. Although cones and asymptomatic shoots from each county yielded D. pinea and D. scrobiculata, D. pinea was detected more frequently. More conidia were obtained from cones from Douglas Co., where there is a history of severe shoot blight damage, than cones from Bayfield Co. In Douglas Co., more conidia were obtained from cones from plantations in areas of more sandy soil and presettlement jack pine dominance than cones from plantations in areas of less sandy soil and presettlement red pine dominance. The numbers of conidia and frequencies of cultural detection of Diplodia species from asymptomatic shoots at a site were positively correlated. These results provide evidence for site-related influences on abundance of pathogen inoculum and asymptomatic persistence on or in red pine crowns that may contribute to differences in frequency and severity of damage from Diplodia shoot blight.     

Analysis of genetic diversity for wild and captive green peafowl populations by random amplified polymorphic DNA technique

The genetic diversity of the populations for 14 wild green peafowls (Pavo muticus) and 18 captive green peafowls was investigated by using the technology of random amplified polymorphic DNA (RAPD). Totally 161 and 166 amplified bands were obtained by using 23 arbitrary primers to amplify the genomic DNA of wild and captive green peafowls respectively. The results showed that the average relative genetic distance of the wild and captive green peafowls populations was 0.0555 and 0.1355, respectively, and difference of the average relative genetic distances between the two populations was 0.1635. The Shannon diversity index for the wild and captive green peafowl populations was 0.4348 and 1.0163, respectively, which means that there exists significant difference in genetic diversity between the two populations, and the genetic diversity of wild green peafowl was low. The two populations originated from two different families according to analysis by the UPGMA method. This research can provide the theoretical basis for supervising genealogies management of peafowl populations.     

Application of random amplified polymorphic DNA (RAPD) to detect genetic variation in Norway spruce

The RAPD assay is a screening method for genetic variation; in Norway spruce biology research, it may find many applications. We have investigated into its suitability for Norway spruce genetics by optimizing the protocol, testing for stability in clones, especially tissue culture clones, assessing the variation in a seed sample, and checking for correct Mendelian segregation in haploid megagametophytes. The RAPD assay produced numerous genetic markers quickly. The data obtained gave insight into the genetic make-up of clones and seed.

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Zusammenfassung Die RAPD Methode, ein Schnelltest für die genetische Variation, wurde für die europäische Fichte getestet, wo viele Aspekte der Biologie der Baumart damit untersucht werden könnten. Das Protokoll wurde optimiert, und genetische Stabilität in Klonen, besonders aus Gewebekultur, die genetische Variation in einer Samenprobe und die Mendel-konforme Aufspaltung in haploiden Megagametophyten wurde untersucht. Zahlreiche genetische Marker konnten gefunden werden, wodurch ein Einblick in die genetische Zusammensetzung des untersuchten Materials gegeben war.

     

文冠果种质资源的RAPD分析

利用RAPD标记技术对文冠果种质资源进行亲缘关系分析。11个引物在6份文冠果种质中共扩增出140条谱带,其中多态性谱带78条,多态性谱带比率为55.7%。UPGMA聚类结果表明,遗传距离为0.47时,6份供试样品可分为2类。     

An analysis of the pattern of genetic variation in Vitellaria paradoxa using RAPD markers

Vitellaria paradoxa C.F.Gaertn. is one of the most economically and socially important tree species in the Sudano-Sahelian region. Little is known of the pattern of variation within its natural range. Eight populations covering most of the natural range from Senegal to Uganda were sampled and leaves of 118 individual trees were collected. An analysis of molecular diversity was carried out using random amplified polymorphic DNA (RAPD) markers. Fifteen random primers generated 67 polymorphic and 15 monomorphic RAPD loci ranging from size 1670 bp to 280 bp. Shannon's diversity index varied from Central Africa/Ndele (0.374) to Uganda/Amoya (0.350) but the differences between populations were smaller than the population standard errors. Correspondence analysis of unrooted neighbour-joining trees suggested that genetic distances between populations were correlated with geographic distances. This trend was confirmed by a Mantel test giving a coefficient of correlation between genetic and geographic distances of R = 0.88 (P = 0.0001). Result of AMOVA (analyses of molecular variance) showed that 14.8% (P = 0.002) of the RAPD variation was distributed among populations. Nested analysis of variance indicated that variance between the western and eastern groups of population represented 8.7% (P = 0.001) of the total variation and the variation amongst populations within group was 9.5% (P = 0.001). Eighty two percent of the variation was explained by variation amongst individuals within populations. The origin of genetic structure and level of diversity may be explained by the glacial refugia, the biological traits of Vitellaria paradoxa and by the impact of semi-domestication. Based on these results, sampling options of the natural populations are suggested for in or ex situ conservation. For the development of Vitellaria paradoxa breeding population, the sampling should consist of many individual trees selected within a few populations to capture a large proportion of variation.This revised version was published online in November 2005 with corrections to the Cover Date.     

Cladistic relationships among the Pleurotus ostreatus complex, the Pleurotus pulmonarius complex, and Pleurotus eryngii based on the mitochondrial small subunit ribosomal DNA sequence analysis

The cladistic analysis of the V4 domain sequences, performed by UPGMA, the neighbor-joining, and parsimony methods, revealed that the 19 Pleurotus strains tested in this study evolved along three lineages, each corresponding to a separate biological species: the Pleurotus ostreatus complex, the Pleurotus pulmonarius complex, and Pleurotus eryngii. Moreover, the cladistic positions of the 3 biological species show that the P. ostreatus complex and P. eryngii were derived from a common ancestor at a later stage of evolution, and that the common ancestor had diverged from the lineage of the P. pulmonarius complex during an earlier evolutionary event. The sequences of the 5 portion of the mt SSU rDNA among the strains of the P. ostreatus complex had 99.2%–99.6% homology. All test strains in the P. pulmonarius complex had completely identical sequences. The homology of the strain sequences between the P. ostreatus complex and the P. pulmonarius complex ranged from 96.0% to 96.3%. The sequence of the strain of P. eryngii showed 97.8%–98.3% and 96.5% homologies with those of the strains in the P. ostreatus and the P. pulmonarius complexes, respectively.     

AFLP analysis of genetic variation of Hyphantria cunea (Drury) populations in Beijing and a nearby site

Hyphantria cunea (Drury) is a severe invasive pest in Beijing. It is important to understand the reason for its successful invasion and outbreak. Accordingly, this study was tailored to assess molecular variation and genetic relationships among three H. cunea populations in Beijing and one population from Sanhe in Hebei Province as control (totally 100 individuals). A silver-labeled amplified fragment length polymorphisms (AFLP) DNA profiling method was used to detect the genetic diversity and relationship among the populations. Five EcoRI and MseI primer combinations produced a total of 171 (91.2% polymorphic) informative fragments. The average Nei’s genetic diversity in four H. cunea populations analyzed by Popgene3.2 software was 0.2287, and the level of genetic diversity ranked in the order BDR (reared population in Daxing, Beijing) > BDN (natural population in Daxing, Beijing) > HSN (natural population in Sanhe, Hebei Province) > BFN (natural population in Fengtai, Beijing). The value of genetic differentiation among populations was 0.3321, and the gene flow was 1.0057. The genetic distance between these populations ranged from 0.0735 to 0.3309, 0.0531 on average. The result of UPGMA (unweighed pair group method with arithmetic averages) clustering showed that populations BFN and HSN were closely related, while BFN and BDN had the largest genetic distance. These results indicate that H. cunea populations in Beijing might originate from several areas.     

Genetic Diversity of RAPD Mark for Natural Davidia involucrata Populations

The genetic diversity and genetic variation within and among populations of five natural Davidia involucrata populations were studied from 13 primers based on random amplified polymorphic DNA (RAPD) analysis. The results show that natural D. involucrata population has a rich genetic diversity, and the differences among populations are significant. Twenty-six percent of genetic variation exists among D. involucrata populations, which is similar to that of the endangered tree species Liriodendron chinense and Cathaya argyrophylla in China, but different from more widely distributed tree species. The analysis of the impacts of sampling method on genetic diversity parameters shows that the number of sampled individuals has little effect on the effective number of alleles and genetic diversity, but has a marked effect on the genetic differentiation among populations and gene flows. This study divides the provenances of D. involucrata into two parts, namely, a southeast and a northwest provenance. Translated from Scientia Silvae Sinicae, 2004, 40(4) (in Chinese)     

Micropropagation of loblolly pine by somatic organogenesis and RAPD analysis of regenerated plantlets

IntroductionOrganogenesis and somatic embryogenesis haveben regarded as the in vitro system of choice withthe Pbtenhal for eventual mass propagation of superior and gen6tically engineering forest tree genotypesIn both conal6rous and hardwood (Gupts et al. 1991,Becwar et al. 1995). Somatic embryogenesis andorganeqenesis have been induced from more than 30ired speCies in conifers, but plant regeneration viaSOmahc embryogenesis and organogenesis remainsdffeutt cd a low rqeneration frequency (A…     

A population genetic study in a scattered forest species, wild service tree [Sorbus torminalis (L.) Crantz], using RAPD markers

Wild service tree [Sorbus torminalis (L.) Crantz] is a forest tree widespread in Europe, characterized by a scattered distribution. Its hermaphroditic flowers are pollinated by insects, and outcrossing is the prominent mating system, also due to the presence of gametophytic self-incompatibility. Genetic diversity and differentiation of 22 populations from northwestern Italy were investigated by means of variation scored at 53 polymorphic RAPD (Random Amplified Polymorphic DNA) loci. Populations differed in genetic variation, with Shannon diversity index ranging from 0.166 to 0.469. According to Analysis of Molecular Variance (AMOVA), most of the genetic variation was found within populations (61.78%) with a significant proportion of variance attributable to genetic differences between regions (23.60%) and between populations (14.62%). Evidence for isolation by distance was found in the set of populations sampled. The effect of habitat fragmentation on genetic variation was also evaluated. The efficacy of RAPD markers in analysing genetic variation, and the contribution of the results in terms of the preservation of biodiversity are discussed.     

Genetic variation among 11 <Emphasis Type="Italic">Abies concolor</Emphasis> populations based on allozyme analysis

In order to obtain information on the genetic structure ofAbies concolor and the genetic variation among 11 populations introduced from America to China, allozyme analysis based on starch gel electrophoresis technology was used. 24 loci of 10 allozyme systems were mensurated, and the genetic structure and genetic diversity of the 11 populations of A. concolor evaluated. The results show that the genetic variation among is significant, and the genetic variation within A. concolor populations is more important. In contrast with other conifers, the variation of A. concolor is above the average level of conifers, and higher than the same level of Abies. The percentage of polymorphic loci (P) was 62.5%, the number of alleles per locus (A) 2.08, the number of effective alleles per locus (Ae) was 1.37, the expected heterozygosity (H) 0.204, and the Shannon information index (/) 0.351 7. There is a short genetic distance (D=0.061) and a low gene flow (Nm=0.839 4) among the 11 introduced populations ofA. concolor with high genetic variation. The genetic differentiation coefficient (Gst) was 0.229 5, which is higher than that of the mean in Abies or Pinus.     

Analyses of the genetic diversity of matsutake isolates collected from different ecological environments in Asia

Matsutake mushrooms are able to grow in two kinds of ecological environments: coniferous forests and broadleaf forests. Japanese matsutake is a form of the matsutake mushroom that mainly appears in coniferous forests in the mid-latitude regions of Asia. In contrast, matsutake from Yunnan and Sichuan Provinces of China grows in broadleaf forests in low-latitude regions. The taxonomic relationship and the genetic distance between these two types of matsutake remain unclear. Here, we compared the random amplification of polymorphic DNA (RAPD) data and the sequences of the internal transcribed spacer (ITS) region and of variable domains V4, V6, and V9 of mitochondrial small subunit ribosomal DNA (mt SSU rDNA) to determine the genetic differences of Tricholoma matsutake strains collected from different ecological and geographic environments in Asia. We found few differences in the sequences of the variable domains, implying that all matsutake strains are the same biological species regardless of their host specificity and geographical differences. This conclusion is also supported by the sequences of the ITS region. On the other hand, RAPD analysis revealed that the matsutake mushrooms collected from different ecological environments have genetic differences and represent distinct varieties.