Lesion formation and host response to infection by Armillaria ostoyae in the roots of western larch and Douglas‐fir

The process of lesion formation and host response to natural infection by Armillaria ostoyae were studied in the roots of western larch (Larix occidentalis) and Douglas‐fir (Pseudotsuga menziesii ssp. glauca) trees in the three age classes, 6–8, 18–19 and 85–95 years. The characteristics of lesions on infected roots were recorded and bark samples were dissected from infection points and lesion margins in the field and stored in liquid nitrogen for macroscopic study in the laboratory. Infection in the roots of 6‐ to 8‐year‐old trees advanced freely, overcoming any host resistance, quickly girdling the root collar and killing the trees. In 18‐ and 19‐year‐old trees, however, 43% of infections on western larch and 27% of the infections on Douglas‐fir roots were confined to lesions bounded by necrophylactic periderms with multiple bands of phellem. Host response was similar in 85‐ to 95‐year‐old trees, but the percentage of confined lesions was higher than in younger trees. The results suggest that larch shows resistance to A. ostoyae at a younger age and with greater frequency than Douglas‐fir.     

Identification of Kenyan Armillaria isolates by cultural morphology,intersterility tests and analysis of isozyme profiles

Three groups of morphologically distinct isolates were observed among nine Kenyan Armillaria isolates based on their mycelium and rhizomorphs characteristics. Seven of the isolates were interfertile with testers of North American biological species III, VII and IX. However, tests with benomyl segregants BEN 433, BEN 157-10 and BEN AVK were intersterile with the same testers and also with the European A. mellea, A. lutea and A. ostoyae. The analysis of isozyme profiles showed that morphologically similar isolates had similar isozyme profiles of esterases. Their profiles however differed from those of the European A. mellea, A. lutea and A. ostoyae. On the basis of intersterility tests and analysis of isozyme profiles, the Kenyan isolates should be considered different.     

Identification of Armillaria species in Japan using PCR-RFLP analysis of rDNA intergenic spacer region and comparisons of Armillaria species in the world

Armillaria species from Japan were characterized using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the intergenic spacer region-1 (IGS-1) of ribosomal DNA (rDNA). Eleven different digestion patterns by restriction endonuclease Alu I were found among 70 isolates of seven Armillaria species in Japan. Isolates within Armillaria nabsnona, A. ostoyae, A. cepistipes, and Japanese biological species E showed the same Alu I digestion patterns. Five Alu I patterns were detected for A. gallica, three patterns for A. mellea, and two patterns for A. tabescens. Seven Armillaria species in Japan were clearly distinguished by using the profiles obtained when PCR products were digested with Alu I, Msp I, and Hae III restriction enzymes. There was considerable variability of Alu I restriction sites within the IGS-1 between the isolates of five Armillaria species, A. gallica, A. nabsnona, A. cepistipes, A. mellea, and A. tabescens, in Japan and those of their European and North American counterparts.     

Use of random amplified polymorphic DNA (RAPD) for detection of genetic variation and proof of the heterothallic mating system in Hypoxylon mediterraneum

The efficiency of RAPD analysis in the evaluation of genetic variability in Hypoxylon mediterraneum has been tested on 49 monoascospore isolates of the fungus from different localities. A large genetic variability was detected within single stromata and among monoascospore isolates in a restricted area. The large variability in restricted populations can be explained by the heterothallic mating system and the life cycle of the fungus and its ecological adaptations. RAPD markers can discriminate monoascospore isolates both in the single stromata and among stromata. The application of RAPD markers for epidemiological studies is discussed.     

Detection of intra- and interspecific variation of the dry rot fungus <Emphasis Type="Italic">Serpula lacrymans</Emphasis> by PCR-RFLP and RAPD analysis

We investigated a genotype-based assay to discriminate the dry rot fungi Serpula lacrymans. DNAs were extracted from 74 isolates from the northern half of Japan, and internal transcribed spacers (ITS) were amplified by polymerase chain reaction. Genotypes of isolates were checked by restriction fragment length polymorphism (RFLP) using two enzymes, Taq I and Hha I. Among the 74 isolates identified as S. lacrymans in terms of morphologic features, 5 isolates were shown to have been misidentified. Random amplified polymorphic DNA (RAPD) analysis was conducted in order to detect the intraspecific diversity of S. lacrymans isolated in Japan. Because no relation between geographical origin and genetic distances was observed, the intraspecific diversity of S. lacrymans is suggested to be small.Part of this report was presented at the 52nd Annual Meeting of the Japan Wood Research Society, Gifu, April 2002     

Agronomic evaluation of Leucaena. Part 2. Productivity of the genus for forage production in subtropical Australia and humid-tropical Philippines

Leucaena leucocephala is an important agroforestry species pan-tropically, but relatively little is known of the forage production potential of other species in the genus. The agronomic potential of 116 accessions, represent ing the 28 species and subspecies of the Leucaena genus and several artificial hybrid accessions, was evaluated at Los Baños, Philippines and Brisbane, Australia over a 2.5–year period. Accessions were planted into replicated line plots, with 10 trees/plot spaced 50 cm apart, and with rows spaced 3 m apart. The L. pallida × L. leuco cephala KX2 F ₁ hybrid accessions were highest yielding at both sites, producing dry matter (DM) yields of over 900 g/m row/month at Los Baños and approximately 320 g/m row/month at Brisbane. In the near-optimal growth conditions at Los Baños, L. leucocephala accessions were highly productive, with the best accessions producing total yields of over 500 g/m row/month. The superiority of KX2 hybrids was most pronounced at Brisbane, where high psyllid pressure during summer, and low temperatures during winter severely constrained growth of L. leu cocephala accessions. In the Brisbane environment, psyllid resistant accessions of L. pallida , L. trichandra and L. diversifolia were more productive than L. leucocephala accessions. Leucaena greggii , L. retusa , L. cuspidata , L. confertiflora , L. pulverulenta , L. pueblana and L. involucrata were of inherently low productivity in both the Brisbane and Los Baños environments. Mortality over the experimental period was very low for most species, particularly for L. leucocephala and KX2 accessions. The KX2 F1 hybrid accessions have considerable agro nomic potential as alternatives to L. leucocephala for use in tropical agroforestry.     

A comparison of UP‐PCR and RAPD markers to study genetic diversity of Fusicladium effusum (G. Winter), cause of pecan scab

Fusicladium effusum infects pecan causing yield loss, but no information is available on the genetic diversity of F. effusum. Randomly amplified polymorphic DNAs (RAPDs) and universally primed polymerase chain reaction (UP‐PCR) were compared to detect polymorphisms on a group of 20 isolates of F. effusum from 11 geographical locations in the southeastern USA. Two tests (run 1 and 2) of both the RAPD and UP‐PCRs were conducted to assess the repeatability of the methods, and the markers scored on agarose gels. In addition, the UP‐PCR markers from run 1 were scored using an automated capillary system. Both RAPDs and UP‐PCR markers detected a high level of polymorphism among the scored markers (92 and 91% of RAPD markers, and 86 and 87% of manually scored UP‐PCR markers in run 1 and 2 were polymorphic, respectively; 93% of UP‐PCR markers were polymorphic when scored using the automated system). Unweighted paired group method of arithmetic averages (UPGMA) analysis showed both RAPDs and UP‐PCR markers individually identified each isolate, producing three groupings, but only the groupings based on run 1 and 2 of the UP‐PCR contained the same isolates. Bootstrap analysis based on the Dice coefficient produced phenograms from the UP‐PCR data with weak to moderate node support (≥54) for the primary branch, but no support for the RAPDs data (≤34). A Mantel test of runs 1 and 2 using RAPDs or UP‐PCR showed good agreement (r = 0.8761 and 0.8289, p < 0.0001), but poor agreement between RAPDs and UP‐PCR. UP‐PCR results based on the interisolate Dice coefficients showed a weak to strong association with distance. Based on these results, both RAPDs and UP‐PCR markers were capable of demonstrating polymorphisms and identifying relationships among isolates of F. effusum; however, UP‐PCR markers appear to be more reliable.     

Molecular analysis of the genus Asparagus based on matK sequences and its application to identify A. racemosus, a medicinally phytoestrogenic species

The plant Asparagus racemosus is one of the most widely used sources of phytoestrogens because of its high content of the steroidal saponins, shatavarins I-IV, in roots. The dry root of A. racemosus, known as "Rak-Sam-Sip" in Thai, is one of the most popular herbal medicines, used as an anti-inflammatory, an aphrodisiac and a galactagogue. Recently, the interest in plant-derived estrogens has increased tremendously, making A. racemosus particularly important and a possible target for fraudulent labeling. However, the identification of A. racemosus is generally difficult due to its similar morphology to other Asparagus spp. Thus, accurate authentication of A. racemosus is essential. In this study, 1557-bp nucleotide sequences of the maturase K (matK) gene of eight Asparagus taxa were analyzed. A phylogenetic relationship based on the matK gene was also constructed. Ten polymorphic sites of nucleotide substitutions were found within the matK sequences. A. racemosus showed different nucleotide substitutions to the other species. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the matK gene was developed to discriminate A. racemosus from others. Only the 650-bp PCR product from A. racemosus could be digested with BssKI into two fragments of 397 and 253-bp while the products of other species remained undigested. Ten commercially crude drugs were analyzed and revealed that eight samples were derived from A. racemosus while two samples of that were not. Thus, the PCR-RFLP analysis of matK gene was shown to be an effective method for authentication of the medicinally phytoestrogenic species, A. racemosus.     

利用RAPD分析药用石斛的遗传多样性及亲缘关系

通过对9种药用石斛进行遗传多样性和亲缘关系的分析,为更好地开发利用石斛资源奠定基础。采用RAPD技术,应用NTSYS软件对9种石斛的RAPD-PCR结果进行分析。利用从103条随机引物中筛选出的70条随机引物对供试材料DNA进行随机扩增,共得到520条带,其中多态性带有491条,多态性百分率为94.42%。采用UPGMA类平均法对扩增出的谱带进行遗传聚类分析,得出反映各种间亲缘关系的树状图。在遗传距离（D）=0.44处,可将供试材料分为2类,RAPD对基因组的分析结果与传统分类学的结果差异不大。试验结果表明,RAPD技术可有效地应用于药用石斛的遗传多样性及亲缘关系的研究。     

Identification and evaluation of reference genes for normalization in quantitative real-time PCR analysis in the premodel tree Betula luminifera

Betula luminifera is a commercial tree species that is emerging as a new model system for tree genomics research. A draft genomic sequence is expected to be publicly available in the near future, which means that an explosion of gene expression studies awaits. Thus, the work of selecting appropriate reference genes for q PCR normalization in different tissues or under various experimental conditions is extremely valuable. In this study, ten candidate genes were analyzed in B. luminifera subjected to different abiotic stresses and at various flowering stages.The expression stability of these genes was evaluated using three distinct algorithms implemented using ge Norm,Norm Finder and Best Keeper. The best-ranked reference genes varied across different sample sets, though RPL39,MDH and EF1 a were determined as the most stable by the three programs among all tested samples. RPL39 and EF1 a should be appropriate for normalization in N-starved roots,while the combination of RPL39 and MDH should be appropriate for N-starved stems and EF1 a and MDH should be appropriate in N-starved leaves. In PEG-treated(osmotic) roots, MDH was the most suitable, whereas EF1 a was suitable for PEG-treated stems and leaves. TUA was also stably expressed levels in PEG-treated plants. The combination of RPL39 and TUB should be appropriate for heat-stressed leaves and flowering stage. For reference gene validation, the expression levels of SOD and NFYA-3were investigated. This work will be beneficial to future studies on gene expression under different abiotic stress conditions and flowering status in B. luminifera.     

Identification of early-flower-related ESTs in an early-flowering mutant of trifoliate orange (Poncirus trifoliata) by suppression subtractive hybridization and macroarray analysis

To increase our understanding of the genes involved in flower development in citrus, we identified genes differentially expressed between juveniles and adults of an early-flowering mutant of trifoliate orange (precocious trifoliate orange, Poncirus trifoliata L. Raf.) by suppressive subtractive hybridization (SSH) and macroarray hybridization analyses. Based on dot blot analysis, we confirmed that the juvenile subtracted cDNA library comprised genes upregulated during floral induction, inflorescence development and flowering. A total of 190 nonredundant expressed sequence tags were identified that were related flower development. The temporal and spatial expression patterns of four SSH-enriched genes were studied in adult precocious trifoliate orange by real-time PCR. Among these differentially expressed genes, the expression pattern of C5-B16 was closely correlated with inflorescence development and flowering. The full-length cDNA of C5-B16 was isolated by 5' and 3' rapid amplification of cDNA ends. Sequence analysis indicated that C5-B16 is a novel gene in citrus.     

应用灰色系统理论预测城峰镇十五期间有林地面积与森林蓄积量变化的研究

以永泰县城峰镇1986～1999年历年有林地面积、森林蓄积量数据为基础,应用灰色系统理论与方法,建立灰色预测GM(1,1)模型,以城峰镇“十五”期间的有林地面积与森林蓄积量进行预测。经检验,有林地面积GM(1,1)模型(1986～1999年)预测值的平均误差2.23％,森林蓄积量GM(1,1)模型(1986～1999年)预测值的平均误差1.16％,预测精度较高,可以对“十五”期间城峰镇有林地面积与森林蓄积量进行预测。     

Development of an efficient protocol for plant regeneration from nodal explants of recalcitrant bamboo (Bambusa arundinacea Retz. Willd) and assessment of genetic fidelity by DNA markers

The present study describes an efficient method for in vitro plant regeneration in B. arundinacea through axillary shoot bud proliferation. Nodal explants were excised, cultured on MS medium containing different concentrations of 6-benzylaminopurine (BAP), kinetin (KIN) (0.5–5.0 mg l^?1) alone and/or in combinations with KIN/BAP (0.5 mg l^?1). The highest frequency (91.5 %) of multiple shoot bud induction with maximum number of shoots (85 shoots/explant) was noticed on MS medium + 3.0 mg l^?1 BAP + 0.5 mg l^?1 KIN. The regenerated multiple shoots were elongated on MS medium + 4.0 mg l^?1 KIN + 2.0 mg l^?1 gibberellic acid (GA₃) with maximum shoot length (4.9 cm). The elongated shoots were transferred to MS medium containing indole-3 butyric acid (IBA; 0.5–5.0 mg l^?1) alone and/or in combination with 0.5 mg l^?1 KIN and BAP. Highest frequency of rooting (75 %) was obtained on half-strength MS medium + 2.0 mg l^?1 IBA + 0.5 mg l^?1 KIN. After hardening, the plantlets were shifted to the green house and subsequently established in the field conditions with 90 % survival rate. random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic stability of the regenerants. RAPD profiles generated from the regenerated plants were found to be monomorphic, similar to the control. Results confirmed that the regenerated plants were true-to-type in nature and the developed micropropagation protocol could be used for large scale plant production of B. arundinacea.     

Climate change and tree genetic resource management: maintaining and enhancing the productivity and value of smallholder tropical agroforestry landscapes. A review

Anthropogenic climate change has significant consequences for the sustainability and productivity of agroforestry ecosystems upon which millions of smallholders in the tropics depend and that provide valuable global services. We here consider the current state of knowledge of the impacts of climate change on tree genetic resources and implications for action in a smallholder setting. Required measures to respond to change include: (1) the facilitated translocation of environmentally-matched germplasm across appropriate geographic scales, (2) the elevation of effective population sizes of tree stands through the promotion of pollinators and other farm management interventions; and (3) the use of a wider range of ??plastic?? species and populations for planting. Key bottlenecks to response that are discussed here include limitations in the international exchange of tree seed and seedlings, and the absence of well-functioning delivery systems to provide smallholders with better-adapted planting material. Greater research on population-level environmental responses in indigenous tree species is important, and more studies of animal pollinators in farm landscapes are required. The development of well-functioning markets for new products that farmers can grow in order to mitigate and adapt to anthropogenic climate change must also consider genetic resource issues, as we describe.     

能源植物膏桐的生产与研究现状

膏桐（Jatropha curcas L.）又称小桐子、麻疯树,作为一种多用途油料植物,具有耐贫瘠、耐干旱、病虫害少、速生、含油率高等优点,目前已被世界各地广泛种植和研究,但还需要进一步研究生产管理技术来确保膏桐产业的长远发展,尤其是不同气候条件下管理和施肥体系。作者力求全面探述国内外生产和研究状况,并指出了膏桐产业发展中需要关注的问题。     

Identification and evaluation of reference genes for normalization in quantitative real-time PCR analysis in the premodel tree <Emphasis Type="Italic">Betula luminifera</Emphasis>

Betula luminifera is a commercial tree species that is emerging as a new model system for tree genomics research. A draft genomic sequence is expected to be publicly available in the near future, which means that an explosion of gene expression studies awaits. Thus, the work of selecting appropriate reference genes for qPCR normalization in different tissues or under various experimental conditions is extremely valuable. In this study, ten candidate genes were analyzed in B. luminifera subjected to different abiotic stresses and at various flowering stages. The expression stability of these genes was evaluated using three distinct algorithms implemented using geNorm, NormFinder and BestKeeper. The best-ranked reference genes varied across different sample sets, though RPL39, MDH and EF1a were determined as the most stable by the three programs among all tested samples. RPL39 and EF1a should be appropriate for normalization in N-starved roots, while the combination of RPL39 and MDH should be appropriate for N-starved stems and EF1a and MDH should be appropriate in N-starved leaves. In PEG-treated (osmotic) roots, MDH was the most suitable, whereas EF1a was suitable for PEG-treated stems and leaves. TUA was also stably expressed levels in PEG-treated plants. The combination of RPL39 and TUB should be appropriate for heat-stressed leaves and flowering stage. For reference gene validation, the expression levels of SOD and NFYA-3 were investigated. This work will be beneficial to future studies on gene expression under different abiotic stress conditions and flowering status in B. luminifera.     

Genetic variation in Picea abies (L.) Karst. and Fomes annosus (Fr.) Cke. shown by growth of the fungus on wood dust

Growth of Fomes annosus is studied by a test based on growth of previously starved hyphae on sawdust of heartwood of Picea abies as the only nutrition. Our observations show that the method can be used to distinguish beween different spruce individuals or clones, between different Fomes annosus strains and between different growing sites of the trees.     

Efficiency of the indirect selection and the evaluation of the genotype by environment interaction using Pilodyn for the genetic improvement of wood density in <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>

The efficiency of the indirect selection for wood density using the Pilodyn in Cryptomeria japonica was studied by comparing the Pilodyn penetration (PP) depth and the direct measurement of wood density in three test sites. The influence of the genotype by environment (G × E) interaction of wood density was estimated using the Pilodyn with common 12 C. japonica clones in 10 test sites in Kanto breeding region in Japan. The PP depth was highly correlated with wood density, and the genetic correlation between them was −0.88. The indirect selection using the Pilodyn realized 87% of the genetic gain obtained by the direct selection of wood density. The G × E interaction in PP depth was small. The ratio of the variance component of the G × E interaction to that of genotype was only 0.096 in the PP depth, whereas it was 0.700 in tree height and 1.410 in diameter at breast height. These findings indicate that the Pilodyn is useful for the genetic improvement of wood density in Cryptomeria japonica. The small G × E interaction in wood density estimated using the Pilodyn indicates that the relative clonal performance in wood density is stable among diverse environments in Kanto breeding region in Japan.     

Interfertile oaks in an island environment: I. High nuclear genetic differentiation and high degree of chloroplast DNA sharing between <Emphasis Type="Italic">Q. alnifolia</Emphasis> and <Emphasis Type="Italic">Q. coccifera</Emphasis> in Cyprus. A multipopulation study

The evergreen Quercus alnifolia and Q. coccifera form the only interfertile pair of oak species growing in Cyprus. Hybridization between the two species has already been observed and studied morphologically. However, little evidence exists about the extent of genetic introgression. In the present study, we aimed to study the effects of introgressive hybridization mutually on both chloroplast and nuclear genomes. We sampled both pure and mixed populations of Q. alnifolia and Q. coccifera from several locations across their distribution area in Cyprus. We analyzed the genetic variation within and between species by conducting analysis of molecular variance (AMOVA) based on nuclear microsatellites. Population genetic structure and levels of admixture were studied by means of a Bayesian analysis (STRUCTURE simulation analysis). Chloroplast DNA microsatellites were used for a spatial analysis of genetic barriers. The main part of the nuclear genetic variation was explained by partition into species groups. High interspecific differentiation and low admixture of nuclear genomes, both in pure and mixed populations, support limited genetic introgression between Q. alnifolia and Q. coccifera in Cyprus. On the contrary, chloroplast DNA haplotypes were shared between the species and were locally structured suggesting cytoplasmic introgression. Occasional hybridization events followed by backcrossings with both parental species might lead to this pattern of genetic differentiation.