绣球属(Hydrangea L.)植物分子系统学及系统发育分析

该研究基于对绣球属(Hydrangea L.)的大尺度取样,选取国内外61种绣球属和近缘属植物,分别基于核基因片段(ITS)和叶绿体基因片段(rbcL,trnL-F,atpB)重建了绣球属及其近缘种属的系统发育关系。结果表明:(1)核基因与叶绿体基因树之间在树形上没有明显的冲突,进而基于核基因和叶绿体基因联合数据重建了绣球属及其近缘种属的系统发育关系。(2)基于联合数据构建的系统树确认了2个大分支,并得到了果实顶端截平与否这一形态学证据的强力支持;每个大分枝又分为4个类群,共确定了8个类群。部分类群也得到了广义宏观形态性状的支持,如第1类群得到了叶形、花粉以及种子形态的支持。因此,该系统发育关系的重建对于全面理解绣球属及其近缘种属的演化关系具有重要的启发。     

Astringent tannins of Viburnum and Hydrangea species

The tannins of the leaves of Viburnum and Hydrangea species consist of proanthocyanidins only, but in each genus the range is very wide. In several species of Hydrangea the proanthocyanidins are of the A type, otherwise they are mostly tri- or tetrameric B type. Tannin content is correlated with evolutionary advancement, the more advanced and more widely dispersed species having the less. Species with most tannin occur in E. Asia and E. N. America, but species with little or no tannin are present in both areas. The occurrence in both genera of globose inflorescences with sterile flowers is correlated neither with morphological nor with chemical characters.     

Deteriorating effects of diapause duration on postdiapause life history traits in the Kanzawa spider mite

Abstract. The aim of this study is to clarify the deleterious effects of long‐term diapause on survival, postdiapause number of offspring and longevity in two populations of Kanzawa spider mite Tetranychus kanzawai Kishida (Acari: Tetranychidae), which were collected from Orixa japonica (Rutaceae) and Hydrangia hirta (Saxifragaceae) in Kyoto Prefecture, Japan. As the chilling period lengthens, the rate of surviving females declines more steeply in the Orixa population than the Hydrangea population. In the Orixa population, the offspring numbers of diapausing females chilled for 30 or 90 days are not significantly different from those of the parental generation, whereas they decrease to approximately one‐half that of the parental generation in the Hydrangea population. The postdiapause longevity in the 30‐ or 90‐day chilling treatments does not shorten significantly compared with that of parental generation in the Orixa population, but the longevity of the Hydrangea population declines significantly after these chilling periods. These results suggest that deteriorating effects of diapause act in a quite different manner in the two populations. A possible mechanism generating such a difference is discussed in view of trade‐off between the maintenance of body conditions and survival during hibernation.     

Sensitive detection of mycoplasmalike organisms in field-collected and in vitro propagated plants of Brassica, Hydrangea and Chrysanthemum by polymerase chain reaction

A polymerase chain reaction (PCR) protocol, previously designed for amplification of a DNA fragment from aster yellows mycoplasmalike organism (MLO), was employed to investigate the detection of MLO DNA in field-collected and in vitro micropropagated plants. PCR with template DNA extracted from symptomatic, naturally-infected samples of Brassica, Chrysanthemum and Hydrangea, each yielded a DNA band corresponding to 1.0 Kbp. However, no DNA product was observed when either infected Ranunculus (with phyllody disease) or Gladiolus with (symptoms of ‘germs fins’) was used as source of template nucleic acid for PCR; further experiments indicated absence of target DNA in the case of Ranunculus and the presence of substances in Gladiolus which inhibited the PCR. The MLO-specific DNA was detected by PCR using less than 95 pg of total nucleic acid (equivalent to total nucleic acid from 1.9, ug tissue) in the case of field-collected Hydrangea and less than 11.4 pg of nucleic acid (equivalent to total nucleic acid from 19 ng of tissue) in the case of field-collected Brassica. The findings illustrate highly sensitive detection of MLOs in both field-grown and in vitro micropropagated infected plants.     

Fatty acids of some cornaceae,hydrangeaceae, aquifoliaceae,hamamelidaceae and styracaceae

Seed, kernel or fruit oils of the Cornaceae (nine species). Hydrangea, Hamamelis, Ilex (Aquifoliaceae) and the Styracaceae (two species) were analysed for fatty acid composition. Special attention was paid to the occurrence of petroselinic acid (18: 1Δ6c). In the species investigated. C₁₈ acids were always present in greater quantities than C₁₆ fatty acids; C₂₀ and C₂₂ acids were only minor components. The Cornaceae show differing fatty acid patterns which correlate well with anatomical, morphological and other chemical data. In Cornus, Curtisia, Mastixia and Corokia linoleic acid predominates, whereas linoleic and linolenic acid form the major components in Davidia and Nyssa. 18 : 1Δ6c, an aralioid type, occurs in large amounts in Aucuba and Griselinia. Hamamelis, Hydrangea and Ilex show a common fatty acid pattern with linoleic acid as the dominant component in all cases. Classification currently based on morphological and anatomical differences between the two species of Styracaceae which were investigated here should include their different fatty acid compositions: in Halesia linoleic acid predominates over oleic acid, whereas in Styrax equal amounts of these two acids are found.     

Facilitating wide hybridization in Hydrangea s. l. cultivars: A phylogenetic and marker-assisted breeding approach

Hydrangea s. l., belonging to the up-market segment of ornamental cultivars, currently faces a renaissance in horticulture. Hence, novel molecular-assisted breeding approaches are timely. Wide hybridization, i.e. crosses between distantly related species, has been shown to be problematic. Recent studies have considerably improved our knowledge of the phylogenetic relationships between the ornamental Hydrangea s. l. species. A fully resolved and highly supported phylogenetic tree is currently available, based on an extensive marker selection including 13 highly variable chloroplast markers. This robust phylogenetic framework includes the majority of widely cultivated Hydrangea s. l. species that have been the center of attention in a number of crossing projects. The present study is based on this highly supported phylogenetic hypothesis. Here, we aim to select the best candidates for future successful breeding projects, involving interspecific crosses of both closely and distantly related Hydrangea s. l. lineages. Therefore, we integrated the phylogenetic relatedness of potential parental lines along with genetic distances calculated from a wide plastid marker selection. Direct crosses between two species were found to be successful up to an average genetic distance of 0.01065, while failure could be expected at an average genetic distance of 0.01385 and higher. In order to overcome this genetic distance threshold, we propose Hydrangea arborescens, H. sargentiana, H. integrifolia, and H. seemannii as the best candidates for future bridge-cross projects with currently available fertile hybrids. We expect that our results will allow breeders to overcome long-standing wide crossing difficulties and motivate breeding initiatives of potential economic value.     

绣球属(Hydrangea L.)及近缘属花粉形态的研究

为深入研究绣球属植物花粉形态的分类学价值和系统学意义,厘清绣球属与近缘属之间的系统发育关系,该文利用扫描电子显微镜(SEM,scanning electron microscope)对国产绣球属及其近缘属41种绣球花科(Hydrangeaceae)植物的花粉形态以及表面纹饰进行了观察。结果表明:绣球属及其近缘属的花粉为三孔沟;形状多数为长圆体形或近球体形;赤道面观为椭圆形或圆形;极面观多为圆形,少数为三角形或圆三角形。花粉外壁纹饰可分为网状和孔穴状。网眼内的三级纹饰可分为光滑和具颗粒状突起。根据花粉形状和外壁纹饰类型将上述物种划分为4个组,即花粉的形状为长圆体形,表面纹饰为孔穴纹饰;花粉的形状为长圆体形,表面纹饰为网状纹饰;花粉的形状为近球体形,表面纹饰为孔穴纹饰;花粉的形状为近球体形,表面纹饰为网状纹饰。以上可进一步细分为8个类型。上述表明花粉形态证据可为绣球属及其近缘属的属下分类和种间界定提供重要佐证;但结合前人的系统发育重建分析该属植物花粉形态的系统学意义相对有限,如花粉形态证据对于该属及其近缘种属系统发育树上大支的界定难以提供有力的证据。     

Relationships and evolution of Hydrangeaceae based on RBCl sequence data

Phylogenetic analyses of rbcL sequences were used to address both systematic and evolutionary questions posed by the angiosperm family Hydrangeaceae. Our analyses suggest the presence of a monophyletic Hydrangeaceae most closely allied with Loasaceae, a finding in agreement with other molecular as well as morphological analyses. Molecular data indicate that Hydrangeaceae comprise Decumaria, Pileostegia, Schizophragma, Hydrangea, Dichroa, Broussaisia, Platycrater, Cardiandra, Deinanthe, Carpenteria, Philadelphus, Deutzia, Fendlerella, Whipplea, Fendlera, Jamesia, and the enigmatic Kirengeshoma. A particularly close relationship of Kirengeshoma and Deutzia is indicated. Analysis of rbcL sequences suggests that Fendlera and Jamesia are sister to the remainder of the family, lending support to the hypothesis that at least some Carpenterieae are basal in the family and that Hydrangeaceae may have originated in xeric habitats. If this phylogenetic placement of Jamesia and Fendlera is correct, the rbcL trees also suggest that the level of epigyny has decreased in these genera, as well as in the Fendlerella- Whipplea clade and Carpenteria when compared to the outgroup taxa, which are wholly epigynous. Furthermore, the rbcL trees support proposed evolutionary trends in wood anatomy, suggesting, for example, that upland tropical taxa have evolved longer vessel elements with more numerous bars on scalariform perforation plates. The xerophytic basal members of Hydrangeaceae, like the closely related Loasaceae, have short, narrow vessel elements with scalariform perforation plates bearing few bars. Following Jamesia and Fendlera, the remaining hydrangeoids are divided into two large subclades that closely parallel the traditional division of the family into Philadelpheae and Hydrangeae. Both rbcL sequences and morphological data suggest close relationships between: 1) Fendlerella and Whipplea; 2) Decumaria, Pileostegia, and Schizophragma; 3) Carpenteria and Philadelphus; 4) Deinanthe and Cardiandra; 5) Dichroa, Broussaisia, and Hydrangea macrophylla. Molecular and morphological data also concur in demonstrating that the large genus Hydrangea is not a monophyletic assemblage.     

Cytogenetic characterization of <Emphasis Type="Italic">Hydrangea involucrata</Emphasis> Sieb. and <Emphasis Type="Italic">H. aspera</Emphasis> D. Don complex (Hydrangeaceae)<Emphasis Type="Italic">:</Emphasis> genetic,evolutional, and taxonomic implications

The subsection Asperae of genus Hydrangea L. (Hydrangeaceae) has been investigated for three reasons: several ambiguous classifications concerning Hydrangea aspera have been published, unexpected differences in genome size among seven accessions have been reported Cerbah et al. (Theor Appl Genet 103:45–51, 2001), and two atypical chromosome numbers (2n = 30 for Hydrangea involucrata and 2n = 34 for H. aspera) have been found when all other species of the genus present 2n = 36. Therefore, these two species and four subspecies of Hydrangea in all 29 accessions were analyzed for their genome size, chromosome number, and karyotype features. This investigation includes flow cytometric measurements of nuclear DNA content and bases composition (GC%), fluorochrome banding for detection of GC- and AT-rich DNA regions, and fluorescent in situ hybridisation (FISH) for chromosome mapping of 5 S and 18 S-5.8 S-26 S rDNA genes. In the H. aspera complex, the genome size ranged from 2.98 (subsp. sargentiana) to 4.67 pg/2C (subsp. aspera), an exceptional intraspecific variation of 1.57-fold. The mean base composition was 40.5% GC. Our report establishes the first karyotype for the species H. involucrata, and for the subspecies of H. aspera which indeed present different formulae, offering an element of discrimination. FISH and fluorochrome banding revealed the important differentiation between these two species (H. involucrata and H. aspera) and among four subspecies of the H. aspera complex. Our results are in agreement with the Chinese classification that places the groups Kawakami and Villosa as two different species: Hydrangea villosa Rehder and Hydrangea kawakami Hayata. This knowledge can contribute to effective germplasm management and horticultural use.     

Quantitative and qualitative changes in carbohydrates associated with spring deacclimation in contrasting Hydrangea species

Cold deacclimation and associated changes in soluble carbohydrates and water status of two Hydrangea species differing in susceptibility to frost injuries was followed under natural conditions. In fully cold hardy plants of H. macrophylla stem freezing tolerance fluctuated in parallel with changes in air temperature, while in a seasonal perspective increased temperatures caused a sigmoid deacclimation pattern in both H. macrophylla and H. paniculata. Timing of deacclimation was approximately synchronized in the two species, but H. paniculata, the hardier species based on mid-winter hardiness, deacclimated faster than H. macrophylla, indicating that deacclimation kinetics were not correlated with mid-winter hardiness. In both species concentrations of soluble sugars decreased during deacclimation and were highly correlated with stem cold hardiness and air temperatures. This suggests that sugar hydrolysis may be an important temperature-driven mechanism of deacclimation in Hydrangea. Accumulation patterns of specific carbohydrates differed between the two species, suggesting that they utilize different strategies to overcome cold. In H. paniculata, deacclimation was associated with an increase in stem water content, which occurred shortly before bud burst and hence may be a prerequisite for leafing out.     

An Efficient Plant Regeneration System of <i>Hydrangea bretschneideri</i> Dipp via Stem Segments as Explants

Hydrangea bretschneideri Dipp is a highly popular ornamental plant for garden decoration. Genetic engineering technology has been successfully used in many plant species, but it is limited in Hydrangea. Here we established an efficient regeneration system by using stem segments as explants for the first time. In our study, the plant growth regulators (PGRs) were evaluated at the different regeneration processes, including axillary shoots regeneration and root induction. We found that the optimal concentration for axillary buds’ induction was 2.0 mgL⁻¹ 6-BA and 0.5 mgL⁻¹ 1 IAA, its highest induction rate was 70%. Moreover, the highest axillary shoots proliferation coefficient was 10.7 on the Murashige and Skoog (MS) medium with 2.0 mgL⁻¹ 6-benzyladenine (BA), 0.2 mgL⁻¹ indole-3-butyric acid (IBA), and 1.0 mgL⁻¹ gibberellin A3 (GA3). The highest frequency of root induction was 80.0 ± 0.06% by culturing the elongated shoots in 1/2 MS medium containing 0.1 mgL⁻¹ IBA. In summary, our study will provide an effective technology for large-scale propagation and important pathway for promoting the popularization and application of Hydrangea bretschneideri Dipp.     

Association of Mycoplasmalike Organisms (MLOs) with Mild Type of Hydrangea virescence: A Study with 60–1000 nm Thick Sections

The study of the agent associated with the mild type of Hydrangea virescence in France involved three steps, with the aid of transmission electron microscope (TEM). In the first step, we observed the presence of polymorphic procaryotes in thephloem sieve tubes of diseased plants and their absence from corresponding healthy plant parts. The procaryotes were detected in the areas suspected in 1000 nm thick sections stained with thionin-acridine orange. In the next step, the ultrastructure of their unit membrane was studied at magnifications higher than 100 000. The two osmiophilic layers of the membrane were 6 nm distant and no preliminary parietal shape was detected. These observations on ultrathin (60 nm) sections allowed us to classify the, particles into the class “Mollicutes”. The third step involving the examination of 350–1000 nm thick sections revealed the absence of spiral forms. The TEM observations are consistent with the hypothesis that the agents associated with the mild type of Hydrangea virescence observed in France should be included within the MLO group. A method specially adjusted to the fixation of MLO inside sieve tubes has been mentioned.     

The influence of local plant growth conditions on non-fastidious bacterial contamination of meristem-tips ofHydrangea culturedin vitro

A high incidence ofin vitro bacterial contamination (69%) has been detected in meristem-tip explants ofHydrangea from widely differing locations in Ireland and the UK. The bacteria were characterised by API 20E biochemical test kits and by fatty acid profile analysis. The results obtained from the different methods were compatible and anomalies were explicable in terms of the limitations of the respective methods. The majority of the isolates were environmental or animal-associated bacteria with clusters ofEnterobacter isolates in Dublin, and ofEscherichia coli in the main Cork location. A cluster of Pseudomonads was detected in the Derby (UK) plants. The main association was between the location and the contaminant clusters. The main finding was that the nature of organic soil amendments may influence inoculum for the contamination of plants and the conclusion was that fertilisation with organic materials should be avoided in the preparation of plants for micropropagation.     

羊毛硫肽SapB类多肽促进链霉菌形态分化作用的研究

【目的】链霉菌属于革兰氏阳性菌,以复杂的形态分化过程和强大的次级代谢产物合成能力为主要特征。链霉菌的形态分化与次级代谢产物的产生密切相关。Ⅲ型羊毛硫肽SapB能够促进天蓝色链霉菌气生菌丝体形成,暗示这类多肽可以作为靶标用于形态分化改造工程开发。本研究表征了SapB类多肽对多种链霉菌形态分化的影响,为该类多肽的工程化应用提供理论基础。【方法】生物信息学分析多个链霉菌基因组中SapB类多肽的生物合成基因簇,构建SapB类多肽的异源表达载体,利用接合转移方法导入不同链霉菌中进行异源表达,探究SapB类多肽对链霉菌形态分化的影响。【结果】SapB类多肽在不同程度上促进了多个链霉菌由营养菌丝向气生菌丝分化,表现为气生菌丝体数量的增多和分化速度的加快,缩短了链霉菌形态分化周期。【结论】SapB类多肽的过表达有助于缩短链霉菌形态分化周期,可用于针对链霉菌形态分化的工程改造。     

DEFINING TAXON BOUNDARIES IN MEMBERS OF THE MORPHOLOGICALLY AND GENETICALLY PLASTIC GENES CAULERPA (CAULERPALES,CHLOROPHYTA)1

Numerous attempts to capture the morphological variability of the genus Caulerpa have resulted in an unstable classification of numerous varieties and formae. In the present study we attempted to test taxon boundaries by investigating morphological and genetic variation within and between seven taxa of Caulerpa, supposedly belonging to four species, sampled at different sites in a Philippine reef system. Using both field and culture observations, we described the relation between the variability of a set of morphological characters and ecological parameters, such as wave exposure, light intensity, and substrate type. Statistical analyses showed that the limits between two (out of three) ecads of the C. racemosa (Forsskål) J. Agardh complex were obscured by the presence of morphological plasticity. Other studied taxa of Caulerpa (i.e. C. cupressoides [Vahl] C. Agardh, C. serrulata [Forsskål] J. Agardh, and two formae of C. sertularioides [S. Gmelin] Howe) could be grouped based on morphology despite the presence of morphological plasticity. Our results indicated a strong association between light intensity and several quantitative morphological variables. Genetic diversity of these taxa was assessed by partial sequencing chloroplast rbcL and tufA genes and the ycf10‐chlB chloroplast spacer. In all phylogenetic analyses, C. serrulata, C. cupressoides, C. sertularioides, and the three ecads of C. racemosa emerged as distinct genetic units. Despite the presence of morphological plasticity and morphological convergence, a subset of morphological characters traditionally used in taxonomic delimitation still had sufficient discriminative power to recognize the terminal phylogenetic clades.     

Can members of the south‐western Gila robusta species complex be distinguished by morphological features?

The goal for this project was to re‐examine key morphological characters hypothesized to differentiate Gila intermedia, Gila robusta and Gila nigra and outline methods better suited for making species designations based on morphology. Using a combination of meristic counts, morphological measurements and geometric morphometrics, morphological dissimilarities were quantified among these three putative species. Traditional meristic counts and morphological measurements (i.e. distances between landmarks) were not useful for species identification. Geometric morphometrics, however, identified differences among species, while also suggesting an effect of geographic location on morphological variation. Using canonical variate analysis for the 441 fish sampled in this study, geometric morphometrics accurately predicted true group membership 100% of the time for G. nigra, 97% of the time for G. intermedia and 91% of the time for G. robusta. These results suggest that geometric morphometric analysis is necessary to identify morphological differences among the three species. Geometric morphometric analysis used in this study can be adopted by management officials as a tool to classify unidentified individuals.     

小叶锦鸡儿天然居群叶形态性状变异研究

为揭示小叶锦鸡儿(Caragana microphylla)天然居群叶形态性状的变异规律及其生态适应性特征,该研究以10个小叶锦鸡儿天然居群为对象,通过多重比较、巢式方差分析、相关性分析、聚类分析和主成分分析等方法,对7个叶形态性状进行分析。结果表明：(1)小叶锦鸡儿叶形态性状在居群内和居群间均存在极显著差异(P < 0.01),平均变异系数为10.13%,不同性状的变异幅度为6.23%~12.78%;平均叶形态性状的表型分化系数为43.62%,居群内变异(30.09%)大于居群间变异(24.91%),说明居群内是其叶形态性状变异的主要来源。(2)相关性分析表明,环境因子对小叶锦鸡儿的叶形态性状变异有很大的影响,在地理空间上主要呈现出沿海拔梯度的变异模式;主成分分析的结果显示,小叶宽、叶柄宽和叶柄长对小叶锦鸡儿叶形态变异起主导作用;利用欧式距离对小叶锦鸡儿居群进行UPGMA聚类分析结果显示,基于叶形态性状和环境因子可分别将小叶锦鸡儿10个居群分为3类和2类,Mantel检验结果表明,小叶锦鸡儿的叶形态性状变异不存在地理连续性。研究结果为小叶锦鸡儿的适应性进化和开发利用提供了理论依据。     

Increasing conspecific density weakens the ability of intermediate predators to develop induced morphological defences to top predators

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MORPHOLOGICAL DIVERSIFICATION AND ADAPTIVE RADIATION: A COMPARISON OF TWO DIVERSE LIZARD CLADES

We compared the morphological diversity (i.e., the amount of morphological space occupied) of two similar clades, the lizard genera Anolis and Sceloporus. These species-rich monophyletic clades are similar in body size, age of origin, and many aspects of their natural history. We examined a number of morphological traits whose variation is likely to represent adaptation to different aspects of the environment, including body size, limb proportions, head dimensions, and tail length. Examination of the position of species in multidimensional space, based on a principal components analysis, indicates that the morphological diversity of Anolis, which we refer to as disparity, is significantly greater than that of Sceloporus. One potential explanation for this pattern is that morphological diversification in Anolis was facilitated by the evolution of subdigital toe-pads, which allow anoles to use the environment in ways not available to Sceloporus. The geographic location of diversification (tropical and subtropical for Anolis, arid for Sceloporus) may also have been important.     

Morphological and phylogenetic analyses of <Emphasis Type="Italic">Uromyces appendiculatus</Emphasis> and <Emphasis Type="Italic">U. vignae</Emphasis> on legumes in Japan

Uromyces appendiculatus, inclusive of three varieties, is distinguished from U. vignae primarily by the position of urediniospore germ pores and putative host specificity. However, opinions concerning these morphological and physiological features as taxonomic characters have varied greatly, and distinction of these species has often been confused. To clarify the taxonomy of these two species, morphological features of urediniospores and teliospores of 225 rust fungus specimens on species of Phaseolus, Vigna, Apios, Lablab, and Dunbaria were examined by light microscopy and scanning electron microscopy. Forty-five specimens were subjected to molecular phylogenetic analyses. As a result, the position of germ pores in urediniospores and the teliospore-wall thickness were considered as good characters to separate three morphological groups. In molecular analyses, the specimens fell into two and three clades based on the nucleotide sequence at D1/D2 domain of LSU rDNA and ITS regions, respectively. One of the D1/D2 clades corresponded to one morphological group whereas another D1/D2 clade included two other morphological groups. In contrast, each of the three ITS clades corresponded to a separate morphological group. Neither morphological groups nor molecular clades were host limited. It is suggested that the three morphological groups that corresponded to three distinct ITS clades constitute distinct species.Contribution no. 186 from the Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba, Japan