Molecular Phylogeny of Magnolia (Magnoliaceae) Inferred from cpDNA Sequences and Evolutionary Divergence of the Floral Scents

Magnolia (disjuncts), however, have similar chemical profiles. A molecular phylogeny of Magnoliaceae was constructed to reveal phylogenetic relationships of taxa by sequencing the trnK intron (including the matK coding region), psbA-trnH, and atpB-rbcL intergenic spacer regions of chloroplast DNA from 25 Magnolia, two Michelia, and two Liriodendron taxa. The psbA-trnH spacer region showed twice the sequence divergence (0.0157) of the trnK intron (0.0073) or the matK coding region (0.0077). The strict consensus tree constructed from the combined data set (ca. 3,700 bp) indicated the genus Magnolia was polyphyletic containing Michelia species as ingroup. The clade of Magnolia liliifera var. obovata, M. coco, and M. delavayi formed the first branch. Among the remaining species, two additional large clades were recognized, i.e., one comprised of American evergreen Magnolia species and another of subgenus Yulania. The relationship among sect. Rytidospermum taxa was not clearly resolved. Parsimonious mapping of the floral scent chemical characters was performed onto the molecular phylogenetic tree to discuss evolutionary trends of the floral scent chemistries. Received 7 December 1998/ Accepted in revised form 18 May 1999     

报春石斛花香气成分及其日变化规律

采用静态顶空气相色谱-质谱(GC-MS)联用技术研究报春石斛(Dendrobium primulinum)花香气成分及香气释放的日变化规律,检测并分析一天内六个时间点(6:00、9:00、12:00、15:00、18:00、21:00)报春石斛花的香气成分。各时间点占总香气释放量排序为：6时(35.26%)>9时(22.78%)>12时(19.35%)>21时(7.86%)>15时(7.59%)>18时(7.17%)。共检测出202种香气物质,包括酯类、烯萜类、醇类、醛类、烷类等,其中6:00有93种,9:00有87种,12:00有68种,15:00有61种,18:00有63种,21:00有65种。各个时间点共有的化合物21种。报春石斛花香气释放量6:00最高,18:00最低,主要香气成分是醋酸辛酯。     

Intraspecific Variation in the Amino Acid Content of Floral Nectar

In order to evaluate the constancy of amino acid composition and concentration, several randomly collected samples of genuine floral nectar from different flowers of several plant species were analysed. Although there seems to exist a rough pattern of species-specific nectar amino acid composition, amino acid concentration varied strongly from sample to sample. Apart from the lack of constancy in concentration of floral amino acids, our samples do not show amino acid concentration values as assigned typical for the pertinent pollination mode by other workers.     

植物花香代谢调节与基因工程研究进展

植物花香在吸引昆虫授粉、提高观赏价值和香精的商业价值方面具有重要的作用。随着分子生物学技术的发展,近年来植物花香基因被大量克隆,对花香化合物的合成与代谢的网络调控机制有了更深刻的认识,基因工程改良花香成为可能。对近年来植物花香的合成途径、花香的释放与基因调节、基因工程的研究进展进行了综述,并就存在的问题进行了分析,为花香的分子育种研究提供参考。     

Reproductive and Pollination Biology of Magnolia and its Allied Genera (Magnoliaceae). I. Floral Volatiles of Several Magnolia and Michelia Species and their Roles in Attracting Insects

Abstract Volatile substances emitted from the flowers of eight Magnolia taxa ( M. sieboldii ssp. japonica, M. praecocissima var. praecocissima and var. borealis, M. tomentosa, M. salicifolia, M. obovata, M. denudata, and M. grandiflora ) and one Michelia species ( M. compressa ) (Magnoliaceae) were examined and identified using GC-MS. Volatile substances of these Magnolia and Michelia species consist primarily of monoterpenoids and sesquiterpenoids produced by the mevalonate pathway, acetogenins by the acetate-malonate pathway, and phenyl-propanoids by the shikimate pathway. These Magnolia and Michelia species all possessed various combinations of volatile monoterpenoids, acetogenins, and phenylpropanoids, except for Magnolia obovata , which emitted primarily sesquiterpenoids. Free amino acids in pollen of 12 Magnolia and one Liriodendron species were also analyzed, and their value as food sources for pollinators evaluated.
Pollinators visiting the flowers of five Magnolia species were collected in their native sites and identified. Their behaviors and roles as pollinating agents were assessed.     

广西木兰属一新种

描述了广西木兰属(木兰科)一新种, 即木论木兰。     

Floral Ontogeny of Two Species in Magnolia L.

Floral ontogeny is described in two species of genus Magnolia （Magnollaceae）, Magnolia alboserlcea Chun et C. Tsoong, and M. amoena Cheng, representing subgenus Magnolia and subgenus Yulani In Magnolia, by using scanning electron microscope （SEM）. The sequence of Initiation of floral organs Is from proximal to distal. The three distinct outermost and middle organs are Initiated in sequence, but ultimately form a single whorl, thus their ontogeny Is consistent with a sepal Interpretation. The last three tepals （petals） alternate with the preceding tepal whorl. The members of androeclum and gynoeclum arise spirally, although the androecium shows some Intermedlacy between a spiral and whorled arrangement. The carpel prlmordia initiate in group of four to five. The order of stamen Initiation within each tier Is not determined. The floral ontogeny Is remarkably homogeneous between the subgenus Magnolia and subgenus Yulani that does not support the resuming of genus Yulani.     

Floral scents of hawkmoth-pollinated flowers in Japan

Similarity among the floral scents of hawkmoth-pollinated plants was investigated with headspace samplings. Six of seven plant species belonging to different families were found to be rich in isoprenoids, among which linalool was the most common compound. Linalool showed rhythmicity with a nocturnal increase inLonicera japonica. These findings suggest that linalool is a common attractant for nocturnal hawkmoths. However, the composition of other isoprenoids, benzenoids and fatty acid derivatives varied markedly among the plant species examined. There was a significant correlation between species composition of flower-visiting hawkmoths and specific floral scents, suggesting that attractiveness to each hawkmoth species is dependent upon floral scent.     

中国木兰科木兰属一新变种

对原产湖北五峰的红花玉兰新变种多瓣红花玉兰（Magnolia wufengensis var. multitepala L.Y.Ma et L.R.Wang）作了描述。新变种与红花玉兰原变种(M. wufengensis var. wufengensis)花色相近,整个花被片内外均为红色,内面略淡。主要区别在于：新变种花被片12、15、18、24;花被片阔倒卵状匙形、倒卵状匙形或窄倒卵状匙形;花色浓淡不同植株间有深红、红、浅红之差别;自然状态下可育,蓇葖果发育良好。     

Evidence for a novel biosynthetic pathway that regulates the ratio of syringyl to guaiacyl residues in lignin in the differentiating xylem of Magnolia kobus DC

The biosynthetic pathways to monolignols in Magnolia kobus were investigated by feeding stems with a deuterium-labeled precursor. Pentadeutero [γ,γ-²H₂, OC²H₃] coniferyl alcohol was supplied to shoots of Magnolia kobus and the incorporation of the labeled precursor into lignin was traced by gas chromatography-mass spectrometry. In addition to the direct incorporation of the labeled precursor into guaiacyl units, we detected a significant amount of pentadeuterium-labeled syringyl units with two γ-deuterium atoms. The relative level of trideuterium-labeled syringyl monomers (the result of conversion via the cinnamic acid pathway, in which two γ-deuterium atoms are removed during enzymatic re-oxidation) was negligible. Our results provide conclusive evidence for a novel alternative pathway for generation of lignin subunits at the monolignol stage and they suggest that this new pathway might be important for regulation of the composition of lignin. Received: 21 August 1998 / Accepted: 30 September 1998     

The F-actin distribution during microsporogenesis in Magnolia soulangeana Soul. (Magnoliaceae)

Summary F-actin distribution during male meiosis in Magnolia soulangeana was studied by means of fluorescence microscopy following staining with rhodaminephalloidin. Actin filaments were observed to persist during all of the developmental stages of meiosis. Four main types of configurations were recognized: (1) peripheral filaments underlying the plasma membrane (cortical network); (2) filaments dispersed throughout the inner cytoplasm (central cytoplasmic network); (3) filaments associated with the meiotic spindles; (4) filaments associated with the phragmoplasts. The cortical and central cytoplasmic filaments exhibited different behaviours. Whereas the cortical network remained present in an apparently unchanged form during all of the meiotic stages, the central cytoplasmic filaments, although they never completely disappeared, were reduced and concentrated around the nucleus at the end of prophase. At metaphase, fluorescent spindles consisting of filament bundles running from pole to pole or being interrupted at the equatorial zone could be seen. At the end of both the first and second division of meiosis, fluorescent bands of filaments (disks) appeared at the level of the cell division planes (equatorial regions) where cleavage furrows were constituted. These cleavage furrows did not form when floral buds were cultivated in a cytochalasin-containing medium. Our results show that during microsporogenesis in M. soulangeana the actin filaments constitute a highly complex and dynamic system that is involved in particular in cytoplasm cleavage of the meiocytes.     

中国木兰科木兰属一新种

描述了中国木兰科木兰属一新种——红花玉兰(Magnolia wufengensis L. Y. Ma et L. R. Wang),并绘了模式图。本种的主要特点是：落叶乔木,花被片9,近相等,整个花被片内外为均匀的红色,叶柄较长,叶片下面沿主脉密被白色柔毛,这些特征与武当木兰(M. sprengeri Pamp.)和玉兰(M. denudata Desr.)存在明显区别。     

Metabolic Regulation of Floral Scent in Petunia axillaris Lines: Biosynthetic Relationship between Dihydroconiferyl Acetate and iso-Eugenol

Aromatic scent-related compounds in flowers were comprehensively analyzed by high-performance liquid chromatography (HPLC) based on their absorption spectra to understand regulation of metabolism leading to floral scent diversity in Petunia axillaris lines. An unknown compound occurring at similar levels to scent compounds in some plant lines was identified to be dihydroconiferyl acetate. Based on the structure, dihydroconiferyl acetate is likely to be a biosynthetically closely related compound to aromatic scent compounds, especially iso-eugenol. Similar time-course changes of the concentrations suggest that the metabolism of dihydroconiferyl acetate is underlaid by the similar regulation to aromatic scent compounds. Dihydroconiferyl acetate and iso-eugenol occurred almost exclusively, implying that metabolism of the common precursors to each compound is selectively regulated in these plants. The branching of the biosynthetic pathway into dihydroconiferyl acetate and iso-eugenol is probably one of regulatory steps leading to scent diversity in P. axillaris lines.     

单性木兰花部综合特征及其传粉适应性

对单性木兰花部综合特征和访花昆虫种类及访花行为进行了研究,结果表明,单性木兰花冠顶端呈半合拢状,花被片之间存在间隙,只有那些能够穿过这些间隙的昆虫才能到达花内为其采粉和授粉,对访花者的要求较为苛刻。于降雨前去掉雄花和雌花花被片,使雄花雄蕊群和雌花柱头充分暴露,另外选择雄花和雌花作对照,降雨后将花取回并测定雄花花粉生活力和雌花柱头上的花粉粒。结果表明,在雨天,单性木兰花冠顶端呈半合拢状的开放方式对雄花和雌花均起到了保护作用,这种花部结构的特化是对花期持续降雨的一种生态适应。从访花昆虫种类上看,单性木兰雄花的昆虫种类是雌花的两倍,显然雄花比雌花更能吸引昆虫。     

Patterns of Floral Variation between Dimorphic and Monomorphic Populations in Distylous Luculia pinceana (Rubiaceae)

Flower morphology plays an important role in the evolution and maintenance of plant mating systems, including disassortative mating of heterostylous species. The transition of mating patterns may be associated with the remodification of intraspecific flower morphology. To determine the functional relationship between floral variation and transition of mating patterns, we conducted a series of morphometric analyses in a distylous species Luculia pinceana, which possesses dimorphic and monomorphic populations. Our results indicate that floral variation was higher between different types of populations than between populations of the same type. Compared to dimorphic populations, some floral characters, reduced stigma anther separation within flowers and increased overlap of stigmas and anthers (illegitimate spatial matching of sexual organs) among individuals in populations containing only the long styled morph may have been modified to increase both selfing and intra morph crossing. The observed patterns of floral variation between dimorphic and monomorphic populations coincide with the transition of mating patterns from disassortative mating to selfing and/or intra morph crossing.     

基于SNP分子标记的凹叶木兰遗传多样性初步研究

凹叶木兰是我国特有的木兰属植物,仅分布于我国四川和云南两省相邻地区,目前已被列入中国物种红皮书名录,属易危物种。该研究以采自四川南部麻咪泽和美姑大风顶两个自然保护区的野生凹叶木兰为材料,利用以PCR和测序为基础的SNP分子标记方法,初步研究了凹叶木兰的遗传多样性。结果表明:在扩增出的4条510bp长的序列上,平均在73bp左右的序列长度上能够检测到一个SNP位点,说明两个自然保护区内不同居群的凹叶木兰具有较高遗传多样性;序列间相似度达97%,与引物所对原序列相似度达36%,表明该序列适宜进行凹叶木兰遗传多样性研究。研究结果为进一步开展凹叶木兰遗传多样性的研究及其保护政策的制定提供了参考。     

Floral scent emission is the highest at the second night of anthesis in Lonicera japonica (Caprifoliaceae)

Floral color change in diverse plants has been thought to be a visual signal reflecting changes in floral rewards, promoting pollinator foraging efficiency as well as plant reproductive success. It remains unclear whether olfactory signals co-vary with floral color change. We investigated the production rhythms of floral scent and nectar associated with floral color change in Lonicera japonica. The flowers generally last 2–3 days. They are white on opening at night (N1) and become light yellow the following day (D1), yellow on the second night (N2), and golden on the second day of flowering (D2). Our measurements in the four stages indicated that nectar production decreased significantly from N1 and D1 to N2 and D2, tracking the floral color change. A total of 34 compounds were detected in floral scent and total scent emission was significantly higher in N2 than in the other three stages. The scent emission of three major compounds, Linalool, cis-3-Hexenyl tiglate, and Germacrene D was also significantly higher in N2, but the relative content of Linalool decreased gradually, cis-3-Hexenyl tiglate increased gradually, and the relative content of Germacrene D did not differ among the four measured stages. Greater scent emission by night than by day suggested a strong olfactory signal to attract nocturnal hawkmoths, the effective pollinators. However, floral scent rhythms in the four stages did not match the color change and nectar secretion, suggesting that floral color (visual) and scent (olfactory) in this species may play different roles in attracting or filtering various visitors.     

Floral biology of Nuphar pumila (Timm) DC. (Nymphaeaceae) in China

Anthesis in individual flowers of Nuphar pumila (Timm) DC. occurs for four consecutive days. First-day flowers are protogynous and functionally female. Flowers can open completely on the first day of anthesis. This contrasts with all previous reports, which state that first-day flowers of Nuphar are characterized by partial expansion of the calyx, leaving a distal small triangular opening just above the stigmatic disc. Flowers close completely on the first night of anthesis and remain partially open on the subsequent three nights. During the entire anthesis period the stigmas emit a sweet, fruity odor and the petal nectaries produce visible nectar drops. The stigma of N. pumila is secretory and unicellular-papillate. Pollen grains are monosulcate with long spines. Our observations on the mating system of N. pumila indicate that neither asexual seed production nor spontaneous self-pollination occurs. Cross-pollination of second-, third- and fourth-day flowers produced few seeds. Flowers of N. pumila were mainly pollinated by sweat bees, with flies playing a minor pollination role. No beetle visits were observed. Our insect-pollination observations substantiate the view that the relative contribution of flies, bees, and beetles to pollination in a single Nuphar population depends on two factors: the relative abundance of the insects, and presence of alternative food sources.     

Floral scent is different between sexual phases within individuals in a synchronously dichogamous shrub (Canella winterana) but there is no distinct female or male scent profile across individuals

Floral scent is a highly complex and variable floral characteristic that is involved in pollinator attraction. One possible cause for variation in floral scent can be sexual identity of the flower. Here, we examine the floral scent bouquet of a synchronously dichogamous shrub, Canella winterana (L.) Gaertn.We used dynamic headspace extraction and gas chromatography – mass spectroscopy, followed by statistical analysis using non-metric multidimensional scaling, SIMPER, MANOVA, and PERMANOVA to identify and compare the scent profiles of Canella winterana in its female and male phase for multiple individuals as well as multiple inflorescences of a single tree over one cycle of its entire sexual phase transition. The scent profile of C. winterana is composed of 49 volatile organic compounds and dominated by five compounds. We found no evidence for distinct male or female scent profiles; however, there were significant differences in scent emission between different sexes within some individuals. Two compounds explained over half of the variation between sexual stages within individuals. Our exploration of a single tree's sexual phase transition, including neuter phase, found that five compounds dominated the female phase scent bouquet and that female phase was distinct from male and neuter phase. This study offers new insight into the role that variability in floral scent between sexual phases might play in variable pollinator behavioral responses. These results suggest partial support for two distinct hypotheses regarding the differences between the sex phases (1) honest signaling and (2) sexual mimicry.