Morpho-physiological and phytohormonal changes during the induction of adventitious root development stimulated by exogenous IBA application in Magnolia biondii Pamp / Alterações morfofisiológicas e fito-hormonais durante a indução do desenvolvimento radicular adventício estimulado pela aplicação exógena do AIB em Magnolia biondii Pamp

Magnolia biondii Pamp is an important ornamental tree species widely grown and used as a rootstock in the propagation of different Magnolia varieties. In the current studies, anatomical, physiological and endogenous hormones were studied to check the effect of IBA 750 mg/L on the adventitious rooting and to provide theoretical and technical support for the propagation of Magnolia biondii Pamp through stem cuttings. Two thousand stem cuttings were prepared and divided into two groups i.e., IBA treated cuttings and water control. For the evaluation of antioxidant enzyme activities, and endogenous hormones levels, samples were collected on the day of planting and each 5th day and further steps were carried out in the laboratory according to the protocols and proper precautions. For the anatomical observations, samples were collected on the 13th, 15th, and 17th day for IBA treated cuttings while 21st, 23rd, and 25th day for control. Collected samples were preserved in the FAA solution and further observations were carried out in the laboratory. Anatomical observations showed that it took 13 days for the differentiation of root primordia to the appearance of young adventitious roots in IBA treated cuttings, while it took 21 days to develop primordia in the control. Antioxidant enzyme activities involved in ROS were significantly higher in the IBA treated cuttings compared to control. POD showed a peak on the 13th day before the emergence of roots in IBA treated cuttings while it showed a peak on the 21st day in the control. PPO showed a peak on the 21st day in the IBA treated cuttings while it showed a peak on the 29th day in the control. SOD showed a peak on the 17th day in IBA treated cuttings, while it showed a peak on the 25th day in the control. Exogenous application of IBA enhanced the endogenous IAA and GA3 levels compared to CK, while it reduced the levels of ABA continuously at the time of rooting and then increased gradually. Inclusively, our study suggests that IBA 750 mg/L is efficient for the rooting of Magnolia biondii Pamp cuttings, as it enhanced the process of antioxidant enzyme activities, endogenous hormones levels and reduced the time of root formation which is evident from the anatomical observations.

Magnolia biondii Pamp é uma importante espécie de árvore ornamental muito cultivada e utilizada como porta-enxerto na propagação de diferentes variedades de Magnolia. Nos estudos atuais, hormônios anatômicos, fisiológicos e endógenos foram estudados para verificar o efeito do AIB na dose de 750 mg / L no enraizamento adventício e fornecer suporte teórico e técnico para a propagação de M. biondii Pamp por meio de estacas. Duas mil estacas foram preparadas e divididas em dois grupos, ou seja, tratadas com AIB e controle de água. Para a avaliação das atividades das enzimas antioxidantes e dos níveis de hormônios endógenos, as amostras foram coletadas no dia do plantio e a cada 5 dias, enquanto as demais etapas foram realizadas em laboratório de acordo com os protocolos e os devidos cuidados. Para as observações anatômicas, as amostras foram coletadas no 13º, 15º e 17º dias para estacas tratadas com AIB e no 21º, 23º e 25º dias para o controle. As amostras coletadas foram preservadas em solução FAA, e outras observações foram realizadas em laboratório. Observações anatômicas mostraram a necessidade de 13 dias para a diferenciação dos primórdios radiculares até o aparecimento de raízes adventícias jovens em estacas tratadas com AIB e de 21 dias para o desenvolvimento dos primórdios no controle. As atividades das enzimas antioxidantes envolvidas nas ROS foram significativamente maiores nas estacas tratadas com AIB em comparação com o controle. A POD apresentou pico no 13º dia antes da emergência das raízes nas estacas tratadas com AIB, enquanto no 21º dia apresentou pico no controle. A PPO teve pico no 21º dia nas estacas tratadas com AIB e no 29º dia no controle. A SOD apresentou pico no 17º dia nas estacas tratadas com AIB e no 25º dia no controle. A aplicação exógena de AIB aumentou os níveis endógenos de IAA e GA3 em relação ao controle, enquanto reduziu os níveis de ABA continuamente no momento do enraizamento e, em seguida, aumentou gradativamente. Inclusive, nosso estudo sugere que o AIB na dose de 750 mg / L é eficiente para o enraizamento de estacas de M. biondii Pamp, visto que potencializou o processo de atividades de enzimas antioxidantes e os níveis de hormônios endógenos, além de reduzir o tempo de formação de raízes, o que fica evidente nas observações anatômicas.

Integrated RNA and miRNA sequencing analysis reveals a complex regulatory network of Magnolia sieboldii seed germination.

Magnolia sieboldii K. Koch (M. sieboldii) is a deciduous Chinese tree species of the Magnoliaceae family with high ornamental, medicinal, and economic benefits. The germination of M. sieboldii seeds under natural conditions is extremely difficult, thereby hindering the cultivation and breeding of this important species. The molecular mechanisms underlying M. sieboldii seed germination remain unclear due to the lack of genomic and transcriptomic resources. Here, we integrated both mRNA and miRNA sequencing to identify the genes and pathways related to M. sieboldii germination. A comprehensive full-length transcriptome containing 158,083 high-quality unigenes was obtained by single-molecule real-time (SMRT) sequencing technology. We identified a total of 13,877 genes that were differentially expressed between non-germinated and germinated seeds. These genes were mainly involved in plant hormone signal transduction and diverse metabolic pathways such as those involving lipids, sugars, and amino acids. Our results also identified a complex regulatory network between miRNAs and their target genes. Taken together, we present the first transcriptome of M. sieboldii and provide key genes and pathways associated with seed germination for further characterization. Future studies of the molecular basis of seed germination will facilitate the genetic improvement M. sieboldii.

A potential endogenous gibberellin-mediated signaling cascade regulated floral transition in Magnolia × soulangeana 'Changchun'.

The floral transition is a critical developmental switch in plants, and has profound effects on the flower production and yield. Magnolia × soulangeana 'Changchun' is known as a woody ornamental plant, which can bloom in spring and summer, respectively. In this study, anatomical observation, physiological measurement, transcriptome, and small RNA sequencing were performed to investigate potential endogenous regulatory mechanisms underlying floral transition in 'Changchun'. Transition of the shoot apical meristem from vegetative to reproductive growth occurred between late April and early May. During this specific developmental process, a total of 161,645 unigenes were identified, of which 73,257 were significantly differentially expressed, while a number of these two categories of miRNAs were 299 and 148, respectively. Further analysis of differentially expressed genes (DEGs) revealed that gibberellin signaling could regulate floral transition in 'Changchun' in a DELLA-dependent manner. In addition, prediction and analysis of miRNA targeted genes suggested that another potential molecular regulatory module was mediated by the miR172 family and other several novel miRNAs (Ms-novel_miR139, Ms-novel_miR229, and Ms-novel_miR232), with the participation of up- or down-regulating genes, including MsSVP, MsAP2, MsTOE3, MsAP1, MsGATA6, MsE2FA, and MsMDS6. Through the integrated analysis of mRNA and miRNA, our research results will facilitate the understanding of the potential molecular mechanism underlying floral transition in 'Changchun', and also provide basic experimental data for the plant germplasm resources innovation in Magnolia.

A New Insight into Flowering Regulation: Molecular Basis of Flowering Initiation in Magnolia × soulangeana 'Changchun'.

Magnolia × soulangeana 'Changchun' are trees that bloom in spring and summer respectively after flower bud differentiation. Here, we use phenological and morphological observation and RNA-seq technology to study the molecular basis of flowering initiation in 'Changchun'. During the process of flowering initiation in spring and summer, the growth of expanded flower buds increased significantly, and their shape was obviously enlarged, which indicated that flowering was initiated. A total of 168,120 expressed genes were identified in spring and summer dormant and expanded flower buds, of which 11,687 genes showed significantly differential expression between spring and summer dormant and expanded flower buds. These differentially expressed genes (DEGs) were mainly involved in plant hormone signal transduction, metabolic processes, cellular components, binding, and catalytic activity. Analysis of differential gene expression patterns revealed that gibberellin signaling, and some transcription factors were closely involved in the regulation of spring and summer flowering initiation in 'Changchun'. A qRT-PCR (quantitative Real Time Polymerase Chain Reaction) analysis showed that BGISEQ-500 sequencing platform could truly reflect gene expression patterns. It also verified that GID1B (GIBBERELLIN INSENSITIVE DWARF1 B), GID1C, SPL8 (SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 8), and GASA (GIBBERELLIC ACID-STIMULATED ARABIDOPSIS) family genes were expressed at high levels, while the expression of SPY (SPINDLY) was low during spring and summer flowering initiation. Meanwhile, the up- and down-regulated expression of, respectively, AGL6 (AGAMOUS-LIKE 6) and DREB3 (DEHYDRATION-RESPONSIVE ELEMENT-BINDING PROTEIN 3), AG15, and CDF1 (CYCLIC DOF FACTOR 1) might also be involved in the specific regulation of spring and summer flowering initiation. Obviously, flowering initiation is an important stage of the flowering process in woody plants, involving the specific regulation of relevant genes and transcription factors. This study provides a new perspective for the regulation of the flowering process in perennial woody plants.

An efficient micropropagation protocol for an endangered ornamental tree species (Magnolia sirindhorniae Noot. & Chalermglin) and assessment of genetic uniformity through DNA markers.

Magnolia sirindhorniae Noot. & Chalermglin is an endangered species with high ornamental and commercial value that needs to be urgently protected and judiciously commercialized. In this study, a protocol for efficient regeneration of this species is standardized. The lateral buds of the M. sirindhorniae plant were used as an explant. Half-strength Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 6-benzyladenine (BA), 0.1 mg/L α-naphthaleneacetic acid (NAA), and 2.0 mg/L gibberellic acid (GA3) was found to be the optimal medium for shoot induction. The maximum shoot multiplication rate (310%) was obtained on Douglas-fir cotyledon revised medium (DCR) fortified with 0.2 mg/L BA, 0.01 mg/L NAA, and additives. The half-strength DCR medium supplemented with 0.5 mg/L NAA and 0.5 mg/L indole-3-butyric acid (IBA) supported the maximum rate (85.0%) of in vitro root induction. After a simple acclimatization process, the survival rate of plantlets in a substrate mixture of sterile perlite and peat soil (1:3; v/v) was 90.2%. DNA markers were used for assessment of genetic uniformity, confirming the genetic uniformity and stability of regenerated plants of M. sirindhorniae. Thus, the described protocol can safely be applied for large scale propagation of this imperative plant.

Tasa relativa de crecimiento en plántulas de dos poblaciones de Magnolia pugana (Magnoliaceae) en distintos niveles de luz y fertilidad del suelo / Relative growth rate in Magnolia pugana (Magnoliaceae) seedlings from two populations at different light levels and soil fertility

Resumen El estudio de la ecofisiología de las especies en peligro de extinción es clave para el éxito de programas de conservación y restauración ecológica. El objetivo de este trabajo fue conocer el efecto de los factores luz, fertilidad de suelo y procedencia de las plántulas en el crecimiento de plántulas de Magnolia pugana. Se estimó la tasa relativa de crecimiento (TRC) y sus componentes (Tasa de Asimilación Neta: TAN y Cociente del Área Foliar: CAF), así como la relación raíz/vástago (C: R/V). Las plántulas fueron obtenidas de semillas recolectadas de dos localidades en Zapopan, Jalisco, México, la primera es una población silvestre en San Nicolás (SN) y la segunda es una plantación ubicada en los jardines del Centro Universitario de Ciencias Biológicas y Agropecuarias (CUCBA). El experimento se llevó a cabo en condiciones de invernadero. En septiembre 2015, 96 plántulas de tres meses de edad fueron sometidas a los siguientes tratamientos con un experimento factorial (2 × 2 × 2): nivel de luz (alta = 1 120 μmol m-2 s-1 y baja = 136.3 μmol m-2 s-1), procedencia de las plántulas (San Nicolás y CUCBA) y fertilidad del suelo (alta = suelo San Nicolás, baja = suelo del CUCBA) con 12 repeticiones por cada combinación de factores y niveles. El crecimiento se estimó con dos cosechas: la primera a los 30 días de establecido el experimento y la segunda a los 60 días. Los niveles de luz, fertilidad del suelo y la procedencia de las plántulas influyeron en el crecimiento a través de ajustes fisiológicos y morfológicos. En general Magnolia pugana mostró mayor TRC y TAN en la luz alta, mientras que el CAF disminuyó. La fertilidad del suelo y la procedencia de las plántulas no afectaron la TRC ni sus componentes. Sin embargo, si fueron afectados por los niveles de luz, las plántulas de SN crecieron más en luz alta. La altura del tallo varió debido a la fertilidad del suelo y a la procedencia de las plántulas, en el suelo con fertilidad baja, en el suelo del CUCBA las plántulas de SN fueron 35 % más altas que las del CUCBA. El suelo de baja fertilidad en la luz alta ocasionó que las plántulas asignaran mayor biomasa a la raíz. El componente fisiológico (TAN) fue el mayor determinante en la variación intraespecífica de la TRC. Las plántulas de SN mostraron mayor plasticidad fenotípica debido a que es una población silvestre, por lo que posiblemente tiene variación genética más alta que la población cultivada del CUCBA. Los resultados sugieren que Magnolia pugana es una especie con capacidad de adaptarse a diversos ambientes debido a su plasticidad fenotípica frente a los distintos niveles de luz y fertilidad del suelo.

Abstract The study of ecophysiology of endangered species is key to the success of conservation and ecological restoration programs. The objective of this work was to know the effect of light, soil fertility and seed origin on the growth of Magnolia pugana seedlings. The relative growth rate (RGR) and its components (Net Assimilation Rate: NAR and Leaf Area Ratio: LAR), as well as the root-shoot ratio (R/S) were estimated. Seedlings were obtained from seeds collected in two localities in Zapopan, Jalisco, Mexico, the first is a wild population in San Nicolás (SN) and the second is a plantation located in the gardens of the Centro Universitario de Ciencias Biológicas y Agropecuarias (CUCBA). The experiment was conducted under greenhouse conditions. In September 2015, 96 seedlings of approximately three months old were subjected to the following treatments in a factorial experiment (2x2x2): light level (high = 1 120 μmol m-2 s-1 and low = 136.3 μmol m-2 s-1), seedling origin (SN and CUCBA) and soil fertility (high fertility = SN soil, low fertility = CUCBA soil) with 12 replicates for each combination of factors and levels. Growth was estimated in two harvests: the first harvest after 30 days of starting the experiment and the second at 60 days. Light and soil fertility levels, as well as seedling origin influenced growth through physiological and morphological adjustments. In general, Magnolia pugana showed higher RGR and NAR in high light, while LAR decreased. Soil fertility and seedling origin did not affect RGR or its components. However, these were affected by the light level, seedlings from SN grew more in high light. Stem height varied due to soil fertility and seedling origin, in low fertility soil (CUCBA) the seedlings of SN were 35 % higher than those of CUCBA. Low fertility soil under high light caused greater investment in seedling root biomass. The physiological component (NAR) was the major determinant of intraspecific variation in RGR. SN seedlings showed greater phenotypic plasticity due to coming from a wild population, which possibly has higher genetic variation than the cultivated population from CUCBA. The results suggest that Magnolia pugana is a species capable of adapting to diverse environments due to its phenotypic plasticity in response to different light and soil fertility levels. Rev. Biol. Trop. 66(2): 622-633. Epub 2018 June 01.

Variations in Volatile Oil Yield and Composition of "Xin-yi" (Magnolia biondii Pamp. Flower Buds) at Different Growth Stages.

Dried flower buds of Magnolia biondii Pamp. are the main ingredient in "Xin-yi" in China, and the volatile oils of M. biondii flower buds are the principal medicinal component. Gas chromatographymass spectrometry (GC-MS) and microscopic techniques were employed to detect the volatile yields of M. biondii flowers at various growth stages. The volatile oil yields of M. biondii flowers differed significantly at different growth stages and were closely related to flower dry weight, oil cell density and degree of oil accumulation. In February 2016, flower buds had the highest dry weight, the maximum percentage of oil cells at the oil saturation stage and the highest density of oil cells, which coincided with the highest oil yield. In March 2016, flower buds had a lower dry weight, a higher percentage of oil cells at the oil-degrading stage and the lowest oil cell density, resulting in decreased oil yields. The total amounts of the major medicinal components in the M. biondii flower also showed regular changes at different growth stages. In January and February of 2016, M. biondii flowers had a higher dry weight, volatile oil yield and total content of medicinal ingredients, which was the best time for harvesting high-quality medicinal components. Our study reveals that volatile oil content and chemical composition are closely related to the growth stage of M. biondii flower buds. The results provide a scientific morphology and composition index for evaluating the medicinal value and harvesting of high-quality M. biondii medicinal herbs.

Developing a Media Moisture Threshold for Nurseries to Reduce Tree Stress and Ambrosia Beetle Attacks.

Exotic ambrosia beetles are among the most damaging pests of trees grown in nurseries. The primary pests Xylosandrus crassiusculus Motschulsky and Xylosandrus germanus Blandford use ethanol to locate vulnerable trees. Research, primarily with X. germanus, has shown that flood-stressed trees emit ethanol and are preferentially attacked by ambrosia beetles. Our goal was to develop a media (also called potting soil) moisture threshold as an integrated pest management (IPM) tactic and assess grower practices that lead to ambrosia beetle attacks. Flooded Cornus florida L., Cornus kousa Burg., and Magnolia grandiflora L. trees incurred more attacks than unflooded trees that were not attacked. To determine optimal media moisture levels, we grew flood-tolerant Acer rubrum L. and flood-intolerant C. florida in containers with 10, 30, 50, 70, or 90% media moisture. No flooded or unflooded A. rubrum were attacked. However, C. florida grown in 70 or 90% moisture were attacked and died, whereas trees at 30 and 50% moisture were not attacked. Thus, we suggest an upper moisture threshold of 50% when growing C. florida and other flood-intolerant trees. However, during peak ambrosia beetle flight activity in spring 2013 and 2014, we found that media moisture levels in commercial nurseries were often between 50 and 90%. Implementing a media moisture threshold, as a new IPM tool, could reduce ambrosia beetle attacks and the need for insecticide applications, which is currently the only available management tactic. Future research should focus on how changes in substrates, irrigation, and other practices could help growers meet this threshold.

Interruption of the semiochemical-based attraction of ambrosia beetles to ethanol-baited traps and ethanol-injected trap trees by verbenone.

We examined the extent to which verbenone, a bark beetle antiaggregation pheromone, interrupted the semiochemical-based attraction of ambrosia beetles. Field trapping studies conducted in Ohio showed that a verbenone dispenser with a release rate of 50 mg/d at 25°C reduced the attraction of Anisandrus sayi Hopkins, Euwallacea validus (Eichhoff), Hypothenemus dissimilis (Zimmermann), Xylosandrus germanus (Blandford), and Xyleborinus saxesenii (Ratzeburg) to ethanol-baited traps. A verbenone dispenser attached to ethanol-injected Magnolia virginiana L. trap trees deployed in Ohio also reduced ambrosia beetle attacks compared to trap trees without a verbenone dispenser. Subsequent field trials demonstrated a direct relationship between distance from a verbenone dispenser and ambrosia beetle attacks on trap trees in Ohio in 2011 and 2012 and Tennessee in 2012, but not in Tennessee and Virginia in 2011. Assessment of the influence of verbenone on the probability of attacks above a density threshold found that although attacks occurred on trap trees regardless of their proximity to a verbenone dispenser, the higher density of attacks per tree occurred on trap trees farthest away from the verbenone source in Ohio and Tennessee. Verbenone alone could be somewhat useful for discouraging ambrosia beetle attacks on individual trees or on a small spatial scale, but deployment of verbenone might be most effective when integrated as part of a "push-pull" strategy.

Pollen flow and effects of population structure on selfing rates and female and male reproductive success in fragmented Magnolia stellata populations.

BACKGROUND: Fragmentation of plant populations may affect mating patterns and female and male reproductive success. To improve understanding of fragmentation effects on plant reproduction, we investigated the pollen flow patterns in six adjacent local populations of Magnolia stellata, an insect-pollinated, threatened tree species in Japan, and assessed effects of maternal plant (genet) size, local genet density, population size and neighboring population size on female reproductive success (seed production rates), and effects of mating distance, paternal genet size, population size and separation of populations on male reproductive success. RESULTS: The seed production rate, i.e. the proportion of ovules that successfully turned into seeds, varied between 1.0 and 6.5%, and increased with increasing population size and neighboring population size, and with decreasing maternal genet size and local genet density. The selfing rate varied between 3.6 and 28.9%, and increased with increasing maternal genet size and with declining local genet density. Male reproductive success increased with increasing paternal genet size, and decreased with increasing mating distance and separation of population. Pollen flow between the populations was low (6.1%) and highly leptocurtic. CONCLUSIONS: Our results indicate that habitat fragmentation, separation and reduced size of populations, affected mating patterns and reproductive success of M. stellata. Local competition for pollinators and plant display size were likely to alter the reproductive success.

[Magnolia liliiflora whole-tree sap flow in response to multiple environmental variables in Beijing].

In order to clarify the environmental factors affecting the water use of typical urban tree species Magnolia liliflora, an investigation was conducted on the responses of M. liliiflora whole-tree sap flow to the air temperature, air relative humidity, radiation, wind speed, soil temperature and water content, and precipitation in Beijing from April to October, 2008. The eight environmental factors affecting M. liliiflora whole-tree sap flow could be divided into three categories, i.e., evaporative demand index, soil index, and precipitation index. The evaporative demand index (air temperature, air relative humidity, total radiation, wind speed, and vapor pressure deficit) could explain 60% of the variation in the sap flow of individual trees, which presented S-type change trend, i.e., the sap flow reached an asymptote where higher light and evaporative demands could not cause sap flow to increase further. Soil index (soil temperature and water content) and precipitation index (precipitation amount) had little influence on the sap flow.

[Seed germinating characteristics of endangered plant Magnolia officinalis].

OBJECTIVE: To find out the optimal condition for the germination and cultivation condition on Magnolia officinalis, and offer the basis for standardized culture of M. officinalis. METHOD: The study was carried out with in-door experiment, using light, temperature, soil water content and seed soaking in different water temperature treatments, the effects of different treatments on seed germination percentage of M. officinalis were investigated. RESULT AND CONCLUSION: The germination percentage between light and dark treatments was significantly difference, it is indicated that the seed is light-sensitive. M. officinalis can germinate in a wide range of temperature, from 20 degrees C to 35 degrees C, and the suitable temperature was 25-30 degrees C, at which germination percentages were 59.2% and 54.6%. Germination percentage at 20/30 degrees C was 65%. The optimum soil water content was 25% for seed germination, at which germination percentages were 66.7%. The suitable soil water content was 20%-25%. In order to find out optimal temperature for soaking, seeds were soaked in water at the temperatures of 40, 60, 80, 90 degrees C for 10 minutes. The optimum water temperature was 60 degrees C for seed germination, and hard coated seed percentage was reduced. Seed germination rate can be inhibited under too high water temperature. Seed germination percentage was significantly difference in different germinating bed.

[Identification of the different cultivated Magnolia biondii].

OBJECTIVE: To distinguish Magnolia biondii of ten different cultivated. METHODS: The original species can be identified by the morphological characteristics and microscopic characteristics. The volatile oil is determinated by the method of Chinese pharmacopoeia. RESULTS: The ten cultivated Mangolia biondii have obviously differents in morphological characteristics, microscopic characteristics and volatile oil. CONCLUSION: It is important for development application of Magnolia biondii and variety improvement.

Giant flowers of Southern magnolia are hydrated by the xylem.

Flowering depends upon long-distance transport to supply water for reproductive mechanisms to function. Previous physiological studies suggested that flowers operated uncoupled from stem xylem transport and received water primarily from the phloem. We demonstrate that the water balance of Southern magnolia (Magnolia grandiflora) flowers is regulated in a manner opposite from that of previously examined flowers. We show that flowers of Southern magnolia rely upon relatively efficient xylem hydraulic transport to support high water demand during anthesis. We measured rapid rates of perianth transpiration ranging from twice to 100 times greater than previous studies. We found that relatively efficient xylem pathways existed between the xylem and flower. Perianth hydraulic conductance and the amount of xylem to transpirational surface area ratios of flowers were both approximately one-third those measured for leafy shoots. Furthermore, we observed that perianth tissues underwent significant diurnal depressions in water status during transpiring conditions. Decreases in water potential observed between flowers and vegetative tissues were consistent with water moving from the stem xylem into the flower during anthesis. Xylem hydraulic coupling of flowers to the stem was supported by experiments showing that transpiring flowers were unaffected by bark girdling. With Southern magnolia being a member of a nearly basal evolutionary lineage, our results suggest that flower water balance represents an important functional dimension that influenced early flower evolution.

[Construction on ISSR fingerprint of Magnolia officinalis from Enshi in Hubei province].

OBJECTIVE: To establish the ISSR fingerprint of the cortex of Magnolia officinalis produced from Enshi. METHODS: Cultivar and habit identification was based on Inter-simple sequence repeats (ISSR). RESULTS: Two ISSR primes could be used to construct the ISSR fingerprint of M. officinalis from Enshi. CONCLUSION: The ISSR marker is an effective method for the cultivar and habit identification of the cortex of M. officinralis.

New cytotoxic phenolic derivatives from matured fruits of Magnolia denudata.

Magnolia species are widely cultivated in Japan as garden plants, and have been found to contain various compounds, including alkaloids, terpenoids, lignans, and neolignans. The constituents of the mature fruits of M. denudata were investigated, and two new phenolic derivatives, named denudalide and denudaquinol, were isolated and characterized, together with a known neolignan compound (denudatin A). Denudalide and denudaquinol showed cytotoxicity against the SFME and r/mHM-SFME-1 cell lines.

Neolignans from North American Magnolia species with cyclooxygenase 2 inhibitory activity.

OBJECTIVES: Based upon reported ethnomedicinal use by Native Americans, extracts and pure isolates from leaves and seeds of Magnolia grandiflora, M. virginiana, M. acuminata and M. macrophylla, all native to the Southeastern United States, were investigated for their anti-inflammatory potential against cyclooxygenase 2 (COX-2). MATERIAL AND METHODS: The extracts and pure compounds from Magnolia species were tested for their production of prostaglandin E(2) (PGE(2)) using a mouse macrophage (RAW 264.7) assay where cells were stimulated by lipopolysaccharide. RESULTS: Leaf extracts were moderately active (44-58% inhibition at 50 microg/ml) whereas seed extracts showed significant activity of 54-88% inhibition, respectively. In the seed extract of M. grandiflora, honokiol, magnolol and 4'-O-methylhonokiol strongly inhibited COX-2 (IC(50): 1.2-2.0 microg/ml), 3-O-methylmagnolol was moderately active while a new compound was inactive towards COX-2. The neolignans were not cytotoxic to macrophages (RAW 264.7) and kidney fibroblast (VERO) cells in vitro. CONCLUSIONS: The results indicate that the reported ethnomedicinal use of the investigated Magnolia species is in agreement with anti-inflammatory activity of their respective compounds.

Machilin G and four neolignans from young fruits of Magnolia denudata show various degrees of inhibitory activity on nitric oxide (NO) production.

Denudatin A and B, denudadione B, fargesone A and machilin G were isolated from Magnolia denudata. These compounds showed inhibitory effects on nitric oxide (NO) production in the lipopolysaccharide plus interferon-gamma activated-murine macrophage cell line, J774.1. Some but not all of the inhibition of NO production by machilin G, and denudatin A and B was apparently through the decreased expression of the inducible NO synthase (iNOS) gene.

[Phenols in seedling cortex of Magnolia officinalis from Enshi].

OBJECTIVE: To study the phenols in Cortex of Magnolia officinalis of biernial seedings produced in Enshi, Hubei. METHODS: The content of magnolol and honokiol in cortexes were determined by HPLC. The chromatograms of 10 samples seedling cortexes were recorded and compared. RESULTS: The content of magnolol and honokiol in Cortex of Magnolia officinalis of the seedlings from Enshi was higher than other samples. There were ten characteristic absorption bands in the HPLC chromatograms, which differed from the cortex of adult trees. CONCLUSION: The results can be used to identify the quality of the seedlings for the breeding.

A phylogenetic evaluation of whether endophytes become saprotrophs at host senescence.

Fungal endophytes and saprotrophs generally play an important ecological role within plant tissues and dead plant material. Several reports based solely on morphological observations have postulated that there is an intimate link between endophytes and saprotrophs. This study aims to provide valuable insight as to whether some endophytic fungi manifest themselves as saprotrophs upon host decay. Ribosomal DNA-based sequence comparison and phylogenetic relationships from 99 fungal isolates (endophytes, mycelia sterilia, and saprotrophs) recovered from leaves and twigs of Magnolia liliifera were investigated in this study. Molecular data suggest there are fungal taxa that possibly exist as endophytes and saprotrophs. Isolates of Colletotrichum, Fusarium, Guignardia, and Phomopsis, which are common plant endophytes, have high sequence similarity and are phylogenetically related to their saprotrophic counterparts. This provides evidence to suggest that some endophytic species change their ecological strategies and adopt a saprotrophic lifestyle. The implication of these findings on fungal biodiversity and host specificity is also discussed.