Study on the variation in and selection of Fraxinus mandshurica provenances and families in northeast China

Fraxinus mandshurica Rupr.is one of the main afforestation species in northeast China,and there is great demand for improved F.mandshurica varieties.The results of an investigation into and analysis of the growth traits of F.mandshurica provenances and families showed that there were significant differences in different periods.However,variations in growth traits decreased year by year.There was a significant negative correlation between tree heights of the provenances and sunshine hours in their areas of origin.The provenances of Xinglong,Hailin and Wuchang were selected based on the volume of 18-year-old trees.The average genetic gain from the selection of fast-growing provenances was 19.4%.Ten superior fast-growing families were selected.The average volume of the selected families was22.6%,higher than that of all families.The correlation coefficient between heights at 6-year-old and at 18a was 0.838for provenances,and between heights at 4-year-old and at 18-year-old was 0.303 for families.These results indicate that early selection for height in provenances or families could be performed at 6 years or 4 years,respectively.Early selection for DBH and volume in families could start at 8 years.

Study on Hardwood Cutting Propagation Technology of Fraxinus

[Objective] Hard-branch cutting propagation technology of Fraxinus was studied under different environmental conditions. [Method] An orthogonal test was designed based on 1-year-old clones of F. pennsylvanica Marsh Lula 5. Under dif- ferent growth hormones, hormone concentrations and processing time, F. velutina Lula 2 and F. pennsy/vanica Marsh Yuanla 2 were treated by vacuum pump for 10 min, [Result] The decreasing order of cutting survival rates of Lula 5 under different etnvironments was small plastic flowerpot （94.4%）, nutrition cup （92.8%）, sand tray （56.1%）, and cuffings treated with 100 mg/L ABT, for 2 h, 200 mg/L ABT, for 4 h and 100 mg/L IBA for 6 h under large nutrition cup environment grew the best, showing a survival rate of 100%. Lula 2 and Yuanla 2 were subjected to vacuum pump treatment, the survival rate of cuttings under the sand tray environment was averagely 29%; and under the large plastic flowerpot environment, the survival rate of cuttings was averagaly 64.4%, and cuttings of Lula 2 treated with vacuum pump for 10 min in 200 mg/L ABTI showed a survival rate up to 95%, which was the highest among all treatments. It could be seen that whether cuttings were treated with vacuum pump, the survival rate of cuttings in sand tray was lower than those under other environments, while there was no big difference in survival rate of cut- tings between the nutrition cup and plastic flowerpot environments. [Conclusion] This study is of great significance to improvement of cutting propagation coefficient.

Influence of main site factors on Fraxinus mandshurica (Oleaceae) plantation

the investigation was carried out on 10-year-old plantation of Fraxinus mandshurica in Mao抏r Mountain Experimental Station of Northeast Forest University. Tree height (H), diameter at breast height (D1.3) and the increment of tree height in 5 years (H5), the thickness of humus layer, as well as the soil moisture were measured for the plantation and the growth indexes (H, D1.3, H5) for different site conditions were analyzed. The results showed that main site factors influencing the growth of Fraxinus mandshurica were soil moisture, gradient and location of slope in order. The growth of Fraxinus mandshurica was better on the middle- or up-slope site than on the down-slope site. Soil moisture and late frost caused by terrain are the main reasons that limit the growth of Fraxinus mandshurica plantation.

Enzyme Solutions and H_2O_2 on Somatic Embryogenesis of Fraxinus mandshurica Rupr. (Oleaceae)

[Objective] This study aimed to explore the effects of treatments with three types of exogenous oxidase solutions and H2O2.solution on the somatic embryogenesis of Fraxinus mandshurica Rupr. （Oleaceae）. [Method] The immature zygotic cotyledons were treated with PPQ （polyphenol oxidase） solution, GQD （glucose oxidase） solution, SOD （superoxide dismutase） solution and H202 （hydrogen peroxide） at different concentrations to explore the effects on the growth, browning and somatic embryogenesis on cotyledon explants in the somatic embryogenesis of F. mandshurica. Through comparative analysis on the effects of different treatments on somatic embryogenesis of F. mandshurica, the relationship between explants browning and somatic embryogenesis was uncovered during the somatic embryogenesis of F. mandshurica. [Result] H2O2 treatment not only advanced the explants browning, but also inhibited the growth and somatic embryogenesis of explants; different concentrations of PPQ promoted the growth and browning of explants, as well as improving the incidence of somatic embryogenesis; both GOD and SOD treatment could raise the explants browning rate; when somatic embryogenesis of explants treated with enzyme solutions advanced, the incidence of somatic embryogenesis was low; however, when the disparity of the incidence of somatic embryogenesis between 30 and 60 d treatments reached its peak, the incidence of somatic embryogenesis was also high. [Conclusion] The results of this study provide basis for raising the incidence and improving the status of somatic embryogenesis of F. mandshurica, as well as optimizing the somatic embryogenesis system of F. mandshurica.

四川梣属(Fraxinus)植物资源

对四川梣属植物11种2变种进行了调查,并编制了13个种和变种的检索表。

Accumulation of heavy metal in Platanus orientalis, Robinia pseudoacacia and Fraxinus rotundifolia

Plants are capable of reducing environmental pollutions through uptaking contaminants in their tissues. In the study site, twenty one-year-old leaves and shoots as well as twenty soil samples were sampled and analyzed by Inductively Coupled Plasma （ICP）. We investigated the uptaking ability of lead （Pb） and cadmium （Cd） by the leaves and shoots of Platanus orientalis, Robinia pseudoacacia and Fraxinus rotundifolia in Karaj city, the western Tehran, Iran. We also evaluated the total metal accumulation capacity by using metal accumulation index （MAI）. Results indicated that there was no significant difference in uptaking contents of Cd and Pb in the leaves among the trees. However accumulation of Cd in shoots of F. rotundifolia and R. pseudoacacia was significantly higher than that of P. orientalis. The accumulation of Pb in shoots of R. pseudoacacia was significantly higher than the other species. The amount of Pb in the soil of the study area was significantly higher than Cd. Concentrations of Cd and Pb in leaves of the three species are in the ranges of 2.4 2.7 mg Kg-1 and 7.1 14.4 mg Kg-1 , respectively. R. pseudoacacia had the highest MAI value for leaves （2.21） and F. rotundifolia had the highest MAI value for shoots （2.4）.

Indirect somatic embryogenesis and regeneration of Fraxinus mandshurica plants via callus tissue

Somatic embryogenesis of Fraxinus mandshurica has the problems of low somatic embryo(SE) yield,unsynchronized SE development,and a high percentage of deformed SEs.We aimed to improve F.mandshurica SE production by synchronizing SE development,improving SE quality,and inducing root formation to obtain complete regenerated plants.Cotyledons of immature zygotic embryos of F.mandshurica were induced to form callus and then SEs.The SE induction percentage from explants differed among 32 mother trees,and the one with the highest SE induction percentage(29.8%) was used for further experiments.The highest callus induction percentage was94.2% on 1/2-strength Murashige and Skoog medium(MS1/2)supplemented with 0.15 mg·L^(-1) naphthalene acetic acid.The highest callus proliferation coefficient(240.5) was obtained on McCown's Woody Plant Medium containing 0.1 mg·L^(-1)6-benzyl adenine and 0.15 mg·L^(-1) 2,4-dichlorophenoxyacetic acid.The highest number of SEs(1020.5 g~(-1) fresh weight) was obtained on MS1/2 medium supplemented with1 mg·L^(-1) 6-benzyladenine.The highest number of cotyledon embryos(397/g fresh weight) was obtained by incubating materials on medium containing 1 mg·L^(-1) abscisic acid and then applying a drying treatment.The cotyledon embryos were milky white,uniformly sized(average length 4.7 mm),and 80% of them were normal.The SE rooting percentage on 1/2 MS medium containing 0.01 mg·L^(-1) NAA was 37.5%.Overall,the germination percentage of SEs was 26.4%,and complete regenerated plants were obtained after transplanting and acclimation.These results provide more possibilities for the preservation and breeding of F.mandshurica.

Molecular cloning and expression under abiotic stresses and hormones of the ethylene response factor Ⅶ gene FmRAP2.12 from Fraxinus mandshurica

RELATED TO AP2.12(RAP2.12)is one of the Ethylene Response Factors(ERF)transcription factor and plays a key role in controlling plant root bending and responding to multiple abiotic stresses including hypoxia stress.In this study,FmRAP2.12 gene was isolated and characterized from Fraxinus mandshurica Rupr.The open reading frame(ORF)of FmRAP2.12 was 1170 bp and encoded a protein of 389 amino acids.The conserved domains,three-dimensional phylogenetic relationship of FmRAP2.12 was also investigated.Quantitative real-time(qRT-PCR)analyzed the expression of FmRAP2.12 in different tissues.The expression level of FmRAP2.12 was highest in roots followed by leaves,and lowest in male flowers.Abiotic stress and hormone signal-induced expression was established using qRT-PCR.Salt stress induced FmRAP2.12 to a double peak pattern:the first peak value was at 6 h and the second at 72 h.Drought stress also induced FmRAP2.12 to a double peak pattern:the first at6 h and the second at 48 h.FmRAP2.12 was up-regulated after initiation of gibberellic acid(GA3)treatment,with a one peak pattern at 24 h.FmRAP2.12 may not respond to cold stress and Abscisic acid(ABA)treatment.The transient overexpression of FmRAP2.12 caused the up-expression of downstream key genes of abiotic stress response and gibberellin pathway.Our research reveals the molecular characteristic and expression patterns under abiotic stress and hormone condition of FmRAP2.12,providing support for the genetic improvement of F.mandshurica at a molecular level.

Fossil-Winged Fruits of Fraxinus(Oleaceae) and Liriodendron(Magnoliaceae) from the Duho Formation,Pohang Basin,Korea

A total of 16 specimens of fossil-winged fruits were found from the Middle Miocene marine deposits, Duho Formation, Pohang Basin, Korea. They were identified into two structurally different groups： 15 specimens into a winged fruit of Fraxinus, and one specimen of Liriodendron. The most samaras （13 specimens） were identified as Fraxinus oishii, which is characterized by narrowly ovate or ovate-elliptic shapes that are 2.7-3.6 cm in length and 0.7-1 cm in width （i/w ratio=3.4-4）. The apexes of the Fraxinus oishii samara are round or slightly emarginated, and a seed of the samara is always located at the base, of which the general shape is narrow rhombic-ellipsoidal. The seed is 1.2-2 cm long and 0.5-0.7 cm wide. Two specimens are different from the samara of Fraxinus oishii. They have a 6.6 length/width ratio （3.3 cm long and 0.5 cm wide）, and thus, are temporarily classified into the Fraxinus sp. One specimen was recognized as a winged seed of Liriodendron meisenense. The wing is broadly lanceolate to elliptic in shape, has a smooth, acute apex, and is approximately 3 cm long and 0.7 cm wide. Samaras of Fraxinus oishii and Liriodendron meisenense were early reported from the Middle Miocene deposits from North Korea, but these specimens are the first discovery in South Korea. Further study of the Duho Formation may connect flora relationships between North and South Korea.

Phylogenetic relationship,biogeography,and conservation genetics of endangered Fraxinus chiisanensis(Oleaceae),endemic to South Korea

Endemic plants are important for understanding phylogenetic relationships,biogeographical history,and genetic variation because of their restricted distribution and their role in conserving biodiversity.Here,we investigated the phylogenetic relationships of the Korean endemic Fraxinus chiisanensis by reconstructing the molecular phylogeny of Fraxinus based on two nuclear DNA(nrITS and phantastica)and two chloroplast DNA(psbA-trnH and rp132-trnL)regions.Within our fossil-calibrated phylogenetic framework,we also inferred the biogeographical history of F.chiisanensis.To provide a scientific basis for the conservation of F.chiisanensis,we determined the levels of genetic diversity and genetic differentiation in this species.Combining information from nuclear and chloroplast DNA sequence data,our molecular phylogenetic analyses identified F.chiisanensis as a genetically distinct unit from its sister group,Fraxinus platypoda from Japan.Our molecular dating analyses using nuclear and chloroplast DNA data sets show F.chiisanensis diverged from its sister F.platypoda in the Early or Middle Miocene and differentiated in the Late Miocene on the Korean Peninsula.Our results suggest that the divergence of F.chiisanensis was associated with the submergence of the East China Sea land bridge and enhanced monsoons in East Asia.When compared to F.platypoda,F.chiisanensis exhibits low genetic diversity within populations and high genetic differentiation among populations.These results help us to understand the evolutionary history of F.chiisanensis and to develop a conservation strategy for this species.

Nutrient retranslocation from the fine roots of Fraxinus mandshurica and Larix olgensis in northeastern China

Nutrient retranslocation in trees is important in nutrient budgets and energy flows in forest ecosystems. We investigated nutrient retranslocation in the fine roots of a Manchurian Ash （Fraxinus mandshurica） and a Larch （Larix olgensis） plantation in northeastern China. Nutrient retranslocation in the fine roots was investigated using three methods, specifically, nutrient concentration, the ratio of Ca to other elements （Ca/other elements ratio） and nutrient content. The method based on nutrient content proved most suitable when investigating nutrient retranslocation from fine roots of the two species. The nutrient-content-based method showed that there were retranslocations of N, P, K and Mg from the fine roots of Manchurian Ash, with retranslocation efficiencies of 13, 25, 65, and 38 %, respectively, whereas there were no Ca retranslocations. There were retranslocations of N, P, K, Ca and Mg from the fine roots of Larch, with retranslocation efficiencies of 31, 40, 52, 23 and 25 %, respectively.

Definition of fine roots on the basis of the root anatomy, diameter,and branch orders of one-year old Fraxinus mandshurica seedlings

Fine roots are important in root absorption of nutrient and water,and in root turnover.Accurate definition of fine roots is a prerequisite to improved estimation of the physiological and ecological functions of forest ecosystems.Root development and physiological functions are reflections of root anatomical structure.In this study,the anatomical structures of different root orders were analyzed by examining paraffin sections of one-year old Fraxinus mandshurica seedlings.One-year-old F.mandshurica seedlings had over five root orders.The root anatomical structures of all orders showed more differences.First and second order roots consisted of four sections： the epidermis,cortex,pericycle,and vascular bundles.Fourth and fifth order roots were mainly composed of the skin and peripheral vascular bundles（including the xylem and phloem）.Third order roots had root epidermal and cortical structures,but the quantity and integrity of the cortical cells were inferior to those of the first and second order roots,and superior to those of the fourth and fifth order roots.All the first and second order roots and some third order roots with discontinuous cork layer（〈0.4 mm in diameter）,but not the fourth and fifth order roots,were the fine roots of one-year old F.mandshurica seedlings.Although they had similar diameters,different portions of root systems had different anatomical structures and therefore,vary in capacity to absorb water and nutrients.Fine roots were accurately defined by root diameter,branch orders,and anatomical structural features of one-year old F.mandshurica seedlings.

Growth and photosynthetic responses of Fraxinus mandshurica seedlings to various light environments

To determine light requirement and adaptability of Fraxinus mandshurica seedlings, the seasonal variations of photosynthetic variables were measured in 3-year-old seedlings grown under four light levels （100%, 60%, 30%, and 15% of full sunlight） with a LI-6400 portable photosynthesis system. The leaf chlorophyll content, special leaf weight, annual height and basal diameter increment of seedlings were also observed. The maximum and minimum values of net photosynthetic rate, maximum rate of carboxylation, and maximum rate of electron transport of F. mandshurica seedlings were detected with 60% and 15% of full sunlight treatments, respectively. With the decrease of light level, both light saturation point and special leaf weight significantly declined （p 0.05）, but leaf chlorophyll content significantly increased （p 0.05）. Annual height and basal diameter increments of seedlings grown under 60% of full sunlight treatment were significantly greater than those of seedlings under other treatments （p 0.05）. It was concluded that F. mandshurica seedlings can adapt to a wide range of light environments from 15% to 100% of full sunlight by adjusting light saturation point, leaf chlorophyll content and special leaf weight. According to the maximum of relative growth, 60% of full sunlight treatment is the optimum light level for the growth of 3-year-old F. mandshurica seedlings.

Relationship between H_(2)O_(2) accumulation and NO synthesis during osmotic stress:promoted somatic embryogenesis of Fraxinus mandshurica

Osmotic stress promotes somatic embryogenesis of Fraxinus mandshurica,which leads to accumulation of reactive oxygen species(ROS).The single pieces of cotyledons of F.mandshurica were used as explants to induce somatic embryogenesis in osmotic-stress medium.Furthermore,the hydrogen peroxide H_(2)O_(2) content of explanted cells was varied by adding exogenous H_(2)O_(2) or catalase solution to assess the effects of the exogenous H_(2)O_(2)on somatic embryogenesis,intracellular H_(2)O_(2)accumulation,and the relationship between signaling mediated by ROS or reactive nitrogen species.The results revealed that exogenous H_(2)O_(2)(100?300μmol L^(–1))increased the number of somatic embryos.On 60th day of exogenous H_(2)O_(2)(200μmol L^(–1))treatment,the number of somatic embryos of explants treated,which was 136.54%,was higher than the control.Moreover,exogenous H_(2)O_(2)(100μmol L^(–1))significantly increased the intracellular H_(2)O_(2)content and enhanced the activities of superoxidase dismutase and peroxidase.Finally,exogenous H_(2)O_(2)(100μmol L^(–1))activated the intracellular non-enzymatic pathway for nitric oxide(NO)synthesis.The somatic embryogenesis in broadleaf trees increases with the change of endogenic ROS content,and depends on the upregulation of antioxidant enzymes.Both H_(2)O_(2)and NO,as signaling molecules,were found to be involved in the process of somatic embryogenesis in broadleaf trees.In the process of exogenous H_(2)O_(2)promoting somatic embryogenesis,NO synthesis depended on non-enzymatic reactions.These results provide a scientific basis for resolving the mechanism by which ROS levels are regulated during somatic embryogenesis of broadleaf trees and establish a reasonable and efficient technology system for regulating somatic embryogenesis of trees.

ENDOGENOUS INHIBITORS IN SEED OF FRAXINUS MANDSHURICA RUPR AND THEIR EFFECT ON GERMINATION AND REGENERATION

Bioassay and GC-MS determination indicated that inhibitors of seeds of Fraxinus mandshurica Rupr were distributed mainly in zones of β-inhibitors (including ABA) and of high and low Rf value. The inhibitory activity of outer seed coat of the dormant seed (control treatment) was higher than that of seeds with only the inner coat attached e.g. ABA was 114 and 54ng/g fresh weight respectively. The inhibitory effect of No. 12 zone was even higher than ABA. The inhibitory activity of all the zones decreased after stratification (St) and burying (AO) treatment, especially that of the outer seed coat, whose ABA content decreased to 7(St) and 11(AO) ng / g(fw). However, the decrease in inhibition of root growth lagged bchind'thc decrease in germination. It is concluded that burial is advantageous to regeneration. Inhibitors of the outer seed coat and seeds were mostly destroyed under the adverse conditions of natural dissemination (Di treatment) indicating that Di was not conducive to regeneration. MS analysis revealed that the largest peaks of No. 6, 7 and 12 zones were linoleic acid, olcic acid and 17-C, 18-C, and 20-C long chain fatty acids. Several coumarins were found in No. 6 and 7 zones, and cadincncs, farncsol and scsquitcrpcncs were found in No. 6 and 12 zones having common bioorigins with ABA.

Progeny Test on the Natural Populations and Selection of Excellent Populations from Fraxinus Mandshurica

Fraxinus mandshurica is one of the ancient relict flora species originated from the Tertiary period, which is mainly produced in the eastern mountains in Northeast China. It has an important scientific value for studying the Tertiary flora and the Quaternary glacial climate. In order to make rational use of the natural genetic resources of F. mandshurica , and to increase the scientific and technological added value of improved varieties, this study used the method of positional observation of fixed standard fields to discuss the genetic variations of tree height and DBH of the natural populations and its open-pollinated progenies, systematically analyze the heritability and genetic gains in growth characters based on the analysis of progeny test, growth dynamic and heredity effects. The populations with good growth, stress resistance and adaptability were selected. The results showed that there were extremely significant difference in the growth characters among the natural populations, and the coefficients of variance for tree height （22.99%） were smaller, while the coefficients of variance for DBH （41.37%） were larger. On the other hand, there were extremely significant differences in the growth characters among the open-pollinated progenies, and the interaction between these progenies and the blocks was significant. Meanwhile, the growth characters had strong heritability, high genetic gain, and remarkable heredity effects. The heritability of tree height and DBH were 0.825 and 0.781, and the genetic gains were 15.16% and 25.85%, respectively. Therefore, Shanhetun （SHT） and Xinglong （XIL） were determined to be excellent natural populations both in the growth, resistance and adaptation with growth traits as the main evaluation index, which could be widely applied in the advanced generation improvement breeding.

STUDY ON PHYSIOLOGICAL CHARACTERISTICS OF LARIX OLGENSIS AND FRAXINUS MANSHURICA UNDER CONTROLLED CONDITIONS

The net photosythetical rate, respiration rate, light compensation point and light saturation point of Larix olgensis and Fraxinus manshurica were measured by Model-865 Co2Infrared Analyzer under controlled conditions (in coviron phytotron). For net photosythetical rate of Larix ogensis and Fraxinus manshurica, the optimum air temperature was 25, 28℃ respectively, the optimum soil water potantial was-20, -10 kPa repectively, the optimum soil water content was 20.31%, 23.42% respectively, and the optimum air humidity was 90%. The optimum soil water potantial of respiration rate was -30 kPa. The light compensation point for the two species was 3.36, 4.8 μmol/m2s respectively and light saturation was 804, 880μmol/m2s.

Fossil Winged Fruits of Fraxinus and Liriodendron from the Duho Formation,Pohang Basin,Korea

The Middle Miocene Duho Formation of the Yeonil Group,Pohang Basin,Korea is one of the best recognized formations for the Cenozoic plant fossils, including various leaves,reproductive organs,and fossil woods.We,in this paper,first report winged fruits(10 specimens) from the Duho Formation.They were identified into two structurally different groups:

Root-shoot relationships of Fraxinus mandchurica

In this paper, root-shoot relationships of seedlings of Manchurican ash (Fraxinus mandchurica) in pots is studied in green house. The results show that roots and shoots have the co-ordination of growth and roots and shoots of ash have the same growth dynamics. There are very close relationships between toall root mass and total shoot mass, between fine root mass and leaf mass in this experiments. At the end growing season, ash root-shoot mass ratio and fine root-leaf mass ratio are 1 .85 and 2.01 respectively. These ratios are difference during growth season. There are larger roots than shoots on seedling stage. Low nitrogen and phosphorus produce higher root-shoot ratio, and high nutrient concentrations decrease the ratios.

Study on the Provenancc of Fraxinus Mandshurica

This paper studied that the significant difference exists among the ten provenances of 9-year-old Fraxinusmandshurica stand. The rule of geographic variation is that dual continuous changes with longitude and latitude, and theprovenance in the southwest range grows rapidlv. There are positive signilicant correlation when the tree growth is influencedby≥ 10℃ accumulated temperature. duration of sunshine and annual precipitation So it is suggested that Lushuihe and Maoershan provenances should be emploved first in Maocrshan Forest Farm and its neighborhoods.