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资助项目
0.05) between wild (AR = 4.651), semi-cultivated (AR = 5.091) and cultivated (AR = 5.132) populations of C. taliensis, which suggested that the genetic background of long-lived woody plant was not easy to be changed, and there were moderate high gene flow between populations. However, there was a significant difference (P < 0.05) between wild (AR = 5.9) and cultivated (AR = 7.1) populations distributed in the same place in Yun county, Yunnan province, which may result from the hybridization and introgression of species in the tea garden and anthropogenic damages to the wild population. The hypothesis of hybrid origin of C. grandibracteata was tested by morphological and microsatellites analyses. Compared with other species, the locules in ovary of C. grandibracteata are variable, which showed a morphological intermediate and mosaic. Except one private allele, Ninety-nine percent alleles of C. grandibracteata were shared with these of C. taliensis and C. sinensis var. assamica. And C. grandibracteata was nested in the cluster of C. taliensis in the UPGMA tree. Conclusively, our results supported the hypothesis of hybrid origin of C. grandibracteata partly. The speciation of C. grandibracteata was derived from hybridization and asymmetrical introgression potentially. It is possible that C. taliensis was one of its parents, but it still needs more evidences to prove that C. sinensis var. assamica was another 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+in%5C+the%5C+UPGMA%5C+tree.%5C+Conclusively%2C%5C+our%5C+results%5C+supported%5C+the%5C+hypothesis%5C+of%5C+hybrid%5C+origin%5C+of%5C+C.%5C+grandibracteata%5C+partly.%5C+The%5C+speciation%5C+of%5C+C.%5C+grandibracteata%5C+was%5C+derived%5C+from%5C+hybridization%5C+and%5C+asymmetrical%5C+introgression%5C+potentially.%5C+It%5C+is%5C+possible%5C+that%5C+C.%5C+taliensis%5C+was%5C+one%5C+of%5C+its%5C+parents%2C%5C+but%5C+it%5C+still%5C+needs%5C+more%5C+evidences%5C+to%5C+prove%5C+that%5C+C.%5C+sinensis%5C+var.%5C+assamica%5C+was%5C+another%5C+parent."},{"jsname":"Chemical constituents and bioactivities research on Cimicifuga species have been became an international hotspot. There are abundant plant resources of this genus in China, however the step of exploitation is far fall behind the western world. In an attempt to fully utilize our domestic resources, our research group has been studying on the chemical constituents and their bioactivities, specially 9,19-cyclolanostanetriterpenes of this genus, since 2003. As a result, several cycloartane triterpenoids with potential anti-tumor activities were obtained for the first time in China. For further searching anti-tumor lead compounds with novel structures and more potent activities, in this thesis，the roots of C. yunnanensis and the aerial parts of C. foetida, which collected from Shangrila County in Yunnan Province, the roots of C. foetida, which collected from Heishitou Town in Guizhou Province and the roots of C. dahurica and C. heracleifolia, which collected from Qingyuan County in Liaoning Province were used as experimental materials and total 105 compounds were isolated and identified by different spectroscopic and chemical methods. Among these 105 compounds, there are 94 9,19-cyclolanostanetriterpenes，including 44 new constituents. All these new compounds can be categorized as Cimigenol-type, Shengmanol-type, Hydroxyshengmanol-type, Acteol-type, Dahurinol-type, Cimicidol-type, and 3,4-Seco-type, 15,16-Seco-type, which were very novel. The left 11 compounds are normal Chromones, sitosterols, oleanane triterpenes and a new Ergostane-type steoid. The isolated 9,19-cyclolanostane compounds were tested for their cytotoxicities against 15 human carcinoma cell lines, which included human hepatoma cell line HepG2, human breast carcinoma cell line MCF-7, human colorectal carcinoma cell line HT-19 and so on. 40 compounds showed activities and three of them, which isolated from C. yunnanensis exhibited as potent activities as positive control Taxol. Mechanism research indicated that these three compounds can increase the expression of p53 and bax, which led to the loss of mitochondrial potential and then resulted in the activation of caspase-7 and induced apoptosis of MCF-7. In addition, a series of compounds which isolated from the roots of C. foetida showed selective significant cytotoxicity (potent than positive controlcisplatin) against the human HepG2 cell line and a domestic patent was applied for these compounds. A conclusion of the structure-activity relationships of Cimigenol-type compounds were proposed: one of the key point for anti-tumor activities is at least one side of the compound has hydrophobic groups such as carbonyl, double bond substitute hydroxyl groups. At the end of this thesis, a comprehensive review about the distribution and utilization of the Cimicifuga genus worldwide, the structures of 9,19-cyclolanostanetriterpenes and their anti-climacteric syndrome, anti-osteoporotic and anti-tumor activities were gave.","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.source_filter&advanced=false&query1=%25E6%258A%2580%25E6%259C%25AF&&fq=dc.project.title_filter%3AChemical%5C+constituents%5C+and%5C+bioactivities%5C+research%5C+on%5C+Cimicifuga%5C+species%5C+have%5C+been%5C+became%5C+an%5C+international%5C+hotspot.%5C+There%5C+are%5C+abundant%5C+plant%5C+resources%5C+of%5C+this%5C+genus%5C+in%5C+China%2C%5C+however%5C+the%5C+step%5C+of%5C+exploitation%5C+is%5C+far%5C+fall%5C+behind%5C+the%5C+western%5C+world.%5C+In%5C+an%5C+attempt%5C+to%5C+fully%5C+utilize%5C+our%5C+domestic%5C+resources%2C%5C+our%5C+research%5C+group%5C+has%5C+been%5C+studying%5C+on%5C+the%5C+chemical%5C+constituents%5C+and%5C+their%5C+bioactivities%2C%5C+specially%5C+9%2C19%5C-cyclolanostanetriterpenes%5C+of%5C+this%5C+genus%2C%5C+since%5C+2003.%5C+As%5C+a%5C+result%2C%5C+several%5C+cycloartane%5C+triterpenoids%5C+with%5C+potential%5C+anti%5C-tumor%5C+activities%5C+were%5C+obtained%5C+for%5C+the%5C+first%5C+time%5C+in%5C+China.%5C+For%5C+further%5C+searching%5C+anti%5C-tumor%5C+lead%5C+compounds%5C+with%5C+novel%5C+structures%5C+and%5C+more%5C+potent%5C+activities%2C%5C+in%5C+this%5C+thesis%EF%BC%8Cthe%5C+roots%5C+of%5C+C.%5C+yunnanensis%5C+and%5C+the%5C+aerial%5C+parts%5C+of%5C+C.%5C+foetida%2C%5C+which%5C+collected%5C+from%5C+Shangrila%5C+County%5C+in%5C+Yunnan%5C+Province%2C%5C+the%5C+roots%5C+of%5C+C.%5C+foetida%2C%5C+which%5C+collected%5C+from%5C+Heishitou%5C+Town%5C+in%5C+Guizhou%5C+Province%5C+and%5C+the%5C+roots%5C+of%5C+C.%5C+dahurica%5C+and%5C+C.%5C+heracleifolia%2C%5C+which%5C+collected%5C+from%5C+Qingyuan%5C+County%5C+in%5C+Liaoning%5C+Province%5C+were%5C+used%5C+as%5C+experimental%5C+materials%5C+and%5C+total%5C+105%5C+compounds%5C+were%5C+isolated%5C+and%5C+identified%5C+by%5C+different%5C+spectroscopic%5C+and%5C+chemical%5C+methods.%5C+Among%5C+these%5C+105%5C+compounds%2C%5C+there%5C+are%5C+94%5C+9%2C19%5C-cyclolanostanetriterpenes%EF%BC%8Cincluding%5C+44%5C+new%5C+constituents.%5C+All%5C+these%5C+new%5C+compounds%5C+can%5C+be%5C+categorized%5C+as%5C+Cimigenol%5C-type%2C%5C+Shengmanol%5C-type%2C%5C+Hydroxyshengmanol%5C-type%2C%5C+Acteol%5C-type%2C%5C+Dahurinol%5C-type%2C%5C+Cimicidol%5C-type%2C%5C+and%5C+3%2C4%5C-Seco%5C-type%2C%5C+15%2C16%5C-Seco%5C-type%2C%5C+which%5C+were%5C+very%5C+novel.%5C+The%5C+left%5C+11%5C+compounds%5C+are%5C+normal%5C+Chromones%2C%5C+sitosterols%2C%5C+oleanane%5C+triterpenes%5C+and%5C+a%5C+new%5C+Ergostane%5C-type%5C+steoid.%5C+The%5C+isolated%5C+9%2C19%5C-cyclolanostane%5C+compounds%5C+were%5C+tested%5C+for%5C+their%5C+cytotoxicities%5C+against%5C+15%5C+human%5C+carcinoma%5C+cell%5C+lines%2C%5C+which%5C+included%5C+human%5C+hepatoma%5C+cell%5C+line%5C+HepG2%2C%5C+human%5C+breast%5C+carcinoma%5C+cell%5C+line%5C+MCF%5C-7%2C%5C+human%5C+colorectal%5C+carcinoma%5C+cell%5C+line%5C+HT%5C-19%5C+and%5C+so%5C+on.%5C+40%5C+compounds%5C+showed%5C+activities%5C+and%5C+three%5C+of%5C+them%2C%5C+which%5C+isolated%5C+from%5C+C.%5C+yunnanensis%5C+exhibited%5C+as%5C+potent%5C+activities%5C+as%5C+positive%5C+control%5C+Taxol.%5C+Mechanism%5C+research%5C+indicated%5C+that%5C+these%5C+three%5C+compounds%5C+can%5C+increase%5C+the%5C+expression%5C+of%5C+p53%5C+and%5C+bax%2C%5C+which%5C+led%5C+to%5C+the%5C+loss%5C+of%5C+mitochondrial%5C+potential%5C+and%5C+then%5C+resulted%5C+in%5C+the%5C+activation%5C+of%5C+caspase%5C-7%5C+and%5C+induced%5C+apoptosis%5C+of%5C+MCF%5C-7.%5C+In%5C+addition%2C%5C+a%5C+series%5C+of%5C+compounds%5C+which%5C+isolated%5C+from%5C+the%5C+roots%5C+of%5C+C.%5C+foetida%5C+showed%5C+selective%5C+significant%5C+cytotoxicity%5C+%5C%28potent%5C+than%5C+positive%5C+controlcisplatin%5C%29%5C+against%5C+the%5C+human%5C+HepG2%5C+cell%5C+line%5C+and%5C+a%5C+domestic%5C+patent%5C+was%5C+applied%5C+for%5C+these%5C+compounds.%5C+A%5C+conclusion%5C+of%5C+the%5C+structure%5C-activity%5C+relationships%5C+of%5C+Cimigenol%5C-type%5C+compounds%5C+were%5C+proposed%5C%3A%5C+one%5C+of%5C+the%5C+key%5C+point%5C+for%5C+anti%5C-tumor%5C+activities%5C+is%5C+at%5C+least%5C+one%5C+side%5C+of%5C+the%5C+compound%5C+has%5C+hydrophobic%5C+groups%5C+such%5C+as%5C+carbonyl%2C%5C+double%5C+bond%5C+substitute%5C+hydroxyl%5C+groups.%5C+At%5C+the%5C+end%5C+of%5C+this%5C+thesis%2C%5C+a%5C+comprehensive%5C+review%5C+about%5C+the%5C+distribution%5C+and%5C+utilization%5C+of%5C+the%5C+Cimicifuga%5C+genus%5C+worldwide%2C%5C+the%5C+structures%5C+of%5C+9%2C19%5C-cyclolanostanetriterpenes%5C+and%5C+their%5C+anti%5C-climacteric%5C+syndrome%2C%5C+anti%5C-osteoporotic%5C+and%5C+anti%5C-tumor%5C+activities%5C+were%5C+gave."},{"jsname":"Chemical investigation of twelve macrofungi and one lichen including Coriolopsis gallica, Conocybe siliginea, Albatrellus confluens, Scutellinia ascoboloides, Lactarius deliciosus, Thelephora terrestris, Collybia acervata, Shiraia bambusicola, Cortinarius alboviolaceus, Mycena galericulata, Polyporus elegans, Trogia sp., and Sulcaria virens were comprehensively reported in this dissertation.113 different compounds have been isolated and elucidated by varied chromatographic methods and extensive spectroscopic analysis, among which 26 compounds were new ones. The classes of these new compounds include acetylenic acids, tremulane-type sesquiterpenes, cleistanthane-type diterpenes, isocoumarin, norleucine-type non-protein amino acid, etc. The highlight of this dissertation is the phytochemical investigation of Trogia sp. leading to two norleucine-type non-protein amino acids, 2R-amino-4S-hydroxy-5-hexynoic acid (104) and 2R-amino-5-hexynoic acid (105), which are responsible for the toxicity of this fungus. Cellulose microcrystalline column chromatography with n-BuOH-EtOH-AcOH-H2O (4:1:1:2, v/v/v) eluting system was extensively used for isolation of amino acids in this study. Comparing to traditional n-BuOH-AcOH-H2O (BAW) solvent system, a time-saving and efficient TLC solvent system OWF (organin layer-water-formic acid) for amino acids detection was developed. TLC solvent system OWF was prepared as below: solvent OW was composed of CHCl3-CH3COCH3-MeOH (10:4:4, v/v/v), which was saturated with water; 1 to 4 drops of formic acid was added to 1 ml solvent OW when used.In the last chapter, the chemical, biological and mycological literature dealing with the isolation, structure elucidation, biological activities, and synthesis of pigments produced by those fungi that produce conspicuous fruiting bodies (macromycetes) was reviewed. 198 references between June 2003 to December 2009 are cited.","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.source_filter&advanced=false&query1=%25E6%258A%2580%25E6%259C%25AF&&fq=dc.project.title_filter%3AChemical%5C+investigation%5C+of%5C+twelve%5C+macrofungi%5C+and%5C+one%5C+lichen%5C+including%5C+Coriolopsis%5C+gallica%2C%5C+Conocybe%5C+siliginea%2C%5C+Albatrellus%5C+confluens%2C%5C+Scutellinia%5C+ascoboloides%2C%5C+Lactarius%5C+deliciosus%2C%5C+Thelephora%5C+terrestris%2C%5C+Collybia%5C+acervata%2C%5C+Shiraia%5C+bambusicola%2C%5C+Cortinarius%5C+alboviolaceus%2C%5C+Mycena%5C+galericulata%2C%5C+Polyporus%5C+elegans%2C%5C+Trogia%5C+sp.%2C%5C+and%5C+Sulcaria%5C+virens%5C+were%5C+comprehensively%5C+reported%5C+in%5C+this%5C+dissertation.113%5C+different%5C+compounds%5C+have%5C+been%5C+isolated%5C+and%5C+elucidated%5C+by%5C+varied%5C+chromatographic%5C+methods%5C+and%5C+extensive%5C+spectroscopic%5C+analysis%2C%5C+among%5C+which%5C+26%5C+compounds%5C+were%5C+new%5C+ones.%5C+The%5C+classes%5C+of%5C+these%5C+new%5C+compounds%5C+include%5C+acetylenic%5C+acids%2C%5C+tremulane%5C-type%5C+sesquiterpenes%2C%5C+cleistanthane%5C-type%5C+diterpenes%2C%5C+isocoumarin%2C%5C+norleucine%5C-type%5C+non%5C-protein%5C+amino%5C+acid%2C%5C+etc.%5C+The%5C+highlight%5C+of%5C+this%5C+dissertation%5C+is%5C+the%5C+phytochemical%5C+investigation%5C+of%5C+Trogia%5C+sp.%5C+leading%5C+to%5C+two%5C+norleucine%5C-type%5C+non%5C-protein%5C+amino%5C+acids%2C%5C+2R%5C-amino%5C-4S%5C-hydroxy%5C-5%5C-hexynoic%5C+acid%5C+%5C%28104%5C%29%5C+and%5C+2R%5C-amino%5C-5%5C-hexynoic%5C+acid%5C+%5C%28105%5C%29%2C%5C+which%5C+are%5C+responsible%5C+for%5C+the%5C+toxicity%5C+of%5C+this%5C+fungus.%5C+Cellulose%5C+microcrystalline%5C+column%5C+chromatography%5C+with%5C+n%5C-BuOH%5C-EtOH%5C-AcOH%5C-H2O%5C+%5C%284%5C%3A1%5C%3A1%5C%3A2%2C%5C+v%5C%2Fv%5C%2Fv%5C%29%5C+eluting%5C+system%5C+was%5C+extensively%5C+used%5C+for%5C+isolation%5C+of%5C+amino%5C+acids%5C+in%5C+this%5C+study.%5C+Comparing%5C+to%5C+traditional%5C+n%5C-BuOH%5C-AcOH%5C-H2O%5C+%5C%28BAW%5C%29%5C+solvent%5C+system%2C%5C+a%5C+time%5C-saving%5C+and%5C+efficient%5C+TLC%5C+solvent%5C+system%5C+OWF%5C+%5C%28organin%5C+layer%5C-water%5C-formic%5C+acid%5C%29%5C+for%5C+amino%5C+acids%5C+detection%5C+was%5C+developed.%5C+TLC%5C+solvent%5C+system%5C+OWF%5C+was%5C+prepared%5C+as%5C+below%5C%3A%5C+solvent%5C+OW%5C+was%5C+composed%5C+of%5C+CHCl3%5C-CH3COCH3%5C-MeOH%5C+%5C%2810%5C%3A4%5C%3A4%2C%5C+v%5C%2Fv%5C%2Fv%5C%29%2C%5C+which%5C+was%5C+saturated%5C+with%5C+water%5C%3B%5C+1%5C+to%5C+4%5C+drops%5C+of%5C+formic%5C+acid%5C+was%5C+added%5C+to%5C+1%5C+ml%5C+solvent%5C+OW%5C+when%5C+used.In%5C+the%5C+last%5C+chapter%2C%5C+the%5C+chemical%2C%5C+biological%5C+and%5C+mycological%5C+literature%5C+dealing%5C+with%5C+the%5C+isolation%2C%5C+structure%5C+elucidation%2C%5C+biological%5C+activities%2C%5C+and%5C+synthesis%5C+of%5C+pigments%5C+produced%5C+by%5C+those%5C+fungi%5C+that%5C+produce%5C+conspicuous%5C+fruiting%5C+bodies%5C+%5C%28macromycetes%5C%29%5C+was%5C+reviewed.%5C+198%5C+references%5C+between%5C+June%5C+2003%5C+to%5C+December%5C+2009%5C+are%5C+cited."},{"jsname":"Chinese Academy of Sciences[2013T2S0030]","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.source_filter&advanced=false&query1=%25E6%258A%2580%25E6%259C%25AF&&fq=dc.project.title_filter%3AChinese%5C+Academy%5C+of%5C+Sciences%5C%5B2013T2S0030%5C%5D"},{"jsname":"Cold stress is one of the major environmental factors that adversely influence plants growth. Cold stress not only limits plants geographic distribution, but also reduces plants yield by shortening growing season, which brought billions of dollars economic losses for global crop. In nature, responses of overwintering plants to low temperature can be divided into three distinct phases: cold acclimation (CA), freezing, and post-freezing recovery (PFR). Until now, plenty intensive study about molecular mechanism of cold stress mainly focused on the above-zero low temperature phase. However, the studies on the freezing phase below zero and the following PFR phase with temperature going up to above-zero were rare. The previous research form our lab hinted that the responses of plants to freezing and PFR were complex and important. Except for passive reflection, there were also crucial active responses during this process. Several special rules were presented at the different levels including gene expression, signal transduction and membrane lipids changes, and fully understanding these rules would be helpful for us to explore the responses of plants to low temperature and then proceed to improve the freezing resistance of plants. In the present study, the mechanisms of respond to freezing and PFR of model plant Arabidopsis thaliana and its close relative Thellungiella halophlia that with extreme tolerance to abiotic stresses were carried out, including regulation of gene expression, signal transduction pathway and membrane lipids changes three levels which were essential for the freezing resistance of plants. Ground on these work, we obtained results from the following five aspects. First, the complete picture of A. thaliana responding to freezing and PFR at transcriptome level was elaborated and three functional genes closely related to the phases were identified. Second, the cis-elements with high frequent presence in differentially expressed genes were elucidated, and the practical binding of one elements among them was experimental verified during freezing and PFR. Moreover, we predicted the new elements which would respond to freezing and PFR. Third, the regulation of freezing stress by microRNA in A. thaliana was preliminarily investigated and 36 functional genes possibly regulated by miRNA during freezing and PFR were gained. Fourth, the negative effect of phytohormone Auxin on A. thaliana subjected to freezing stress was identified. Fifth, for the freezing-resistant plant T. halophlia, the rules of membrane lipids composition changes under freezing stress were uncovered.","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.source_filter&advanced=false&query1=%25E6%258A%2580%25E6%259C%25AF&&fq=dc.project.title_filter%3ACold%5C+stress%5C+is%5C+one%5C+of%5C+the%5C+major%5C+environmental%5C+factors%5C+that%5C+adversely%5C+influence%5C+plants%5C+growth.%5C+Cold%5C+stress%5C+not%5C+only%5C+limits%5C+plants%5C+geographic%5C+distribution%2C%5C+but%5C+also%5C+reduces%5C+plants%5C+yield%5C+by%5C+shortening%5C+growing%5C+season%2C%5C+which%5C+brought%5C+billions%5C+of%5C+dollars%5C+economic%5C+losses%5C+for%5C+global%5C+crop.%5C+In%5C+nature%2C%5C+responses%5C+of%5C+overwintering%5C+plants%5C+to%5C+low%5C+temperature%5C+can%5C+be%5C+divided%5C+into%5C+three%5C+distinct%5C+phases%5C%3A%5C+cold%5C+acclimation%5C+%5C%28CA%5C%29%2C%5C+freezing%2C%5C+and%5C+post%5C-freezing%5C+recovery%5C+%5C%28PFR%5C%29.%5C+Until%5C+now%2C%5C+plenty%5C+intensive%5C+study%5C+about%5C+molecular%5C+mechanism%5C+of%5C+cold%5C+stress%5C+mainly%5C+focused%5C+on%5C+the%5C+above%5C-zero%5C+low%5C+temperature%5C+phase.%5C+However%2C%5C+the%5C+studies%5C+on%5C+the%5C+freezing%5C+phase%5C+below%5C+zero%5C+and%5C+the%5C+following%5C+PFR%5C+phase%5C+with%5C+temperature%5C+going%5C+up%5C+to%5C+above%5C-zero%5C+were%5C+rare.%5C+The%5C+previous%5C+research%5C+form%5C+our%5C+lab%5C+hinted%5C+that%5C+the%5C+responses%5C+of%5C+plants%5C+to%5C+freezing%5C+and%5C+PFR%5C+were%5C+complex%5C+and%5C+important.%5C+Except%5C+for%5C+passive%5C+reflection%2C%5C+there%5C+were%5C+also%5C+crucial%5C+active%5C+responses%5C+during%5C+this%5C+process.%5C+Several%5C+special%5C+rules%5C+were%5C+presented%5C+at%5C+the%5C+different%5C+levels%5C+including%5C+gene%5C+expression%2C%5C+signal%5C+transduction%5C+and%5C+membrane%5C+lipids%5C+changes%2C%5C+and%5C+fully%5C+understanding%5C+these%5C+rules%5C+would%5C+be%5C+helpful%5C+for%5C+us%5C+to%5C+explore%5C+the%5C+responses%5C+of%5C+plants%5C+to%5C+low%5C+temperature%5C+and%5C+then%5C+proceed%5C+to%5C+improve%5C+the%5C+freezing%5C+resistance%5C+of%5C+plants.%5C+In%5C+the%5C+present%5C+study%2C%5C+the%5C+mechanisms%5C+of%5C+respond%5C+to%5C+freezing%5C+and%5C+PFR%5C+of%5C+model%5C+plant%5C+Arabidopsis%5C+thaliana%5C+and%5C+its%5C+close%5C+relative%5C+Thellungiella%5C+halophlia%5C+that%5C+with%5C+extreme%5C+tolerance%5C+to%5C+abiotic%5C+stresses%5C+were%5C+carried%5C+out%2C%5C+including%5C+regulation%5C+of%5C+gene%5C+expression%2C%5C+signal%5C+transduction%5C+pathway%5C+and%5C+membrane%5C+lipids%5C+changes%5C+three%5C+levels%5C+which%5C+were%5C+essential%5C+for%5C+the%5C+freezing%5C+resistance%5C+of%5C+plants.%5C+Ground%5C+on%5C+these%5C+work%2C%5C+we%5C+obtained%5C+results%5C+from%5C+the%5C+following%5C+five%5C+aspects.%5C+First%2C%5C+the%5C+complete%5C+picture%5C+of%5C+A.%5C+thaliana%5C+responding%5C+to%5C+freezing%5C+and%5C+PFR%5C+at%5C+transcriptome%5C+level%5C+was%5C+elaborated%5C+and%5C+three%5C+functional%5C+genes%5C+closely%5C+related%5C+to%5C+the%5C+phases%5C+were%5C+identified.%5C+Second%2C%5C+the%5C+cis%5C-elements%5C+with%5C+high%5C+frequent%5C+presence%5C+in%5C+differentially%5C+expressed%5C+genes%5C+were%5C+elucidated%2C%5C+and%5C+the%5C+practical%5C+binding%5C+of%5C+one%5C+elements%5C+among%5C+them%5C+was%5C+experimental%5C+verified%5C+during%5C+freezing%5C+and%5C+PFR.%5C+Moreover%2C%5C+we%5C+predicted%5C+the%5C+new%5C+elements%5C+which%5C+would%5C+respond%5C+to%5C+freezing%5C+and%5C+PFR.%5C+Third%2C%5C+the%5C+regulation%5C+of%5C+freezing%5C+stress%5C+by%5C+microRNA%5C+in%5C+A.%5C+thaliana%5C+was%5C+preliminarily%5C+investigated%5C+and%5C+36%5C+functional%5C+genes%5C+possibly%5C+regulated%5C+by%5C+miRNA%5C+during%5C+freezing%5C+and%5C+PFR%5C+were%5C+gained.%5C+Fourth%2C%5C+the%5C+negative%5C+effect%5C+of%5C+phytohormone%5C+Auxin%5C+on%5C+A.%5C+thaliana%5C+subjected%5C+to%5C+freezing%5C+stress%5C+was%5C+identified.%5C+Fifth%2C%5C+for%5C+the%5C+freezing%5C-resistant%5C+plant%5C+T.%5C+halophlia%2C%5C+the%5C+rules%5C+of%5C+membrane%5C+lipids%5C+composition%5C+changes%5C+under%5C+freezing%5C+stress%5C+were%5C+uncovered."},{"jsname":"Craigia yunnanensis W. W. Smith & W. E. Evans (Tiliaceae) is an endangered deciduous tree species which has high scientific and economic value. C. yunnanensis is seriously threatened and has been pushed to the verge of extinction due to vegetation destruction in China and consequent contraction of its distribution. Hence, it was listed as a nationally rare and endangered plant in 1999 and has also been proposed as a second-ranked plant for national protection in China and included in IUCN red list. As a scientifically important and valued tree species with endangered status, the wild populations of C. yunnanensis therefore represent is a genetic resource that must be conserved. To provide basic information for its conservation, the population dynamics and population size structures, pollination biology and breeding system, eleven fitness-related characters and the genetic variability based on AFLP were comprehensively studied. The main results are summarized as follows: A total of six wild populations of C. yunnanensis were found in two disjunct regions of Yunnan, i.e. WenShan (SE Yunnan) and DeHong (SW Yunnan), from 2005 to 2007. Additionally, in all but one of the populations we detected, mature trees were felled between 2005 and 2007, so destruction of most of these populations is ongoing. Across the six populations of extant C. yunnanensis found during our study, the total number of mature (reproductive) individuals detected was 584 in 2007，plus larger numbers of seedling and resprouts from cut trunks. The result of surveying Population structure showed that there are two regeneration types which are seedlings and sprouts. Seedlings occurred abundantly in gaps or open areas and the size class frequency distributions were often discontinuous, and the same general pattern occurred in all the investigated populations for juveniles and adults. The numbers of seed-origin individuals did however decline sharply with increasing size, indicating a high mortality rate going from seedling to sapling stage may be a problem for this species. Additionally, the cash crop cultivation and logging seriously threaten the survival of the species. We conducted field observations and artificial pollination experiments on the floral biology, pollination process and breeding system of Craigia yunnanensis in Fadou, Xichou county of Yunnan province. The lifespan of a single hermaphrodite flower is approximately 3-4 days. A cyme has 2-9 flowered. The flowering period of an inflorescence is usually 5-14 days. The flowers of C. yunnanensis were protandrous. The stamens were within petal-like staminodes in the opening flowers until the flower withered. Without touchment, the bractlike staminodes can’t open. Self-pollination was partially avoided by temporal and spatial isolation of male and female organs within the same flower. However, autogamous and geitonogamous pollination is unavoidable because of the large number of flowers on a single tree and the action of pollinators. The values of both OCI (≥4) and P/O (1381) and the results of bagging tests indicated there was no apomixes in C. yunnanensis and the breeding system of the species was outcrossing with partial self-compatibility and the pollinators were required during the pollination process. The most frequent effective floral visitor was only beautiful fly (Chrysomyia megacephala). Fruit set and seed set in natural condition were 56.67±3.85％ and 6.26±0.75％, respectively. Therefore, lack of pollinators, low pollination efficiency, unavoidable geitonogamous pollination and partial self-compatibility and inbreeding in small populations may account for the low fruit set, especially seed set.Variations in seed traits, seed germination, and seedling growth characters among six Craigia yunnanensis populations were evaluated. All seed and seedling traits exhibited significant differences among populations (P < 0.05). The fitness of seed as assessed by seed size, seed germination and seedling trait was independent of population size, except for the number of seeds per capsule (r = 0.93，P < 0.01). Correlations between geo-climatic variables of seed origin and seed and seedling related characters were insignificant (P > 0.05). For some populations, germination capacity in 12-h photoperiod was significantly higher than that in completed darkness（W-FD: P < 0.01, W-JD: P < 0.05）.Genetic variation within and among six populations was assessed using AFLP markers. Genetic diversity was higher at species level (PPL = 69.19%, HE = 0.221) than at population level (PPL = 26.22%, HE = 0.095, Is =0.140), and populations in southeast Yunnan were strongly differentiated from those in southwest Yunnan (Nei’s GST = 0.575; FST = 0.655). UPGMA analysis demonstrated a clear genetic division between the two populations from DeHong (SW Yunnan; D-JD and D-HG) and the four from WenShan (SE Yunnan; W-FD, W-LH, W-ML, and W-MG). Within-population genetic variation was significantly correlated with population isolation (r(PPL) = -0.94, P = 0.006; r(HE) = -0.85, P = 0.032; r(Is) = -0.87, P = 0.025), but not with population size (r(PPL) = 0.63, P = 0.178; r(HE) = 0.54, P = 0.268; r(Is) = 0.56, P = 0.249).","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.source_filter&advanced=false&query1=%25E6%258A%2580%25E6%259C%25AF&&fq=dc.project.title_filter%3ACraigia%5C+yunnanensis%5C+W.%5C+W.%5C+Smith%5C+%5C%26%5C+W.%5C+E.%5C+Evans%5C+%5C%28Tiliaceae%5C%29%5C+is%5C+an%5C+endangered%5C+deciduous%5C+tree%5C+species%5C+which%5C+has%5C+high%5C+scientific%5C+and%5C+economic%5C+value.%5C+C.%5C+yunnanensis%5C+is%5C+seriously%5C+threatened%5C+and%5C+has%5C+been%5C+pushed%5C+to%5C+the%5C+verge%5C+of%5C+extinction%5C+due%5C+to%5C+vegetation%5C+destruction%5C+in%5C+China%5C+and%5C+consequent%5C+contraction%5C+of%5C+its%5C+distribution.%5C+Hence%2C%5C+it%5C+was%5C+listed%5C+as%5C+a%5C+nationally%5C+rare%5C+and%5C+endangered%5C+plant%5C+in%5C+1999%5C+and%5C+has%5C+also%5C+been%5C+proposed%5C+as%5C+a%5C+second%5C-ranked%5C+plant%5C+for%5C+national%5C+protection%5C+in%5C+China%5C+and%5C+included%5C+in%5C+IUCN%5C+red%5C+list.%5C+As%5C+a%5C+scientifically%5C+important%5C+and%5C+valued%5C+tree%5C+species%5C+with%5C+endangered%5C+status%2C%5C+the%5C+wild%5C+populations%5C+of%5C+C.%5C+yunnanensis%5C+therefore%5C+represent%5C+is%5C+a%5C+genetic%5C+resource%5C+that%5C+must%5C+be%5C+conserved.%5C+To%5C+provide%5C+basic%5C+information%5C+for%5C+its%5C+conservation%2C%5C+the%5C+population%5C+dynamics%5C+and%5C+population%5C+size%5C+structures%2C%5C+pollination%5C+biology%5C+and%5C+breeding%5C+system%2C%5C+eleven%5C+fitness%5C-related%5C+characters%5C+and%5C+the%5C+genetic%5C+variability%5C+based%5C+on%5C+AFLP%5C+were%5C+comprehensively%5C+studied.%5C+The%5C+main%5C+results%5C+are%5C+summarized%5C+as%5C+follows%5C%3A%5C+A%5C+total%5C+of%5C+six%5C+wild%5C+populations%5C+of%5C+C.%5C+yunnanensis%5C+were%5C+found%5C+in%5C+two%5C+disjunct%5C+regions%5C+of%5C+Yunnan%2C%5C+i.e.%5C+WenShan%5C+%5C%28SE%5C+Yunnan%5C%29%5C+and%5C+DeHong%5C+%5C%28SW%5C+Yunnan%5C%29%2C%5C+from%5C+2005%5C+to%5C+2007.%5C+Additionally%2C%5C+in%5C+all%5C+but%5C+one%5C+of%5C+the%5C+populations%5C+we%5C+detected%2C%5C+mature%5C+trees%5C+were%5C+felled%5C+between%5C+2005%5C+and%5C+2007%2C%5C+so%5C+destruction%5C+of%5C+most%5C+of%5C+these%5C+populations%5C+is%5C+ongoing.%5C+Across%5C+the%5C+six%5C+populations%5C+of%5C+extant%5C+C.%5C+yunnanensis%5C+found%5C+during%5C+our%5C+study%2C%5C+the%5C+total%5C+number%5C+of%5C+mature%5C+%5C%28reproductive%5C%29%5C+individuals%5C+detected%5C+was%5C+584%5C+in%5C+2007%EF%BC%8Cplus%5C+larger%5C+numbers%5C+of%5C+seedling%5C+and%5C+resprouts%5C+from%5C+cut%5C+trunks.%5C+The%5C+result%5C+of%5C+surveying%5C+Population%5C+structure%5C+showed%5C+that%5C+there%5C+are%5C+two%5C+regeneration%5C+types%5C+which%5C+are%5C+seedlings%5C+and%5C+sprouts.%5C+Seedlings%5C+occurred%5C+abundantly%5C+in%5C+gaps%5C+or%5C+open%5C+areas%5C+and%5C+the%5C+size%5C+class%5C+frequency%5C+distributions%5C+were%5C+often%5C+discontinuous%2C%5C+and%5C+the%5C+same%5C+general%5C+pattern%5C+occurred%5C+in%5C+all%5C+the%5C+investigated%5C+populations%5C+for%5C+juveniles%5C+and%5C+adults.%5C+The%5C+numbers%5C+of%5C+seed%5C-origin%5C+individuals%5C+did%5C+however%5C+decline%5C+sharply%5C+with%5C+increasing%5C+size%2C%5C+indicating%5C+a%5C+high%5C+mortality%5C+rate%5C+going%5C+from%5C+seedling%5C+to%5C+sapling%5C+stage%5C+may%5C+be%5C+a%5C+problem%5C+for%5C+this%5C+species.%5C+Additionally%2C%5C+the%5C+cash%5C+crop%5C+cultivation%5C+and%5C+logging%5C+seriously%5C+threaten%5C+the%5C+survival%5C+of%5C+the%5C+species.%5C+We%5C+conducted%5C+field%5C+observations%5C+and%5C+artificial%5C+pollination%5C+experiments%5C+on%5C+the%5C+floral%5C+biology%2C%5C+pollination%5C+process%5C+and%5C+breeding%5C+system%5C+of%5C+Craigia%5C+yunnanensis%5C+in%5C+Fadou%2C%5C+Xichou%5C+county%5C+of%5C+Yunnan%5C+province.%5C+The%5C+lifespan%5C+of%5C+a%5C+single%5C+hermaphrodite%5C+flower%5C+is%5C+approximately%5C+3%5C-4%5C+days.%5C+A%5C+cyme%5C+has%5C+2%5C-9%5C+flowered.%5C+The%5C+flowering%5C+period%5C+of%5C+an%5C+inflorescence%5C+is%5C+usually%5C+5%5C-14%5C+days.%5C+The%5C+flowers%5C+of%5C+C.%5C+yunnanensis%5C+were%5C+protandrous.%5C+The%5C+stamens%5C+were%5C+within%5C+petal%5C-like%5C+staminodes%5C+in%5C+the%5C+opening%5C+flowers%5C+until%5C+the%5C+flower%5C+withered.%5C+Without%5C+touchment%2C%5C+the%5C+bractlike%5C+staminodes%5C+can%E2%80%99t%5C+open.%5C+Self%5C-pollination%5C+was%5C+partially%5C+avoided%5C+by%5C+temporal%5C+and%5C+spatial%5C+isolation%5C+of%5C+male%5C+and%5C+female%5C+organs%5C+within%5C+the%5C+same%5C+flower.%5C+However%2C%5C+autogamous%5C+and%5C+geitonogamous%5C+pollination%5C+is%5C+unavoidable%5C+because%5C+of%5C+the%5C+large%5C+number%5C+of%5C+flowers%5C+on%5C+a%5C+single%5C+tree%5C+and%5C+the%5C+action%5C+of%5C+pollinators.%5C+The%5C+values%5C+of%5C+both%5C+OCI%5C+%5C%28%E2%89%A54%5C%29%5C+and%5C+P%5C%2FO%5C+%5C%281381%5C%29%5C+and%5C+the%5C+results%5C+of%5C+bagging%5C+tests%5C+indicated%5C+there%5C+was%5C+no%5C+apomixes%5C+in%5C+C.%5C+yunnanensis%5C+and%5C+the%5C+breeding%5C+system%5C+of%5C+the%5C+species%5C+was%5C+outcrossing%5C+with%5C+partial%5C+self%5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species, usually called tree ferns, are considered as relicts of a time when dinosaurs were common. In recent several decades, the number of Cyatheaceae plants decreases dramatically. In order to find the reasons and provide directions for protecting these endangered plants, the biological characteristics of Cyatheaceae were surveyed. Using AFLP and cpDNA sequence variations, the genetic diversity and phylogeography of Sphaeropteris brunoniana were also analyzed. Based on these findings, implications for conservation strategies were discussed for this relict tree fern. Main results of the dissertation were summarized as follows, (1) Cyatheaceae plants have extensive distribution in Yunnan, China, and most of them distribute in southeast of Yunnan. In southeast, they usually inhabit margins of evergreen broad-leaved forests or secondary coniferous forests; however, the population update is very different and the age structure is unscientific. The spore of Cyatheaceae is trilete, radially symmetrical, and perinous. The spores of Alsophila species feature a ridged perine and a granular, verrucate or smooth exine. The spores of S. brunoniana are characterized by an incipient granular outermost layer and a verrucate exine. The metaphase chromosome numbers of gametophytes in the three examined species, viz. A. podophylla, A. gigantea and A. austro-yunnanensis, are 69, indicating that they are diploid and do not display variety in chromosome number. The chemical constituents of S. brunoniana are main simple and familiar compounds, such as saccharides, fatty acids and alcohols, and stigmasterols. (2) An unexpectedly high level of nDNA genetic diversity and low cpDNA diversity were detected in S. brunoniana. (3) This study showed that the genetic differentiation among populations within regions was low and between regions was significant. (4) There were several refugia of S. brunoniana in Yunnan during glacial periods. The Hainan populations were likely new colonizations and originated from Southeast Asia. (5) To retain existing genetic diversity, whether in situ or ex situ conservation or collection of germplasm is used, the populations of the two regions should be considered equally. Furthermore, ex situ conservation of this species should be preferably conducted on large populations.","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.source_filter&advanced=false&query1=%25E6%258A%2580%25E6%259C%25AF&&fq=dc.project.title_filter%3ACyatheaceae%5C+species%2C%5C+usually%5C+called%5C+tree%5C+ferns%2C%5C+are%5C+considered%5C+as%5C+relicts%5C+of%5C+a%5C+time%5C+when%5C+dinosaurs%5C+were%5C+common.%5C+In%5C+recent%5C+several%5C+decades%2C%5C+the%5C+number%5C+of%5C+Cyatheaceae%5C+plants%5C+decreases%5C+dramatically.%5C+In%5C+order%5C+to%5C+find%5C+the%5C+reasons%5C+and%5C+provide%5C+directions%5C+for%5C+protecting%5C+these%5C+endangered%5C+plants%2C%5C+the%5C+biological%5C+characteristics%5C+of%5C+Cyatheaceae%5C+were%5C+surveyed.%5C+Using%5C+AFLP%5C+and%5C+cpDNA%5C+sequence%5C+variations%2C%5C+the%5C+genetic%5C+diversity%5C+and%5C+phylogeography%5C+of%5C+Sphaeropteris%5C+brunoniana%5C+were%5C+also%5C+analyzed.%5C+Based%5C+on%5C+these%5C+findings%2C%5C+implications%5C+for%5C+conservation%5C+strategies%5C+were%5C+discussed%5C+for%5C+this%5C+relict%5C+tree%5C+fern.%5C+Main%5C+results%5C+of%5C+the%5C+dissertation%5C+were%5C+summarized%5C+as%5C+follows%2C%5C+%5C%281%5C%29%5C+Cyatheaceae%5C+plants%5C+have%5C+extensive%5C+distribution%5C+in%5C+Yunnan%2C%5C+China%2C%5C+and%5C+most%5C+of%5C+them%5C+distribute%5C+in%5C+southeast%5C+of%5C+Yunnan.%5C+In%5C+southeast%2C%5C+they%5C+usually%5C+inhabit%5C+margins%5C+of%5C+evergreen%5C+broad%5C-leaved%5C+forests%5C+or%5C+secondary%5C+coniferous%5C+forests%5C%3B%5C+however%2C%5C+the%5C+population%5C+update%5C+is%5C+very%5C+different%5C+and%5C+the%5C+age%5C+structure%5C+is%5C+unscientific.%5C+The%5C+spore%5C+of%5C+Cyatheaceae%5C+is%5C+trilete%2C%5C+radially%5C+symmetrical%2C%5C+and%5C+perinous.%5C+The%5C+spores%5C+of%5C+Alsophila%5C+species%5C+feature%5C+a%5C+ridged%5C+perine%5C+and%5C+a%5C+granular%2C%5C+verrucate%5C+or%5C+smooth%5C+exine.%5C+The%5C+spores%5C+of%5C+S.%5C+brunoniana%5C+are%5C+characterized%5C+by%5C+an%5C+incipient%5C+granular%5C+outermost%5C+layer%5C+and%5C+a%5C+verrucate%5C+exine.%5C+The%5C+metaphase%5C+chromosome%5C+numbers%5C+of%5C+gametophytes%5C+in%5C+the%5C+three%5C+examined%5C+species%2C%5C+viz.%5C+A.%5C+podophylla%2C%5C+A.%5C+gigantea%5C+and%5C+A.%5C+austro%5C-yunnanensis%2C%5C+are%5C+69%2C%5C+indicating%5C+that%5C+they%5C+are%5C+diploid%5C+and%5C+do%5C+not%5C+display%5C+variety%5C+in%5C+chromosome%5C+number.%5C+The%5C+chemical%5C+constituents%5C+of%5C+S.%5C+brunoniana%5C+are%5C+main%5C+simple%5C+and%5C+familiar%5C+compounds%2C%5C+such%5C+as%5C+saccharides%2C%5C+fatty%5C+acids%5C+and%5C+alcohols%2C%5C+and%5C+stigmasterols.%5C+%5C%282%5C%29%5C+An%5C+unexpectedly%5C+high%5C+level%5C+of%5C+nDNA%5C+genetic%5C+diversity%5C+and%5C+low%5C+cpDNA%5C+diversity%5C+were%5C+detected%5C+in%5C+S.%5C+brunoniana.%5C+%5C%283%5C%29%5C+This%5C+study%5C+showed%5C+that%5C+the%5C+genetic%5C+differentiation%5C+among%5C+populations%5C+within%5C+regions%5C+was%5C+low%5C+and%5C+between%5C+regions%5C+was%5C+significant.%5C+%5C%284%5C%29%5C+There%5C+were%5C+several%5C+refugia%5C+of%5C+S.%5C+brunoniana%5C+in%5C+Yunnan%5C+during%5C+glacial%5C+periods.%5C+The%5C+Hainan%5C+populations%5C+were%5C+likely%5C+new%5C+colonizations%5C+and%5C+originated%5C+from%5C+Southeast%5C+Asia.%5C+%5C%285%5C%29%5C+To%5C+retain%5C+existing%5C+genetic%5C+diversity%2C%5C+whether%5C+in%5C+situ%5C+or%5C+ex%5C+situ%5C+conservation%5C+or%5C+collection%5C+of%5C+germplasm%5C+is%5C+used%2C%5C+the%5C+populations%5C+of%5C+the%5C+two%5C+regions%5C+should%5C+be%5C+considered%5C+equally.%5C+Furthermore%2C%5C+ex%5C+situ%5C+conservation%5C+of%5C+this%5C+species%5C+should%5C+be%5C+preferably%5C+conducted%5C+on%5C+large%5C+populations."},{"jsname":"Cycas micholitzii complex is composed of 5 species: C. micholitzii Dyer, C. bifida (Dyer) K. D. Hill,C. longipetiolula D. Y. Wang, C. debaoensis Y. C. Zhong et C J. Chen, C. multipinnata C J. Chen et S. Y. Yang，and distributed from southwest China to central Vietnam and eastern Laos. Based on sequence data from two maternally inherited cpDNA and one biparentally nuclear DNA fragments, our study revealed the population genetic structure of C. micholitzii complex and explored the potential causes. The evolutionary and demographic histories were investigated. The genetic relationship among species in the complex was also clarified.The results were summarized as follows: 1. Phylogeographic analysis based on chloroplast sequences，We examined chloroplast sequence variation of the atpB-rbcLand psbA-trnHintergenic spacers in 27 populations of C. micholitzii complex, recovering 26 haplotypes. The average within-population diversity (HS = 0.140) was low while total diversity (HT = 0.911) was high. Population differentiation was also high(GST = 0.846, NST = 0.919), indicating significant phylogeographical structure (NST > GST,p < 0.001) and low levels of seed-based gene flow. C. debaoensis (Cycadaceae) is an endangered species restricted to the border of Guangxi and Yunnan province in southwest China. This species has been classified into two types: sand and karst, according to the soil matrix they grow on. We examined chloroplast sequence variation of the cpDNA sequences from 11 populations of this species. Significant population genetic differentiation was detected (GST= 0.684 and FST = 0.74160). There was marked genetic differentiation between populations in the sand and karst regions and no expansion was detected. Climate changes during glacial periods have had significant effects on the current distribution of cycads. The molecular phylogenetic data, together with the geographic distribution of the haplotypes, suggest that C. debaoensis experienced range contraction during glacial periods, and that the current populations are still confined to the original refugia in southwest China which have favorable habitats in glacial period. These results imply that small refugia were maintained in both sand and karst regions during the LGM (last glacial maximum). This species had no postglacial recolonization and only stayed in these refugia up to now. The low within-population diversity of C. debaoensis suggests that there were strong bottleneck events or founder effects within each separate region during the Quaternary climatic oscillations. Relatively high genetic and haplotype diversities were detected in the newly discovered populations, which located at intermediate locality of sand regions and had morphological variation; this is probably the consequence of the admixture of different haplotypes colonizing the area from separate sources. C. micholitzii occurs in the Annan Highlands in central Vietnam near the Laos border. C. bifida occurs in North Vietnam; its distribution extends across the border into adjacent localities in Guangxi and Yunnan in China. For the comparability between them,theywere considered as the same species C. micholitzii by many academicians. The cpDNA sequences from 11 populations showed that these very controversial species, C. micholitzii and C. bifida, is paraphyletic and should belong to the same species C. micholitzii. AMOVA analysis showed that the component of among-population within region/species (76.46%) was unexpectedly larger than the among-species/region component (14.97%), which also indicates that there is no justification for recognizing two species as C. micholitzii and C. bifida. This hypothesis was also supported by the geological data, especially the neotectonic history of the indo-china block, which started to move south since Oligocene and cause the geographic isolation of these two groups. Therefore, the most likely explanation to the phenotypic similarities between these two groups may be the retention of ancestral polymorphisms in the paraphyletic group due to incomplete lineage sorting. Furthermore, the similarities may also be ascribed to pollen-mediated gene flow among geographically proximate populations and/or phenotypic convergence under similar selection schemes in the same region. C.micholitzi had the higest genetic diversity (HT = 0.980,) and genetic differentiation (GST = 0.830, NST = 0.915) among the C. micholitzii complex. The high genetic diversity might be attributed to its long evolutionary history, highly diverse habitats. The ineffective mode of seed dispersal and dramatic neotectonic movement in the distribution range of this species could result in the high genetic differentiation. 2. Phylogeographic analysis based on nuclear ribosomal sequences, We sequenced the nrDNA ITS in all 27 populations sampled, 7 haplotypes were identified, among which C. micholitzii had 6, while C. multipinnata, C. longipetiolula and C. debaoensis shared the remaining one. Compared to chloroplast genes, nuclear genes had higher correlation between genetic and geographical distance, but lower interspecies differentiation (54.42% vs 25.24%). Phylogeographical structure of C. micholitzii and C.bifida based on ITS Variation was consistent with the morphology differentiation. This similar in nuclear gene should be ascribed to pollen-mediated gene flow among geographically proximate populations.Long-distance gene flow over the two groups was clearly interrupted, which brought on the nrDNA genetic differenciation between the geographically isolated groups, to a certain extent affected the morphological variation. 3. Interspecies relationships among Cycas micholitzii complex, We analysed chloroplast sequence variation of the atpB-rbcL and psbA-trnH intergenic spacers in 27 populations sampled of C. micholitzii complex, AMOVA analysis showed that the component of among-species/region component (59.21%). However, phylogenic analysis showed that the haplotypes of C. micholitzii complex couldn`t grouped into four clusters closely corresponding to the narrowly defined C. micholitzi, C. multipinnata, C. debaoensis and C. longipetiolula. We concluded that the conflict may result from several factors: firstly incomplete lineage sorting of C. micholitzii; secondly hybridization/introgression of sympatrically cycads, which would be supported by evidence base on nrDNA ITS sequences; thirdly intramolecular recombination in cpDNA of cycads; eventually the neotectonic movement in the distribution range of this 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