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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 parent.","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.indexedby_filter&advanced=false&query1=Molecular%2BMarker&&fq=dc.project.title_filter%3ACamellia%5C+taliensis%5C+%5C%28W.%5C+W.%5C+Smith%5C%29%5C+Melchior%2C%5C+a%5C+member%5C+of%5C+Camellia%5C+sect.%5C+Thea%2C%5C+is%5C+an%5C+indigenous%5C+species%5C+in%5C+local%5C+natural%5C+forest%5C+and%5C+has%5C+a%5C+long%5C+cultivative%5C+history%5C+in%5C+western%5C+Yunnan%5C+and%5C+its%5C+neighborhood%2C%5C+where%5C+the%5C+domestications%5C+of%5C+this%5C+species%5C+in%5C+different%5C+historical%5C+periods%5C+and%5C+in%5C+different%5C+ways%5C+can%5C+be%5C+found.%5C+C.%5C+taliensis%5C+is%5C+an%5C+important%5C+contributor%5C+to%5C+the%5C+formations%5C+of%5C+tea%5C+landraces%5C+by%5C+hybridization%5C+and%5C+introgression.%5C+In%5C+the%5C+present%5C+study%2C%5C+14%5C+microsatellite%5C+loci%5C+screened%5C+from%5C+37%5C+loci%5C+were%5C+used%5C+to%5C+explore%5C+the%5C+genetic%5C+diversity%5C+about%5C+this%5C+species%5C+with%5C+579%5C+samples%5C+from%5C+25%5C+populations%5C+%5C%2816%5C+wild%5C+populations%2C%5C+4%5C+semi%5C-cultivated%5C+populations%5C+and%5C+5%5C+cultivated%5C+populations%5C%29.%5C+At%5C+the%5C+same%5C+time%2C%5C+the%5C+potential%5C+hybrid%5C+speciation%5C+of%5C+C.%5C+grandibracteata%2C%5C+was%5C+investigated%5C+using%5C+39%5C+individuals%5C+from%5C+2%5C+populations%2C%5C+along%5C+with%5C+C.%5C+taliensis%5C+and%5C+C.%5C+sinensis%5C+var.%5C+assamica%5C+%5C%2883%5C+individuals%5C+from%5C+4%5C+populations%5C%29%5C+by%5C+the%5C+same%5C+microsatellite%5C+markers.%5C+C.%5C+taliensis%5C+had%5C+a%5C+moderate%5C+high%5C+level%5C+of%5C+genetic%5C+diversity%5C+%5C%28A%5C+%3D%5C+14.3%2C%5C+Ne%3D%5C+5.7%2C%5C+HE%5C+%3D%5C+0.666%2C%5C+I%5C+%3D%5C+1.753%2C%5C+AR%5C+%3D%5C+7.2%2C%5C+PPB%5C+%3D%5C+100%25%5C%29.%5C+This%5C+may%5C+result%5C+from%5C+several%5C+factors%5C+including%5C+K%5C-strategy%2C%5C+genetic%5C+background%2C%5C+gene%5C+flow%5C+between%5C+populations%2C%5C+hybridization%5C+and%5C+introgression%5C+among%5C+species.%5C+Between%5C+wild%5C+populations%5C+of%5C+C.%5C+taliensis%2C%5C+the%5C+gene%5C+flow%5C+was%5C+moderate%5C+high%5C+%5C%28Nm%5C+%3D%5C+1.197%5C%29%2C%5C+and%5C+genetic%5C+variation%5C+was%5C+less%5C+than%5C+20%25%5C+%5C%28GST%5C+%3D%5C+0.147%2C%5C+FST%5C+%3D%5C+0.173%5C%29%2C%5C+which%5C+was%5C+similar%5C+to%5C+other%5C+research%5C+results%5C+of%5C+long%5C-lived%5C+woody%5C+plants%2C%5C+and%5C+reflected%5C+the%5C+genetic%5C+structure%5C+of%5C+its%5C+ancestry%5C+to%5C+same%5C+extent.%5C+There%5C+was%5C+a%5C+high%5C+significant%5C+correlation%5C+between%5C+geographic%5C+distance%5C+and%5C+Nei%E2%80%99s%5C+genetic%5C+distance%5C+%5C%28r%5C+%3D%5C+0.372%2C%5C+P%5C+%3D%5C+0.001%5C%29%5C+of%5C+populations%2C%5C+which%5C+accorded%5C+with%5C+isolation%5C+by%5C+distance%5C+model.%5C+Inferring%5C+from%5C+Bayesian%5C+clustering%5C+of%5C+genotypes%2C%5C+all%5C+individuals%5C+of%5C+C.%5C+taliensis%5C+were%5C+divided%5C+into%5C+two%5C+groups%2C%5C+conflicting%5C+with%5C+the%5C+result%5C+based%5C+on%5C+Nei%E2%80%99s%5C+genetic%5C+distance%5C+and%5C+real%5C+geographic%5C+distribution%2C%5C+which%5C+suggested%5C+there%5C+were%5C+heavy%5C+and%5C+non%5C-random%5C+influences%5C+by%5C+human%5C+practices.%5C+According%5C+to%5C+allelic%5C+richness%2C%5C+there%5C+were%5C+no%5C+significant%5C+differences%5C+%5C%28P%5C+%3E%5C+0.05%5C%29%5C+between%5C+wild%5C+%5C%28AR%5C+%3D%5C+4.651%5C%29%2C%5C+semi%5C-cultivated%5C+%5C%28AR%5C+%3D%5C+5.091%5C%29%5C+and%5C+cultivated%5C+%5C%28AR%5C+%3D%5C+5.132%5C%29%5C+populations%5C+of%5C+C.%5C+taliensis%2C%5C+which%5C+suggested%5C+that%5C+the%5C+genetic%5C+background%5C+of%5C+long%5C-lived%5C+woody%5C+plant%5C+was%5C+not%5C+easy%5C+to%5C+be%5C+changed%2C%5C+and%5C+there%5C+were%5C+moderate%5C+high%5C+gene%5C+flow%5C+between%5C+populations.%5C+However%2C%5C+there%5C+was%5C+a%5C+significant%5C+difference%5C+%5C%28P%5C+%3C%5C+0.05%5C%29%5C+between%5C+wild%5C+%5C%28AR%5C+%3D%5C+5.9%5C%29%5C+and%5C+cultivated%5C+%5C%28AR%5C+%3D%5C+7.1%5C%29%5C+populations%5C+distributed%5C+in%5C+the%5C+same%5C+place%5C+in%5C+Yun%5C+county%2C%5C+Yunnan%5C+province%2C%5C+which%5C+may%5C+result%5C+from%5C+the%5C+hybridization%5C+and%5C+introgression%5C+of%5C+species%5C+in%5C+the%5C+tea%5C+garden%5C+and%5C+anthropogenic%5C+damages%5C+to%5C+the%5C+wild%5C+population.%5C+The%5C+hypothesis%5C+of%5C+hybrid%5C+origin%5C+of%5C+C.%5C+grandibracteata%5C+was%5C+tested%5C+by%5C+morphological%5C+and%5C+microsatellites%5C+analyses.%5C+Compared%5C+with%5C+other%5C+species%2C%5C+the%5C+locules%5C+in%5C+ovary%5C+of%5C+C.%5C+grandibracteata%5C+are%5C+variable%2C%5C+which%5C+showed%5C+a%5C+morphological%5C+intermediate%5C+and%5C+mosaic.%5C+Except%5C+one%5C+private%5C+allele%2C%5C+Ninety%5C-nine%5C+percent%5C+alleles%5C+of%5C+C.%5C+grandibracteata%5C+were%5C+shared%5C+with%5C+these%5C+of%5C+C.%5C+taliensis%5C+and%5C+C.%5C+sinensis%5C+var.%5C+assamica.%5C+And%5C+C.%5C+grandibracteata%5C+was%5C+nested%5C+in%5C+the%5C+cluster%5C+of%5C+C.%5C+taliensis%5C+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 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.indexedby_filter&advanced=false&query1=Molecular%2BMarker&&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":"China Postdoctoral Science Foundation","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.indexedby_filter&advanced=false&query1=Molecular%2BMarker&&fq=dc.project.title_filter%3AChina%5C+Postdoctoral%5C+Science%5C+Foundation"},{"jsname":"China Scholarship Council","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.indexedby_filter&advanced=false&query1=Molecular%2BMarker&&fq=dc.project.title_filter%3AChina%5C+Scholarship%5C+Council"},{"jsname":"China Scholarship Council[201504910423]","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.indexedby_filter&advanced=false&query1=Molecular%2BMarker&&fq=dc.project.title_filter%3AChina%5C+Scholarship%5C+Council%5C%5B201504910423%5C%5D"},{"jsname":"Chinese Academy of Medical Sciences Central Public-interest Scientific Institution Basal Research Fund[2016ZX310196]","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.indexedby_filter&advanced=false&query1=Molecular%2BMarker&&fq=dc.project.title_filter%3AChinese%5C+Academy%5C+of%5C+Medical%5C+Sciences%5C+Central%5C+Public%5C-interest%5C+Scientific%5C+Institution%5C+Basal%5C+Research%5C+Fund%5C%5B2016ZX310196%5C%5D"},{"jsname":"Chinese Academy of Sciences[2013T2S003]","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.indexedby_filter&advanced=false&query1=Molecular%2BMarker&&fq=dc.project.title_filter%3AChinese%5C+Academy%5C+of%5C+Sciences%5C%5B2013T2S003%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.indexedby_filter&advanced=false&query1=Molecular%2BMarker&&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":"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 species.","jscount":"1","jsurl":"/simple-search?field1=all&field=dc.citation.indexedby_filter&advanced=false&query1=Molecular%2BMarker&&fq=dc.project.title_filter%3ACycas%5C+micholitzii%5C+complex%5C+is%5C+composed%5C+of%5C+5%5C+species%5C%3A%5C+C.%5C+micholitzii%5C+Dyer%2C%5C+C.%5C+bifida%5C+%5C%28Dyer%5C%29%5C+K.%5C+D.%5C+Hill%2CC.%5C+longipetiolula%5C+D.%5C+Y.%5C+Wang%2C%5C+C.%5C+debaoensis%5C+Y.%5C+C.%5C+Zhong%5C+et%5C+C%5C+J.%5C+Chen%2C%5C+C.%5C+multipinnata%5C+C%5C+J.%5C+Chen%5C+et%5C+S.%5C+Y.%5C+Yang%EF%BC%8Cand%5C+distributed%5C+from%5C+southwest%5C+China%5C+to%5C+central%5C+Vietnam%5C+and%5C+eastern%5C+Laos.%5C+Based%5C+on%5C+sequence%5C+data%5C+from%5C+two%5C+maternally%5C+inherited%5C+cpDNA%5C+and%5C+one%5C+biparentally%5C+nuclear%5C+DNA%5C+fragments%2C%5C+our%5C+study%5C+revealed%5C+the%5C+population%5C+genetic%5C+structure%5C+of%5C+C.%5C+micholitzii%5C+complex%5C+and%5C+explored%5C+the%5C+potential%5C+causes.%5C+The%5C+evolutionary%5C+and%5C+demographic%5C+histories%5C+were%5C+investigated.%5C+The%5C+genetic%5C+relationship%5C+among%5C+species%5C+in%5C+the%5C+complex%5C+was%5C+also%5C+clarified.The%5C+results%5C+were%5C+summarized%5C+as%5C+follows%5C%3A%5C+1.%5C+Phylogeographic%5C+analysis%5C+based%5C+on%5C+chloroplast%5C+sequences%EF%BC%8CWe%5C+examined%5C+chloroplast%5C+sequence%5C+variation%5C+of%5C+the%5C+atpB%5C-rbcLand%5C+psbA%5C-trnHintergenic%5C+spacers%5C+in%5C+27%5C+populations%5C+of%5C+C.%5C+micholitzii%5C+complex%2C%5C+recovering%5C+26%5C+haplotypes.%5C+The%5C+average%5C+within%5C-population%5C+diversity%5C+%5C%28HS%5C+%3D%5C+0.140%5C%29%5C+was%5C+low%5C+while%5C+total%5C+diversity%5C+%5C%28HT%5C+%3D%5C+0.911%5C%29%5C+was%5C+high.%5C+Population%5C+differentiation%5C+was%5C+also%5C+high%5C%28GST%5C+%3D%5C+0.846%2C%5C+NST%5C+%3D%5C+0.919%5C%29%2C%5C+indicating%5C+significant%5C+phylogeographical%5C+structure%5C+%5C%28NST%5C+%3E%5C+GST%2Cp%5C+%3C%5C+0.001%5C%29%5C+and%5C+low%5C+levels%5C+of%5C+seed%5C-based%5C+gene%5C+flow.%5C+C.%5C+debaoensis%5C+%5C%28Cycadaceae%5C%29%5C+is%5C+an%5C+endangered%5C+species%5C+restricted%5C+to%5C+the%5C+border%5C+of%5C+Guangxi%5C+and%5C+Yunnan%5C+province%5C+in%5C+southwest%5C+China.%5C+This%5C+species%5C+has%5C+been%5C+classified%5C+into%5C+two%5C+types%5C%3A%5C+sand%5C+and%5C+karst%2C%5C+according%5C+to%5C+the%5C+soil%5C+matrix%5C+they%5C+grow%5C+on.%5C+We%5C+examined%5C+chloroplast%5C+sequence%5C+variation%5C+of%5C+the%5C+cpDNA%5C+sequences%5C+from%5C+11%5C+populations%5C+of%5C+this%5C+species.%5C+Significant%5C+population%5C+genetic%5C+differentiation%5C+was%5C+detected%5C+%5C%28GST%3D%5C+0.684%5C+and%5C+FST%5C+%3D%5C+0.74160%5C%29.%5C+There%5C+was%5C+marked%5C+genetic%5C+differentiation%5C+between%5C+populations%5C+in%5C+the%5C+sand%5C+and%5C+karst%5C+regions%5C+and%5C+no%5C+expansion%5C+was%5C+detected.%5C+Climate%5C+changes%5C+during%5C+glacial%5C+periods%5C+have%5C+had%5C+significant%5C+effects%5C+on%5C+the%5C+current%5C+distribution%5C+of%5C+cycads.%5C+The%5C+molecular%5C+phylogenetic%5C+data%2C%5C+together%5C+with%5C+the%5C+geographic%5C+distribution%5C+of%5C+the%5C+haplotypes%2C%5C+suggest%5C+that%5C+C.%5C+debaoensis%5C+experienced%5C+range%5C+contraction%5C+during%5C+glacial%5C+periods%2C%5C+and%5C+that%5C+the%5C+current%5C+populations%5C+are%5C+still%5C+confined%5C+to%5C+the%5C+original%5C+refugia%5C+in%5C+southwest%5C+China%5C+which%5C+have%5C+favorable%5C+habitats%5C+in%5C+glacial%5C+period.%5C+These%5C+results%5C+imply%5C+that%5C+small%5C+refugia%5C+were%5C+maintained%5C+in%5C+both%5C+sand%5C+and%5C+karst%5C+regions%5C+during%5C+the%5C+LGM%5C+%5C%28last%5C+glacial%5C+maximum%5C%29.%5C+This%5C+species%5C+had%5C+no%5C+postglacial%5C+recolonization%5C+and%5C+only%5C+stayed%5C+in%5C+these%5C+refugia%5C+up%5C+to%5C+now.%5C+The%5C+low%5C+within%5C-population%5C+diversity%5C+of%5C+C.%5C+debaoensis%5C+suggests%5C+that%5C+there%5C+were%5C+strong%5C+bottleneck%5C+events%5C+or%5C+founder%5C+effects%5C+within%5C+each%5C+separate%5C+region%5C+during%5C+the%5C+Quaternary%5C+climatic%5C+oscillations.%5C+Relatively%5C+high%5C+genetic%5C+and%5C+haplotype%5C+diversities%5C+were%5C+detected%5C+in%5C+the%5C+newly%5C+discovered%5C+populations%2C%5C+which%5C+located%5C+at%5C+intermediate%5C+locality%5C+of%5C+sand%5C+regions%5C+and%5C+had%5C+morphological%5C+variation%5C%3B%5C+this%5C+is%5C+probably%5C+the%5C+consequence%5C+of%5C+the%5C+admixture%5C+of%5C+different%5C+haplotypes%5C+colonizing%5C+the%5C+area%5C+from%5C+separate%5C+sources.%5C+%5C+C.%5C+micholitzii%5C+occurs%5C+in%5C+the%5C+Annan%5C+Highlands%5C+in%5C+central%5C+Vietnam%5C+near%5C+the%5C+Laos%5C+border.%5C+C.%5C+bifida%5C+occurs%5C+in%5C+North%5C+Vietnam%5C%3B%5C+its%5C+distribution%5C+extends%5C+across%5C+the%5C+border%5C+into%5C+adjacent%5C+localities%5C+in%5C+Guangxi%5C+and%5C+Yunnan%5C+in%5C+China.%5C+For%5C+the%5C+comparability%5C+between%5C+them%2Ctheywere%5C+considered%5C+as%5C+the%5C+same%5C+species%5C+C.%5C+micholitzii%5C+by%5C+many%5C+academicians.%5C+The%5C+cpDNA%5C+sequences%5C+from%5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scent is a very important character in rose breeding. However, many of 25,000 rose cultivars have no scent or weak scent. The tea scent of modern roses mainly originated from Rosa odorata (Andrews) Sweet, which is one of the most important ancestors of modern cultivated roses and the very important rose breeding resource. Due to the land expanding, habitat fragmentation and so on, R. odorata has been listed as an endangered species in ‘Chinese Plant Red Data Book—Rare and Endangered Plants’ and as the third-category endangered species in ‘Chinese Rare and Endangered Protective Plants List’. Therefore, it is urgent to protect this species and studying the conservation genetics of R. odorata is essentially important to work out a strategy of conservation.R. odorata comprises three double-petaled varieties (R. odorata var. odorata, R. odorata var. erubescens, and R. odorata var. pseudindica) and one single-petaled variety (R. odorata var. gigantea). The taxonomy of the three double-petaled varieties of R. odorata has been disputed for a long time. They have been treated as intraspecific taxa of R. odorata var. gigantea or R. chinensis by different botanist. According to the morphological analyses, Hurst (1941) inferred that R. odorata var. odorata was the hybrid between R. odorata var. gigantea and R. chinensis. Therefore, in order to clarify the right protective units, two single-copy nuclear genes (GAPDH and ncpGS), together with two plastid loci (trnL-F and psbA-trnH) were applied to study the hybrid origin of the three double-petaled varieties and to identify their possible parents. Our data suggested the hybrid origin of the three double-petaled varieties. We inferred that R. odorata var. gigantea could be the maternal parent and R. chinensis cultivars be the paternal parent. It is strongly suggested that the conservation of R. odorata is the conservation of its wild type, R. odorata var. gigantea. We first applied seven microsatellite loci (SSR) coupled with a single-copy nuclear gene GAPDH to study the genetic diversity and genetic structure of R. odorata var. gigantea. The main results are shown as follows:1. Genetic diversity：R. odorata var. gigantea maintains high degree of genetic diversity within and among populations (SSR: HT = 0.738, HS = 0.569, AR = 5.583, PPB = 97.35%, I = 1.703; GAPDH: HT = 0.739, HS = 0.540). We inferred that, outcrossing, long-lived tree species, clonal reproduction and general intraspecies hybridization between individuals, have contributed to the high degree of genetic diversity in R. odorata var. gigantea.2. Genetic differentiation and genetic structure：There was some degree of genetic differentiation among populations (SSR: GST = 0.229, FST = 0.240; GAPDH: GST = 0.269). The geographic isolation limited the dispersal of pollen or seeds, which resulted in the limitation of gene flow (Nm = 0.792). Then, the limited gene flow should be accounted for the genetic differentiation. Both the results of SSR data and haplotype analysis of GAPDH indicated that, the studied populations were divided into two distinct groups by Honghe River. These two groups showed significant genetic differentiation and represented two separate evolutionary lineages, which should be recognized as two evolutionary significant units (ESUs) for conservation concerns.3. Conservation of R. odorata：R. odorata var. gigantea has been listed in the ‘National Key Protective Wild Species List (II)’. Therefore, the conservation of this species is urgent. We inferred that, the main endangered reasons should be the habitat fragmentation and the reduction of populations and individuals per population resulted from environmental damage and human activities. We proposed that the strategy of in-situ conservation combining with ex-situ conservation should be carried 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the rapid uplift of the Himalaya, the reorganization of the major river drainages was primarily caused by river capture events，e.g. those of the Jinshajiang River (comprising the Upper, Middle and Lower Jinshajiang) and its tributaries (Yalongjiang, Daduhe, Jialingjiang), the Nujiang, the Lancangjiang, and the Honghe. We selected Terminalia franchetii var. franchetii and T. franchetii var. intricata in the Sino-Himalayan region to study the relationship with Honghe diversion events. The distribution of this species is predicted to have retained genetic signatures of past hydrological landscape structures. The major result as flowing:1. Chloroplast phylogeography of T. franchetii based on haplotype analysis，Based on a range-wide sampling comprising 28 populations and 258 individuals, and using chloroplast DNA sequences (trnL-trnF, petL-psbE), we detected 12 haplotypes. Terminalia franchetii was found to harbour high haplotype diversity (hT = 0.784) but low average within-population diversity (hS = 0.124). The analysis of genetic structure using SAMOVA showed that the number of population groups equaled five, and all the haplotypes can be divided into five groups. Group B and C identified exhibited a disjunctive distribution of dominant haplotypes between northern and southern valleys, corresponding to the geography of past rather than modern drainage systems.Mismatch distribution (multimodal curve) and neutral tests provided no evidence of recent demographic population growth. We suggest that the modern disjunctive distribution of T. franchetii, and associated patterns of cpDNA haplotype variation, result from vicariance caused by several historical river separation and capture events. By assuming a common mutation rate of the cpDNA-IGS regions, our inferred timings of these events (0.82-4.39 Mya) broadly agrees with both previous geological and molecular estimated time of drainage rearrangements in this region. So we conclude that there were several historical vicariance events play a major role for the distribution of T. franchetii in this region.2. Genetic diversity and structure of T. franchetii var. franchetii based on AFLP analysis，We determined the genotype of 251 individuals of T. franchetii var. franchetii from 21 populations using amplified fragment length polymorphism (AFLP), for our aim is only investigated the relationship between the modern distribution of T. franchetii and geological changes in drainage patterns. The overall estimate of genetic structure (Gst) was 0.249, indicating that clear genetic differentiation existed among the populations. Estimates of gene flow (Nm = 0.754) between populations based on the Gst value revealed that the number of migrants per generation is not frequently.Using Neighbor-Joining tree, Principal Coordinates Analysis, STRUCTURE and network methods, Analyses of AFLP markers identified two main population groups (I and II) and four subgroups (A – D) of T. franchetii. Genetic diversity was lower in Group I than in Group II. The results show that Groups I and II probably once occupied continuous areas respectively along ancient drainage systems and there were several historical separation and capture events that can account for the distribution of T. franchetii in this region. After all，these are good examples of the way in which historical events can change a species’ distribution from continuous to fragmented (Jinshajiang/ Yalongjiang and Honghe), and a disjunct distribution to a continuous one (Upper/Lower Jinshajiang and Yalongjiang). The results provide new insights into the phylogeographic pattern of plants in southwest China.3. Relationships between T. franchetii var. franchetii and T. franchetii var. intricata ，While T. franchetii var. Franchetii and var. intricata slightly differ in overall size and leaf hairiness, these taxa did not exhibit reciprocal monophyly. As results show, the genetic difference between the two varieties is much smaller than that within var. franchetii (Salween population vs. other populationsof this variety). It is also revealed in a phylogenetic analysis of ITS region of Combretoideae. The habitats of var. franchetii and var. intricata have obviously difference. Thus, the differences between the two varieties in overall size and leaf hairiness might reflect different phenotypic responses to environmental changes and the divergent environmental niche spaces they occupy. Based on the reasoning above, we agree with Flora of China that “T. intricata” represents a variety of T. franchetii rather than a separate 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