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资助项目
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=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&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%5C+11%5C+populatio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a field trip at a brule in Shangri-La, a mixed population of Ligularia Cass. was found, which including L. subspicata (Bur. et Franch.) Hand.-Mazz., L. nelumbifolia (Bur. et Franch.) Hand.-Mazz., L. tongolensis (Franch.) Hand.-Mazz., L. cymbulifera (W.W.Smith) Hand.-Mazz., L. lingiana S.W.Liu, and also some individuals morphologically intermediate between L. subspicata and L. nelumbifolia. Hence, these intermediate individuals were preliminarily assumed as natural hybrids of the two Ligularia. According to their morphology, they’re assumed to form hybrids A and B. Through careful comparison of specimens in herbarium and those we collected, the inflorescence of putative hybrid A is close to L. nelumbifolia, but the shape of laminae are intergradation of L. subspicata and L. nelumbifolia; overall morphology of putative hybrids B is similar to L. nelumbifolia, but inflorescence color is as same as L. subspicata. Compared to L. nelumbifolia (39%) and L. subspicata (36.8%), the germination rate of putative hybrid B (45.7%) slightly higher than the two; but that of hybrid A is extraordinarily low (0.3%). One possible interpretation of the low rate is hybridization. 60 individuals were collected, including putative parents, other 4 species of Ligularia nearby, putative hybrid A and B. They were all direct sequenced of four cpDNA fragments, and direct sequenced or cloning sequenced of nrDNA ITS4-5. The results support that L. nelumbifolia and L. subspicata are parents of putative hybrid A, and the majority female parent is L. subspicata, L. vellerea may also be involved in the hybridization in some degree; the nuclear sequences of putative hybrid B have no superposition, and its chloroplast DNA sequences are identical with L. nelumbifolia, so putative hybrid B could not be hybrid; and there are backcross individuals exist among the putative parent L. subspicata. NewHybrids analysis of ISSR markers indicated that, the individuals of putative hybrid A are almost L. nelumbifolia and L. subspicata F1 hybrid generation (10/11), only 1/11 possibly backcross or other forms; all individuals of hybrid B are L. nelumbifolia; except one individual of L. subspicata as backcrossed, the other parent individuals are 100% reliable. This study focused on molecular evidence, complemented by ecological, reproductive and other characteristics, we demonstrated that the morphologically intermediate individuals’ origin, and the probability of belonging to each parental or hybrid class. And concluded that L. nelumbifolia and L. subspicata are the parents of putative hybrid A, L. vellerea may also be involved in the hybridization in some degree, hybrids mainly are the first generation, a few individuals may be involved in backcross, and most probably backcross with L. subspicata according to the anthesis, while the assumption of hybrid B is not supported.","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3ADuring%5C+a%5C+field%5C+trip%5C+at%5C+a%5C+brule%5C+in%5C+Shangri%5C-La%2C%5C+a%5C+mixed%5C+population%5C+of%5C+Ligularia%5C+Cass.%5C+was%5C+found%2C%5C+which%5C+including%5C+L.%5C+subspicata%5C+%5C%28Bur.%5C+et%5C+Franch.%5C%29%5C+Hand.%5C-Mazz.%2C%5C+L.%5C+nelumbifolia%5C+%5C%28Bur.%5C+et%5C+Franch.%5C%29%5C+Hand.%5C-Mazz.%2C%5C+L.%5C+tongolensis%5C+%5C%28Franch.%5C%29%5C+Hand.%5C-Mazz.%2C%5C+L.%5C+cymbulifera%5C+%5C%28W.W.Smith%5C%29%5C+Hand.%5C-Mazz.%2C%5C+L.%5C+lingiana%5C+S.W.Liu%2C%5C+and%5C+also%5C+some%5C+individuals%5C+morphologically%5C+intermediate%5C+between%5C+L.%5C+subspicata%5C+and%5C+L.%5C+nelumbifolia.%5C+Hence%2C%5C+these%5C+intermediate%5C+individuals%5C+were%5C+preliminarily%5C+assumed%5C+as%5C+natural%5C+hybrids%5C+of%5C+the%5C+two%5C+Ligularia.%5C+According%5C+to%5C+their%5C+morphology%2C%5C+they%E2%80%99re%5C+assumed%5C+to%5C+form%5C+hybrids%5C+A%5C+and%5C+B.%5C+Through%5C+careful%5C+comparison%5C+of%5C+specimens%5C+in%5C+herbarium%5C+and%5C+those%5C+we%5C+collected%2C%5C+the%5C+inflorescence%5C+of%5C+putative%5C+hybrid%5C+A%5C+is%5C+close%5C+to%5C+L.%5C+nelumbifolia%2C%5C+but%5C+the%5C+shape%5C+of%5C+laminae%5C+are%5C+intergradation%C2%A0of%5C+L.%5C+subspicata%5C+and%5C+L.%5C+nelumbifolia%5C%3B%5C+overall%5C+morphology%5C+of%5C+putative%5C+hybrids%5C+B%5C+is%5C+similar%5C+to%5C+L.%5C+nelumbifolia%2C%5C+but%5C+inflorescence%5C+color%5C+is%5C+as%5C+same%5C+as%5C+L.%5C+subspicata.%5C+Compared%5C+to%5C+L.%5C+nelumbifolia%5C+%5C%2839%25%5C%29%5C+and%5C+L.%5C+subspicata%5C+%5C%2836.8%25%5C%29%2C%5C+the%5C+germination%5C+rate%5C+of%5C+putative%5C+hybrid%5C+B%5C+%5C%2845.7%25%5C%29%5C+slightly%5C+higher%5C+than%5C+the%5C+two%5C%3B%5C+but%5C+that%5C+of%5C+hybrid%5C+A%5C+is%5C+extraordinarily%5C+low%5C+%5C%280.3%25%5C%29.%5C+One%5C+possible%5C+interpretation%5C+of%5C+the%5C+low%5C+rate%5C+is%5C+hybridization.%5C+60%5C+individuals%5C+were%5C+collected%2C%5C+including%5C+putative%5C+parents%2C%5C+other%5C+4%5C+species%5C+of%5C+Ligularia%5C+nearby%2C%5C+putative%5C+hybrid%5C+A%5C+and%5C+B.%5C+They%5C+were%5C+all%5C+direct%5C+sequenced%5C+of%5C+four%5C+cpDNA%5C+fragments%2C%5C+and%5C+direct%5C+sequenced%5C+or%5C+cloning%5C+sequenced%5C+of%5C+nrDNA%5C+ITS4%5C-5.%5C+The%5C+results%5C+support%5C+that%5C+L.%5C+nelumbifolia%5C+and%5C+L.%5C+subspicata%5C+are%5C+parents%5C+of%5C+putative%5C+hybrid%5C+A%2C%5C+and%5C+the%5C+majority%5C+female%5C+parent%5C+is%5C+L.%5C+subspicata%2C%5C+L.%5C+vellerea%5C+may%5C+also%5C+be%5C+involved%5C+in%5C+the%5C+hybridization%5C+in%5C+some%5C+degree%5C%3B%5C+the%5C+nuclear%5C+sequences%5C+of%5C+putative%5C+hybrid%5C+B%5C+have%5C+no%5C+superposition%2C%5C+and%5C+its%5C+chloroplast%5C+DNA%5C+sequences%5C+are%5C+identical%5C+with%5C+L.%5C+nelumbifolia%2C%5C+so%5C+putative%5C+hybrid%5C+B%5C+could%5C+not%5C+be%5C+hybrid%5C%3B%5C+and%5C+there%5C+are%5C+backcross%5C+individuals%5C+exist%5C+among%5C+the%5C+putative%5C+parent%5C+L.%5C+subspicata.%5C+NewHybrids%5C+analysis%5C+of%5C+ISSR%5C+markers%5C+indicated%5C+that%2C%5C+the%5C+individuals%5C+of%5C+putative%5C+hybrid%5C+A%5C+are%5C+almost%5C+L.%5C+nelumbifolia%5C+and%5C+L.%5C+subspicata%5C+F1%5C+hybrid%5C+generation%5C+%5C%2810%5C%2F11%5C%29%2C%5C+only%5C+1%5C%2F11%5C+possibly%5C+backcross%5C+or%5C+other%5C+forms%5C%3B%5C+all%5C+individuals%5C+of%5C+hybrid%5C+B%5C+are%5C+L.%5C+nelumbifolia%5C%3B%5C+except%5C+one%5C+individual%5C+of%5C+L.%5C+subspicata%5C+as%5C+backcrossed%2C%5C+the%5C+other%5C+parent%5C+individuals%5C+are%5C+100%25%5C+reliable.%5C+This%5C+study%5C+focused%5C+on%5C+molecular%5C+evidence%2C%5C+complemented%5C+by%5C+ecological%2C%5C+reproductive%5C+and%5C+other%5C+characteristics%2C%5C+we%5C+demonstrated%5C+that%5C+the%5C+morphologically%5C+intermediate%5C+individuals%E2%80%99%5C+origin%2C%5C+and%5C+the%5C+probability%5C+of%5C+belonging%5C+to%5C+each%5C+parental%5C+or%5C+hybrid%5C+class.%5C+And%5C+concluded%5C+that%5C+L.%5C+nelumbifolia%5C+and%5C+L.%5C+subspicata%5C+are%5C+the%5C+parents%5C+of%5C+putative%5C+hybrid%5C+A%2C%5C+L.%5C+vellerea%5C+may%5C+also%5C+be%5C+involved%5C+in%5C+the%5C+hybridization%5C+in%5C+some%5C+degree%2C%5C+hybrids%5C+mainly%5C+are%5C+the%5C+first%5C+generation%2C%5C+a%5C+few%5C+individuals%5C+may%5C+be%5C+involved%5C+in%5C+backcross%2C%5C+and%5C+most%5C+probably%5C+backcross%5C+with%5C+L.%5C+subspicata%5C+according%5C+to%5C+the%5C+anthesis%2C%5C+while%5C+the%5C+assumption%5C+of%5C+hybrid%5C+B%5C+is%5C+not%5C+supported."},{"jsname":"Kunming Institute of Botany[KIB2017003]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3AKunming%5C+Institute%5C+of%5C+Botany%5C%5BKIB2017003%5C%5D"},{"jsname":"Ministry of Science and Technology, China[2013FY112600]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3AMinistry%5C+of%5C+Science%5C+and%5C+Technology%2C%5C+China%5C%5B2013FY112600%5C%5D"},{"jsname":"National Key Basic Research Program of China[2014CB954100]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3ANational%5C+Key%5C+Basic%5C+Research%5C+Program%5C+of%5C+China%5C%5B2014CB954100%5C%5D"},{"jsname":"National Natural Science Foundation of China[31200182]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3ANational%5C+Natural%5C+Science%5C+Foundation%5C+of%5C+China%5C%5B31200182%5C%5D"},{"jsname":"National Natural Science Foundation of China[31370252]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3ANational%5C+Natural%5C+Science%5C+Foundation%5C+of%5C+China%5C%5B31370252%5C%5D"},{"jsname":"National Natural Science Foundation of China[31470336]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3ANational%5C+Natural%5C+Science%5C+Foundation%5C+of%5C+China%5C%5B31470336%5C%5D"},{"jsname":"National Natural Science Foundation of China[31600178]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3ANational%5C+Natural%5C+Science%5C+Foundation%5C+of%5C+China%5C%5B31600178%5C%5D"},{"jsname":"National Natural Science Foundation of China[41571059]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3ANational%5C+Natural%5C+Science%5C+Foundation%5C+of%5C+China%5C%5B41571059%5C%5D"},{"jsname":"National Research Council of Thailand (Mae Fah Luang University)[592010200112]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3ANational%5C+Research%5C+Council%5C+of%5C+Thailand%5C+%5C%28Mae%5C+Fah%5C+Luang%5C+University%5C%29%5C%5B592010200112%5C%5D"},{"jsname":"National Research Council of Thailand (Mae Fah Luang University)[60201000201]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3ANational%5C+Research%5C+Council%5C+of%5C+Thailand%5C+%5C%28Mae%5C+Fah%5C+Luang%5C+University%5C%29%5C%5B60201000201%5C%5D"},{"jsname":"National Science Foundation of China (NSFC)[31750110478]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3ANational%5C+Science%5C+Foundation%5C+of%5C+China%5C+%5C%28NSFC%5C%29%5C%5B31750110478%5C%5D"},{"jsname":"National Science Foundation of China[31470336]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3ANational%5C+Science%5C+Foundation%5C+of%5C+China%5C%5B31470336%5C%5D"},{"jsname":"National Science Foundation of China[31600178]","jscount":"1","jsurl":"/simple-search?field1=all&field=location.comm.id&advanced=false&query1=Hybrid%2Bzone&&fq=dc.project.title_filter%3ANational%5C+Science%5C+Foundation%5C+of%5C+China%5C%5B31600178%5C%5D"},{"jsname":"Natural hybridization has many evolutionary consequences, such as the origin of new ecotypes or species and the maintenance of reproductive barriers. However, few studies have involved in and quantified the potential threat to rare species by natural hybridization. In this study, we focused on the hybridization between a narrow endemic species R. cyanocarpum Franch. ex W.W. Sm and the widespread species R. delavayi Franch.. We revealed this process of natural hybridization, composition and possible significance of the detected single hybrid zone and comparative analysis of fitness among parental species and the hybrids. These significant results were summarized as follows: 1. Field observation and comparative analysis of morphological characters, We investigated 6 sites where R. cyanocarpum distributed, and found the single putative hybrid zone only in one site. 22 morphological characters were selected and measured, according to which the putative parental species were determined. Among the examined morphological characters, intermediate characters accounted for 45%, whereas the transgressive characters accounted for 14%. These results showed that intermediate characters were predominant. In addition, the ecological adaptations of these transgressive characters remain unknown. 2. Reproductive biology and reproductive evidence for natural hybridization, Three main results were achieved via the study of pollination biology of R. cyanocarpum: (1) from the results of floral color experiments, we found that the pink flowers have preponderant adaptation over the white ones. Because either proportions of individuals with pink flowers within and between populations, or visitation rate of pollinators were all significant higher than white types; (2) the effective pollinators of R. cyanocarpum were solely the bumblebees (Bombus festivces and B. richardsi), and the pollination is primarily due to geitonogamy that is mediated by bumblebees; (3) most of fruits were either composed of 5 or 6 carpels. Moreover, the proportions of carpel number per fruit were significantly related to the altitudes ((r=0.807, p<0.01) and the positions of inflorescence (r=0.62, p<0.01). Meanwhile, seed weight, seed number and seed germination rate were significant difference to the fruits with different carpel number; For R. delavayi, the experiment showed that the associated function of nectary and spots promote reproductive success in R. delavayi, because of significantly higher fruit production was examined in half petal deprived with degenerative nectarines than in half petal deprived with normal ones. Reciprocal hand pollination treatments showed that either species, as pollen donor or pollen receiver, could produce fruits indicating weak reproductive barriers between species. It was noted that fruit set varied among treatments. The same pollinators (bumblebees) were shared in both parental species. Therefore, flowering period overlapping, weak reproductive barriers and sharing the same pollinators, all contributed to reproductive evidence for the possible occurrence of natural hybridization between R. cyanocarpum and R. delavayi. 3. Molecular evidence for natural hybridization, On the basis of two nuclear DNA (ITS and Waxy) and three chloroplast DNA (trnC-F, trnH-psbA and trnL-rpl32), hybrids and parental species were clearly identified and individuals with intermediate morphologies are indeed of hybrid origin from natural hybridization between R. cyanocarpum and R. delavayi. Hybridization occurred in both directions, but was asymmetrical, with R. delavayi as the major maternal parent (90%) whereas only one hybrid had identical chloroplast to R. cyanocarpum (10%). 4. The study of hybrid zone, Two molecular methods were employed to successfully disclosed characters of the single hybrid zone: (1) this hybrid zone was composed of parental species, F1s, F2s and backcross derivatives; (2) F2s outnumbered either F1s or backcross derivatives; (3) the direction of gene flow was from R. cyanocarpum to R. delavayi. Based on the one directional gene flow and fewer hybrids formation in the single hybrid zone, no threat to R. cyanocarpum posed by natural hybridization was observed. However, such a threat is likely to arise if more hybrids are produced via serious habitat disturbance. 5. Comparative analysis of fitness among parental species and hybrids, We made a preliminary result that the fitness of hybrids’ offspring is not always lower than their parental offsprings, after comparative analysis of some detected fitness index in experimental condition, such as the lengths of radicle and hypocotyls, and the seedling height. Therefore, it is reasonable to presume that these hybrids can have similar or higher fitness than parental species only if natural habitat is changed substantially in a short 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