Pollinator System And The Fitness Of Filial Generation In Different Controlled Pollination Of Sinocalycanthus Chinensis

The low genetic diversity of small population is attribute to the mating system,under which the control of passive inbreeding,and the pollinators play an important role.Study the effects of the floral traits of Sinocalycanthus chinensis on pollinators,combine with the research of offspring fitness under different mating methods(selfing,inbreeding,outbreeding),to provide theoretical basis for rejuvenation populations of Sinocalycanthus chinensis.(1)Manual manipulation of floral tissues to alter the floral structures and traits of Sinocalycanthus chinensis(Cheng et S.Y.Chang)Cheng et S.Y.Chang in Damin mountain and C.floridus var.glaucus at an introduced location(Lin'an,Zhejiang,China)was performed to investigate the effects of the various floral traits on pollination.The results showed that the wild population of Sinocalycanthus chinensis,seven types of insects(Apis cerana,Ceratina sp.,Ferdinandus cupea,Bombus,Cotalpa lanigera,Disteniinae and Trongylium albopilosum)were observed to be the pollinators.In addition,five types of beetles(Carpophilus humeralis,Carpophilus marginellus,Pinophilus sp.,Formicomus sp.and Thaumaglossa sp.)were observed to be the pollinators of C.floridus var.glaucus under natural conditions.Both of them are pollinated by beetles(coleoptera).Bees are another major pollination groups of Sinocalycanthus chinensis.In C.floridus var.glaucus,The central cone formed by the inner petals interacts with the floral fragrance,which mainly originates in the succulent bodies at the tops of the stamens and staminodes,to determine pollinator behavior.Sinocalycanthus chinensis use the brilliant colors(yellow inner tepals impression foiled on the white outer tepals)to attract pollinators.(2)The result of controlled pollination ways among different populations of Sinocalycanthus chinensis showed that the fecundity and phenotype of filial generations by distant pollination(population in Tantai and Linan),had a more stronger fitness than the other population way.The genetic diversity and the mating system of the filial generation of different populations and different controlled pollination ways need to be analyzed subsequently.The filial generations by pollination in neighboring populations who's fruit phenotypes are larger and seed germination rate is higher.Inbreeding depression occurs to geitonogamy-pollination offspring on the germination rate and seedling growth potential.The natural pollinated progeny exhibited the same fitness to sympatric population and geitonogamy-pollination,thus speculated that the populations may mainly exist those two mating ways.The results show that the species can be protected by mixed pollination outbreeding in different populations.(3)To develop expressed sequence tag(EST)-simple sequence repeat(SSR)primers based on transcriptome sequences of Sinocalycanthus chinensis,26564 SSRs were detected from 125014 unigene sequences,which were assembled from transcriptome sequencing data.Among the 125014 unigene sequences,the average density is 5.18 kb.(AG/CT)noccupies the largest number of these di-nucleotiderepeats.The best SSR-PCR technique system of Sinocalycanthus chinensis was gained by L16(45)orthogonal experimental design: 20 ?L SSR-PCR systemcontaining 0.6 U Taq enzyme,Mg2+ 1.5 mmol·L-1,dNTP 0.25 mmol·L-1,Primer 0.2 ?mol·L-1,DNA 75 ng.15585 primer pairs that conform to the request are designed with all EST-SSR locus.Among these primer pairs,it is amplificated successfully with 120 primer pairs(54.55%)and polymorphism is amplificated with 14 primer pairs in different populations.The average allele number is 2.64 and polymorphism is amplificated with 7 primer pairs in the same population.These polymorphic primers can provide more abundant marks for the genetic analysis of Sinocalycanthus chinensis.