| Luculia yunnanensis Hu is a member of the genus Luculia Sweet,and belongs to the family Rubiaceae.It is a heterostyly species,which is endemic to Southwestern China.Fragrance,long flowering period and nice plant shape make L.yunnanensis a versatile floriculture crop.It is small trees or shrub and a kind of excellent wild ornamental plant which has been applied in garden landscape and potted plant.However,the related references and information about L.yunnanensis are scarce.Current research indicated the optimum conditions of germination of L.yunnanensis seeds and 21 microsatellite primers were developed to analysis the genetic diversity in L.yunnanensis.Based upon previous studies on Luculia,this study employed to explore the present resources of L.yunnanensis.An all-round analysis of the result of L.yunnanensis resources survey was made,objective to know the resources current situation and the threat level of L.yunnanensis.Analysis of genetic diversity aimed to explore the genetic structure.Analysis of volatile composition L.yunnanensis,L.pinceana and the hybrid's flowers,aimed to find special compounds in L.yunnanensis and make a base for breeding.The tactics of protective development were proposed based on the Analysis of the reason of vulnerable.The aim is to lay some foundation to cultivate new varieties and pave the way for protection and utilization of this rare plant.There are 7 main conclusions of this research:1.Resources survey results showed that L.yunnanensis currently exists only in Lu shui County,Fu gong County and Gong shan County in Nu jiang,Yunnan Province,the distribution range only has 10465.48 km2 area.Compared with flora of China and flora of Yunnan,the distribution range was shrink from 7 counties in 5 cities to 3 counties in 1 city.The area of the existing distribution range was only 20% of the historical record.The result of Threat Level Assessment showed that L.yunnanensis was vulnerable plant,need protection.2.The phenotypic characteristics of 6 L.yunnanensis populations were investigated.The results of variance analysis showed that there were significant differences among populationsand among individuals within populations,petals reflexed degree varied most.The correlations between distyly with some characters related flower were significant.The variation among populations was slightly higher than that within populations,which indicated that the variance within populations was the main part of the phenotypic variation of L.yunnanensis populations.Principal component analytical results indicated that anthotaxy diameter,flower number in an anthotaxy,the width and length of leaf,flower color,the length of corolla tube,the width of anther have great effect on surface L.yunnanensis.The UPGMA clustering results showed that the five populations from the east region of Gaoligong mountains clustered together different from the GDX population from the west region of Gaoligong mountains.3.The transcriptome sequencing of short style & long style flowers of L.yunnanensis and L.pinceana flowers.A total of 16.74 Gb Clean Data and 98 389 Unigenes were obtained.Between L.pinceana and L.yunnanensis,954 unigenes identified genes expressed differently,including 655 up-regulated and 299 down-regulated in which 17 were related to Phenylpropanoid biosynthesis,2 were related to Sesquiterpenoid and triterpenoid biosynthesis,and 3 were related to Flavonoid biosynthesis.Between short style and long style flowers of L.yunnanensis,133 unigenes identified genes expressed differently,including 73 up-regulated and 60 down-regulated.The SSR sites in transcriptome were exploited with high-through.There were 15 384 SSRs,the occurrence frequency of SSRs was 15.63%.In L.yunnanensis,there were 14 772 SSRs,the occurrence frequency of SSRs was 16.50%.The effective primers in L.yunnanensis SSR amplification were screened,in include 81 arbitrary primers.A total of44 primer sets were identified in 6 populations,26 displayed polymorphisms and 18 were monomorphic.4.We analyzed that the genetic structure of all populations collected in the whole distribution areas using 17 pairs of SSR makers.Seventy-four alleles were found.The average of effective number of alleles was 2.326.Genetic diversity among 6 populations were LLO>LCM>FMM>FPK>GCG>GDX,according toN_E,H_E and Shannon'? that were detected,the genetic diversity increases from north to south.Hierarchical AMOVA analysis showed that the genetic variation was mainly found within population,the genetic differentiation amongpopulations was low(t=0.129,=2.830,Percentage variation =2.62%).The UPGMAclustering results showed that the two populations from the north region clustered together,the four populations from the south region clustered together.5.On altitude,geographic distance and genetic diversity for correlation analysis by Mantel test.The results showed that high relativity between phenotypic character in the flower and leaf with the altitude or genotype.No geographic isolation has appeared,but geographic distance great influence the genetic structure.There was not remarkable correlational dependence between phenotype and genotype,but the correlation coefficient between phenotype and genotype was very high.6.Headspace-solid phase microextraction-gas chromatography-mass spectrometry(HS-SPME-GC-MS)was used to identify the volatile organic compounds(VOCs)of the different flower development stages of L.yunnanensis,L.pinceana and the hybrid for the evaluation of floral volatile polymorphism.By comparing with L.pinceana and the hybrid,the composition and its relative content of floral scent were different.The hybrid has much more in common with L.pinceana than L.yunnanensis.L.yunnanensis possessed 12 special compounds(8.29~16.71 %)such as cis-Muurola-3,5diene,Humulen,(-)-Aristolene,Isopropyl myristate,et al.7.The reason of vulnerable plant L.yunnanensis can be divided into the external and internal.External was caused by man-made factors and climate change.Man-made factors lead to habitat fragmentation phenomenon.Climate change control the growth and reproduction of the L.yunnanensis.Internal reason was the poor capacity for natural regeneration and deficiency of genetic diversity.Through specific analysis we get a preliminary idea of protection measure,the effective protection of the population resources in-situ protection,ex-situ protection and reintroduction of these three methods are more suitable for L.yunnanensis. |
PDF Full Text Request