The Chromosome And ITS, TrnL-F Sequence Studies On Eurya Obtusifolia H. T. Chang And Its Related Species

Eurya belonging to Theaceae Ternstroemoideae, about130species in the world and83species in China, is a very important ornamental plants, nectar source plants and medicinal plants. But, so far, the researches about Eurya ismostly in the classical taxonomy and nectar plant resources investigation.In this study, we made cytology producer of Eurya obtusifolia H. T. Chang and its related species to observe their chromosome. DNA extraction, PCR amplication and sequences cloning of Eurya obtusifolia H. T. Chang was finished, Eurya loquaiana Dunn and Eurya groffii Merr.The sequencing results are pieced together and compared in the software. Download ITS sequence and trnL-F sequence of the Eurya from GenBank, then we analyze their homology and calculate the length, G+C(or A+T) content. In Adinandra bockiana E. Pritz. ex Diels as alien species, analysis of the evolutionary relationship was get by N-J method. The result also contain the phylogenetic relations of three species, the location in the Eurya evolution as well as the inherent differences between the male and female.The main results are as follows:1. The ITS sequence and trnL-F sequence on the Eurya obtusifolia, Eurya loquaiana and Eurya groffii were cloned. ITS sequence is about700bp in size, trnL-F sequence is about1000bp in size.2. Compared the ITS sequence of Eurya loquaiana and Eurya groffii with it downloaded from GenBank, there are three differences obtained from male and female plants on the ITS sequence of Eurya groffii. There are located285,479and416sites in the cloned sequence. There are six differences between female and male plants of ITS sequences on the GenBank ITS and cloning sequencing Eurya loquaianasequences obtaind,115,353,505,672,679sites were located in cloning sequence. Another difference of a triple base located at the site of197,198and199.3. Compared the trnL-F sequence of Eurya loquaiana with it downloaded from GenBank, there are six differences between female and male plants of trnL-F sequences on the GenBank trnL-F and cloning sequencing Eurya loquaiana sequences obtaind,29,30,579,699,864,973sites were located in cloning sequence.4. Compare ITS and trnL-F sequences of male and female plants of Eurya obtusifolia, Eurya loquaiana and Eurya groffii, ITS sequences of Eurya groffii between male and female, we could find three differences. ITS sequence of Eurya loquaiana between male and female also existed three differences. Eurya obtusifolia female plants of ITS sequences were similar to male basically. TrnL-F sequence between male and female of the Eurya groffii only existed one difference, there are five different trnL-F sequences of Eurya loquaiana between male and female, trnL-F sequences of male and female plants of Eurya obtusifolia only exists between two differences.5.5.8SrDNA of Different sex Eurya obtusifolia, Eurya loquaiana and Eurya groffii plants were the same length with164bp, GC%is53.66%-54.27%. Three different sexual plants are equalin ITS1and ITS2length, ITS1length was302bp, GC%was53.31%-54.64%.Lengthof ITS2is226bp, GC%was58.41%-59.73%. TrnL-F sequence cloned length was1010and1066bp, the percent of GC is35.83-36.44%.6. Phylogenetic analysis based on ITS sequences of Eurya, we found that the Eurya groffii and Eurya loquaiana belong to the Ser. Longistylae Hu et L.K.Ling, but Eurya loquaiana has a closer genetic relationship with Eurya obtusifolia of Ser. Brevistylae Hu et L.K.Ling.Based on trnL-F sequence of Eurya, phylogenetic analysis was finished and obtained that most of the other species into two groups except Eurya macartneyi Champ. in involving plant materials. One group consisted of Eurya obtusifolia, Eurya loquaiana and Eurya groffii, the other groupincluded Eurya alata Kobuski, Eurya ciliata Mem, Eurya brevistyla Kobuski.Meanwhile, analysis of selection pressure was executed based on two sequences of Eurya, Euryawas relatively stable in the long process of evolution, and rarelyresponse to external evolutionary pressures arising from mutations of the behavior.Analysis results were roughly consistent based on the two different sequences.7. The chromosome image of Eurya obtusifolia, Eurya loquaiana and Eurya groffii can be get, so that the chromosome morphology of them can be more clearly seen.In summary, this study observed chromosome morphology of Eurya obtusifolia and its related species.The inherent differences of ITS and trnL-F sequence on three inter-related species of male and female plants were got. And it put forward some new ideas for identification of male and female gender to distinguish new ideas.Drawed the phylogenetic tree of ITS sequences and trnL-F sequence, it can make clear that phylogenetic distance between Eurya obtusifolia and its related species and position in Eurya evolution. It determine the genetic relationship of Eurya species and provide further molecular level basis.