The Study On DNA Molecular Identification Of 8 Kinds Of Jiangxi Native Plants Named After Lushan Or Kuling

DNA barcode technology can quickly and accurately identify species,which makes it an important complement in the traditional plant identification system,but also as a new tool to a good protection of plants to accurately identify the classification.In this experiment,eight kinds of plants named after Lushan or Kuling were used to explore whether DNA barcode technology could be used for species identification in this area.Objectives:Using DNA bar code technology to identify eight kinds of native plant species and their closely related species,in order to lay the foundation for the following research on Management of Rural Plant Classification System in Jiangxi local plants.Methods:In this study,DNA was extracted from DNA samples of eight native plant samples named after Lushan or Kuling in Lushan,and amplified and sequenced by reverse transcription-polymerase chain reaction(RT-PCR).In order to broaden the scope of the study,the DNA sequence of several closely related species was downloaded from GenBank.All sequences were constructed by CodonCode Aligner V software splicing,Clustalw 4.0 software comparison analysis,Using MEGA 7.10 to Calculate the relevant data,Based on K2 P Model to construct the NJ and ML Adjacent Cluster Trees to carry out identification analysis.The main conclusions are as follows:?.The ITS and psbA-trnH sequences were screened by PCR in eight native plants named after Lushan or Kuling,and the success rate was 100%.The length of the five plant sequences obtained from ITS sequences was between 440 and 724 bp,and the length was 316 ~ 661 bp.The mutation rate was 31.9% ~ 61.3% and the simple locus rate reached 49.1% ~ 54.5%.The length of the three plant sequences obtained from the psbA-trnH sequence was between 475 and 581 bp,and the length was 351 ~ 481 bp.The mutation rate was 35.5% ~ 52.9% and the simple locus rate was 27.8% ~ 51.6%.The results show that the two sequences have not only largevariability,but also have more information sites to reconstruct the reliable phylogenetic relationship.?.The average GC content in ITS + PsbA-trnH sequence was 29.8% ~ 62.5%in eight native plants and related species named after Lushan or Kuling.The K2 P genetic distance among the eight native plant species named after Lushan or Kuling was 0.The results showed that the ITS + PsbA-trnH sequence of eight native plants named after Lushan or Kuling had a large difference,The average distance of K2 P in the eight kinds of native species was 0.0744 ~ 0.4148,so the K2 P genetic distance of the eight native plants and related species was greater than that of the species.?.The NJ and ML methods were used to construct the phylogenetic tree.The results showed that the branches of the eight native plants named after Lushan or Kuling were similar to each other.Structure of its high credibility.This result confirms the high reliability of the topology used in the constructed NJ and ML system trees.?.The results showed that ITS + psbA-trnH sequence could provide molecular basis for the development of species interspecies system:(1)To support the?Chinese Flora?Hibuscus sinosyriacus Bailey belonging to the view of Hibiscus L,and has the recent relationship with Hibuscus syriacus;(2)In this study the Philadelphus sericanthus Koehne var.kulingensis and the Philadelphus sericanthus individuals were clustered together,and other closely related species were significantly different to them.To support the?Chinese Flora?will regard the Philadelphus sericanthus Koehne var.kulingensis as a Philadelphus sericanthus;(3)Corylopsis sinensis Hemsl var.calvescens Rend.et wils.,?Chinese flora?and some local flora has been changed to Corylopsis willottiae Rend.et wils..In this study Corylopsis Sinensis var.and Corylopsis Sinensis and Corylopsis glanduifera petals were classified as one kind.? Chinese Flora ? thirty-fifth volume of the Corylopsis Sinensis var.belong to the genus of the Ser.Spicatae Harms of the CorylopsisL..(4)In this study we found that there were significant differences in the interspecies and variation of ITS sequence between Sedum species and the success rate was 100%.In this study we can accurately separate Sedum aizoon L.,SedumTosaense and Sedum Japonicum;so the ITS sequence can be regarded as the universal sequence of DNA barcode of Sedum.(5)Based on the psbA-trnH sequence can accurately identify Pyrrosia and its relatives indicating that the psbA-trnH sequence is feasible for the identification of DNA barcodes of ferns.(6)The ITS sequence has a high amplification efficiency and a success rate in Lonicera Linn..In the experiment,we solved the problem of polyA / T structure which existed in the sequencing process by the two-way sequencing of the two ends and got the information needed for species identification.The sequence agrees with the?Chinese Flora?(seventy-second volume)will be Lonicera modesta var.as a variety of Lonicera modesta classified as safflower subgroup.(7)In this study we found that the ITS sequence can accurately distinguish among Rubus kulingensis,Rubus articua,Rubus idaeus and Rubus saxatilis;therefore the ITS sequence can be used as an accurate method for the identification of Rubus L.and its relatives.(8)In this study it was found that there were significant differences in the interspecies and variation of the ITS sequence between the Tongoloa L.;we can accurately distinguish the Umbelliferae in the Middle Tongoloa L.,Pimpinella L.,Sinocarum L.and Sinolimprichtia L.;therefore the ITS sequence can be regarded as a universal sequence of DNA bar codes for Umbelliferae.The innovation of this study: the first attempt at the DNA molecular level on the Lushan area of local plants to study.Based on the theory of molecular system theory,based on the principle of DNA barcode in plant identification,the phylogenetic tree was constructed by identifying the optimal DNA bar code of the plant species to determine the system position of the plant species to be identified in the whole genus.