Studies On Genetic Diversity Of Impatiens L. Germplasm Resources In Western Yunnan

The plants of Impatiens L.belong to annual or perennial ornamental herbaceous flowers with large and right-colored flower,and particular flower shape whose petals are made of petalum,petala and sepalum.They can be used not only as cut flowers and potted plants,but also as the plants of flower beds and flower borders.In this study,24 species of Impatiens L.(includes 23 species,1 varietas)were systematically collected,including Ailao Mountain,Nujiang River,Gong Moutain,Fu Gong,Li Jiang,Shangri-La,Tengchong and Yingjiang,and their geographical distribution and morphology were investigated 24 species of Impatiens L.resources were evaluated by using both qualitative and quantitative analysis(Analytic Hierarchy Process,in short AHP)methods comprehensively,meanwhile,their genetic diversity and relationship were studie by using ISSR,trnL-F,atpB-rbcL,and AGAMOUS(AG)molecular markers,which provided new basal data and scientific basis for their genetic breeding research in this region.The main results were as follows:1.24 species of Impatiens L.plants were collected by on-the-spot investigation from 8 places in the western Yunnan,such as Impatiens mengtzeana,I.delavayi,I.amplexicaulis,I.amplexicaulis,I.rubrostriata etc.They were mainly distributed at the altitude from 1300 m to 3600 m,and grew in some places with high relative humidity e.g.besides the roads,grassland and wetlands under the forest etc.,whose height ranged from 15 cm to 89 cm,flower colors were rich(e.g.yellow,purple,white,pink etc.),leaf length varied from 2.0cm to 16.5cm,leaf width ranged from 0.5cm to 5.6cm,leaf area were from 5.53cm2 to 20.23cm2,seed diameter varied from 0.68 mm to 2.79 mm,and 1000 grains weight reached from 1.124 g to 5.900 g.The petala of many species were dorsale orbiculare,some with costa dorso medio protrusion;the petala of inferiora showed different shapes,e.g.cycloied,cuneiform,and blunt triangle,etc.,while their superiora paullo of petala were teniform or dolabriformis;most sepala mainly displayed cymbiformis vel infundibularis and inferius breviter saccatum;leaf edge with crenation,serration,and satae or not.2.Based on the investigation of ornamental values of Imaptiens L.plants in the western Yunnan,4 level evaluation layers,including target layer,restraint layer,standard layer and bottom layer and 17 evaluation indices(e.g.flower color,flower shape,inflorescence type,etc.)were primarily determined to establish the analytic hierarchy process(AHP)evaluation model about their ornamental values and sustainable utilization,by which all the resources collected in this study were evaluated.The results showed that 10 species of Impatiens L.were provided with high ornamental values,e.g.I.mengtzeana,I.rubrostriata,I.polyceras etc.,meanwhile,their applications in landscape architecture were preliminarily analyzed and evaluated according to their different characteristics.3.Modified SDS method was used to extracted total DNA from 24 species of Impatiens L.in western Yunnan,furthermore,L9(34)orthogonal experimental method at four different levels of three different factors(i.e.dNTPs,template DNA Taq and Mg2+concentration)was utilized to optimize the ISSR-PCR system of Impatiens L.plants in western Yunnan.The result indicated that the optimal ISSR-PCR treatment was the reaction system contained 45 ng template DNA,187.5umol/L dNTPs,3.0mmol/L Mg2+?0.3ul 5U/ul Taq DNA polymerase,2.5ul 10×PCR Buffer,1ul primer and 13.1ul ddH2 O.27 ISSR primers were tested in this study,234 polymorphic bands were amplified and the polymorphic ratio(PPB)reached 94.1%,among which PPB of 13 pieces of primers reached 100% took up 48%;and the amplification bands of each primer were 10.17.The results showed that genetic similarity(GS)among all tested 24 species of Impatiens L.ranged from 0.512 to 0.951,which suggested that their genetic diversity was rich.4.Based on polygenetic analysis of 24 species of Impatiens L.in western Yunnan by using ISSR,atpB-rbcL,trnL-F and AG gene,the result showed that I.weihsiensis,I.mengtzeana,I.racemosa,I.radiate and I.rubrostriata were clustered in one subbranch of the phylogenetic tree by using 4 different molecular marks which suggested that their genetic similarity was high and genetic relationship was very closer;that the morphological characteristics of flowers e.g.flower type,petalum shape,petala shape and sepalum shape etc.could be regarded as the important indices of this classification in this genus,especially,petalum shape.The results showed that the genetic diversity of Impatiens L.plants in western Yunnan by using ISSR and AG gene could be consistent with classification according to their flower morphological characteristics.