| Impatiens has belong to genus of Impatiens L.,family of Balsaminaceae.Most of the plants in genus of Impatiens L.are annual or perennial herbaceous with large and colorful flowers and ornamental value,particularly whose flower petals include banner petal,wing petal and lip petals.The Impatiens spp.can be applied not only as cut flowers and potted plants,but also as the plants growing in flower beds and flower borders.In this study,32 species or varieties of Impatiens L.(includes 30 species,2varieties)were systematically collected from Zhaotong,Qujing,Wenshan,Honghe and some areas around Kunming,the geographical distribution of the resources and morphological features of the 32 species or varieties were surveyed and comprehensively evaluated by using both qualitative and quantitative analysis(Analytic Hierarchy Process,AHP)methods.Meanwhile,the genetic diversity and relationship of these resources were studied by using ISSR,trnL-F,atpB-rbcL,and DEF molecular markers,which could have provided the genetic breeding research in this region with new fundamental data and scientific reference.The main results were as follows:1.32 species or varieties of Impatiens L.plants were collected by on-the-spot survey from some places in the eastern part of Yunnan Province.They were mainly distributed at the altitude from 1300m to 3500m,along the roads,on the grassland and wetlands under the forest and in other humid places.There were great differences in the morphological characteristics,i.e.,the plant height ranged from 10cm to 90cm,the flower color changed from yellow,purple,white,to pink etc.,the leaf length varied from 3.0cm to 13.2cm,leaf width ranged from 1.6cm to 3.9cm,leaf area differed from3.88cm~2 to 25.22cm~2,seed diameter varied from 1.04mm to 2.81mm,and 1000 grains weight changed from 0.793g to 6.144g.The banner petal of many species is usually round in shape,some species have bulging median band in the banner petal.The shape of the basal lobes of the wing petal varied from round,cuneiform to obtuse triangle,whereas the upper lobes of the wing petal were in banding or securiform shape.The lip petal is mainly in infundibular or vesicular form.The leaf edge of Imaptiens spp.plants changed from crenate to serrate,with bristle or not within the teeth.2.Based on the survey and observation on the ornamental values of Imaptiens L.plants collected in east Yunnan,the analytic hierarchy process(AHP)evaluation model about their ornamental values and sustainable utilization of the Imaptiens L.plant resources in the studied area was established with 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.),by which all the 32species or varieties collected were evaluated.The results showed that 13 species of Impatiens L.were of high ornamental values,the application of these species to landscape architecture was preliminarily analyzed and evaluated according to their different characteristics.3.Modified kit method was adopted to extract total DNA from samples of the 32species or varieties of Impatiens L.collected in east Yunnan.23 ISSR primers were applied to PCR amplification in this study,223 polymorphic bands were amplified and the polymorphic ratio(PPB)reached 94.9%,among which PPB of 14 primers reached100%,accounting to 60.9%of the 23 primers tested.Averagely the amplification bands of each primer were 9.70.The results showed that the genetic similarity(GS)among all the 32 species or varieties of Impatiens L.ranged from 0.540 to 0.990,indicating that there was rich genetic diversity among the tested plant resources.4.The systematic development of the 32 species or varieties of Impatiens L.in east Yunnan was analyzed by using ISSR,atpB-rbcL,trnL-F and DEF gene segments.The results showed that Impatiens radiata,Impatiens rubrostriata,Impatiens racemosa were clustered in one branch of the phylogenetic tree by using 4 different molecular marks which suggested that their genetic similarity was high and the genetic relationship was very close.The clustering analyses by the 4 gene segments showed that the morphological characteristics of flowers including the flower size,shape of the banner petal,wing petal and lip petal could be regarded as the important classification indices of this genus.The shape feature of the wing petal could play an especially important role in the classification for Impatiens L.plant species It was showed that the results from the genetic diversity analyses of Impatiens L.plants in east Yunnan by using ISSR and DEF gene segments was consistent with the classification according to their flower morphological characteristics.5.A pair of degenerate primers was designed and synthesized according to AG gene sequence of the known species reported in Genbank,and the whole AG gene sequence of I.uliginosa was cloned through RT-PCR method.Having been analyzed by the related software and compared to the AG gene sequences reported in Genbank,it was found that the cloned genes were highly homologized with those of the former.The analysis on structure of the amino acid sequence of the cloned AG gene indicated that there were very conservative MADS-box and K-box in the cloned AG gene sequence.The AG gene homology comparison and the system evolution analyses on the newly cloned I.uliginosa and the species in some other genera and families reported in Genbank indicated that there was a high homology between the cloned AG genes and the AG genes of I.uliginosa,which showed these two were most closely related. |
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