Classification Of Cultivar Flower Colors And The Function Identification Of Anthocyanin Synthase Gene(IsANS)in Iris Sanguinea

Iris sanguinea is a kind of cold-tolerant perennial flower with excellent ornamental effect in China.The flower is blue,and the variant is white.After natural hybridizations,the two varieties form rich intermediate flower color.In this study,the special flower color of I.sanguinea was selected by self-cross breeding,which further enriched the flower color type.On the basis of the color chart,colorimetric meter was used to quantify each color to provide reference for parent selection and color classification in cross breeding.At the same time,the expression vector of anthocyanin synthesis key enzyme gene IsANS was constructed,and the color regulation function of Nicotiana tabacum was studied by agrobacterium-mediated genetic transformation.The main findings are as follows:1.A self-cross-pollination experiment was conducted on 12 flower flowerweeds,and it was found that there were some differences in pollen viability and fruit set rate among different flower colors:the worst pollen viability was FZ(55%)and LZ(59%),while the best was QO(90%)and SL(89%).The lowest fruit setting rates were LZ(29%)and FZ(33%),while the highest were QO(94%)and SL(0.96).To the difficult of self-crossing,FZ and LZ are more difficult while QO and SL are easily.Pollen viability is not the only factor that determines the fruit set rate in different flowers.2.The germination rate and germination potential of self-crossing seeds of different colors were determined.It was found that the lowest germination rate was TH(44%)and LZ(57%),while the highest was SO(87%)and SF(94%).The lowest germination rates were TH(14%)and BS(22%),while the highest were SF(29%)and SO(30%percent).The quality of seeds in TH is the worst,while the quality of seeds in SO and SF is the best.3.Among the 132 self-crossing descendant plants(1165 single plants),30 flowering descendant plants(76 single plants)in the second year were selected to select 6 special single plants,and the genetic background analysis showed that the two-generation parents of 5 plants contained QO,and the two-generation parents of 3 lines were all QO.To sum up,it can be considered that the hybrids of inbred or inbred progeny of QO is rich in variation.4.On the basis of compared with the Royal Horticultural Society Color Chart,there are 32 different colors with their L*,a*,b*parameters have been measured by colorimetric meter CR-10 Plus,from inner perianths,outer perianths and styles of 36 types of I.sanguinea.These colors belong to white group and four coterminous groups:purple?purple-violet?violet and violet-blue,however,the L*,a*,b*parameters have large difference.It was found that all the colors were the middle color of two wild types,the blue-violte and the white flower,by calculation AE parameters.The colors obtained by hybridization have certain limitation.5.Using these parameters to classificate,32 colors can be divided into 7 small classes,of which three classes with special color properties,can be used as parents to breeding special color varieties:category A1-1(L*?45;10<a*?30;b*?-30)has extremely dark color;category A2-1(L*?45;L*?45;-20<b*?0)has a red property,category B2-1(L*>45;a*?10;b*>0)class has a yellow property.6.The recombinant plasmid,pCXSN-IsANS,was successfully constructed,and IsANS gene was successfully transferred into N.tabacum with stable expression.the color of flower organs in transgenic tobacco was significantly deepper(63B,Royal Horticultural Society Color Chart,and the wild type is 63C)through observation.The color difference(? E)between two colors is 8.46.It was thought that IsANS participates in flower pigment synthesis.7.In the growth process of transgenic tobacco and contrast,the transgenic plants appeared a variety of changes:they increased 19.22 cm in high,0.21 cm in plant diameter,8.79 cm in inflorescence length,6.12 cm in blade length,4.28 cm in blade width.The inflorescence branchs increased 4.5 branchs,and 27.33 more flowers during the full-blossom period.The flower increased 3.48 mm in crown diameter and 3.61 mm in corolla tube length.Cultivating time from transplanting to the beginning of blooming was shortened 22.33 days.Therefore,it is speculated that IsANS gene has other functions,which may play a direct or indirect role in vegetative and reproductive growth of plants.