Mustard (brassica Juncea Coss) Plants In Situ Vacuum Infiltration Genetic Transformation System,

MUSnd Hicajbecea Coss．) is widely planed and it's a nutritive vegetable． But there existserious plant diseases and insect Pests, Low yield and bad quality Only resolving method is throughcultUring new cultivatOL Conventional breeding demands longer time and eqet gene aPpere too late, WhilethrOUgh thesgenetic technology W gene is inserted plant cell, So breeding efficiency is linproved raPidlyHowever the genetic transformation technology in mustards had not gained breakthrugh． The vacuuminfiltraion genetic bosformation with unique advantages is aPplied and stUdied． Thrugh StUdying on theeffeCts of the tusformatic facors in order to establish in sitU vacuum infiboion genehc tranSformationsystem of mUSeds and provide the theOreicaI basis． Main conclusions are as follow:l．After MtJStards seeds at 50C hot watr were warmedl0min, at23C soaked for 24h, at 23oC sProuted for20h or at 50C hot waer were wanned l0min, at 23C soaked 5h, then aMC treated for 25days, then plantedin soil, they were eaSy to form optiInum plant ShaPe for vacuum whhatio ~c taformallon during lOngday light． Cv YUanyejie seeds being sowed in oPen field on 20 SePtember, on 30 SePtember and on l7 Febmpand CvXuelthong seeds being sowed in open fieId on 30 SePtember and on 27 FebrUary are eaSy to formoPtimum plat shaPe for in situ vacuum infiIthaon genetic tranSformation．2． Through studying on the factors of the effects on genetic traflsformation． On Mustards plant in sittIvacuum infiltration high efficiency genetic transformation was established． The vacuum infiItration timeis 2minj intermission30s/30s or 2minjintermissiom 1．5minj30s． Agrobacterium concentration is OD00o0．8． Infiltration medium is l/2MS inorganic composition and B5 organic composition． In bud stage thebuds are treated by the vacuum infiltTation．3． Primarily a method of identifitalion of triformatic plant was consmictedWwth point selection． Onscreening CvXuelthong and YUanyeie seeds, opdritim conCetaion is 800 mgh and oPtimurn time ofscreening is l0 days afle being inoculated． It does not reqube medium, not POllute, save medicinamen andgain high survive． The hygromycin resiStance seedling: leaf buds appear become gree and grOW normally4． On screening CvXuelihong and Yuanyejie seedsthrough medium with hygromycin, optimumconcentration is 25 mg/L and optimum time of screening is l6 days5． The genome DNA was ertted bough microbial etating method and ethelished PCR reaction system6． Transgenetic plants were obtained． There were 386 Hyg resistance seedlings, and at random sevenseedlings were selected among them, and then their leaves were eXtr8cted into DNA and made PCRidentification． It proved that the foreign-gene was collocated into genome．...