Establishment The High Efficient Regeneration System Of Camelina Sativa (L.) Crantz And Studing On Transforming Of The KLU Gene Into Camelina Sativa(L.) Crantz

Camelina （Camelina sativa （L.）Crantz）belongs to the tribe Sisymbrieae of the familyBrassicaceae. It was extensively grown in the past. However, Camelina growing area andtotal output fell sharply and almost vanished after World War â…¡because of the introductionof rapeseeds. It has attracted increasing attentions in recent years as an alternative oil cropbecause of its excellent agronomy characteristics, such as drought resistance、pest resistance、saline-alkali tolerance, and excellent oil quality, its seed oil contained abundant unsaturatedfatty acid for human health, having polar altitude food value different from the most marketblending edible oil, and its seed oil also can be used as an new type of biofuel.This study chosen hybridized Camelina between Camelina microcarpa and Camelinasativa as experimental material. We detailed researched the impact on Camelina regenerationcapacity of many factor as explant、plant hormone、different days’ explant、AgNO₃. The resultshowed that selecting the cotyledon petiole from5-6days’ aseptic seedling as explant,cultured in MS medium containing1.0mg/L BA and0.3mg/L NAA regenerated shoots at thehighest frequency（30%）, accompanied by the highest number of shoots per explants （4.3）.Moreover, the addition of AgNO₃had no obvious impact on the Camelina regenerationcapacity, the addition of2.5mg/L AgNO₃could limited improve the regeneration capacity,but the regeneration capacity declined with the raising of AgNO₃concentration.To further improving the oil content of Camelina, in this report, we construct plantexpression vector pCambia2301::pINO::KLU, the ovule endothelial specific promoter pINOdriving the KLU gene expressing, transformed into Agrobacterium tumefaciens GV3101byelectroporation and then transformed into Camelina by Floral dipping. This researth detailedanalysises of the impact of many factors, such as Agrobacterium bacterial concentration、permeating buffer on the transforming efficiency. then screening the transgenic seeds, PCRdetecting the resistant plants. The results shows that Kan selection marker optimumconcentration is50mg/L. The highest transforming efficiency can be obtained using thismethod: Agrobacterium grown to OD₆₀₀=0.8, then resuspended in equal volume ofpermeating buffer（1/2MS，Sucrose5%，Silwet L-77200μl/L, pH5.7）, transforming Camelina.A total of107resistant plants were identified from707T0seeds, PCR detected13positivetransgenic plants, the transformation efficiency is1.8%. This study laid the experimentalfoundation for improving the quality and yield of Camelina, as well as provide reference forthe other plant transgenic technology.