LIUChun-xiang,LENGChuan-yuan,LIUFu-yan.StudyontheTransformationofCs-FAD7GeneintoNicotianatabacum[J].Northern Horticulture,2012,36(11):120-123.
CsFAD7基因转化烟草的研究
- Title:
- StudyontheTransformationofCs-FAD7GeneintoNicotianatabacum
- 文章编号:
- 1001-0009(2012)11-0120-04
- Keywords:
- Nicotianatabacum; Agrobacteriumtumefaciens; cucumber; FAD7gene
- 分类号:
- S572.035.2
- 文献标志码:
- A
- 摘要:
- 以“革新一号”烟草种子为试材，采用根癌农杆菌介导的烟草叶盘转化法将来源于黄瓜的脂肪酸去饱和酶基因(FAD7)转入烟草，并在附加300mg/L氨苄青霉素(Amp)的MS+0.1mg/LNAA+1.0mg/L6-BA的培养基上诱导植株分化，再生芽在附加300mg/LAmp和100mg/L卡那霉素(Kan)的MS培养基上生根，获得再生植株。结果表明：Kan阳性植株经PCR检测筛选，得到转基因阳性植株，证明异源FAD基因已转入烟草基因组中。T1代经PCR-Southern检测证明转化基因可以遗传；转基因烟草在低温胁迫下生长较好，叶绿素荧光参数F0值下降不大，Fv/Fm值保持较高，表现了对低温的耐受能力，脂肪酸分析表明，该基因可以使烟草叶片的亚麻酸含量比例升高。
- Abstract:
- Using‘GexinNo.1’tobaccoseedsastestmaterials,FAD7genewastransferredintotobaccoleaveswiththehelpofAgrobacteriumtumefaciens.ShootswereregeneratedonMSmediumsupplementedwith300mg/LAmpicillin，1.0mg/L6-BAand0.1mg/LNAAandrootedonMSmediumwith300mg/LAmpicillinand100mg/LKanamycin.PCRanalysisprovedFAD7geneintegrationintoKanamycinresistanceplants.TheresultsofPCRanalysisinT1generationtransgenicplantsshowedthatthetransformedgenecouldinherittonextgeneration，itgrewwellunderlowtemperature.TheparametersofleavechlorophyllfluorescenceF0decreasedslightly，andthevaluesofFv/FmkepthighlevelafterCsFAD7genetransformedintotobacco.fattyacidanalysisshowedthatthecontentsratiooflinolenicacidinleaveswerehigherintransgenictobaccothaninwildtypetobacco.
参考文献/References:
[1]ArondelV,LemieuxB,HwangI,etal.Map-basedcloningofagenecontrollingomega-3fattyaciddesaturationinArabidopsis[J].Science,1992,258:1353-1355.
[2]MurakamiY,TsuyamaM,KobayashiY,etal.Trienoicfattyacidsandplanttoleranceofhightemperature[J].Science，2000，287(5452):476-479.
[3]IbaK,GibsonS,NishiuchiT,etal.Ageneencodingachloroplastomega-3fattyaciddesaturasecomplementsalterationsinfattyaciddesaturationandchloroplastcopynumberofthefad7mutantofArabidopsisthaliana[J].JBiolChem,1993,268：24099-24105.
[4]刘春香,何启伟,赵光强,等.黄瓜ω-3脂肪酸去饱和酶基因cDNA的克隆及表达[J].园艺学报,2008，35(9):1357-1362.
[5]刘进元,张淑平,武耀庭.分子生物学实验指导[M].2版.北京:高等教育出版社,2006:7-8.
[6]张一波,王丽红,周青.Tb（Ⅲ）对辣根叶片膜脂脂肪酸构成的影响[J].环境化学,2009,28(3):392-394.
[7]KodamaH,HamadaT,HoriguchiG,etal.GeneticenhancementofcoldTolerancebyexpressionofageneforchloroplast[omega]-3fattyaciddesaturaseintransgenictobacco[J].PlantPhysiol,1994,105(2):601-605.
[8]KodamaH，HoriguchiG,NishiuchiT,etal.Fattyaciddesaturationduringchillingacclimationisoneofthefactorsinvolvedinconferringlow-temperaturetolerancetoyoungtobaccoleaves[J].PlantPhysiol,1995,107(4):1177-1185.
[9]刘训言.番茄叶绿体-3脂肪酸去饱和酶（LeFAD7）的克隆及其在低温逆境下的功能分析[D].泰安:山东农业大学,2006.
相似文献/References:
[1]颜秦峰,李科友,石振楠,等.过表达miR398对烟草抗旱性的影响[J].北方园艺,2013,37(13):111.
YAN Qin-feng,LI Ke-you,SHI Zhen-nan,et al.Effect of Over expressed miR398 on the Drought Resistance of Nicotiana tabacum[J].Northern Horticulture,2013,37(11):111.
[2]高金秋,于丽杰,陶 雷.白细胞介素-4基因转化胡萝卜的研究[J].北方园艺,2012,36(24):115.
GAO Jin-qiu,YU Li-jie,TAO Lei.Study on Transferred Interleukin-4 Gene in Daucus carota L. by Cell Transformation[J].Northern Horticulture,2012,36(11):115.
[3]胡焕焕,李卫东,姬国杰,等.根癌农杆菌介导IFS基因对岷山红三叶遗传转化研究[J].北方园艺,2012,36(21):110.
HU Huan-huan,LI Wei-dong,JI Guo-jie,et al.Genetic Transformation Isoflavones Synthase Gene from Trifolium pratense Minshan Callus Mediated by Agrobacterium Tumefaciens[J].Northern Horticulture,2012,36(11):110.
[4]许灵杰,蒋岁寒,赵二卫,等.竹醋液复合洋葱、鱼腥草提取物对烟草青枯病菌的室内抑制效果[J].北方园艺,2015,39(05):120.[doi:10.11937/bfyy.201505035]
XU Ling-jie,JIANG Sui-han,ZHAO Er-wei,et al.Inhibitory Effect of Vinegar Composite Onion and Houttuynia Extracts on Ralstonia solanacearum in Laboratory[J].Northern Horticulture,2015,39(11):120.[doi:10.11937/bfyy.201505035]
[5]喻晓敏,吴雷,王魁,等.根癌农杆菌介导的萝卜遗传转化方法研究[J].北方园艺,2016,40(06):99.[doi:10.11937/bfyy.201606024]
YU Xiaomin,WU Lei,WANG Kui,et al.Study on Genetic Transformation in Radish by Agrobacterium tumefaciens[J].Northern Horticulture,2016,40(11):99.[doi:10.11937/bfyy.201606024]
[6]王 茜,王 颉,石 磊.不同萃取条件对分光光度法测定烟草中尼古丁含量的影响[J].北方园艺,2009,33(01):0.[doi:10.11937/bfyy.200901006]
WANG Qian,WANG Jie,SHI Lei.Different Extract Conditions of Spectrophotometric Method to Influence the Content of Nicotine in Tobacco[J].Northern Horticulture,2009,33(11):0.[doi:10.11937/bfyy.200901006]
[7]车建美.,刘 波,张 彦l.,等.青枯雷尔氏菌胁迫对烟草植株抗氧化酶系活性的影响[J].北方园艺,2011,35(02):0.[doi:10.11937/bfyy.201102046]
CHE Jian-mei.,LIU Bo,ZHANG Yan.,et al.Effects of Avirulent Ralstonia solanacearum Strain on Some Antioxidant Enzymes Activities in Tobacco Plants[J].Northern Horticulture,2011,35(11):0.[doi:10.11937/bfyy.201102046]
[8]赵 湛,李 文 生.不同根癌农杆菌菌株类型对木霉菌遗传转化效率的影响[J].北方园艺,2006,30(03):0.[doi:10.11937/bfyy.200603008]
Zhao Zhan,Li Wen-sheng.Influence of Different Strains on Agrobacterium tumefaciens - mediated Transformation Efficiencyof Trichoderma harzianum[J].Northern Horticulture,2006,30(11):0.[doi:10.11937/bfyy.200603008]
[9]王雷,孙尧,李瑶,等.罗布麻eIF-5A基因功能[J].北方园艺,2017,41(23):190.[doi:10.11937/bfyy.20172087]
WANG Lei,SUN Yao,LI Yao,et al.Function of eIF-5A Gene in Apocynum venetum L．[J].Northern Horticulture,2017,41(11):190.[doi:10.11937/bfyy.20172087]
[10]常怀成,倪蕾,孙涌栋,等.农杆菌介导的黄瓜遗传转化的影响因素[J].北方园艺,2018,42(02):9.[doi:10.11937/bfyy.20173043]
CHANG Huaicheng,NI Lei,SUN Yongdong,et al.Factors Influencing Agrobacterium-mediated Genetic Transformation of Cucumber[J].Northern Horticulture,2018,42(11):9.[doi:10.11937/bfyy.20173043]
备注/Memo
第一作者简介：刘春香（1974-）,女,博士,副教授,研究方向为蔬菜育种及分子克隆。
基金项目：山东省自然科学基金资助项目（ZR2009DQ003）。
收稿日期：2012-03-09