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1、HUAZHONGAGRICULTURALUNIVERSITY烟草叶片愈伤组织诱导与植株再生的研究ResearchonCallusInductionandPlantRegenerationofTobaccoLeaves姓名:黄金波CANDIDATE:Huang-JinBo学号:2012304200510STUDENTID:2012304200510课程:植物细胞工程实验CURRICULUM:PlantCellEngineeringExperiment班级:MAJOR:生技1201班Biotechnology1201指导老师:SUPERVISOR:柳俊齐迎春陈浩L
2、iu-JunQi-YingChunChen-Hao中国武汉WUHAN,CHINA二○一四年十二月DEC,20149烟草叶片愈伤组织诱导与植株再生的研究黄金波华中农业大学生命科学技术学院生技1201班[摘要][目的]以烟草叶片为材料,通过这次实验,基本掌握植物愈伤组织诱导与植株再生的原理和方法,熟练掌握相关实验操作技术;另外,通过本次实验,完成烟草植株再生,并对比光照和黑暗等不同方法,对诱导愈伤组织状态和诱导率的影响,进行结果分析。[方法]在无菌条件下,把烟草叶片剪成1cm2大小的小方块,先通过含0.25mg/LNAA和0.25mg/LBA的MS诱导培养基在光照
3、和黑暗条件下诱导培养;三周后,转接到含0.25mg/LNAA和1.0mg/LBA的MS分化培养基中光照培养;两个月后,观察结果,统计分析。[结果]诱导培养后,在光照条件下和在黑暗条件下培养的均有67.7%的被污染了;用剩下的33.3%全部接种到分化培养基上培养。结果表明分化率在光照条件下和在黑暗条件下无差别;每个愈伤组织分化芽数在光照条件下明显多于在黑暗条件下诱导;分化芽状态在黑暗和光照条件有一定的区别,分化植株苗在光照和黑暗条件下诱导没有明显区别。[结论]光照条件和在黑暗条件下诱导可以影响愈伤分化芽数和分化芽状态。[关键词]烟草叶片;愈伤组织;细胞工程;分化
4、;诱导;植株再生ResearchonCallusInductionandPlantRegenerationofTobaccoLeavesHuang-JinBoHuazhongAgricultureUniversity,CollegeofLifeScienceandTechnology,Biotechnology1201[Abstract][Objective]Usingtobaccoleavesasmaterials,throughtheexperiment,weshouldhandlethebasicprincipleandmethodofplantcall
5、usinductionandplantregeneration,masterrelatedexperiments’technology;Inaddition,throughthisexperiment,tocompletethetobaccoplantregeneration,andcompareddifferentmethodsofcallusinduction,suchaslightanddarkstate,andtheeffectstothestateandinductionrate,thenanalyzingtheresults.[Method]The
6、tobaccoleafwascutinto1cm2sizesmallsquaresunderasepticconditions.FirstlyinducedintheMSinductionmediumwith0.25mg/LNAAand0.25mg/LBA,undertheconditionoflightanddark,respectively;Threeweekslater,transferredtoMSdifferentiationmediumwith0.25mg/LNAAand1.0mg/LBAculturingunderlight;Twomonthsl
7、ater,observingtheresults,andanalyzingthestatistics.[Results]Aftertheinductionculturing,67.7%materialswerepollutedbothundertheconditionoflightanddark;rest33.3%materialswereinoculatedtothedifferentiationmedium,continuingtocultivate.Asaresult,itwassuccessfully.Theresultsshowthattherear
8、enoobviousdifferenc