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ID:53270207
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时间:2020-04-17
《杂交转育植酸酶phyA大豆阳性材料筛选研究-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、植物遗传资源学报2013,14(1):124—131JournalofPlantGeneticResources杂交转育植酸酶phyA大豆阳性材料筛选研究李喜焕,刘渊,闫瑞叶,孔佑宾,李桂兰,常文锁,张彩英(’教育部华北作物种质资源研究与利用重点实验室/河北农业大学,保定071001;河北科技师范学院,昌黎066000)摘要:植酸及其盐类占土壤非有效态磷30%~40%,利用转基因技术结合常规育种手段培育能够分解利用植酸磷的作物新品种是解决这一问题的最新途径。本研究以农杆茵转化子叶节所获得的JL35-phyA为试材,采用PCR与RT—PCR进行目的基因检测,获得转基因阳性材料;随后将这些阳性材料
2、与38个常规大豆杂交,实现p厅"向不同大豆品种的转育。结果表明,利用农杆菌转化技术已将p转入吉林35,且基因在大豆根系能够正常转录表达,转基因株系的单株荚数、粒数、粒重及百粒重显著高于野生型,蛋白质和脂肪含量与野生型差异不显著;利用这些转基因株系,通过杂交转育获得F阳性单株427个,涉及上述38个不同组合,说明目标基因phyA已转移到杂交后代;将F.阳性单株自交后筛选得到部分组合的阳性F,植株及F子粒,经农艺性状考察,这些后代材料中存在丰富的遗传变异,并在杂交后代中选育出一些转有目标基因的优良株系,为今后培育转p大豆新品种(系)提供了一批重要的遗传资源。关键词:植酸酶基因;农杆菌转化;大豆;杂
3、交转育;遗传资源IntroducingPhytaseGenephyAintoDiferentSoybeanVarietiesfromTransgenicLineUtilizingSexualHybridizationLIxi.huan,LIUYuan,YANRui—ye,KONGYou—bin,LIGui—lan,CHANGWen—SUO,ZHANGCai—ying(NorthChinaKeyLaboratoryofCropGermplasmResources,EducationministryofChina,AgricultureUniversityofHebei,Baoding071001
4、;HebeiNormalUniversityofScience&Technology,Changli066000)Abstract:Phosphorus(P)isoneofthemostimportantinorganicnutrientsthatcansignificantlyaffectplantgrowthandmetabolism.However,30to40percentoftheunavailablePinagriculturalsoilsexistsasphytate,whichcannotdirectlybeabsorbedbyplantexceptforresolvedbyp
5、hytase.Soselectingordevelopingnewvarietiesthatcanresolvephytate—Pthroughthemoderntransgenicbreedingapproachprovidedanewopportunitytoimprovetheeficiencyofphosphorusbyplants.Inthispaper,aphytasegene(phyA),isolatedfromAspergilluscMm,wasintro—ducedintosoybeanbyAgrobacterium-mediatedtransformation,andthe
6、nthirty—eightsoybeancrosseshadbeenmadebetweentransgeniclines(JL35-phyA,maleparent)andothernon—transgenicvarieties(femaleparent).PCRandRT—PCRresultsshowedthatthephyAwassuccessfulllyincorporatedintosoybeangenomeandexpressedintransgeniclineJL35-phyA.Futhermore,thepods,seeds,andweightsperplantandweight1
7、00seedsofJL35-phyawerehigherthanwildtypeJL35significantly.Theresultsalsoshowedthat427PCRpositivetransgenicF1plantsfromthethirty—eightcrosscombinationsaboveand377PCRpositivetransgenicF2plantsfromthepro
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