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1、植物遺傳資源學(xué)報2013，14(1)：124—131JournalofPlantGeneticResources雜交轉(zhuǎn)育植酸酶phyA大豆陽性材料篩選研究李喜煥，劉淵，閆瑞葉，孔佑賓，李桂蘭，常文鎖，張彩英(’教育部華北作物種質(zhì)資源研究與利用重點實驗室／河北農(nóng)業(yè)大學(xué)，保定071001；河北科技師范學(xué)院，昌黎066000)摘要：植酸及其鹽類占土壤非有效態(tài)磷30％～40％，利用轉(zhuǎn)基因技術(shù)結(jié)合常規(guī)育種手段培育能夠分解利用植酸磷的作物新品種是解決這一問題的最新途徑。本研究以農(nóng)桿茵轉(zhuǎn)化子葉節(jié)所獲得的JL35-phyA為試材，采用PCR與RT—PCR進行目的基因檢測，獲得轉(zhuǎn)基因陽性材料；隨后將這些陽性材料
2、與38個常規(guī)大豆雜交，實現(xiàn)p廳"向不同大豆品種的轉(zhuǎn)育。結(jié)果表明，利用農(nóng)桿菌轉(zhuǎn)化技術(shù)已將p轉(zhuǎn)入吉林35，且基因在大豆根系能夠正常轉(zhuǎn)錄表達，轉(zhuǎn)基因株系的單株莢數(shù)、粒數(shù)、粒重及百粒重顯著高于野生型，蛋白質(zhì)和脂肪含量與野生型差異不顯著；利用這些轉(zhuǎn)基因株系，通過雜交轉(zhuǎn)育獲得F陽性單株427個，涉及上述38個不同組合，說明目標基因phyA已轉(zhuǎn)移到雜交后代；將F．陽性單株自交后篩選得到部分組合的陽性F，植株及F子粒，經(jīng)農(nóng)藝性狀考察，這些后代材料中存在豐富的遺傳變異，并在雜交后代中選育出一些轉(zhuǎn)有目標基因的優(yōu)良株系，為今后培育轉(zhuǎn)p大豆新品種(系)提供了一批重要的遺傳資源。關(guān)鍵詞：植酸酶基因；農(nóng)桿菌轉(zhuǎn)化；大豆；雜
3、交轉(zhuǎn)育；遺傳資源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
