分子植物育种
分子植物育種
분자식물육충
MOLECULAR PLANT BREEDING
2013年
2期
217-224
,共8页
番茄%转化酶抑制子%瞬时表达%实时荧光定量PCR
番茄%轉化酶抑製子%瞬時錶達%實時熒光定量PCR
번가%전화매억제자%순시표체%실시형광정량PCR
Tomato%Invertase inhibitor%Transient expression%Real-time fluorescence quantitative PCR
利用含有过表达抑制子载体的根癌农杆菌分别在番茄源叶和果实中进行瞬时表达,快速验证转化酶抑制子(inhibitors of invertases, INH)转基因的表达情况及对果实发育过程中转化酶活性的影响. Micro-Tom在源叶注菌处理后,Lin6的表达含量增加,表明转化酶在植株中响应病菌侵染;但转INH在源叶几乎没有明显表达变化.果实注射后,注射空菌(EHA105)菌液和空载体(pBI121-2A11)菌液后,转化酶(invertases, Inv)基因Lin5、Lin6表达量没有明显改变,但Inv的活性明显升高1倍左右,这表明转录数量并不能反映翻译后的蛋白质量;在注射转基因(p1300-2A11-INH)菌液后3 d,INH的表达明显增加,是其它处理的数倍,这是由于在果实特异性启动子2A11的作用下,INH在果实中特异地表达.而且注射p1300-2A11-INH菌液3~5 d后, Inv的活性显著下降,尤其胶质胎座中最为明显.这一结果表明INH主要在翻译后水平调控Inv的活性.
利用含有過錶達抑製子載體的根癌農桿菌分彆在番茄源葉和果實中進行瞬時錶達,快速驗證轉化酶抑製子(inhibitors of invertases, INH)轉基因的錶達情況及對果實髮育過程中轉化酶活性的影響. Micro-Tom在源葉註菌處理後,Lin6的錶達含量增加,錶明轉化酶在植株中響應病菌侵染;但轉INH在源葉幾乎沒有明顯錶達變化.果實註射後,註射空菌(EHA105)菌液和空載體(pBI121-2A11)菌液後,轉化酶(invertases, Inv)基因Lin5、Lin6錶達量沒有明顯改變,但Inv的活性明顯升高1倍左右,這錶明轉錄數量併不能反映翻譯後的蛋白質量;在註射轉基因(p1300-2A11-INH)菌液後3 d,INH的錶達明顯增加,是其它處理的數倍,這是由于在果實特異性啟動子2A11的作用下,INH在果實中特異地錶達.而且註射p1300-2A11-INH菌液3~5 d後, Inv的活性顯著下降,尤其膠質胎座中最為明顯.這一結果錶明INH主要在翻譯後水平調控Inv的活性.
이용함유과표체억제자재체적근암농간균분별재번가원협화과실중진행순시표체,쾌속험증전화매억제자(inhibitors of invertases, INH)전기인적표체정황급대과실발육과정중전화매활성적영향. Micro-Tom재원협주균처리후,Lin6적표체함량증가,표명전화매재식주중향응병균침염;단전INH재원협궤호몰유명현표체변화.과실주사후,주사공균(EHA105)균액화공재체(pBI121-2A11)균액후,전화매(invertases, Inv)기인Lin5、Lin6표체량몰유명현개변,단Inv적활성명현승고1배좌우,저표명전록수량병불능반영번역후적단백질량;재주사전기인(p1300-2A11-INH)균액후3 d,INH적표체명현증가,시기타처리적수배,저시유우재과실특이성계동자2A11적작용하,INH재과실중특이지표체.이차주사p1300-2A11-INH균액3~5 d후, Inv적활성현저하강,우기효질태좌중최위명현.저일결과표명INH주요재번역후수평조공Inv적활성.
@@@@The purpose of this study was to use an Agrobacterium-mediated transient expression system contai-ning the expression carrier inhibitor to prove the effect of INH gene expression and the invertase activities in the process of tomato fruit development. Micro-Tom treatment after Agroinfiltration in source leaves, the expression of Lin6 was increased, indicating that cell wall invertase should response to pathogen infection in plants. Micro-Tom were injected with EHA105 or injected that contains pBI121-2A11 plasmid, expression of the invertase gene (Lin5, Lin6) did not change obviously. Whereas tomato was injected strain containing p1300-2A11-INH plasmid after 3 d, expression of INH was significantly increased several times. This would be contributed by the function of the fruit specific promoter 2A11, resulting in INH specific expression in the fruit, while activity of Inv decreased significantly after 3 d to 5 d infection which injected strain containing p1300-2A11-INH plasmid, especially expression in pectinic placenta was most obvious. The results in this research presented that the activity of inverase was regulated by inhibitor in the post-translational level.
