高等植物中的转化酶报告人:王爱华•转化酶的概念•转化酶的分类分布•转化酶在植物糖代谢中的作用•转化酶基因的进化1概念转化酶(Invertase),又称蔗糖酶或β-呋喃果糖苷酶。是与蔗糖代谢密切相关的一种酶,催化蔗糖不可逆水解成葡萄糖和果糖。Sucrose+H2O-----Glucose+Fructose与糖代谢相关的酶蔗糖磷酸合成酶(SPS)磷酸蔗糖磷酸化酶(SPP)蔗糖合成酶(SuSy)转化酶(Inv)2.1转化酶的分类转化酶类型最适pH范围溶解性底物是否糖基化细胞壁类型(酸性,细胞外)4.5-5.0不溶或可溶蔗糖/棉籽糖…是液泡类型(酸性)4.5-5.0可溶蔗糖/棉籽糖…是细胞质类型(中性/碱性)7.0-7.5可溶蔗糖否2.2转化酶在源和库细胞中的分布淀粉VACINV质外体细胞质液泡CYTINVVACINV叶绿体CALVIN循环丙糖蔗糖葡萄糖果糖葡萄糖果糖蔗糖液泡蔗糖CYTINVCWINV库细胞源细胞蔗糖葡萄糖果糖果糖葡萄糖葡萄糖果糖蔗糖调节植物的生长发育和器官建成细胞壁转化酶在蔗糖的卸载中发挥作用液泡转化酶促进成熟器官液泡的蔗糖积累调节淀粉合成一些酸性转化酶基因作用于病原防治细胞壁转化酶与抗衰老有关酸性转化酶与植物的非生物(干旱)抗性有关转化酶在信号传导中的作用3转化酶在植物糖代谢中的作用3.1调节植物生长发育和器官建成a.一般在植物的分生组织和快速生长的幼嫩的组织和器官中(如幼嫩的叶片、茎、根尖和果实等)细胞壁转化酶以及液泡转化酶的活性较高(Estruch和Beltran,1991;Xu等,1989)b.细胞壁转化酶和单糖转运蛋白互作控制细胞分裂和生长(Weschke等,2003)c.抑制转化酶基因表达植物生长异常(反义RNA技术)PhenotypicChangesinTransgenicPlantletsExpressingAntisensemRNAforVacuolarInvertaseorCellWallInvertaseAllplantletsweregrownonhalf-strengthMurashigeandSkoogmediumcontainingonlymacroelementsandmicroelementsand3%sucrose.ThetransgeniccontrolplantletsharborthepromoterofthegeneforcarrotcellwallinvertasefusedtotheGUSreportergene.(A)Antisenseplantletsforcellwallinvertase(Acwi-59,Acwi-65,andAcwi-76)andwild-type(WT)andtransgeniccontrolplantlets(B)Antisenseplantletsforvacuolarinvertase(Avi-34,Avi-47,andAvi-65)andwild-type(WT)andtransgeniccontrolplantlets反义mRNA技术研究转化酶功能(1)Source:PlantCell,Vol.11,177-190,February1999,Copyright©1999,AmericanSocietyofPlantPhysiologistsPhenotypicChangesinMatureTransgenicPlantsPlantsweregrowninsoilinagrowthchamber.ThecontrolplantsaretransgenicandharborthepromoterofthegeneforcarrotcellwallinvertasefusedtotheGUSreportergene.(A)AnAcwiplantandacontrolplant(B)AnAviplantandacontrolplantSource:PlantCell,Vol.11,177-190,February1999,Copyright©1999,AmericanSocietyofPlantPhysiologists反义mRNA技术研究转化酶功能(2)Electronmicroscopicpicturesofpollenfromwild-type(AandB)andtransgenicplants(C-F)(A)SEMpictureofwild-typepollen.(Bar=5µm.)(B)TransmissionEM(TEM)pictureofwild-typepollen.(Bar=5µm.)(C)SEMpictureofpollenfromNT23-4.(Bar=5µm.)(D)TEMpictureofpollenfromNT23-4.(Bar=5µm.)(E)SEMpictureofpollenfromNT23-6.(Bar=5µm.)(F)TEMpictureofpollenfromNT23-6.(Bar=5µm.)NT23-4representsoneoftheplantswhosepollenshowsgerminationefficienciesbetween25%and40%;NT23-6isoneoftheplantswithgerminationefficiency<2%Source:MarcGoetz,PNAS2001;98:6522-6527反义mRNA技术研究转化酶功能(3)3.2细胞壁转化酶在蔗糖的卸载中发挥作用Bachelier等(1997)认为处于细胞壁的转化酶能调节蔗糖从韧皮部卸出,并且控制蔗糖吸收速度。Sturm和Tang(1999)认为细胞壁转化酶使蔗糖在源和库之间形成了浓度梯度,是蔗糖卸载的关键酶。细胞壁转化酶主要作用于质外体卸载(Wobus和Weber,1999;Weschke等,2003)3.3液泡转化酶有利于成熟器官液泡积累蔗糖a.蔗糖进入液泡具有代谢上的双重作用(Tauberger等,1999)b.液泡转化酶通过液泡的蔗糖循环,调节蔗糖从叶片输入液泡的有效性(Scholes等,...