为了研究番茄LYC-B干扰对类胡萝卜素合成主要酶和主要代谢产物的影响，构建了果实特异性的番茄红素β-环化酶LYC-B干扰载体，并验证了其在不同颜色番茄果实中的有效性。依据X13437.1扩增番茄果实特异启动子E8，构建了果实特异性载体E8-pBI121，其在粉色、红色、绿色和紫色的番茄果实中均能表达。依据X86452.1扩增番茄LYC-B从61～861 bp 间长度为801 bp的片段LYC-B1和从480～781 bp 间长度为302 bp的片段 LYC-B2，构建了以CaMV 35S为启动子的LYC-B干扰表达载体pBI121-B1B2，以E8替换CaMV 35S，构建了果实特异性干扰载体E8-pBI121-B1B2。采用农杆菌注射法分别侵染番茄叶片和果实，GUS染色显示，pBI121-B1B2在叶片、果实和种子中均表达，E8-pBI121-B1B2只在果实和种子中表达。%The experiment will be conducted to research the specific impacts of lycopene β-cyclase interference on carotenoid metabolism,cherry tomato(Solanum lycopersicum L. var. cerasiforme Alef.) fruit-specific lycopene β-cyclase interference vector was constructed,and the validity in different color fruits were verified. Cherry tomato fruit-specific promoter E8 was amplified according to X13437.1 and fruit-specific vector of E8-pBI121 was constructed,which could be expressed in cherry tomato fruits of pink,red,green and purple colors. According to X86452.1,two different lycopene β-cyclase sequences B1 and B2 were amplified,LYC-B1 with 801 bp from 61 bp to 861 bp and LYC-B2 with 302 bp from 480 bp to 781 bp of LYC-B. The interference vector of pBI121-B1B2 with CaMV 35S promotor and the fruit-specific expression interference vectors of E8- pBI121-B1B2 with E8 promotor were constructed. E8-pBI121,pBI121-B1B2 and E8-pBI121-B1B2 were transferred into Agrobacterium tumefaciens GV3101 with the freeze-thaw method,then transformed into living cherry tomato fruits and leaves by injection. The results of GUS staining 7 days after injection showed that E8-pBI121 expressed in pink,red,green and purple cherry tomato fruits;pBI121-B1B2 expressed in leaves,fruits and seeds,while E8- pBI121-B1B2 expressed just in fruits and seeds.
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