番茄果实特异性LYC-B 干扰载体的构建及在不同颜色果实中的表达特异性验证

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。

Construction of Cherry Tomato Fruit Specific LYC-B Interference Vector and Validation of Its Specific Expression in Fruits of Discrepant Colors

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.） fruitspecific
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.