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对反义MLPK基因在甘蓝柱头特异启动子SLR的驱动下,通过农杆菌介导转化法将其导入高度自交不亲和甘蓝材料‘TF’。转基因甘蓝T0代植株定量PCR分析结果显示,不同的转MLPK反义基因单株内源MLPK m RNA积累量具有明显差异,其中转基因植株花期柱头内源MLPK m RNA积累量明显低于野生型对照。花粉原位萌发的荧光显微镜观察结果显示,转基因甘蓝植株花期自交后,吸附在柱头上的花粉粒大量萌发,且穿过柱头的花粉管明显增加,并导致花期自交结籽数上升,转基因植株花期和蕾期自交亲和指数均明显高于野生型对照植株。结果表明,下调MLPK基因表达能部分打破甘蓝自交不亲和,提高其花期自交结籽能力。
The antisense MLPK gene was introduced into highly self-incompatibile cabbage material ’TF’ by Agrobacterium-mediated transformation under the drive of SLR of cabbage stigma promoter. Quantitative PCR analysis of transgenic T0 plants showed that there was a significant difference in the accumulation of endogenous MLPK m RNA among different transgenic MLPK antisense plants, in which the accumulation of endogenous MLPK m RNA in the flowering stigma of transgenic plants was significantly lower than that of the wild-type control. Fluorescence microscope observation of in situ germination of pollen showed that the pollen grains adsorbed on the stigma germinated significantly and the pollen grains that passed through the stigma increased significantly after selfing of transgenic cabbage plants and resulted in the increase of the number of self- The anthesis compatibility index at flowering stage and bud stage were significantly higher than that of wild-type control plants. The results showed that down-regulation of MLPK gene expression could partly break the self-incompatibility of cabbage and increase its self-fertility and seed-setting ability.