PtAP3不育结构转化毛白杨的研究
Genetic transformation of Populus tomentosa Carr. with sterility construct of PtAP3
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摘要: 毛白杨是优良的绿化和用材树种，但是毛白杨雌株在蒴果成熟时有飞絮现象，给人们的生产和生活带来诸多不便。为获得不飞絮的不育毛白杨，将利用不育结构35S--PtAP3--IR--NOS对毛白杨进行转化，经PCR验证，共获得阳性转化植株12株，通过荧光定量检测转基因植株中外源基因AP3的表达量，发现不同株系间表达量一致，差异较小，相对表达量最高的株系为内参的4.96倍，最低的株系为内参的3.02倍。转基因植株中的上游基因LFY、PI的相对表达量与野生型对照相比下降明显，而下游基因SEP2、SEP3、SEP4的相对表达量与野生型对照比较没有太大变化。这些研究结果表明：转35S--PtAP3--IR--NOS不育结构的毛白杨阳性植株中AP3、LFY、PI基因的表达受到明显抑制，对毛白杨花发育将起到一定的抑制作用。本研究为利用转基因手段获得不育材料提供了依据。Abstract: Populus tomentosa Carr. is a native species in section Leuce with fast-growing and high wood quality in northern China. However, its catkin pollution brings a lot of inconvenience to people’s life and production activity. To obtain sterile poplar, one sterility construct 35S-PtAP3-IR-NOS was transformed into P. tomentosa. The positive results verified by PCR included twelve 35S-PtAP3-IR-NOS transgenic poplar plants. RT-qPCR showed little difference in expression among transgenic lines. The highest level of expression was 4.96 times and the lowest was 3.02 times of that of the reference gene. The expression of upstream gene LFY and PI in transgenic plants relatively declined compared with the wild type, there were no dramatic changes of downstream gene SEP2,SEP3 and SEP4 between transgenic poplar plants and wide type control. Our research suggests that the expression of AP3，LFY and PI in transgenic poplar plants is obviously suppressed, which provides reference for genetically engineered sterility.
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Keywords:
- PtAP3 /
- Populus tomentosa Carr. /
- genetic transformation /
- suppressed flowering
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