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The Internalization of Cell-penetrating Peptide in Chinese Cabbage Microspores and Its Efficiency
Qin Ying1,2, Wang Guixiang2, Guo Ning2, Zong Mei2, Liu Fan2*
1School of Life Sciences, Capital Normal University, Beijing 100089, China;
2Vegetable Research Centre, Beijing Academy of Agricultural and Forestry Sciences, Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (North China),Ministry of Agriculture, Beijing 100097, China
Abstract: In this study， our results proved that the cpp-penetrating peptide (CPP) Tat2 had the ability to penetrate the pollen wall and carry the cargoes including DNA (circular and linear plasmid) and GUS enzyme into the microspores of Chinese cabbage ‘94-323’. The influence factors of internalization efficiency were also researched. The results showed that internalization efficiency of CPP into Chinese cabbage microspores had positive relationship with incubation time of Tat2/DNA and microspores. Meanwhile， the internalization efficiency increased with the increasing of complexes concentration and 33 °C was the optimal incubation temperature. The internalization efficiency had no obvious relationship with DNA fragment length. This study first proved that Tat2 could act as a nanocarrier to deliver macromolecules into microspores cells of dicotyledonous plants and the internalization frequency of Tat2/GUS enzyme could reach to 18%. The results proved a direct experimental basis for establishment of a new transgenic technology mediated by CPP in microspore of Brassica crops.
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