Cloning and RNAi construction of a LEAFY homologous gene from Populus tomentosa and preliminary study in tobacco |
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Authors: | An Xin-min Wang Dong-mei Zhang Zhi-yi Li Shan-wen and He Cheng-zhong |
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Institution: | (1) Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education, Beijing Forestry University, 100083 Beijing, P. R. China |
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Abstract: | PtLFY, a LEAFY (LFY) gene, was cloned from Populus tomentosa (LM50) by PCR. Sequencing analysis indicated that PtLFY was 2 629 bp long, composed of three exons and two introns and encoded 378 amino acids. The splice donor sites and the splice
acceptor sites were in identical positions to the LFY and its homologues. The amino acid sequence inferred was 68%-99% homologous to those of LFY and its homologues by blast analysis in GenBank. The Southern blot analysis indicated that there was a single copy of the
PtLFY gene in genomic DNA of male and female P. tomentosa (LM50 and 5082). The pBI121-Ptalfy (reverse)-intron-Ptlfy-GUS-nos was constructed using RNA interference (RNAi) technique
and verified by PCR and digestion identification and transformed into tobacco. Some transgenic tobacco plants were obtained
by PCR and PCR-Southern identification. The growth was generally repressed in transgenic tobacco plants compared with wild-type
ones and some phenotypic differences were observed.
Supported by the National Natural Science Foundation of China (Grant No. 30371175) and Postdoctoral Foundation of China (Grant
No. 2002032041)] |
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Keywords: | Populus tomentosa PtLFY cloning RNAi construction transformation |
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