| Two main works were conducted in this research. Firstly, using an antisense of seed-setting-related (ssr) gene, which was cloned from rice recently by our laboratory and constructed into a mono-T-DNA vector, to re-transform a marker-free indica transgenic insertion mutant line MH63/Bt. Secondly, using a Bt gene, which was kindly provided by Biotechnology Center of Chinese Academia of Agriculture Science, to transform two newly-bred indica thermo-sensitive genic male sterile (TGMS) line 03B198S and 03B199S. The purpose of this research is to set up a material base to confirm biology function of the recently-cloned ssr gene and to create a better Lepidoptera insect pest-resistant germplasm. Both target genes were transformed using Agrobacterium-mediated method and selected with hygromycin-resistant gene hpt. The main results achieved are as follows:1) A number of putative independent transformants as well as their regenerated green plants positive for hygromycin bioassay were attained from each recipient variety;2) Further PCR and Southern blot analysis revealed that for the mono-vector-transformed anti-ssr gene, majority of the independent transformants have its intact copy integrated in the recipient genome, although there was an exception;while for the super binary-vector transformed Bt gene, the co-integration efficiency was approximately 50%;3) RT-PCR analysis of positive Bt-transgenic plants indicated that the target transgene could express in the recipient genome but the expression levels were varied among the different independent transformants, this perhaps due to positional effect;4) The transgenic plants were normally sterile in the local high-temperature season. However, when brought to Hainan reproduction base and transplanted in a well-isolated paddy filed, the main parts of the survival transgenic plants turned fertile and showed highly resistant to the nature-outbreaks of Lepidoptera insects such as stem borers as well as leaf folders.
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