| In this study Pleurotus ostreatus was cultured in the media containing materials of transgenic cottonseeds. The degradation of Bt protein and DNA in the process of culture was detected. The effects of the transgenic cottonseeds on the growth, the esterase isozyme, the peroxidase isozyme, the DNA in the cells of P. ostreatus were investigated. The impact of the transgenic cottonseeds on the main pests of P. ostreatus was also investigated. The ecological safety of the cultivation of P. ostreatus in the cultures containing transgenic cottonseeds was evaluated.1. The contents of Bt protein in the storage process of CrylA(c) transgenic cottonseeds, cottonseed kernels and cottonseed shell (NuCTON 99B and DP33B) were determined using Envirologix kit and Agdia kit. The content of Bt protein in cottonseed shells was one fourth after one year's storage. The contents of Bt protein in cottonseeds and cottonseed kernels were half after one year's storage.2. The cultures containing 99B and 33B cottonseeds were used to cultivate P. ostreatus. The optical density of Bt protein in the cultures in the process of cultivation was detected using Agdia kit. The net optical density of Bt protein in the culture with 3% and 20% cottonseeds could not be detected after 35d and 45d cultivation respectively with inoculation of P. ostreatus.3. The DNA fragment of the NOS terminator gene from Agrobacterium tumefaciens in the cultures was detected using nest PCR. The NOS 180 fragment could not be detected in the cultures with 3% cottonseeds after 160d's cultivation and the following storage, and could not be detected in the cultures with 20% and 30% cottonseeds after 190d's cultivation and the following storage.4. The liquid media with Bt protein and the liquid media with extraction of cottonseeds were used to culture the mycelia of P. ostreatus. The undried weight of the mycelia increased with the increase of the Bt protein or the extraction of cottonseeds in the media. The effects of the extraction of transgenic cottonseeds on the growth of the mycelia were positive in some cases and negative in other cases. The growth characteristics of some cultivars of P ostreatus were different between in the liquid media with extractions of the CrylA(c) transgenic cottonseed CR130 and its parental cottonseed CRI16. The growth of mycelia in the cultures, the time for the mycelia colonizing the whole culture bag and the time for the emergence of the primordia was not affected with the transgenic or non-transgenic cottonseeds in the cultures.5. The isozyme patterns in the fruit bodies cultivated in the 3% cottonseed cultures with different cultivars of cottonseeds were similar, while those cultivated in the 20% and 30% cottonseeds cultures with different cultivars of cottonseeds were different. The isozyme patterns in some mycelia cultured in the liquid media with extractions of different cultivars of cottonseeds were also different. The effects on the isozyme in some mycelia of different cultivars of P. ostreatus varied with the CRI16 and CRI30 extraction in the liquid media. The isozyme pattern whether in the fruit bodies or in the mycelia cultured in the media with high content of transgenic cottonseeds and their parental cottonseeds were different.6. The 35S165, 35S195 and 35S205 fragments identical with the CaMV 35S promoter gene were detected in P. ostreatus using PCR method, while the 35S101, 35S162 and 35S273 fragments were not detected. The Nosl80 fragment identical with the NOS terminator gene from Agrobacterium tumefaciens was also found in some cultivars of P. ostreats, while the Nosl65 fragment was not found. The Nos 180 and Nos 165 fragments were not detected in the mycelia (cultivars 5.511 and MGPG) cultured in the liquid media with the extraction of transgenic cottonseed and in the fruit bodies (cultivar P40) cultivated in the cultures with transgenic cottonseeds.7. DNA extracted from mycelia cultured in PD (the liquid medium with potato extraction and dextrose), DPD (the PD liquid medium containing DNA solution), C30PD (the PD liquid medium cont... |
