Identification And Resistance Detection Of Future Generation CBF4 Transgenic Wheatgrass

Self cross future generation(T1 generation)of CBF4 transgenic wheatgrass were used as material in this study. By molecular and resistence determination, stably transferred wheatgrass were obtained and the integration and expression of exogenous gene CBF4 in T1-generation transgenic wheatgrass were confirmed. Transgenic wheatgrasses with strong resistance were identified by detecting there drought tolerance, as basic materials screening for high tolerance varieties and breeding new varieties of transgenic wheatgrasses with drought resistance that are suitable for China's western region.The preliminary findings of this study are as follows:1. The exogenous gene CBF4 have integrated into the genomic DNA of T1-generation transgenic wheatgrass and the integration number is 2～5, which confirmed by PCR and Southern blot analysis. Northern blot analysis showed that some of the exogenous gene in the T1-generation transgenic wheatgrass expressed at transcriptional level. There are 61 plants presented positive result in a total of 144 T1 generation transgenic wheatgrass in PCR analysis.2. Under continuous drought stress(first drought stress and then watering), the moisture capacity in the leaves of transgenic wheatgrasses transformed by CBF4 gene decreased slowly, compared with the leaves of non-transgenic plants; Membrane permeability and MDA content increased less; Proline content increased greatly.In summary, drought resistance of CBF4 transgenic wheatgrasses is higher than non-transgenic wheat grasses.3. Under 15% PEG stress, compared with the leaves of non-transgenic wheatgrasses, the moisture capacity in the leaves of transgenic wheatgrasses transformed by CBF4 gene decreased slowly; Permeability of membrane and MDA content increased less; Chlorphyll content is relatively high; Pro and Soluble protein content increased greatly; SOD, POD and CAT activity is comparatively higher.4. Under continuous drought stress and PEG stress with wheatgrasses, the drought tolerance of CBF4 transgenic wheatgrasses were significantly higher than non-transgenic wheatgrasses, that proved CBf4 gene have passed into T1-generation transgenic wheatgrass, and expressed.