Tetraploid Induction And Identification Of Vaccinium Australe

Vaccinium L., a kind of perennial shrub plants belonging to Vaccinium spp, Ericaceae, is originated in the North American perennial fruit trees. It has alternate simple leaves, the leaves are often ovoid. The most kinds of northern trees are fallen leaves and a few of the evergreen leaves in south. Because of different species or genus, the sizes and colours of the fruits are different. In the natural conditions, Vaccinium grows in rich organic matter micro acid soil. Vaccinium australe "V3"diploid of experiment had been introduced into Chongqing for many years. The speed of its growth is fast, but the size of its fruits and production is in poor condition.Polyploid plants generally grow strongly, fruit is huge, and the resistance is strong. This study attempts to get Vaccinium australe "V3" polyploid by chemicals colchicine mutagenic, in order to cultivate a new variety with Chongqing large fruits with powerful adaptability in the tropical and subtropical areas.The test of Vaccinium australe "V3" used cultivation no vaccine as material, treated shoot tips of Vaccinium for tetraploid mutation by chemicals colchicine and studied induction effect between the outdoor drop method and vitro induction method when using different treating way processed on shoot tips of Vaccinium australe "V3", to determine the best treatment concentration and time of colchicine in different method. Through the morphological observation and chromosome counting, we can get variant plants from ploidy identification. The variant plants were chimaeras that could get tetraploid with separating. Then by comparing the diversity of biological characteristics, resistant, anthocyanin content between diploid and tetraploid that were identificated by chromosome, we can provide basis theorey for breeding new varieties of huge fruit, strong stress resistance. The main research results are as follows:1. In outdoor drop method, we had tried to treat shoot tips of Vaccinium australe "V3"with colchicine. The results of the study show that in the range of concentration from0.05%to0.20%, the sensitivity colchicine of shoot tips of Vaccinium australe "V3" is higher, especially24h at0.20%concentration of colchicine treatment worked best.The survival rate was83.33%, the mutation rate was6.67%.2. In vitro induction method(1)By selecting the multiplication coefficient as different hormone concentrations proliferation ability index, and determining the most appropriate ZT hormone concentration, we could find that the improved WPM medium add concentration is0.20mg/L ZT hormone, and the maximum of multiplication coefficient is8.0.(2) The soaking shoot tip method mutagenic effect is better than that of medium add method. It showed that:immersion method in0.10%colchicine treatment24h effect is best. The mutation rate was16.67%, the medium add method in0.30%colchicine processing shoo tip20d, and the mutation rate is the highest, about6.67%.3. Through the external morphology of the plants that treated with colchicine, we could observe that the stem of variant became stronger, the color became darker, the leaf number of surrounding new shoot tip increased, shoot tip color redden, table fur increasing, leaf became more thick, leaf color deepened, guard cells increased, and the number of stomata per unit area reduced, as well as wider palisade and sponge tissue.4. The observation of the transverse slice of Vaccinium stem observation found that variant stem section skin cells, horny film, and cortical thickness. Primary phloem increased significantly than diploid. As the primary xylem and pith ray quantity of Variant increased, the area of the pulp increased significantly. Variant primary phloem thickness and pulp diameter was328%,218%of diploid.5. The chromosome number count results show that the chromosome number of diploid plant is24,2n=2x=24, variant plants are chimaeras whose chromosome number is24or48. The chromosome number of tetraploid plant is48.6. The results from the comparison between tetraploid of Vaccinium Australe "V3" and plant chlorophyll, MDA, SOD, soluble sugar, proline physiological indexes showed that tetraploid plants had a significant change than diploid plants in physiological characteristics. Chlorophyll content, SOD, soluble sugar, praline of tetraploid was higher than diploid, but the malondialdehyde was lower in tetraploid. Leaf water content of tetraploid was slightly above the diploid. 7. To distill anthocyanidin of Vaccinium australe "V3" leaves and stem rod by solvent and enzymatic extraction. The results showed that the solvent extraction is superior to enzymatic extraction. In the solvent extraction, anthocyanin content of tetraploid stems and leaves was higher than diploid, and the anthocyanin content in leaves was higher than stem.