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The widely cultivated Chongqing Riverside II Landrace, jasper three bitter gourd varieties for material tissue culture, complete balsam pear tissue culture. In tissue culture basis, bitter gourd adventitious buds with colchicine induced tetraploid successfully induced bitter gourd the tetraploid and long into whole plants. The comparative analysis of the physiological and biochemical characteristics of the bitter gourd diploid and tetraploid. Its main contents are as follows: 1. Material selection and pretreatment screening, we finalized Chongqing commonly cultivated representative of Riverside II, Landrace, jasper three bitter gourd varieties tissue culture material. Riverside II and jasper varieties of the Green Paper, Changbai white varieties. Hard and bitter gourd seed species of thick-skinned, impede water absorption, it is sterile seed germination before removal should testa. The embryo is attached to the surface of a layer of mucous membrane, impede water absorption, and is likely to cause pollution, it should be washed. And then the embryo is hot water soaking treatment to improve the germination rate and uniformity. 2. Establishment of appropriate disinfection methods solution of sodium hypochlorite disinfection effect, and 0.1% mercuric chloride solution should strictly control the disinfection time. Below 6min disinfection is not complete, higher than 8min seed being poisoned, the germination rate is much lower. Suitable disinfection method was 0.1% mercuric chloride solution is disinfected 6 ~ 8min. 3. The seeds sterile germination under aseptic conditions, the Riverside II germination rate of 66.7%, by Changbai germination rate was 73.3%, jasper germination rate of 100%. And Jasper germinating, stout, is the ideal tissue culture material. 4. Callus induction and adventitious bud differentiation medium MS 6-BA NAA, bitter melon can easily form a callus. Callus was 100%, but is not sensitive to the hormone concentration. Adventitious bud differentiation is extremely fast, usually 5 ~ 6d can differentiate Worse still 2 ~ 3d can be differentiated. The hypocotyl differentiation medium for MS 6-BA3 ~ 5mg / L NAA0.5mg / L, but lower differentiation rate, an average of only 9.37%. Riverside II differentiation average rate of 7.33% in the MS 6-BA4mg / L NAA0.5mg / L in the highest, 14.30%; Changbai differentiation average rate of 9.55%, the in MS of 6-BA3mg / L NAA0.5mg / L in highest 14.30%: jasper differentiation rate of 11.23% on average in MS 6-BA3mg / L NAA 0.5 mg / L in the highest, 16.70%. The cotyledons differentiation is not ideal. 5. Proliferation of culture and subculture to get a lot of bitter melon bud adventitious buds, sterile seedlings of the terminal bud and lateral buds growing parts hypocotyl segment culture in the proliferation medium MS-BA3mg / L NAA0.5mg / L in proliferation. Each explant 3 to 4 may be formed buds. Stem segments, then multiple shoots cut into with buds subculture, the subculture medium in order to achieve the rapid propagation of balsam pear buds. 6. Polyploid jasper bitter melon bud culture induced tetraploid tetraploid induction medium subculture medium recovery culture, then rooting culture in the rooting medium. The cut apical growing point chromosome tablet, observed with a microscope, found that the induced bitter gourd tetraploid colchicine concentration of 0.2%, the processing time of 48h, the mutation rate up to 14.30%. 7. Tetraploid identified as compared with homozygous tetraploid variant material repeatedly subculture. Rooting culture, chromosome tablet observation, found that the material can be obtained following the 4th generation cultured than homozygous tetraploid by colchicine treatment. 8. Rooting and transplanting tetraploid rooting medium 1/2MS NAA0.2mg / L rooting culture, rooting rate was 100%. Transplanting seedlings up to 4 ~ 5cm, hardened, and the survival rate was 60%. 9. Diploid and tetraploid physiological and biochemical characteristics of comparative analysis of stem diameter and plant height measuring the 30d of bitter gourd seedlings after planting, and take the middle of the leaves of bitter gourd diploid and tetraploid comparative analysis of the physiological and biochemical characteristics, found tetraploid stem diameter, plant height, soluble protein, soluble sugar, chlorophyll, carotenoids, malondialdehyde, proline, peroxidase, superoxide dismutase indicators are better than the diploid, results show that The bitter gourd tetraploid physiological and biochemical function better than diploid. This study used a new combination of hormone tissue culture of the bitter gourd, bitter melon in vitro rapid propagation, perfect balsam pear tissue culture, and to lay a solid theoretical foundation for the the bitter melon tissue culture and other aspects of the research. In tissue culture successfully induced with colchicine the bitter gourd tetraploid homozygous basis, bitter gourd germplasm and intermediate materials for other aspects of the breeding work (such as bitter gourd triploid breeding) . We further comparative analysis of the physiological and biochemical characteristics of a the bitter gourd diploid and tetraploid, and provide a reference for the study of the physiological and biochemical functions of the bitter gourd diploid and tetraploid.
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