In Vitro Production And Biology Characteristics Research Of Autoteraploids Of Rehmannia Glutinosa F. Hueichingensis (Chan Et Sehih) Hsiao

Rehmannia glutinosa f. hueichingensis (Chan et Sehih) Hsiao is one of the Chinese famous "FourHuaiqing Chinese medicines", produced in Henan wenxian and wuzhi. Long-term asexual propagation isleaded to its decline of production and quality. So to develop new varieties with high yielding, excellentquality, resilience has the great significance to the production of Rehmannia glutinosa f. hueichingensis(Chan et Sehih) Hsiao. We have detailed studied of the stimulation and identification of the Rehmanniatetraploid plants,the comparison of the difference between diploid and tetraploid plants in isoenzyme, theresistance of cold,the form in field,physiology,Yield and quality.The results are as follows:1.The induction of tetraploid plants. We treat the Rehmannia stem segment with buds through themethods of soaking and medium adding autumn grain,which included in the mutagenesis techniques oftissue culture combined with colchicine induces Rehmannia glutinosa to tetraploid.These methods all caninduce tetraploid. The best combination is that soaking Rehmannia stem with a bud in0.1%colchicinesolution for24h, and the induction rate is12.9%.Culturing the stem in the medium supplemented with0.05%colchicine for7d and the induction rate is12.5%.2. Identification of tetraploid.We use the direct identification method(chromosome counting method)and indirect identification method (counting the number and guard cell size in the unit field of view area)which allow us to reach the goal of ploidy identification of diploid and tetraploid.The results show that thenumber of diploid chromosome is56and tetraplont is112.Average density within stoma in the lowerepidermal unit vision of diploid and tetraploid, respectively,66.00and49.20,and the guard cell length,respectively is24.00μm, and43.56μm, and width is8.10μm and13.11μm.3. Comparison of diploid and tetraploid biology(1)The comparison of isoenzyme. The results of comparison of POD and SOD isozyme in diploid andtetraploid test-tube seedlings show that tetraploid plantlets POD isozyme has one more B bands thandiploid,the color of A band is deeper and tetraploid plantlets SOD isozyme has one more E bands thandiploid,D band is brighter than the diploid, and C band is darker than diploid.(2)The comparison of the resistance of low temperature stress. Under10°C/5°C (day/night) lowtemperature stress,tetraploid plantlets relative conductivity and MDA content is less than diploid. The content of chlorophyll, soluble protein and POD and SOD is higher than diploid. It shows that theRehmannia tetraploid plantlets can reduce the damage of low temperature stress on cell, and the Heatpretreatment is higher than the diploid.(3)Comparison of morphological and physiological characteristics. The comparison of diploid andtetraploid morphological and physiological indicators in the field shows that tetraploid plant canopy, plantheight, maximum length and width of leaf and number of leaves are less than the diploid.The content ofchlorophyll, soluble protein and POD and SOD of tetraploid is higher than diploid.(4)Comparison of yield and quality. tetraploid No.1and No.2lines fresh weight has been increasedby26.6%and56.7%and catalpol increased by15.0%and13.8%,and verbascoside increased by28.6%and157.1%per plant than diploid. It shows that chromosome doubling can effectively improve the yield andquality of Rehmannia.