Evaluation Of Different Genotypes And Establishment Of Plant Regeneration System In Vitro Of Schisandra Chinensis (Turcz.) Baill

Schisandra chinensis （Turcz.） Baill. is the precious Chinese drugs in northeast China which favored by consumers in domestic and overseas. With the deepening of the development and utilization of S. chinensis, the demand for raw materials also increased in recent years. However, because of late start for artificial cultivation, less fine improved varieties, biological characteristics and quality diversity among the plants and poor yield stability seriously restrict the cultivation industry development of S. chinensis. In order to better utilize the resource advantage of S. chinensis, this research also studies the fruit phenotype and main lignan content in fruit of different genotypes, which provides foundation for the germplasm resources evaluation of S. chinensis. To breed improved variety, this study established a stable and efficient plant regeneration system for S. chinensis with good fruit characters via somatic embryogenesis, and the dynamic changes of endogenous hormones at various stages in this process are researched comprehensively. The main research results are as follows:1. Comprehensive comparison for11samples of S. chinensis, the fruit phenotype were extensive variation including size, average fresh and dry weight of fruit, dry-fresh ratio, cluster length, number of fruits and fruits compactness, moreover, these fruit phenotype were instable between two years. The average fres hweight, dry weight, the dry-fresh ratio and the compactness were greater in pink fruits than that in red fruits and white fruits. The size of red fruits was larger. The fruit phenotypes of B, D, E, G, P1and P2were better.2. A Symmetry（?） C18column （4.6×250mm,5μm） was maintained at35℃. Detection wavelength was set at254nm, The mobile phase consisted of acetonitrile （A） and water （B） at a flow-rate of0.5mL·min-1. The gradient programme was as follows:0～15min,40:60â†'50:50;15～20min,50:50â†'75:25;20～25min,75:25â†'95:5; injection volume:20μ L. schisandrin, schisantherin A, deoxyschizandrin, γ-schissandrin, schisandrol B and schisandrin C could be detected at the same time. This method was simple, repeatability and precision.3. We determined the contents of six lignans in11samples of S. chinensis, the results showed that most of the lignan contents varied between two years except the contents of deoxyschizandrin were relatively stable. The total contents of lignans in fruits with different colors presented large differences, which was found highest in red fruits and lowest in pink fruits. High contents of schizandrol, γ-schizandrin and schisandrin C in red fruits were observed while white fruit was rich in schisandrol B and deoxyschizandrin. The contents of schisantherin A in pink fruits were higher than that in W and the average contents in red fruits. Considering the factors of phenotype and fruit quality, fruit character of samples B, D and E were good and suitable for breeding.4. This study used4pairs of SRAP primer for PCR amplification of seven samples of S. chinensis, which produced22amplification bands and13bands were polymorphism. The results showed that the genotypes of these seven samples of S. chinensis were difference.5. This research adopted the combination of solid and liquid cultivation for the establishment of stable and efficient plant regeneration system for S. chinensis. Different genotype influenced the callus induction rate of S. chinensis. The callus induction rate was highest when the sterilized dormant bud of S. chinensis cultured on MS+3.0mg·L-12,4-D+0.2mg·L-1TDZ. MS+1.0mg·L-1TDZ+0.2mg·L-1ZT+5.0mgL-1AgN₃was beneficial to embryonic callus induction.3g embryonic callus cultured in50mLl/2MS liquid medium was suitable for the formation of globular embryo.1/2MS+0.2mg·L-1IBA was most favorable for the roots formation of the regeneration plant of S. chinensi and the survival rate was90%after2months transplanting. This study used4pairs of SRAP primer for PCR amplification of seven S. chinensis female parent and the regeneration plant via somatic embryogenesis system, and there were no specific bands produced by four pairs of primers, suggesting that the plant regeneration system we established in this study was stable and reliable, which could be applied to mass production for plant regeneration of S. chinensis.6. The effect of dynamic changes of endogenous hormones on the formation and development of somatic embryos was studied.The contents of endogenous IAA, ABA and GA3related to the embryonic ability,which were highest in embryonic callus among several different types of callus of S. chinensis. During the development of somatic embryos of S. chinensis, the content of endogenous IAA peaked in globular embryo period and the content of endogenous ABA was higher in cotyledon embryo phase. In the whole process, the content of endogenous GA3peaked at heart shaped embryo stage, but no significant difference with the content in globular embryo stage. The contents of endogenous IAA and GA3were significantly higher in malformed embryo of S. chinensis than that in normal somatic embryo, but the content of ABA in malformed embryo were significantly lower than that in normal somatic embryo. The content of IAA was increased significantly when the contents of ABA and GA3declined in rooted plant treated with0.2mg·L-1IBA.