| Ziziphora bungeana Juz.belongs to the category of Labiatae.It is an endemic medicinal plant in Xinjiang that can be applied to a great variety of illness.Flavonoids are the main medicinal ingredients of Ziziphora bungeana Juz.Based on the experimental foundation of cell totipotency,the technique of Plant tissue culture can cultivate the new plants or secondary metabolites in vitro propagation.Its main application includes the rapid propagation of plants or suspension culture of cells.With the stem segments of Ziziphora bungeana Juz.'s aseptic seedlings as the experimental material,this study is to determine the changes of cell growth and total flavonoids content under stress conditions by inducing and subculturing axillary buds and callus.The rapid propagation and cell suspension culture system of the Ziziphora bungeana Juz.were preliminarily studied.It provides technical support for large-scale and rapid production of Ziziphora bungeana Juz.,and lays the foundation for mass production of effective medicinal secondary metabolites at the cellular level,and provides a new source of medicine.The results were as follows:?1?The suitable medium for seed germination was MS medium;2%NaClO disinfection time was 4 min;the optimum concentration of GA3,6-BA and NAA were200 mg·L-1,0.8 mg·L-1 and 0.25 mg·L-1,and the germination rate were 50.44%,55.56%and 57.78%,which increased significantly?P<0.05?.?2?Determine that the optimal medium for stem segments inducing adventitious buds was MS+0.5 mg·L-11 6-BA+0.1 mg·L-11 NAA,with an average induction rate of90.47%.The suitable combination of bud proliferation was MS+0.5 mg·L-11 6-BA+0.1mg·L-11 NAA+0.3 mg·L-11 GA3+35 g·L-11 sucrose.The multiplication coefficient was 1.95after 20 days.The most optimum rooting medium was 1/2MS+0.5 mg·L-11 NAA+0.2%AC,with a rooting rate of 60%,the average rooting numbers of 3.5 and plant length of4.5 cm for 20 days.Plantlets were transplanted to a potting mixture?1:1:1,nutritive soil:perlite:vermiculite?in trays with about 80.45%survival percentage.?3?The suitable materials for callus induction were stem segments,and the cutting environment was wet;the appropriate for callus induction were MS+1 mg·L-11 6-BA+1mg·L-11 2,4-D+30 g·L-1 sucrose;and the feasible environment for subculture was MS+0.6 mg·L-11 6-BA+0.9 mg·L-11 2,4-D+0.5 mg·L-11 GA3+0.15 mg·L-11 AgNO3;the suitable light intensity was 4.5 klux;there were no significant effect on total flavonoids?P>0.05?but had a significant effect on callus proliferation?P<0.05?with the conditions of NaCl.SA was significantly effect the growth of callus and the accumulation of total flavonoids?P<0.05?;MeJA had a significant effect on the fresh weight of callus?P<0.05?,but not had a significant effect on the content of total flavonoids?P>0.05?.?4?With the loose yellowish-green callus as material,the optimum inoculation amount was 0.1 g·mL-1,and the hormone ratio was MS+1.5 mg·L-11 2,4-D+1 mg·L-1NAA+0.2 mg·L-11 6-BA+1 mg·L-11 KT.The growth of cells and the content of total flavonoids increased significantly?P<0.05?under the suitable culture conditions;the optimum concentration of sucrose was 30 g·L-1;the optimum speed was 100 r/min;The best generation days were 67 days.;the growth of suspension cells and the accumulation of total flavonoids were significantly promoted?P<0.05?with 10 uM SA;However,the higher concentration of SA has a significant inhibitory effect on them?P<0.05?;the 10 uM MeJA has boost the growth of cells?P>0.05?and the accumulation of total flavonoids?P<0.05?. |
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