| This research took tissue culture seedling obtained from Schisandra chinensis superior varieties“Zaohong”somatic embryo as test subject,explored suitable conditions for Schisandra chinensis tissue culture seedling from strong seedling to transplanting,studied Schisandra chinensis tissue culture seedling's morphology and rhizome's occurrence characterization,implemented preliminary stem tip transcriptome difference research between terrestrial stem and rhizome;explored photosynthetic ability difference among tissue culture seedling,seedling and grafted seedling;carried out polyploid induction through acting colchicine on Schisandra chinensis embryonic callus,screened out suitable induction conditions about colchicine concentration and time for Schisandra chinensis polyploid induction,and induced somatic embryo polyploidy.Main research results are as follows:1.At strong seedling stage of tissue culture seedling obtained from Schisandra chinensis somatic embryo,different basal culture medium,light quality and illumination time all have significant influence on growth of the tissue culture seedling.The promotion effect of 1/2MS medium on plant height,root length and root number was significantly better than that of MS and 1/3MS mediums,the promotion effect of 1/2MS and 1/3MS mediums on the growth of stem diameter was significantly better than that of MS medium;The effect on plant height,stem diameter,root length and root number of tissue culture seedling exposed to white light for 12 hours per day were significantly better than any other treatments.2.At transplanting stage of tissue culture seedling obtained from Schisandra chinensis somatic embryo,different transplanting medium,ambient humidity and moisturizing time all have significant influence on transplanting survival rate and tissue culture seedling's growth.When using turfy soil:Vermiculite=3:1 as substrate,the plant height and root length were significantly better than any other treatments,stem diameter and root number were not significantly different from tissue culture seedling using turfy soil as substrate,but were significantly higher than any other treatments,there was no significant difference between transplanting survival rate and seedling raising substrate,but significantly higher than any other treatments;80%-90%ambient humidity and 12 day moisturizing time were better for Schisandra chinensis tissue culture seedling's growth,the transplanting survival rate could reach 100%.3.Compared with seedling,tissue culture seedling obtained from Schisandra chinensis somatic embryo,have characteristics of short hypocotyledonary axis,mostly colyliform growth dicots,densely growth plant base leaf bud,and low position and large density for rhizome's occurrence.After carrying out transcriptome difference expression analysis between rhizome and terrestrial stem of Schisandra chinensis tissue culture seedling,results showed that there are 102 genes?E3 ubiquitin protein ligase,cytochrome P450,indole-3-acetic acid induced protein ARG7,gibberellin regulatory protein,etc.?overexpressed specifically in rhizome,there are 21 genes?Isoprene plant protein,ethylene reactive transcription factor,flowering promoting factor,etc.?expressed with significant difference in rhizome,16 genes related with root,root hair,branch root and expressed with significant difference.4.Took Schisandra chinensis tissue culture seedling,seedling and grafted seedling with similar growth situation in one year as test subject to study their photosynthetic characteristics.It was found that three kinds of Schisandra chinensis seedlings'net photosynthetic rate?Pn?diurnal variation curves are bimodal type;under the same light intensity and CO2 concentration,three kinds of Schisandra chinensis seedlings'photosynthesis rank from strong to weak is tissue culture seedling,grafted seedling and seedling;the light compensation point rank from high to low is grafted seedling,seedling,tissue culture seedling,light saturation point rank is tissue culture seedling,grafted seedling and seedling;the CO2 compensation point rank from high to low was seedling,grafted seedling,tissue culture seedling,the CO2 saturation point rank from high to low was tissue culture seedling,grafted seedling and seedling.5.Acted colchicine with different concentrations on Schisandra chinensis embryonic callus,carried out ploidy identification on produced somatic embryo and regeneration plant using FCM?flow cytometry?.The results showed that Schisandra chinensis embryonic callus can produce somatic embryo polyploidy by treated with colchicine 200 mg·L-1 for 48 hour and 400 mg·L-1 for 24 hour;colchicine treatment made Schisandra chinensis embryonic callus somatic embryo's occurrence rate significantly better than control group,but produced inhibiting effect on later development and germination of somatic embryo. |
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