Construction of an in vitro regeneration system for the leaves and stem segments of Daphne genkwa
[Objective]Daphne genkwa is a well-known native flower and traditional medicinal plant with significant ecological,ornamental,and medicinal value.Researching and addressing the rapid propagation methods for D.genkwa is an essential requirement for its conservation and utilization.[Method]Using leaves and stem segments of robust D.genkwa plants as explants,we investigated the effects of different sterilization methods on D.genkwa explants.We also studied the effects of different hormone ratios on the induction of callus tissues,the proliferation of adventitious shoots,and the rooting of D.genkwa Our aim was to construct a highly efficient regeneration system for D.genkwa explant tissues.[Result]The explants of D.genkwa were disinfected with 75%ethanol solution for 25-35 second,2%NaClO solution and 0.1%HgCl2 solution for 15 min,The contamination rate of D.genkwa stem segment explants was 52.7%,while the leaf explants was 49.0%.MS+0.8 mg/L 6-BA+0.35 mg/L NAA was the best primary induction medium for D.genkwa stem segment explants,with an 79.7%tissue induction rate and 96.4%induced adventitious shoots.MS+0.8 mg/L IBA+0.85 mg/L NAA was the best primary induction medium for D.genkwa leaf explants,with a 99.8%tissue induction rate and 94.5%induced adventitious shoots.MS+0.35 mg/L 6-BA+0.35 mg/L IBA was the best successional proliferation medium for D.genkwa stem segments and leaves,with the highest proliferation coefficient of 7.36 and the highest average shoot height of 2.47 cm.MS+0.6 mg/L IBA+1.0 mg/L NAA+0.08 g/L ascorbic acid was the best rooting medium for D.genkwa stem explants,with an average rooting of 3.67 and root length of 2.16 cm;MS+0.6 mg/L IBA+0.6 mg/L NAA+0.07 g/L ascorbic acid was the best rooting medium for leaf explants of D.genkwa,with an average of 1.33 roots and a root length of 1.95 cm.[Conclusion]An isolated tissue regeneration system was developed using D.genkwa leaves and stem segments as explants.This system showed high rates of tissue healing induction,stimulated adventitious shoots,proliferation coefficients,and rooting rates.It effectively addressed the technical challenges related to efficient D.genkwa seedling propagation.
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