Somatic Embryo Induction and Histocytological Observation of Isodon rubescens
Establishing the somatic embryogenesis system of Isodon rubescens to give full play to its value in traditional Chinese medicine,and promote the stability and utilization of its resources.The leaves of I.rubescens were used as explants to dedifferentiate into embryonic callus and further proliferate to induce somatic embryo.The histological and cytological observation of embryonic callus and somatic embryo at each stage was made by paraffin section,and the effect of hormones on plant regeneration was tested.The results showed that in B5+1.0 mg·L-1 2,4-D medium,leaves could dedifferentiate into light yellow and granular embryonic callus.In MS+1.0 mg·L-1 2,4-D+0.2 mg·L-1 6-BA medium,the embryogenic callus had the best proliferation effect,and the maximum net proliferation was 2.81 g.The optimal medium for somatic embryo induction was MS+1.5 mg·L-1 TDZ+0.2 mg·L-1 NAA,and the highest induction rate was 91.33%.In MS+0.5 mg·L-1 6-BA+0.5 mg·L-1 IBA medium,embryogenic callus containing somatic embryos could differentiate into small plants.In histological observation,the embryonic callus was yellowish or yellowish white granules with large and obvious nuclei.Two kinds of somatic embryogenesis were observed in the inner and outer origin of the callus,and the later developed into spherical,heart-shaped,torpedo-shaped,cotyledon-shaped embryo or cluster embryo structures containing starch granules.This study successfully established the indirect somatic embryogenesis system of I.rubescens,which laid a foundation for optimizing the"quality"and"quantity"of embryogenic callus,and also provided the theoretical basis for the research of molecular breeding and somatic embryo technology.
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