Regeneration System Establishment Of Kalanchoe Daigremontiana And The Somatic Embryogenesis

Kalanchoe daigremontiana is model plant in researching stonecrop acid metabolism and plant cell totipotency,studying the somatic embryogenesis has what molecular mechanism for plant cytology research has important significance.At the same time,the establishment of Kalanchoe daigremontiana regeneration system can be difficult to draw,difficult to bud plant to provide reference and system basis.In this study,we investigated the mechanism of somatic embryogenesis of rhizome and the regenerated system,and established the regeneration system of rooting.The establishment of regeneration system was divided into three steps:induction of callus,induction of buds and induction of rooting.Three kinds of experiments were carried out.Three kinds of different concentrations of 6-BA:2,4-D=10:1,6-BA:IAA=10:1,KT:2,4-D=10:1.At the same time,the callus of the leaves was induced to take the callus for subculture,and the external morphology was observed.The browning and water swelling rate were recorded.The method was quantitatively determined by real-time fluorescence quantitative PCR.Changes of Callus Gene Expression in Roots and Roots.In order to study the mechanism of somatic ernbryogenesis in the process of viviparous growth and regeneration of the leaves,the real-time fluorescence quantitative PCR was used to determine the genes of the three important periocds in the regeneration system.The results showed that four genes,CUC1,STM,PIN1 and CYCBl,whicj had an important role in the development of somatic embryos,were used as the internal,reference gene,and three representative periods were selected.The callus was induced by callus and callus for 30 days,and the callus was induced for 60 days.The leaves were selected for the three periods,which were leaves that are not yet engraved,give birth to missing leaves and leaves of births.The results showed that the browning and water swelling rate increased to 34.3%in the third generation,and the browning and water swelling rate increased gradually during the subculture.The sixth generation up to 97%.Although the external morphology of the third generation was measured,the relative expression of the four genes was not obvious.In the induction of callus,some of the shoots were explants,and the shoots were 17.9%.The stem segments of the shoots were placed in 1/2Ms medium for 1 week to take root.The callus of the callus was only 6-BA:IAA=10:1.The callus could shoot at about 2 months,and the germination rate was 27%.The germination rate was 30%at 4 months after the induction of the callus.The callus after budding should be rooted in the rooting induction medium for 1 week.The results of real-time quantitative PCR showed that the relative expression of the genes could be different in the buds and buds,and the three genes selected from the reproductive system The relative expression was significantly different.