| The petiole of Pinellia ternata were used as test materials to induce callus by using the single factor experimental study on the different basic media,sucrose concentration,plant growth regulators,determined suitable key factors of Pinellia ternate forming loose callus,and compared the characteristics of different callus with combined microstructure observation,chemical component location and constituent content determination.Preliminary clarify the mechanism of formation of the Pinellia loose callus.The main results and conclusions are as follows:?1?The effects of different concentration combinations,sucrose concentration and culture conditions on the induction of loose callus were studied.The results showed that the growth of callus was better and more porous with MS as the basic medium,with the addition of 2,4-D1.0 mg·L-1 and 6-BA 1.5 m g·L-1.All of induced callus grew well and have high degree of osteoporosis by MS+2,4-D1.0 mg·L-1+6-BA 1.5 mg·L-1+30 g·L-1sucrose induced medium,setting single factor experiment of different concentration of sucrose(0?15?25?30?35g·L-1).Setting different concentrations of 6-BA(0.5?1.0?1.5 mg·L-1)and 2,4-D(1.0?1.5?2.0 mg·L-1)combination test,select the Pinellia loose callus induction of best plant growth regulating substance combination,namely add 1 mg·L-1 2,4-D and 1 mg·L-1 6-BA to the minimal medium.It could be concluded that the key factors for the formation of the loose callus were the basic medium?MS?,sucrose(30 g·L-1),plant growth regulating substance(1mg·L-1 2,4-D+1 mg·L-1 6-BA).?2?Compared with dense callus,the structure of loose callus of Pinellia was loose,the cell and intercellular space were larger,and the cell wall was thinner.The chemical composition of the dense callus indicated that both the cell wall and the intercellular space contain both a cellulose part that was green and a mixture of pectin and cellulose that were colored purple.The dark green color indicated that the cellulose content of the cell wall was relatively high.while the purple part was lighter,indicating that its pectin content is low,that was,cell glue pectin in the mesial layer rarely hydrolyzes the resulting glue and the cells were tightly connected.The location of the chemical components of the loose callus revealed that there was almost only a mixture of pectin and cellulose colored in purple in the cell wall and intercellular space.The darker color indicated that the pectin content was higher,that was,the cell adhesion in the mesangial layer.The fruit pectin hydrolyzes the lotion,and the intercellular connection was relieved,making the callus cells loose;while the green is rare,indicating that the cellulose synthesis of the cell wall is less,the content was very little,the rigidity of the cell wall was weakened,and the cell growth was accelerated.?3?The degree of looseness or compactness of Pinellia callus was mainly related to pectin and cellulose.Pectin was mainly located in the intercellular mesangial layer and was an important component that binds adjacent cells and made the tissue firm.Cellulose was the main component of the cell wall and was a substance that maintains the morphology of the plant cells and enhances the mechanical strength of the cells because of its rigidity.And to a certain extent,limiting the growth of cells. |
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