Establishment Of In Vitro Regeneration System Of Narcissus Tazetta Var. Chinesis 'Yulinglong'

In this study,the bulb disc of‘Yulinglong'from three provenances?Fujian Zhangzhou,Shanghai Chongming,Zhoushan Putuo?was used as the explants to induce the regeneration of small bulbs,so as to find out the best disinfection method of bulb disc and obtain the best medium,laying a foundation for the establishment of in vitro rapid propagation system.In order to obtain the best callus culture medium,the regeneration small bulb of‘Yulinglong'from three provenances and the pedicel and ovary of‘Yulinglong'from Shanghai provenance were induced as explants.Different explants were induced on the same medium,and the endogenous hormone content and physiological changes were measured during the callus process,providing some basic physiological data for the Narcissus tazetta var.chinensis.The main research results are as follows:1.Establishment of in vitro rapid propagation system of Narcissus tazetta var.chinensis?1?The best disinfection methods for bulb plates of‘Yulinglong'from the three provenances sites are different.Zhangzhou and Putuo:soak 3%carbendazim solution for 5h+75%alcohol 50 s+20%sodium hypochlorite for 20 min+0.1%PPM culture medium;Chongming:soak 3%carbendazim solution for 5 h+75%alcohol 50 s+20%sodium hypochlorite for 20 min.?2?The higher 6-BA was beneficial to the proliferation of bulb disc.Optimal proliferation medium:Zhangzhou and Chongming:MS+9 mg/L 6-BA+1.0 mg/L2,4-D+0.5 mg/L NAA;Putuo:MS+9 mg/L 6-BA+0.5 mg/L 2,4-D?3?The best small bulb induction medium:Zhangzhou MS+3 mg/L 6-BA+0.1 mg/L 2,4-D;Chongming and Putuo MS+3 mg/L 6-BA+0.1 mg/L 2,4-D+0.5 mg/L NAA.?4?The rooting rate could reach 100%.The optimal rooting medium was 1/2 MS+15 g/L sucrose+4.6 g/L carrageenan+1.0 mg/L NAA+2 g/L AC.2.Callus induction?1?The callus induction rate was 56.67-58.89%for the regeneration of small bulbs from three provenances The best medium:Zhangzhou:MS+3.0 mg/L 6-BA+3.0 mg/L 2,4-D+1.0mg/L NAA.Chongming:MS+3.0 mg/L 6-BA+3.0 mg/L 2,4-D+3.0 mg/L NAA;Putuo:MS+3.0mg/L 6-BA+9.0 mg/L 2,4-D+1.0 mg/L NAA.?2?Pedicel induction rate was up to 100%,and the optimal medium was MS+3.0 mg/L 6-BA+2.0 mg/L 2,4-D+1.5-2.5 mg/L NAA+1.0 mg/L KT.The rate of induction of ovary in scapes stage was improved.The induction rate of ovary was not significantly increased after cutting.The optimal induction medium for ovary at the stage of scape respectively was MS+2.0 mg/L 6-BA+6.0 mg/L 2,4-D+2.5 mg/L NAA+1.0mg/L KT;MS+1.0mg/L 6-B+6.0mg/L 2,4-D+2.5mg/L NAA+0.5mg/L KT.The optimal induction medium for ovary at flowering stage respectively was MS+3.0 mg/L 6-BA+6.0mg/L 2,4-D+3.5 mg/L NAA+0.5 mg/L KT;MS+3.0 mg/L 6-BA+6.0 mg/L 2,4-D+1.5-3.5 mg/L NAA+0.5 mg/L KT.3.Endogenous hormone changes during callus induction at different sites?1?A culture index was introduced to evaluate the comprehensive effect of callus induced by different explants in the same medium.?2?During callus induction,ABA and IAA contents were lower in the initiation stage and the fission stage,and higher in the formation stage,and the change of IAA was different in the formation stage.Among them,the content of young leaves and anthers decreased,while the content of pedicels and ovaries increased.?3?ABA/GA₃callus induction in young leaves increased,the other three explants decreased,the division stage decreased,the other three explants increased;ABA/GA₃ of ovary increased in the formative stage,while the other three explants decreased.Pedicel,IAA/ZR of anther and ovary all decreased during callus induction.The IAA/GA₃ changes in leaves and pedicels were opposite at the initiation stage of callus,and the changes were the same at the division stage and the formation stage.ABA/ZR in anther and ovary decreased in the initiation stage,and in contrast to the change trend of young leaves,and all three decreased in the division stage and formation stage.4.Physiological changes of callus differentiation in different partsThe changes of SOD,POD and CAT activity in pedicel and leaf showed an inverted"V"pattern during callus induction.The variation trends of anther POD,SOD and CAT were roughly the same,and the variation trends of ovary POD,SOD and CAT were also roughly the same.In the callus induction process,the protein content of the four explants,such as leaves,decreased at first,but the consumption of pedicels and leaves reached the lowest on the 7th day,the consumption of ovaries reached the maximum on the 14th day,and the anther reached the first low point on the 3rd day and then increased.The protein content of leaf and pedicel increased after 7 days and ovary increased after 14 days.The changes of soluble sugar in the four types of explants were all decreasing.