| The research studied the rooting differences of the cutting,grafting and seed breeding of the peony,the three of which were the most common propagation in the peony,to solve the rooting difficulties of the tissue culture.To filter and master the cutting materials and methods of the peony,we chose different growth regulators under different treating time on different peony cutting varieties.,Studing the fusion and rooting of anvil under different grafting anvil ear combinations,to selecte the best peony grafting anvil combination.By mearsuring in the process of enzyme activity of POD,PPO,IAAO,select the peony is most suitable for cutting varieties of anvil spike and grafting combination;And measure the content of endogenous hormone IAA,ABA,GA,ZR,the content of soluble sugar,soluble protein,and mineral elements N,P,K content of the phloem of the grafted scion,cutting incision parts of the interface and the west stem base root,and integrating the dynamic change of each physiological indexes to judge the period of the cutting rooting in different stages and the main influence factors,to determine the different fusion and rooting stages of the peony grafting.To chose the differences of the related gene in the key rooting period under the three methods of breeding and study the application of the related gene to the peony tissue culture.All were expected to lay a solid foundation for further study to solve the difficulties of rooting in peony issue culture.Test of the main results were as the following:1.Effects of different periods and material on the rooting of cutting: selecting the soil buds,shoots,annual hard branch of the peony as cutting materials,the rates of survival and rooting of the annual hard branch cut in mid-september were the best.As different varieties of tree peony cut dip by ABT-1 1800 mg · L-1 of 2 h,the average rootting number,rootting length and rootting rate of feng Dan were the highest,and the rooting rate reached to 57.14%,mortality was 0.And it could be found that the treatment of Feng Dan had completed the development of the adventitious rootting on the 25 d.and luoyanghong were about on the 29 d,but the control were about on the 32 d.2.The change of phloem physiological indexes in peony cutting during the rooting2.1 the change of enzyme activity: as the cutting dip in 800 mg · L-1 ABT-1 for 2 h,the three kinds of enzyme activity POD,PPO,IAAO in the incision phloem changed with different varieties,but the overall trend was consistent.The activity of POD and PPO increased first and then decreased,the activity of IAAO rose first and then reduced,and rose again.The results showed that promoting effect on the Feng Dan treatment was better than that of luoyanghong,and the both treatment werebetter than that of the control.2.2 the change of the endogenous hormone content: the content of IAA,ABA,GA,ZR in Feng Dan cuttings treated with ABT-1 during the rooting reduced first then rose and reduced at last.The content of GA and ZR were significantly lower than that of IAA and ABA.The trend of IAA and ZR was consistent,and the trend of ABA and GA are basically identical.In the period of the root primordium incubation,the content of IAA and ZR rose steadily,but the content of ABA and GA reduced continuously.In the stage of adventitious roots formating mainly,the content of IAA and ZR gradually reduce,the content of ABA and GA began to rose,and kept rising after the formation of adventitious.2.3 the change of soluble sugar and protein: the content of soluble sugar and soluble protein of Feng Dan in the cutting process were the same,which reduced after rising first,then steadily rose.at the period of 0-10 d,both the two kinds of nutrients were rising,declined 10 d later,and began to rise steadily from 15 d.2.4 the change of the mineral elements N,P,K content: the three nutrients all were rising overall in the process of rooting in Feng Dan cutting,during the period of 0-20 d,the three elements increased first and then decreased,jumped sharply in 20-27 d and sustained high concentration in 25-29 d.The trend of K was similar to that of P.3.The change of phloem physiological indexes of peony grafting during the process of anvil ear fusion and rooting3.1 The change of enzyme activity: significant difference was found in the three kinds of enzyme activity of the peony grafting,the three enzyme activitys of the combination of Fengdan and white peony was higher than the two others.It could be inferred that the combination of Fengdan and white peony was the best of the three,followed by Luoyanghong and white peony,Fengdan and Luoyanghong was the lowest combination.3.2 The change of endogenous hormone content: The content of the four endogenous hormone generally rose first and then reduced,rose again at last.The content of GA and ZR were significantly lower than that of IAA and ABA.The change tendency of IAA and ZR was consistent in certain period of time.ABA and GA in certain period of time the change tendency.3.3 The change of soluble sugar and protein,soluble sugar and soluble protein content increased and are generally first to rise again.Grafting,the two nutrients are increased,soluble sugar fell sharply after 30 d peak gradually recovered during the 40-70-d.Soluble protein for the first time in 20 d after peak slash,30 to 45 d sharply after the stable state.Anvil spike in 30-60 d soluble sugar increased gradually after the first slash,and soluble protein increased before they slow down.3.4 The change of the content of mineral elements N,P,K: the change P elements was severer than that of K and N in the process of rooting,which decreased first then rose and decreased at last,gradually reduced in the period of 0-40 d,jumped sharply during 40-55 d,decreased slowly and sustained higher concentration at 45-60 d;K element continue to rise in 0-30 d,gradual change in 30-45 d,rose to the peak at 55 d,and then presented the decrease tendency;N elements gradually rose in 40-60 d.4.The change of phloem physiological indexes of peony in the process of rooting4.1 The change of enzyme activity: The three enzymes overall rose first and then decreased and rose at last.The variation changed severly in the period of 0-20 d,then flatten.It could be concluded that the rooting period concentrated in 0-20 d.4.2 The change of endogenous hormone content: endogenous hormone changes of IAA,ABA,GA,ZR is different.The content of GA and ZR and change significantly lower than that of IAA and ABA.The content of IAA decreased slightly in 0-5 d,and began rising at 5-10 d,reached to themaximum at 10 d,and gradually declined during 10-20 d,then kept rising;The content of ABA continued to drop dramatically in 0-10 d,went to minimum at 10 d,20 d after slowly rising;ZR fallen slightly in 0-5 d,then began rising,and reached to the peak at 20 d,then gradually reduced and kept stable state;GA took out a modest rise in the period of 0-10 d,and declined slowly during10-20 d,then kept rising.4.3 The change of soluble sugar and protein content: the change of soluble sugar content was not obvious,which presented a small upward trend.Soluble protein content was not stable,and rose at the rooting stage of 5-20 d4.4 The change of the mineral elements N,P,K content: the content of P element was significantly higher than that of N and K in the rooting process of feng Dan,the three kinds of nutrients incerased first and then decresased in 0-20 d overall,and kept stable state.And P element content changed severer than K and N in the process of rooting.Based on the above physiological parameters trends,it could be concluded that the active rooting period was approximately during 10-29 d of the Fengdan cutting.The induction and expression period of the adventitious root primordia in Fengdan cutting was about 15-25 d,while the elongation growth of the adventitious root was during 25-29 d.The active rooting period of Luoyanghong cutting werer approximately in 25-34 d,while the control’s was substantially at the period of 25-40 d.In the process of grafting,it could be initially speculated that the integration activity times of peony and Fengdan was about 30-45 d,the active of integration in combination of Luoyanghong and peony was approximately at 60-70 d,the active of integration in combination of Fengdan and Luoyanghong was approximately during 55-65 d.The rooting period of perennial seedlings Feng Dan concentrated in 0-20 d,and the root primordia formatted at 10 d,the elongation growth of the adventitious root completed substantially at 20 d,after which the peony entered the gently stage in preparation for the budding of the aerial parts. |
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