| In this study,two varieties of Blue Honeysuckle were selected as experiment material.buds stems sprouted with dormant buds were used as explants.A rapid propagation system for tissue culture was established by primary culture and subculture.The leaves and stem segments of the aseptic seedlings were used as explants which used to establish an efficient simple in vitro plant regeneration system for Blue Honeysuckle.This study aims to lay the foundation for the rapid propagation of seedlings,preservation of resources,and breeding of biotechnology.The results was as follows:(1)Different growth regulators have different effects on the proliferation and culture of the two varieties,and the proliferation culture medium for the two varieties is also different.MS+2.0 mg/L 6-BA+0.15 mg/L NAA can be used for the proliferation culture of “Qiuleem”;3/4 MS+1.0 mg/L 6-BA+0.2 mg/L NAA can be used for the proliferation culture of “Narem”.The multiplication factor was 5.06 and 5.30 respectively.(2)Different growth regulators have different effects on callus induction of Blue Honeysuckle leaves.The results showed that 2,4-D was the best growth regulator of five to induce callus from leaves.The highest callus inductivity(100%)was achieved on medium with 0.5-2.0 mg/L 2,4-D,when supplemented with 6-BA(1.0?2.0 mg/L),the callus induction rate and the amount of callus increased accordingly,and the callus becomes dense.Addition of 6-BA and TDZ alone did not induce Callus.The combination of 6-BA and 2,4-D is a more suitable solution for inducing the callus of Blue Honeysuckle;With the addition of three growth regulators of 6-BA,IBA,and KT,Qiu Leimu's induction rate and amount of callus were better than that of “Nareim.”(3)Among the three growth regulators including 6-BA,IBA,and KT,IBA had a significant effect on callus induction from stems of both varieties.KT had a significant effect on the callus induction of Nareim stems,but had no significant effect on the callus induction of “Qiuleem”.For “Qiuleem”,6-BA is more influential than KT.The F-numbers of 6-BA and KT were 10.188,and 3.245,respectively For “Narem”,KT is more important than 6-BA.The F-numbers of 6-BA and KT are 20.663 and 33.027,respectively.MS+6-BA 1.0mg/L+IBA 0.3mg/L+KT 0.5mg/L can be used for the callus induction from stems of “Qiuleem”.MS+6-BA 0.5 mg/L+IBA 0.2 mg/L+KT 1.0 mg/L can be used for the callus induction from stems of “Nareim” the induction rate was 98% and 77% respectively.(4)When callus was induced by 6-BA,IBA and KT combinations,the induction rate of the leaves of the two varieties was greater than the induction rate of the stem segments,and the callus of the leaves was smaller than that of the stem segments.The inducing rate of Qiulemu stems is 49%-98%,and the induction rate of leaves is 60%-100%.Nareim stem induction rate is 1%-77%,leaf induction rate is 54%-95%.Both Stems and leaves are suitable as explants for inducing callus.Qiulem's induction effect is better than Narem's.(5)Cytokinins play a leading role in the regeneration of Lonicera japonica callus,and auxins play a supporting role.For the regeneration of “Qiulem”,KT is better than 6-BA,For the regeneration of “Nareim.”,6-BA is better than KT,MS+6-BA 1.5 mg/L+IBA 0.3 mg/L+KT 1.0 mg/L Can be used as regeneration media for both two varieties,and the frequency of adventitious shoot regeneration in “Qiuleem” and “Nareim” was 83% and 99%,respectively.(6)Minimal medium and IBA were the dominant factors in the induction of rooting,and the basic medium had greater influence than IBA,and NAA was a secondary factor.3/4 MS+0.5 mg/L IBA can be used as rooting medium for “Qiuleem”.3/4 MS+1.0 mg/L IBA can be used as rooting medium for “Nareim”.(7)Proper dark treatment time can shorten the time of initiation of calli and accelerate the growth of callus.The appropriate dark treatment time for both varieties is 15 days. |
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