| The Clematis L. genera is subordinate to Ranunculaceae family, which is theworld famous ornamental with wide distribution, known as the queen of climbing plants.Because of its high ornamental value, with a variety of anti-resistance, its introductioncultivation and breeding research has a long history in foreign research. Its breeding,cultivation and application have been developed to a very high level, and breedingtechnology has been mature. Hundreds of different colors, types, blooming seasonhorticultural varieties have been cultivated through the Plant Regeneration in vitro culture,which, together with the production of a large number of new varieties each year, haslaunched a mature commercial operation. The domestic research focuses on systematictaxonomy, germplasm investigation, chemical composition, medicinal value, cultivationand so on. At the same, it also discusses its landscape application. But the introductioncultivation and breeding are rarely touched by researchers. Although many gardenvariety’s main parents are originated from China, but our utilization rate of native plants isvery low,approximately ten kinds of species were studied on introduction anddomestication. After the introduction, its breeding as a good cross-breeding materials wasnot well proceed, and has not been used to increase the species diversity of landscape’svertical greening and provide material species for landscape richness.Based on948application project of the State Forestry Administration, used15cultivars of Clematis（‘Gipsy Queen’,‘Warszawska Nike’,‘Hagley Hybrid’,‘MarieBoisselot’,‘Niobe’,‘Ville de Lyon’,‘Viticella GˋMme Julia Correvonˊ’,‘Westerplatte’,‘Rouge Cardinal’,‘Huldine’,‘Henryi’,‘General Slkorshy’,‘Ramona’,‘Lagonbleu’,‘Bagatelle’） which from United Kingdom, France, Netherlands and other Europeancountries introduction to Kunming, the reasearch used annual tender stem of15cultivars astest material for in vitro culture study to establish the system of induction on6cultivars,the system of induction and proliferation on8cultivars, and the system of plantletregeneration for ‘Gipsy Queen’, so as to explore the effect of plant hormones, seasons andthe difference between various cultivars to induction, proliferation, anti-browning androoting for explants in vitro culture process. Selected culture medium formula of induction,proliferation for each cultivars, and anti-browning and rooting for ‘Gipsy Queen’. Toestablish rapid propagation system, thus multiply seedlings, and provide the theoreticalbasis for rapid propagation introduction of other Clematis, and to provide the scientificbasis for future promotion and application. The results were as follows:（1） The best induced medium for ‘Gipsy Queen’ was MS+2mg·L-16-BA+0.05mg·L-1NAA, the induction rate was89.5%, the best bud differentiation medium was MS+2mg·L-16-BA+0.01mg·L-1NAA, budding percentage was111%.There has a peak ofinduction for callus after10d of inoculation, and then significantly proliferated, during thewhole induction culture the explants without callus only showed enlargement and no otherchanges occur.The best proliferation medium for ‘Gipsy Queen’ was MS+1mg·L-16-BA+0.1mg·L-1NAA+1mg·L-1KT+35g·L-1S，proliferate multiple up to6.6, If theformula used for factory production, the large seedlings can be propagation and the costscan be saved. Added2g·L-1AC in anti-browning medium for ‘Gipsy Queen’ the browning rate related to concentration level of6-BA and NAA. When the rate between them wasequal, the hormone concentration will increase. The browning rate increases inevitablely.The best rooting medium for ‘Gipsy Queen’ was1/2MS+0.2mg·L-1NAA，the rhizogenesisrate reaches to66.7%, the root growing point appears on5d. The total rooting bands is30and the average rooting bands is5, the longest is5cm; The more suitable NAAconcentration is0.1～0.2mg·L-1over0.2mg·L-1f, it could be suppressed without root.Add0.5mg·L-16-BA in1/2MS+0.3mg·L-1NAA medium the NAA concentration limitationcould be broken; the rhizogenesis rate reaches to54.6%. It not only induces rooting butalso makes roots thick.（2） The best induced medium and bud differentiation for ‘Warszawska Nike’wasMS+1mg·L-16-BA+0.05mg·L-1NAA, the induction rate and buds rate was81%and83.3%,the best proliferation medium was MS+1mg·L-16-BA+0.01mg·L-1NAA proliferativemultiple reached2.8during subculture, the proliferation rate and multiple shoot both arenot high. The highest are36.4%and46.7%.（3） The best induced medium for ‘Hagley Hybrid’ was MS+2mg·L-16-BA+0.05mg·L-1NAA, the induction rate was83.9%, the best bud differentiation medium was MS+1mg·L-16-BA+0.05mg·L-1NAA buds rate was86.7%. The best proliferation medium wasMS+2mg·L-16-BA+0.05mg·L-1NAA proliferate multiple reached2during subculture theproliferation rate and multiple shoot both are low. Appeared check batter than treatments,this phenomenon has to further study.（4） The best proliferation medium for ‘Marie Boisselot’、‘Niobe’、‘Ville de Lyon’、‘Viticella GˋMme Julia Correvonˊ’and ‘Westerplatte’ was MS+3mg·L-16-BA+0.1mg·L-1NAA+1.5mg·L-1KT，proliferate multiple of ‘Marie Boisselot’and‘Niobe’ reached2.4and2.9. At5d post inoculation, the multiple shoot Germinating. The subcultureproliferation rate was up to115.4%and103.8%. During the proliferation, multiple shootappears to Germinating and callus proliferating; proliferate multiple was1.9and2.1, At5dpost inoculation,‘Viticella GˋMme Julia Correvonˊ’ appear multiple shoot Germinating,At10d all callus proliferating, proliferation rate up to100%,‘Ville de Lyon’ Germinatinguntil8d, multiple shoot reached89.5%, callus proliferating till10d, proliferation rate over96%;‘Westerplatte’ proliferate multiple up to3.2, only multiple shoot Germinating with nocallus proliferating after post inoculation, proliferation rate up to93.9%.（5） More comfortable multiplicative medium for ‘Rouge Cardinal’ was MS+3mg·L-16-BA+0.1mg·L-1NAA+2mg·L-1KT, proliferation multiples was1.6, lower thanwhen KT was1.5mg·L-1, but multiple clumps rate and proliferation rate is higher,especially rate of multiple clumps, reached up to30.8%, while when KT was1.5mg·L-1without the emergence of multiple clumps.（6） The optimal induced medium for ‘Huldine’ was MS+2mg·L-16-BA+0.01mg·L-1NAA, training120d induction rate reached as high as96%, were all callus.Although in70d occurs adventitious buds, but eventually failed to differentiate into buds,compared with induction for0. Confirmed the basic medium only guarantee the explants survival and the lowest physiological activity, only adding appropriate plant hormones caninduce callus growth and cell division.（7） More comfortable induced medium for ‘General Slkorshy’ was MS+0.01～0.05mg·L-1NAA+0.125mg·L-1KT, The best medium for shoot differentiation wasMS+0.05mg·L-1NAA+1mg·L-1KT, budding rate was up to100%. Although from the first8d start adventitious buds appeared, but induction time extended as long as42d. The20dappeared callus, but after60d induction rates was only42.3%and42.9%. Judging fromthe current formula several groups of induced the effect not good, need to be furtherstudied.（8） More comfortable induced medium for ‘Ramona’ was MS+0.05mg·L-1NAA+1mg·L-1KT. From the8d began to have in vitro shoot germination, but ultimately nodifferentiation into bud, appeared small callus until42d and the induction rate was only40%, including four treatment and control of induction rate were all0, currently thehormone type and concentration had no significant effect on induction for ‘Ramona’.（9） In Inductive medium of MS+2mg·L-16-BA+0.05mg·L-1NAA+1mg·L-1KT,‘Marie Boisselot’>‘Gipsy Queen’>‘Niobe’>‘Westerplatte’, the highest rate of inductionwas168.1%and the lowest was110%. The callus was texture loose, appeared transparentfor pale yellow to yellow-green. While‘Viticella GˋMme Julia Correvonˊ’,‘Ville deLyon’and‘Westerplatte’, the callus was a yellowish-white, texture brittle, easy browning,the worst was ‘Viticella GˋMme Julia Correvonˊ’, nearly1/3explants happenedbrowning.‘Ville de Lyon’and‘Westerplatte’ Slightly better, only about2.7%happenedbrowning.（10） According the induction rate and the overall performance in different seasons of‘Gipsy Queen’ and ‘Warszawska Nike’, picking seasons of explants have direct effects.Explants picked in spring the observations on test index is significantly better than theautumn, the more suitable for induction training. If in autumn or winter for training, thoughthe season is the main factors of influence for proliferation, but added certain concentrationKT in the medium can break season restrictions, allowing the proliferation multiplesmultiplied. |
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