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Objective: To optimize the culture methods and conditions for the rapid propagation of Lycium ruthenicum in vitro, explore effective proliferation methods, and screen suitable plant regeneration pathways, so as to establish an artificial high-efficiency breeding technology system. Methods: Stem segments with one or two nodes of aseptic seedlings were used as materials. MS, modified MS1 and modified MS2 were used as basic media. The effects of different plant hormones and their concentrations on callus induction, axillary bud germination, basal stem adventitious cluster bud induction and plant regeneration were studied by single factor, complete combination and L9(34) orthogonal experiments. Results: A large number of callus were induced in MS + NAA 1.0 mg/L + 6-BA 0.1 mg/L medium, but the ability of redifferentiation was weak, and the highest multiplication coefficient was only 4.36 after 35 d of culture; While in MS + NAA 0.1 mg/L + 6-BA 0.05 mg/L + KT 0.5 mg/L medium, with axillary buds starting to germinate, the node of stem segment contacted with culture medium began to swell and appeared adventitious bud points, which sprouted basal stem cluster budswith the incidence rate of 100%: And the highest multiplication coefficient could up to 42.84 after 45 d of culture. The suitable medium (MS1 + NAA 1.0 mg/L) for rooting of test-tube seedlings was modified. After 40 d of culture, the rooting rate reached 98.9%; And the survival rate of transplanted tube seedlings after refining was over 90%. Conclusion: In this study, the basal stem cluster buds were used as a new way for proliferation, and an efficient propagation system of L. barbarum in vitro was successfully established, which not only greatly improved the yield and quality of test-tube seedlings, but also provided another idea for the rapid propagation of other Lycium plants in vitro.
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Objective To establish an efficient tissue culture and rapid propagation system, and study the protocorm proliferation and regeneration conditions using seeds as explants in Dendrobium officinale. Methods Seeds of D. officinale were used as explants, protocorm was induced on inducement medium. After proliferation, the protocorm were transferred to the regeneration medium. Then the regenerated shoots were transferred into rooting medium to induce rooting of plantlets, and developing complete plant. Results The basic culture medium for D. officinale growth was 1/2 MS; The best culture medium formula for inducing protocorms was 1/2 MS +1.0 mg/L 6-BA + 0.2 mg/L NAA + 50 g/L mashed potato; The optimal proliferation medium for protocorm was 1/2 MS + 1.0 mg/L 6-BA + 0.5 mg/L NAA, and the maximum multiplication factor could reach 23. The optimal regeneration medium was 1/2 MS + 2.0 mg/L 6-BA + 0.2 mg/L NAA, regeneration rate can reach 95%; The best culture medium for seedling rooting was 1/2 MS + 0.3 mg/L NAA + 50 g/L potato juice, and rooting rate reached 100%. Conclusion This research provides an effective way for keeping good varieties of features and rapid propagation of D. officinale, at the same time helps to solve some theoretical problems in factory production of D. officinale.
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Objective: To establish a rapid propagation system of Magnolia officinalis subsp. biloba. Methods: Using peeling seeds of M. officinalis subsp. biloba as the initial explants, different media and various combinations of plant growth regulators (6-BA, NAA, IBA, and IAA) on the seeds germination, the seedlings subculture, and rooting culture were studied by single factor test and orthogonal test. Results: The best culture medium of seeds germination was B5 + 6-BA 0.5 mg/L + NAA 0.1 mg/L, and the budding percentage was 87.0%; The effective medium for cluster buds-inducing and subculture was MS + 6-BA 2.5 mg/L + NAA 0.5 mg/L, and the propagation coefficient was 6.2; The best rooting medium was 1/2 MS + NAA 0.5 mg/L, and the rooting rate was over 84% after 30 d. Conclusion: Rapid propagation technique system is established in order to lay technique foundation for the industrial production of M. officinalis subsp. biloba.
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Objective:To seek the best part and optimal culture medium for the tissue culture of Bletilla striata. Methods: The effects of hormone concentration and proportion on the induced differentiation, propagation and rootage of Bletilla striata were investiga-ted. Results:The radicle of Bletilla striata was the best part to induce the clustered shoots with the optimal culture medium of 1/2MS+1. 0 mg·L-1 6-BA+2. 0 mg·L-1 NAA. The best hormone concentration for inducing the clustered shoots was MS+1. 0 mg·-1 L 6-BA+0. 05 mg·L-1 NAA, and the optimal rooting medium was 1/2MS+0. 5 mg·L-1 NAA. Conclusion: The tissue culture system for Bletilla striata is established.
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Objective: To establish the rapid propagation system and tissue culture techniques of Typhonium flagelliforme for large-scale seedlings. Methods: Using media with different hormones proportions, the tuber tissue was found to be a good explant for inducing asexual propagation system. Results: The callus culture medium consisted of MS+NAA 0.2 mg/L+KT 1.0 mg/L+sucrose 3%+agar 6 g/L could generate callus successfully. Medium consisted of MS+NAA 0.2 mg/L+6-BA 2 mg/L+sucrose 3% + agar 6 g/L was a superlative proportion of hormones concentration to generate adventitious buds efficiently, especially for higher proliferated multiples. The rooting medium consisted of 1/2 MS+NAA 0.2 mg/L+IB A 0.4 mg/L+KT 0.2 mg/L+sucrose 3%+agar 6 g/L+AC 0.3 g/L was conducive to generate roots rapidly. The expiants could be acclimatized after a period of 12 to 14 weeks. Experiments of one-step-seedling formation indicated that the one-step-seedling formation medium, containing MS+NAA 0.2 mg/L+6-BA 1.5 mg/L+sucrose 3%+agar 6 g/L was the best proportion of hormone concentration, for 5 to 6 weeks, new plantlets could be developed, which will be acclimatized in 10 weeks. Conclusion: The tissue culture techniques and rapid propagation system of T. flagelliforme could be used for large-scale seedlings and lay a foundation for its improved breeds.
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Objective: To develop a rapid propagation of Sehisandra chinensis. Methods: The tissue culture of S. chinensis was studied by orthogonal test and the media for the rapid propagation of S. chinensis were optimized. Results: When NAA 0.3, ZT 0.1, and 6-BA 1.5 mg/L were added in the MS, the best rate of bud proliferation was obtained, while the NAA 0.2 mg/L was added, the best rooting was got. The optimal cultural contion is mat temperature is 25 °C, light intensity 2 000 lx, and photoperiod 12 h. Conclusion: The optimal media and cultural condition in the test could be used to improve the yield and output, as well as quality of S. chinensis.
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Object To study the suitable conditions on tissue culture of stem apex and shoot rapid propagation of ginger. Methods The ginger stem apex ((?)
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Object To establish and optimize the technology of clonig rapid propagation by tissue culture in Scutellaria baicalensis Georgi for the protection and development of nature resource as well as seeking for new breeding way of S. baicalensis by biotechnology. Methods Research on the optimized technology of tissue culture including callus inducing, differentiation, plantlet propagation, and rooting were designed. Results Callus could be induced from nodes and internodes easily with 100% inducing rate compared with that from leaves. PP 333 has significant stocky effect to the plantlets. Conclusion The nodes are good explant source in inducing callus. PP 333 in certain concentration which is added to culture media could improve and regulate the differentiation of callus and growth of plantlets as well as increase the survival rate of cultivated seeding.
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Objective Effects of different factors on proliferation and rooting of stems with a bud were studied by using Dioscorea bulbifera as test material to optimize the rapid propagation system of D.bulbifera virus-free plantlets.Methods Plant tissue culture method was used in shoot tip culture and rapid propagation study,and RT-PCR method was used in virus detection of virus-free plantlets.Results The best proliferation medium of D.bulbifera stems with a bud was MS+KT 2 mg/L+6-BA 1 mg/L+NAA 0.5 mg/L;The best sucrose and agar concentration of D.bulbifera stems with a bud was 30 g/L and 0 g/L,respectively;The best rooting medium of D.bulbifera stems with a bud was 1/2 MS+IBA 0.1 mg/L+NAA 0.5 mg/L+PP_(333) 1 mg/L;The best transplanting matrix of regeneration plantlets from D. bulbifera stems with a bud was perlite-vermiculite(2:1);The best PP_(333) concentration of D.bulbifera regeneration plantlets for transplanting was 50 mg/L.Conclusion The rapid propagation system of D. bulbifera virus-free plantlets is established successfully for the first time,which provides a technological basis for factory production of D.bulbifera virus-free plantlets.
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Objective To provide a scientific basis for the creation and promotion of the rapid propagation of excellent Bupleurum chinense and study on botanical characters of test-tube plantlets and seed plantlets in B.chinense.Methods B.chinense with high effective medicinal ingredients and great stability of genetic characters was selected for rapid propagation,and some of botaninal characters between test-tube plantlets and seed plantlets were compared.Results The tiller number,the branch stem number,and the inflorescence number of the test-tube plantlets were significantly greater than those in seed plantlets.Conclusion Through the rapid propagation and the large-scale cultivation of test-tube plantlets of B.chinense,much more roots and seeds are obtained comparing with those of seed plantlets,while maintaining the excellent traits of B.chinense.
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Objective To study the condition for induction and differentiation of callus and propagation of adventitious buds in lamina,stem,and bud of Bupleurum chinese and establish a new method for rapid propagation.Methods In MS media added with different phytohormones,calli were induced from explants of lamina,stem,and floral bud of B.chinese,adventitious buds and adventitious roots were differentiated from calli of stem and floral bud,test-tube plantlets were formed.Results MS Medium added with 2,4-D 1.0 mg/L,KT 0.5 mg/L,and 6-BA 0.5 mg/L was suitable for calli induction of the lamina,stems,and floral buds.In medium added with 6-BA 1.0 mg/L,NAA 0.03 mg/L,CM 15% and CH 500 mg/L,the differentiation rate of floral buds callus was the highest.MS Medium added with 6-BA 1.5 mg/L,NAA 0.05 mg/L and CH 250 mg/L was suitable for propagation of test-tube plantlets,1/2 MS medium added with NAA 0.5 mg/L was suitable for rooting.Conclusion A great deal of test-tube plantlets could be differentiated and propagated rapidly by calli induced from stems and floral buds of B.chinese.Then the regeneration plantlets with normal growth and development are obtained.
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Objective For selecting and developing the excellent Bupleurum chinense to mass-produce seedlings and seeds in high quality.Methods B.chinense was picked from eight different areas,such as Lingchuan and Wanrong county in Shanxi Province,Longxi county in Gansu Province,and Shangluo in Shaanxi Province,etc.Rapid propagation was done.The testa of Zhongchai No.1 was scrapped or seeds were soaked in different phytohormones.The effects on germination rates of seeds were compared.ResultsThe optimum medium for bud propagation was B5supplemented with 1.0 mg/L 6-BA and 0.2 mg/L KT.The optimum medium for root induction of test-tube plantlets was 1/2 MS added with 0.1 mg/L NAA,0.5 mg/L IBA,and 1.0 mg/L DSC.The annual propagation coefficient of B.chinense plants was more than 1?108,and the survival rate of transplantation reached to 94%—97%.The phytohormone has little effect on seed germination of B.chinense,but scraping the testa could increase the germination percentage of seeds to 20% and shorten the germinating time greatly.Conclusion By tissue culture of excellent B.chinense,a great deal of plants and seeds could be produced in short time.By scrapping testa in a certain extent,the germination of seeds could be increased.
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Object To establish a calli culture system for the rapid propagation of Speiraea japonica L. f.. Methods Callus and shoot induction were carried out on MS, 6, 7 V or B 5 media with different parts of the plant such as stem tip, tender leaves and petiole as explants. Results A calli culture system was established for the rapid propagation of S. japonica. Conclusion MS cultural medium was found to be most suitable for calli induction. MS with 2.0 mg/L 2,4 D+0.3 mg/L KT can induce calli when the explants were used for the induction, with stem tips being the most satisfactory explant. Clusters of seedlings can be induced on MS+2 mg/L BA+0.1 mg/L NAA and when these seedlings were transferred to 1/2 MS +0.25 mg/L BA+0.5 mg/L NAA medium, root were developed to give young seedlings.
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Object\ To establish a plantlet regeneration system of Rubus idaeus L for the purpose to obtain a large number of high quality seedling in a short time Methods\ Stem apex and part of the stem were used as the explant and the optimal culture media and conditions were selected by orthogonal design Results\ An optimum culture medium for the induction of callus, adventitious bud and root was obtained which can be carried out in the laboratory with comparative ease and good repeatability Conclusion\ A basic medium + BA 0 2 mg/L+NAA 1 0～1 5 mg/L was most suitable for the induction of callus; a medium + BA 1 mg/L+NAA 0 1～0 2 mg/L+GA 3 6～8 mg/L+CH 300 mg/L was good for the induction of bud; a medium +BA 1 mg/L+NAA 0 1 mg/L+GA 3 2 mg/L was suitable for propagation of the bud; and the basic medium+IBA 0 1 mg/L+NAA 0 5 mg/L was good for the induction of root