Establishment of an in vitro Tissue Culture and Rapid Propagation System using Lonicera hypoglauca and Content Assessment of Total Flavonoids and Chlorogenic Acid

Lonicera hypoglauca is a traditional Chinese medicinal plant.In this study,the tender young leaves of L.hypoglauca were used for the first time as the explants to establish a rapid in vitro propagation and regeneration system.The results revealed that the optimal time for disinfection of the explants was 8 min and the optimal medium for callus induction was MS+2,4-D 4.0 mg·L^(-1)+sucrose 30 g·L^(-1),with an average callus induction rate of 86.67%.The optimal medium to induce differentiation of callus to bud was MS+6-BA 1.0 mg·L^(-1)+NAA 0.10 mg·L^(-1)+sucrose 30 g·L^(-1),with an average germination rate of 83.33%.The optimal medium to induce multiplication was MS+6-BA 1.5 mg·L^(-1)+NAA 0.05 mg·L^(-1)+sucrose 30 g·L^(-1),with a multiplication coefficient of 5.42.The optimal medium for root induction was 1/2 MS+NAA 0.15 mg·L^(-1)+activated carbon 0.3 g·L^(-1)+sucrose 15 g·L^(-1),with an average rooting rate of 91.11%.The survival rate of tissue-cultured seedlings in nutrient soil cultivation medium was as high as 100%.The total flavonoid content and chlorogenic acid content in the explant,callus tissue and regenerated plant were 1.83%,2.27%,1.33%and 2.77%,1.83%,1.74%respectively.This study provides novel insights into the rapid propagation and mass production of L.hypoglauca seedlings at an industrial scale and that it exhibits important application value and future prospects.

Tissue Culture and Rapid Propagation of Spathiphyllum kochii Engl. et Krause

[Objectives] This study was conducted on tissue-culture rapid propagation techniques of Spathiphyllum kochii Engl. et Krause. [Methods] With lateral buds of S. kochii as explants, the effects of such four basic media as MS, B5, Nitsch and Wpm and the ratio of two hormones(1, 2, 3 mg/L 6-BA and 0.1, 0.3, 0.5 mg/L NAA) on bud proliferation of S. kochii were studied by the complete test method, and the effect of the ratio of the two hormones(0.25, 0.50, 1.00 mg/L NAA and 1.0, 1.5 mg/L IBA) on rooting of S. kochii as well as the effect of substrate ratio(perlite and peat soil at 1∶9, 2∶8, 3∶7, 4∶6 and 5∶5) on its transplanting survival rate were also studied. [Results] The best basic medium for the rapid propagation of S. kochii was MS. The best hormone ratio suitable for bud proliferation was 2 mg/L6-BA+0.1 mg/L NAA, and the average number of buds proliferated at 45 d was 3.04. The average bud height was 2.05 cm; the most suitable medium for rooting was 1/2 MS+1 mg/L IBA+0.25 mg/L NAA, and its rooting rate was 100%; and the best transplanting substrate was 5∶5 perlite and peat. The soil ratio had a survival rate of 94%. [Conclusions] This study provides a theoretical basis for the improvement of the transplanting survival rate of test-tube S. kochii plantlets.

Tissue Culture Rapid Propagation and Transplantation Techniques of Anoectochilus roxburghii (Wall) Lind.

[Objectives]To screen the tissue culture rapid propagation formula suitable for each tissue culture stage of Anoectochilus roxburghii( Wall) Lind. [Methods] The stem segments of Fujian A. roxburghii were used as explant to study the tissue culture rapid propagation and transplantation techniques. The comparative experiment was carried out to study the effects of different hormone concentrations on the induction of stem segments,proliferation of cluster buds,rooting and seedling hardening of A. roxburghii,and study the effects of transplantation matrix on the transplantation of A. roxburghii. [Results]MS + 0. 5 mg/L NAA + 2 mg/L BA + 20 g/L sucrose + 6 g/L agar was suitable for induction of stem segments of A. roxburghii; MS + 0. 5 mg/L NAA + 2 mg/L BA + 1 mg/L KT + 25 g/L sucrose + 6 g/L agar was most suitable for proliferation of cluster buds of A. roxburghii; MS + 1. 0 mg/L IBA + 1. 0 mg/L NAA + 1 g/L activated carbon + 50 g/L mashed banana + 25 g/L sucrose + 6 g/L agar was most suitable for rooting and seedling hardening of A. roxburghii; using peat soil: fine sand( 3∶ 1) as transplantation matrix,the survival rate was the highest. [Conclusions] The experiment results are expected to provide references for factory production of A. roxburghii.

A Study on Tissue Culture and Rapid Propagation ofGinger

The buds of ginger grown in Tongdao Dong Autonomous County were used as explants, the tissue culture and rapid propagation of ginger were studied and the CMV (Cucumber mosaic virus) and TMV (Tobacco mosaic virus) of ginger tissue culture seedlings were detected. The results showed that MS+6-BA 4.0 mg/L+NAA 0.2 mg/L was the suitable medium for the proliferation induction and differentiation of ginger seedlings. This medium can realize the synchronous culture of ginger seedling proliferation and rooting, and one-step seedling re fining and transplanting, and the proliferation multiple can reach 3.00. The suitable rooting medium was 1/2MS+0.4 mg/L NAA. The survival rate of coir and vermiculite was the highest among the 4 culture mediums, reaching 100%. The survival rate of peat and cottonseed shell was very low, among which the ginger seedling transplanted with vermiculite grew more robustly than that transplanted with coir, with developed root system, more adventitious roots and less yellowing of leaves. No CMV and TMV were found in the tissue culture seedlings of ginger. The tissue culture seedlings can be used for the production of non-toxic ginger seedlings.

Research on Rapid Propagation of Gongshui Pomelo by Tissue Culture

[Objective] This study aimed to investigate the optimal medium and hor-mone combinations for efficient rapid propagation of Gongshui pomelo and analyze key technical measures in the tissue culture process. [Method] Stem tips and stem segments with buds were col ected from four varieties of pomelo adult trees as explants, to investigate the main effect and key regulatory factors of vegetative organs and tissue culture explants and to propose a series of measures to prevent and control microbial contamination. Final y, an efficient rapid propagation technology system of Gongshui pomelo was established. [Result] Spring shoot explants contained large amounts of auxin, cytokinins, gibberel ins and other growth regulators, which could be used for tissue culture with high bud generation rate and rapid growth. Different conditions led to various culture results. Specifical y, mature pomelo seeds should be generated on semisolid 1/2MS medium and transferred to solid MS medium for incubation. The propagation coefficient of stem segments with axillary buds was greater than that of stem tips, exhibiting significant differences. In ad-dition, the optimal hormone combination was 6-BA 0.5 mg/L ＋ NAA 0.5 mg/L, which significantly promoted the induction and differentiation of adventitious buds. [Conclusion] This study provided basis for basic research, production and application of pomelo germplasm resources.

Study on Tissue Culture and Rapid Propagation of Tilia amurensis

To explore the establishment of a tissue culture and rapid propagation system of Tilia amurensis,the effects of basic medium and concentrations and ratios of plant growth regulators on tissue culture and rapid propagation of T.amurensis were studied.The results showed that 1/2 MS medium was the most suitable proliferation medium,and the proliferation coefficient could reach 13.5 after adding 0.05 mg/L 6-BA and 0.03 mg/L IBA;MS medium was the most suitable medium for strong plantlets and rooting,and the best medium for strong plantlets was MS+0.1 mg/L 6-BA+0.1 mg/L IBA+0.03 mg/L GA_(3),with which the average plantlet height reached 5.15 cm;and the best rooting medium was MS+1.0 mg/L6-BA+0.05 mg/L NAA,with which the rooting rate was 93.3%and the number of roots was 5.7 roots.

Overview of Research on Tissue Culture and Rapid Propagation Technology of Pholidota

The genus Pholidota has good medicinal value,and is often over-excavated by humans.Coupled with its low natural reproduction rate,Pholidota is almost endangered.This paper summarized the tissue culture and rapid propagation technology of Pholidota in recent years,aiming to provide key technical support for resource protection and development of Pholidota and preliminary foundation and technical support for follow-up related research.

Study on Virus-free Culture and Rapid Propagation Technology of Xiangshu Series Varieties of Sweet Potato

[Objective] Sweet potato virus disease had a significant harm to the yield and quality of sweet potato, directly causing the degradation of sweet potato vari- eties and even the harvest failure. Therefore, the detection and removal of sweet potato virus and the establishment of rapid propagation method of sweet potato is of great significance to ensure the stable inheritance of excellent characters of sweet potato, prevent the spread of sweet potato virus and develop sweet potato industry. [Method] With Xiangshu series varieties of sweet potato, Xiangshu 15 and Xiangshu 19 as the research materials, a virus-free culture program was established for meristem tip apex tissue culture of different cultivars, and a rapid propagation method was developed for virus-free seedlings. [Result｜ On the basis of analysis on seedling emergence rate, the optimal addition scheme of plant hormones in the MS culture medium of Xiangshu 15 was 6-BA 3.0 mg/L ＋ NAA 1.0 mg/L, and the opti- mal plant hormone addition scheme for Xiangshu 19 was 6-BA 2.0 mg/L ＋ NAA 0.67 mg/L Under the developed rapid propagation system, the annual reproductive coefficient was up to 49 152, far higher than that （20 000） in field. IConclusionl Based on the actual production, combined with the meristem tip apex tissue culture, a comprehensive prevention and control measure was put forward, which included virus detection, early warning, removal and virus-free seedlings breeding, tt was of great strategic significance to improve the yield and quality of high-quality sweet potato and ensure the healthy development of sweet potato industry in China.

Study on Application Technology for Tissue Culture and Propagation of Tagetes patula L.

[ Objective] This study aimed to investigate the tissue culture and propagation technology in Tagetes patu/a L. [ Method] By using tissue culture tech- nology, different mass fractions of 6-BA and NAA were added to MS medium to compare the effect of different culture medium on the rapid propagation of T. patu/a L. [Result] Shoot tips or stem segments of T. patu/a L. were used as explants for tissue culture with an appropriate sterilization time of 8 min; differentiation effect of shoot tips was better than that of stem segments; callus generation rate was high with the high content of growth regulators; MS medium containing O. 1 mg/L NAA and 1.5 rag/L 6-BA was used for subculture proliferation with a subculture period of 4 weeks; rooting rate of plantlet was the maximum （97%） in 1/2MS medium containing 0.2 mg/L NAA, and the root system was relatively developed. [ Conclusion] This study provided technical support for the industrialized seedling breeding of T. patula L.

Primary Establishment of the Tissue Culture Technique and Regeneration System for Ornamental Lupinns polyphyllus

The purpose of this paper is to develop a system for tissue culture and rapid propagation of two ornamental lupins, Minaretie and Russell Prize. In view of screening out the better explant regeneration and suitable culture medium, through adding hormone 6-BA, NAA and 2, 4-D into MS and B5 basic culture medium, a series of experiments were carried out with the shoot tips, leaves, leaf petioles and stems from the asepsis seedling. The results showed that the shoot tips had favorableness on the rapidly propagation; MS＋6-BA 0.5 mg. L-1 for first generation, the induction rate of Minaretie and Russell Prize was 90.5% and 95.86% respectivdy; Minaretie had the highest propagation index （6.35） on MS＋6-BA 0.5 mg.L^-1＋NAA 0 mg-L^-1＋GA 30.8 mg. L^1＋AC 2 g. L^-1, but Russell Prize had the highest propagation index （7.24） on MS＋6-BA 0.5 mg.L^-1＋NAA 0.15 mg.L^-1＋GA3 1.0 mg.L^-1＋AC 0.5 g.L^-1; 1/2 MS＋NAA 0.25 mg.L^-1 was the best rooting medium. The ratios of getting roots of Minaretie and Russell Prize were 94.78% and 96.32%, respectively.

贵州野生葛枣猕猴桃组培快繁体系的建立

【目的】探明贵州野生葛枣猕猴桃组培快繁体系,为优异猕猴桃野生资源繁殖推广和保存利用提供参考。【方法】以葛枣猕猴桃单芽茎段为外植体,对比分析不同激素浓度组合下葛枣猕猴桃单芽茎段离体后诱导培养、增殖培养、生根培养和植株移栽情况,建立组培快繁技术。【结果】葛枣猕猴桃单芽茎段诱导和增殖培养均以MS培养基为基本培养基,添加不同种类和浓度的植物激素组合后腋芽生长情况不同,诱导率、增殖率和增殖系数呈先升后降趋势或下降趋势;生根培养以1/2 MS培养基为基本培养基,随植物激素的种类和浓度不同其生根情况存在一定差异性,在相同浓度(0.25 mg/L)不同种类植物生长素的生根培养基上,猕猴桃单芽茎段在添加IBA的培养基上生根情况最好。野生葛枣猕猴桃最佳单芽诱导培养基为MS+1.0 mg/L 6-BA+0.2 mg/L IBA,诱导率达87.33%;最佳增殖培养基为MS+1.0 mg/L 6-BA+0.2 mg/L NAA,增殖系数达4.37;最佳生根培养基为1/2 MS+0.25 mg/L IBA,生根率可达74.00%,主根数8.57条/株,根长3.93 cm;将生根≥3且带有3~5片叶的组培苗进行移栽,移栽成活率达88.60%。【结论】以贵州野生葛枣猕猴桃的带芽茎段为外植体,成功构建葛枣猕猴桃茎段组培快繁体系:诱导培养(MS+1.0 mg/L 6-BA+0.2mg/L IBA)、增殖培养(MS+6-BA 1.0 mg/L+NAA 0.2 mg/L)、生根培养(1/2 MS+0.25 mg/L IBA)和移栽培养(生根≥3且带有3~5片叶的组培苗进行移栽),为葛枣猕猴桃组培苗工厂化快速繁育种苗、苗圃资源保存和利用奠定了基础。

珠芽魔芋组织诱导快速繁殖技术研究

以珠芽魔芋球茎为外植体,开展组织诱导快速繁殖技术研究,以期为珠芽魔芋的规模化种植提供技术支撑。结果表明:最佳萌发培养基为MS+0.5 mg/L 6-BA+0.1 mg/L 2,4-D+30 g/L蔗糖+5 g/L琼脂,萌发率达100%;最佳愈伤组织诱导培养基为MS+2.0 mg/L 6-BA+0.5 mg/L GA3+30 g/L蔗糖+5 g/L琼脂,叶片和叶柄形成愈伤组织的诱导率分别为99.17%、92.50%;最佳不定芽诱导培养基为MS+1.0 mg/L 6-BA+0.4 mg/L NAA+25 g/L蔗糖+5 g/L琼脂,叶片和叶柄不定芽诱导率分别为94.17%、65.83%,单位愈伤组织分化芽数分别为3、1.87个;最佳生根培养基为1/2 MS+0.4 mg/L NAA+0.2 mg/L IBA+0.5 g/L碳粉+20 g/L蔗糖+5 g/L琼脂,生根率为97.50%,平均生根数为3.0条,平均根长为3.0 cm;炼苗10 d后,移栽到配比为表土∶椰砖营养土∶黑沙土=1∶2∶1的混合基质中,移栽成活率达到91.7%。

油橄榄幼嫩茎段组培快繁技术研究

以阿贝基纳优良单株幼嫩茎段为试验材料,通过对油橄榄外植体灭菌、基本培养基、生长调节剂等因素对油橄榄组织培养技术影响的研究,试验结果表明:使用75%乙醇30 s、0.1%HgCl_(2) 10 min进行外植体消毒处理时获得率最高,达36.15%;改良MS培养基适宜外植体诱导芽萌发,最佳增殖培养基为改良MS+6-BA 4.0 mg·L^(-1)+NAA 0.5 mg·L^(-1)+TDZ 0.5 mg·L^(-1),诱导率为65.3%,最佳生根培养基为1/2MS+ABT 0.6 mg·L^(-1)+IBA 0.2 mg·L^(-1),生根率90%,活性炭浓度最佳添加浓度0.5 g·L^(-1),蔗糖最佳添加浓度30 g·L^(-1)。

元宝枫组织培养与快速繁殖技术

【目的】元宝枫是我国特有的木本油料植物,非常适合开发利用,但其组培离体繁殖生根非常困难,建立合理的完整组培离体再生体系,在短时期内获得大量种苗和当年生带芽茎段,为该树种优良种苗规模化生产提供新途径,为选育元宝枫良种、种质资源保存与推广等提供研究基础。【方法】以元宝枫当年生带芽茎段和种子为试材,通过正交设计筛选法和单因素筛选法对其灭菌方式及腋芽诱导、继代增殖、生根培养基进行设计筛选。在MS、1/2MS和NN69基础培养基中分别添加不同质量浓度的6-BA、NAA和TDZ激素,通过不同配比找出元宝枫各阶段的最适培养基。【结果】全年中4月份为带芽茎段最佳采集时间,经洗衣粉溶液轻刷表面并冲洗半小时后,以75%酒精30 s+0.1%HgCl_(2) 10 min浸泡处理的灭菌效果最佳。再生体系各阶段最适培养基均为MS基础培养基,在其基础上再添加不同质量浓度的激素可以诱导嫩芽分化成不同器官。启动培养阶段,6-BA对腋芽诱导影响最大,最适培养基诱导率可达73.33%;继代增殖阶段,TDZ对不定芽分化影响最大,增殖系数最高可达4.31;生根阶段,采用单因素筛选法,发现添加0.2 mg·L^(-1) IBA元宝枫生根率和生根条数均为最高,达93.33%和4.3。【结论】元宝枫最适培养基为:1)启动培养:MS+0.06 mg·L^(-1) 6-BA和0.06 mg·L^(-1) NAA,诱导率为73.33%;2)增殖培养:MS+0.30 mg·L^(-1) TDZ和0.05 mg·L^(-1) NAA;3)生根培养:MS+0.2 mg·L^(-1) IBA。

连续继代培养下培养基降本措施对甘薯组培苗生长的影响

为降低甘薯(Dioscorea esculenta(Lour.)Burkill)组培苗快繁培养的生产成本,提高工厂化育苗的经济效益,在MS基本培养基的基础上,通过简化培养基组成、以自来水代替去离子水、以白砂糖代替蔗糖等降本措施,研究了连续继代培养下培养基组成简化或替代措施对甘薯组培苗生长的影响。结果表明,无论是初代培养还是继代培养,以白砂糖代替蔗糖,对甘薯组培苗生长无显著影响,但去除培养基中的微量元素、有机物质或以自来水代替去离子水,均对组培苗产生不利影响,导致株高降低、增殖系数减小、根系生长变差,且这种不利影响在经历长时间的继代培养后进一步加剧,组培苗生长更差。因此,在甘薯组培苗的工厂化生产中,可以用白砂糖代替蔗糖,降低组培生产成本,而其他降本措施不宜采用。

霞妃睡莲的组织培养及快繁技术研究

文章以霞妃睡莲(Nymphaea cv.Sunshine Princess)的根状茎为外植体,通过全面系统研究其初代培养、增殖、生根及移栽等不同阶段的培养条件,首次成功建立了睡莲的组培快繁技术体系。研究结果显示:固体培养基更适合霞妃睡莲的组织培养,且有利于提高睡莲的增殖率和人工操作效率;噻二唑苯基脲(TDZ)对睡莲不定芽的增殖诱导效果优于苄基腺嘌呤(BA),且两者一起使用具有协同效应,增殖率达到3.44且保持较好的生长状态;吲哚丁酸(IBA)对睡莲的生根诱导效果优于相同浓度的萘乙酸(NAA),活性炭的添加也有利于睡莲的生根诱导;适量的多菌灵能够提高睡莲组培苗的移栽成活率。该研究为睡莲种质资源的无菌保存及新品种的快速推广提供了有力的技术支持,对于加速睡莲种苗的规模化生产和推动国内外睡莲产业发展具有重要的现实意义。

亚洲百合‘穿梭’组培快繁体系的建立

以亚洲百合‘穿梭’为试材,采用植物组织培养的方法,研究了不同浓度激素对百合鳞片诱导、不定芽增殖、小鳞茎生根培养的影响,并筛选出适宜百合组培苗生长的移栽基质,以期为亚洲百合‘穿梭’的组培快繁体系的建立提供参考依据。结果表明:百合鳞片最佳诱导培养基为MS+0.5 mg·L^(-1)NAA+0.5 mg·L^(-1)6-BA+30 g·L^(-1)蔗糖+6 g·L^(-1)琼脂,诱导率最高可达74.44%,显著高于其他处理;最佳增殖培养基为MS+0.1 mg·L^(-1)NAA+1.0 mg·L^(-1)6-BA+30 g·L^(-1)蔗糖+6 g·L^(-1)琼脂,不定芽增殖数量最多,增殖系数为3.27;适合小鳞茎生根培养基为1/2MS+0.1 mg·L^(-1)NAA+0.3 mg·L^(-1)IBA+40 g·L^(-1)蔗糖+6 g·L^(-1)琼脂,平均生根数量为14.33,是其他处理的1.15~2.02倍;移栽后组培苗在草炭与珍珠岩为1∶1的基质配比中成活率高达87.50%,且组培苗长势最好。

尖叶茶藨子种子无菌苗再生体系的建立

以尖叶茶藨子成熟种子为试材,采用不同处理及消毒方式获得无菌苗,再以幼苗茎段为外植体,研究不同植物生长调节剂对丛芽诱导、增殖、壮苗及生根的影响。结果表明:500 mg/L GA_(3)浸泡处理有利于种子的萌发,发芽率可达66.7%,最佳消毒处理为75%乙醇30 s+0.1%HgCl_(2)消毒8 min,污染率为23.3%;最佳丛芽诱导培养基为MS+2.0 mg/L 6-BA+0.05 mg/L NAA,增殖倍数为7.5;最佳壮苗培养基为MS+0.1 mg/L 6-BA+0.05 mg/L NAA+50 g马铃薯泥;最佳生根培养基为1/2MS+2.0 mg/L IBA,生根率为100%,平均生根条数为6.33。炼苗后,移栽到泥炭土∶椰糠∶珍珠岩体积比为3∶1∶1的基质中,成活率达87.5%。该研究建立了尖叶茶藨子种子组培快速繁育技术体系,适用于尖叶茶藨子种苗的规模化生产和应用。

台湾榕组培快繁技术研究

【目的】建立台湾榕(Ficus formosana)组培高效繁育技术体系。【方法】以台湾榕茎段为试验材料,研究不同基础培养基对台湾榕茎段诱导率和萌芽数的影响;利用正交试验研究不同植物生长调节剂、活性炭及其质量浓度配比对丛生芽增殖系数、生根率和生长发育的影响;最后通过设置不同移栽基质对生根的组培苗开展移栽驯化研究,筛选能提高组培苗移栽成活率和质量的基质。【结果】同样添加1.0 mg/L 6-苄基腺嘌呤(6-BA)、0.5 mg/L激动素(KT)、0.2 mg/L萘乙酸(NAA),以WPM为基础培养基对台湾榕茎段的诱导效果优于MS培养基,培养30 d后其诱导率达97.13%,萌芽数为3.38。正交试验的极差结果显示,NAA是影响丛生芽增殖的主要植物生长调节剂,其次是6-BA,丛生芽增殖的最佳培养基为WPM+2.0 mg/L 6-BA+0.5 mg/L KT+0.1 mg/L NAA,接种60 d后,增殖系数达到5.77,丛生芽健壮;同时NAA也是影响丛生芽生根的主要因素,吲哚丁酸(IBA)、活性炭对生根率的影响差异不显著,丛生芽生根的最佳培养基为WPM+0.1 mg/L IBA+0.1 mg/L NAA+1 g/L活性炭,接种60 d后,生根率达到100%,组培苗健壮、根系发达、不易折断;组培苗最适移栽基质是河沙∶蛭石∶珍珠岩=1∶1∶1的组合基质,移栽30 d后,苗株成活率97.78%,长势很好。【结论】台湾榕茎段经过WPM+1.0 mg/L 6-BA+0.5 mg/L KT+0.2 mg/L NAA诱导,再利用WPM+2.0 mg/L 6-BA+0.5 mg/L KT+0.1 mg/L NAA和WPM+0.1 mg/L IBA+0.1 mg/L NAA+1 g/L活性炭进行丛生芽的增殖和生根,最后由河沙∶蛭石∶珍珠岩=1∶1∶1的组合基质进行移栽驯化,是最适合台湾榕组培高效繁育的技术体系。

梵净山石斛快速繁殖技术研究

为研究梵净山石斛快速繁殖技术,对其保育提供基础资料,本实验分别以梵净山石斛茎段、种子为外植体进行组培快速繁殖技术研究。结果表明,以茎段为外植体,最佳消毒方法为:75%乙醇消毒30 s,0.1%HgCl_(2)消毒13 min,腋芽诱导最佳组合为:1/2 MS/B_(5)+0.5 mg/L 6-BA+0.2 mg/L NAA,不定芽诱导和增殖最佳组合为:1/2 MS+0.5 mg/L NAA+1 mg/L 6-BA,以种子为外植体,种子萌发和原球茎诱导最佳组合为:MS+1.0 mg/L NAA+1.0 mg/L 6-BA+1 g/L活性炭,原球茎增殖最佳组合为:MS+0.5 mg/L NAA+2 mg/L 6-BA,无菌小苗增殖最佳为:MS+0.5 mg/L NAA+1.0/2.0/3.0 mg/L 6-BA。生根壮苗最佳组合为:MS+0.5 mg/L NAA+1/0.1/0.5 mg/L IBA。