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.

Tissue Culture and Rapid Propagation of Pedicels of Hemerocallis hybrida

[ Objective] This study aimed to develop tissue culture and rapid propagation methods for pedicels of HemerocaUis hybrida. [ Method] Tender pedicels of dwarf H. hybrida were used as experimental materials to sdect callus induction, subculture and rooting media for rapid propagation of H. hybrida.[ Result ] MS ＋ 2.0 - 3.0 mg/L 6-BA ＋ 0.2 - 0.3 mg/L NAA, MS ＋ 1.0 - 2.0 mg/L 6-BA ＋ 0.1 - 0.2 mg/L NAA, MS and 1/2MS ＋ 0.2 mg/L NAA were the appropriate me- dium for callus induction, subculture and rooting, respectively. [ Conclusion] The in vitro culture and clustered seedling rooting technology used in this study are effective methods for rapid propagation of H. hybrida, which provide technieal reference for industrialized production of H. hybrida.

Effect of Different Plant Growth Regulators on Callus Induction and in vitro Rapid Propagation of Wild Petunia Juss.

In this study,the seeds of wild Petunia Juss.were used as explants to investigate the optimal condition for tissue culture.Several different kinds and concentrations of growth regulators were adopted to produce more multiple bud clumps,callus or roots in this study.The experiments may provide experimental foundation for the rapid propagation technology and establishment of tissue culture system for wild Petunia Juss.

Study on the rapid propagation of Phalaenopsis hybrid

This paper studied the rapid propagation technology of Phalaenopsis hybrid by using the peduncle buds as the explants, and screened out a kind of culture medium, which was simple, rapid, available and high rate of propagation.

Cutting Rapid Propagation Technology for Antigonon leptopus in Tropical Areas

The shoots with good functional leaves were selected, from which cutting slips with the length of about 20 cm were cut, and 2 nodes were left for the ones with long sections. All leaves could be kept or 1/3 of each leaf could be cut. The cutting depth was that making the bottom node touch the substrate, and the cutting slips with short nodes should have 1/3 of the length insert in the soil. Each pot was planted with 2 - 3 cutting slips, and the substrate around the cutting slips was pressed by hands, followed by enough watering. And then white plastic bags were used to cover the pots, which were then placed in the place with the shading rate of 75%. The method had low costs but good landscape effect, promising with good development prospects, which could provide references for the promotion of out- of-season rapid propagation technology of Antigonon leptopus in tropical areas.

Rapid Propagation of Rhynchostylis retusa in Vitro

An efficient regeneration system of Rhynchostylis retusa was established to provide technical reference for the application of tissue culture tube seedlings in production.The mixtures of callus and protocorm from aseptic germination were used as explants.The optimal media of each stage was selected for callus proliferation,protocorm occurrence and growth,rejuvenation and rooting via a single,complete combination and orthogonal experiment.The results showed that the optimal medium for callus proliferation,protocorms occurrence and growth was 1/2 Murashige and Skoog(MS)medium adding 50 g·L^(−1) banana puree,0.1 mg·L^(−1)α-naphthaleneacetic acid(NAA),1.5 mg·L^(−1)6-benzylaminopurine(6-BA)and 1.0 mg·L^(−1) kinetin(KT)with 17.33 proliferation coefficient of callus and 19.63 occurrence coefficient of buds after 90 days.Then the buds occurred from protocorm were cultured on 1/2 MS medium including 100 g·L^(−1) banana puree,1.0 mg·L^(−1) NAA,2.0 mg·L^(−1)6-BA and 0.05 mg·L^(−1) KT,in which the proliferation coefficient of callus was 10.32 and occurrence coefficient of buds reached 17.87.In the further subculture,the same medium was simultaneously used for callus proliferation,protocorm occurrence and bud growth.The plantlets developed roots in 1/2 MS medium containing 70 mL·L^(−1) coconut water and 1.5 mg·L^(−1) NAA with 100%rooting rates after 90 days.The survival rate was more than 90%after domestication and transplantation.This regeneration protocol will provide technique foundation for protecting wild resource and developing artificial cultivation.

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 and Rapid Propagation Technology of Artemisia selengensis sp.in Yangxin County

[Objectives]This study was conducted to culture virus-free tissue culture plantlets of mid-maturing green-stalk Artemisia selengensis sp.varieties in Yangxin County.[Methods]With bud tips of A.selengensis in Yangxin as explants and MS as the basal medium,screening and proportioning of plant growth regulators were performed to establish a virus-free and rapid propagation system for mid-maturing green-stalk varieties of A.selengensis in Yangxin.[Results]The optimal callus induction medium,adventitious shoot differentiation medium,adventitious shoot elongation medium and rooting medium for explants were MS+6-BA 1.0 mg/L+NAA 0.5 mg/L+sucrose 25 g/L+agar 7 g/L,MS+6-BA 0.1 mg/L+NAA 0.1 mg/L+sucrose 25 g/L+agar 7 g/L,MS+6-BA 0.02 mg/L+NAA 0.02 mg/L+sucrose 25 g/L+agar 7 g/L and 1/2 MS+NAA 0.05 mg/L+sucrose 25 g/L+agar 7 g/L,respectively,and the seedling rate reached more than 95%.The culture conditions were as follows:temperature(25±2)℃,relative humidity 85%,illumination intensity 3000 lx,and illumination time 14 h/d.[Conclusions]This study has important practical significance for the purification and rejuvenation and large-scale industrial breeding of Yangxin A.selengensis seedlings.

Research on Rapid Propagation of Gongshui Pomelo by Tissue Culture

[Objective]This study aimed to investigate the optimal medium and hormone 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 collected 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.Finally,an efficient rapid propagation technology system of Gongshui pomelo was established.[Result]Spring shoot explants contained large amounts of auxin,cytokinins,gibberellins 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.Specifically,mature pomelo seeds should be generated on semi-solid 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 addition,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.

A SUPPLEMENTARY STUDY OF ANISOTROPIC PLASTIC FIELDS AT A RAPIDLY PROPAGATING PLANE－STRESS CRACK－TIP（I）

The results in Ref. [1] are not suitable for the cases of β≥2. For this reason, byusing the methods in Ref [1] and Ref [2], we derive the general expressions ofamsotropic plastic fields at a rapidly propagating plane-stress crack-tip for both thecases of β=2 and β>2 .

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.

In vitro Propagation of Codonopsis pilosula"Fengdang"

[Objectives]The paper was to establish in vitro propagation of an excellent germplasm of Codonopsis pilosula"Fengdang".[Methods]The effects of different plant growth regulators on callus induction and redifferentiation were tested by using sterile stem explants in vitro that germinated from seeds of superior"Fengdang"plant.[Results]The optimum medium for callus induction was MS+0.5 mg/L NAA+1.0 mg/L 16-BA.The optimal medium for bud differentiation was MS+0.1 mg/L NAA+0.5 mg/L 6-BA,and that for rooting was 1/2 MS+0.2 mg/L NAA+0.2 mg/L IBA.By using the media mentioned above,the rates of callus induction,bud differentiation and rooting reached 91%,100%and 97%,respectively.[Conclusions]The rapid propagation system of regenerated plants in vitro established in this study lays a foundation for the popularization and industrialized seedling of excellent germplasm of"Fengdang".

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。

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。

玫红百合组培小鳞茎瓶内膨大影响因子的研究

玫红百合(Lilium amoenum)是具有玫瑰甜香味的珍稀百合资源。为探索玫红百合组培小鳞茎瓶内膨大的最适培养条件及培养基配方,以直径0.4~0.6 cm的玫红百合组培小鳞茎为材料,将其接种于添加不同质量浓度蔗糖或蔗糖与萘乙酸(naphthalene acetic acid,NAA)、6-苄氨基嘌呤[N-(phenylmethyl)-9H-purin-6-amine,6-BA]、吲哚丁酸(indole butyric acid,IBA)、多效唑(paclobutrazol,PP333)组合的Murashige&Skoog(MS)培养基中,设置16 h光照/8 h黑暗和全黑暗2种光照环境,培养45 d后统计小鳞茎的膨大系数和直径增大倍数。结果表明,在本试验条件下,随着蔗糖及激素质量浓度的升高,玫红百合组培小鳞茎膨大系数及直径增大倍数基本呈先增加后下降的趋势,其中:在添加蔗糖质量浓度为50~120 g/L处理中,以90 g/L蔗糖处理的膨大效果最好,小鳞茎嫩绿有光泽,抱合度好;当50~120 g/L蔗糖与0.1~0.7 mg/L NAA/6-BA、0.5~4.0 mg/L IBA或5~40 mg/L PP333协同作用时,以70 g/L蔗糖和1.0 mg/L IBA处理的小鳞茎膨大效果最好,其膨大系数与直径增大倍数最高,分别为3.63和1.83;高质量浓度的蔗糖与PP333协同作用会显著抑制小鳞茎的膨大。在16 h光照/8 h黑暗环境下培养的玫红百合组培小鳞茎较全黑暗环境下长势好,其膨大系数及直径增大倍数高,但黑暗环境下生根效果更好。本研究可缩短玫红百合田间培育成球时间,为种质资源保护提供了科学依据。

铁皮石斛和红龙石斛杂交后代繁育技术研究

对铁皮石斛和红龙石斛杂交后代的繁育技术进行了研究。结果表明:以铁皮石斛为母本与红龙石斛杂交,成功率较高,达96.7%;其杂交后代的最佳播种培养基为HB01(含花宝1号3.0 g)+6-BA 0.5 mg/L+NAA 0.1 mg/L,最佳增殖培养基为HB01+BA 0.6 mg/L+NAA 0.1 mg/L+香蕉50 g/L,最佳壮苗培养基为HB01+BA 0.3 mg/L+NAA 0.2 mg/L+香蕉50 g/L,最佳生根培养基为HB01+NAA 0.1 mg/L+IBA 0.1 mg/L+蛋白胨0.25 g/L;杂交后代组培苗的较佳移栽基质为刨花和5号树皮(直径0.8~1.2 cm);将稳定生长的杂交后代种苗连同基质在室外栽培,可营造热带园林景观,且种苗生性强健,较耐强光和干旱;扩繁杂交后代的优秀单株时宜选用幼嫩茎段作为外植体,并用0.1%氯化汞溶液对外植体消毒5 min,外植体萌芽的适宜培养基为HB01+6-BA 0.6 mg/L+NAA 0.1 mg/L。

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。

金柑组织培养与快速繁殖体系的建立

为了建立金柑组织培养与快速繁殖体系,以金柑胚根、子叶、茎段、叶片等不同外植体为材料,采用器官发生型以及短枝扦插型为主要培养途径,进行金柑组织培养,研究不同浓度植物生长调节剂对金柑快速繁殖体系建立的影响以及愈伤组织诱导和分化的最适条件,筛选出诱导愈伤组织形成不定芽的最佳外植体与最适培养基。结果表明,金柑诱导愈伤组织形成不定芽的最佳外植体为幼嫩的茎段,诱导金柑子叶形成愈伤组织的最佳培养基为MS+6-BA 1.5 mg/L+NAA 1.0 mg/L+3%蔗糖+0.7%琼脂,诱导子叶愈伤组织分化不定芽的最佳培养基为MS+6-BA 1.0 mg/L+IBA 0.1 mg/L+3%蔗糖+0.7%琼脂。