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 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.

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.

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.

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.

Rapid Propagation of Three Herbal Species in a Newly Developed Temporary Immersion System

[ Objeelive] This study aimed to develop a rapid propagation method in a novel temporary immersion bioreactor system （TIS） for herbal plantlets com- pared with solid culture method. [ Method ] Three herbal species, including Dendrobium candidum Wall. Ex IJndl （D. candidum）, Anoectochilus roxburghii （Wall.）Lindl. （A. roxburghii） and Lilium davidii var. unicolor （L. davidii）, were used and tested by TIS against solid culture method. [Result] When the two culture methods were compared, the multiplication rate of D. candidum in TIS was found to be 1 ： 24.71, which was 6.55 times to those of solid culture. The multiplication rate of A. roxburghii was higher than those of the solid culture, but the plantlets was poorer than those of solid culture at the last phase in bioreactor culture, under the culture condition tested. The multiplication rate in TIS ofL. davidii was 1：17.23 whilst the rate was only 1：4.45 on solid culture, resulting larger bulbs than those in the solid culture. [ Conclusion] The TIS designed in our study could provide a potential mean for industrial production of plandets. However, the parameters vary greatly among different species, and it is to be optimized according to plant species.

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。

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。

中药知母组培快繁体系的建立

为了建立知母高效组织培养及快速繁殖体系,将知母的叶片、根茎和幼嫩花葶分别接种于9种组合培养基上,筛选最佳的外植体材料;以MS培养基为基本培养基,通过不同种类激素和浓度的配比试验,筛选出知母离体组培快繁不同阶段的培养基组合;测定不同种类激素和浓度组合培养基下炼苗移栽成功的知母组培苗芒果苷含量。结果表明,幼嫩花葶为诱导愈伤组织最佳外植体,诱导知母幼嫩花葶产生愈伤组织的培养基组合为MS+1.0 mg/L KT+0.5 mg/L NAA,出愈率为79.40%;诱导愈伤组织产生不定芽的培养基组合为MS+0.5 mg/L 6-BA+0.5 mg/L NAA,生芽率为100%;诱导不定芽生根培养的培养基组合为MS+0.5 mg/L NAA,生根率为100%,且在此培养基组合下炼苗移栽的知母组培苗芒果苷含量最高。

粉葛赣葛2号组培快繁体系建立与优化

为了加快横峰县本地粉葛品种赣葛2号优质种苗的繁育与推广,本研究以带芽茎段为材料,探讨无菌体系建立、不定芽诱导及增殖、试管苗生根、驯化移栽等过程,建立并优化了粉葛赣葛2号组培快繁体系。结果表明,初代芽诱导效果最优培养基为MS+6.5 g/L琼脂+30 g/L蔗糖+1.5 mg/L 6-BA+1 mg/L NAA,平均诱导率为80.55%;继代培养效果最优培养基为MS+6.5 g/L琼脂+30 g/L蔗糖+1 mg/L 6-BA+0.5 mg/L NAA,平均诱导率为91.66%;适合生根的培养基为MS+6.5 g/L琼脂+30 g/L蔗糖+0.5 mg/L NAA,生根率达84.38%。移栽基质配方为V泥炭土∶V蛭石∶V珍珠岩=3∶2∶1,组培苗成活率达100%。

向海一号桑组培快繁技术优化

为提高桑树育苗效率,以向海一号桑茎段为外植体,开展了腋芽诱导培养、增殖培养、生根培养研究,优化了其组培快繁体系。结果表明:腋芽诱导培养最适培养基为MS+6-BA3.0 mg·L^(-1)+IAA0.1 mg·L^(-1),诱导率达78.3%;增殖培养的最适培养基为4/5MS(大量)+6-BA0.1 mg·L^(-1)+NAA0.15 mg·L^(-1),30 d左右平均增殖倍数达5.12;生根培养的最适培养基为1/2MS(大量)+1/4MS(铁盐)+IBA1.0 mg·L^(-1),生根率达82.5%左右,8 d开始生根,28 d后即可移栽炼苗。

尤加利茎尖组培快繁体系的建立

尤加利是当前国内外鲜切花市场上重要的切叶材料,筛选其茎尖生长及生根的最佳培养基,建立其组培快繁体系,可为尤加利组培苗工厂化生产提供参考。以尤加利无菌茎尖为试验材料,分析不同基础培养基、6-BA、NAA组合及其浓度处理下尤加利组培生长和生根情况,并对组培苗进行移栽试验。结果表明:基本培养基是影响尤加利组培苗褐化和增殖的主要因子,NAA是影响尤加利组培苗生根的主要因子;1/4 WPM+6-BA1.3 mg/L+NAA0.1 mg/L组合处理下尤加利茎尖组培褐化率(2.63%)较低,增殖率(115.79%)和生根率(97.37%)均达到最大,移栽后幼苗健壮,叶色浓绿,移栽成活率可达73.71%。可见,1/4 WPM+6-BA1.3 mg/L+NAA0.1 mg/L+蔗糖15 g/L+琼脂6.0 g/L作为最佳培养基,可建立高效的尤加利茎尖组培快繁体系。

基于茎段外植体的茶树组培苗快繁技术研究

【目的】目前,茶树组培苗繁育存在诱导出芽效率不高、繁殖系数较低等问题。因此,为提高茶树种质资源保存繁育效率,丰富保存手段与技术,建立一种高效、快速、实用的基于茎段外植体的茶树组织培养高效快繁技术体系非常必要。【方法】选用来自安徽省六安市舒城县鲍家村群体种的一个国家级品种‘报春1号’作为研究试材,对该品种进行组织培养快繁技术的探索,包括消毒、定芽诱导、不定芽诱导(I期和II期)、芽伸长、以及生根等阶段。【结果】离体快繁:最佳定芽诱导培养基为MS+3 mg·L^(-1)6-苄基腺嘌呤(6-BA),诱导率达82.2%;不定芽增殖I期最佳培养基为MS基础培养基(MS)+2 mg·L^(-1)6-BA+0.2 mg·L^(-1)萘乙酸(NAA),诱导率为77.78%,平均芽数为5.4个;不定芽增殖II期最佳培养基为MS+1 mg·L^(-1)6-BA+0.3 mg·L^(-1)NAA+2 mg·L^(-1)噻苯隆(TDZ),诱导芽数最多(平均值为10.43个),芽苗呈绿色,叶片舒展;不定芽伸长最佳培养基为MS+0.8 mg·L^(-1)6-BA+0.08 mg·L^(-1)NAA,伸长率为63.33%;不定根诱导培养基以1/2MS培养基+3 mg·L^(-1)IBA效果较佳。【结论】本研究结果建立了一种基于茎段外植体的茶树组织培养高效快繁技术,将对优良茶树品种的保存繁育及其高效利用提供理论基础。

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。

粉防己组织培养体系的建立和细胞悬浮培养初探

为促进粉防己的资源保护和利用,以茎段和愈伤组织为材料建立植物组织培养体系,并利用愈伤组织细胞进行悬浮培养。结果表明,以茎段为外植体直接分化不定芽的最佳培养基为MS+6-BA 1.5 mg/L+NAA 0.2 mg/L,60 d不定芽诱导率为100%,单个外植体分化不定芽数16个,90 d时分化数可达28个,该配方同时适用于诱导愈伤组织分化不定芽;不定芽生根以培养基MS+IAA 0.2 mg/L为佳,30 d的生根率达92.5%,平均主根长7.78 cm,炼苗移栽15 d的成活率达88.00%;利用愈伤组织细胞进行悬浮培养,可在较短的培养周期获得可观的生物积累量,干培养物提取液生物碱定性检测为阳性。

乌奴龙胆组培快繁体系研究

为了建立乌奴龙胆的组培快繁体系,该研究选用采自西藏自治区拉萨市堆龙德庆区羊达乡境内的乌奴龙胆植株进行组培快繁实验。实验对乌奴龙胆的合适外植体、HgCl2溶液最佳消毒时间、最佳不定芽诱导分化培养基、最佳生根培养基进行了筛选。结果表明:带节茎段适合作为乌奴龙胆组培实验中的外植体,叶片块不适合作为外植体;HgCl2溶液最佳消毒时间为6min,平均未污染率为66.67%±3.33%,平均成活率为63.33%±3.33%;最佳不定芽诱导分化培养基为:P7处理组(MS+0.1g/L肌醇+30g/L蔗糖+5g/L琼脂粉+2.0mg/L 6-BA+1.0mg/L IAA pH5.6),平均芽增殖倍数为2.47±0.12,平均芽长度为17.07±1.56mm,平均未玻璃化率为80.00±5.57%;最佳生根培养基为:T5处理组(1/2MS+0.1g/L肌醇+30g/L蔗糖+5g/L琼脂粉+0.3mg/L NAA+0.3mg/L IBA pH5.6),平均生根率为100%,平均生根数为3.63±0.09条,平均根长度为39.03±0.30mm,平均根健康率为85.27%±1.22%.

马大杂交相思组培快繁技术研究

选取马大杂交相思优树的当年生带嫩芽的枝条进行腋芽诱导、增殖、生根等研究,在保持马大杂交相思较高的增殖倍数前提下,缩短增殖周期,提高移栽存活率,繁育出健壮的相思组培苗,为杂交相思快速繁殖和推广奠定良好的基础。研究表明,最适宜的外植体消毒方式为75%酒精浸泡10 s,并用0.1%氯化汞处理10 min,存活率达到83%;最佳诱导培养基为MS+6-BA 0.2 mg/L+蔗糖30 g/L,萌芽率达93.17%;最佳增殖培养基为改良MS+6-BA 0.5 mg/L+NAA 0.1 mg/L+蔗糖30 g/L,增殖倍数可达3.32,增殖周期缩短至25d;最佳生根培养基为MS+IBA 0.5 mg/L+IAA 0.1 mg/L+蔗糖15 g/L,生根率达91.33%;苗木驯化移栽30 d后成活率为97.8%。