大薯类原球茎的离体诱导及再生体系的建立

采用正交试验设计方法,以大薯带节茎段为外植体,离体诱导类原球茎并建立大薯类原球茎的再生体系,以解决愈伤组织分化成苗和试管苗移栽成活率低的难题。结果表明：以带节茎段为外植体诱导类原球茎的最适培养基为MS（含3×Ca2＋）＋1.0 mg·L-1 6-BA＋0.2 mg·L-1 NAA＋0.1%PVP＋3%蔗糖,诱导率高达93.33%;类原球茎增殖的最适培养基为MS＋4mg·L-1 6-BA＋80 mg·L-1 Ad＋0.1%PVP＋3%蔗糖;类原球茎生根的最适培养基：1/2MS＋0.10 mg·L-1 NAA＋0.1%PVP＋3%蔗糖。将诱导得到的生根类原球茎植株进行炼苗,移栽基质珍珠岩：蛭石=2：1,移栽成活率可达到95%。     

Rapid Multiplication and Induction of Early In Vitro Flowering in Dendrobium Primulinum Lindl

Dendrobium primulinum is an important epiphytic orchid. A successful protocol for mass multiplication and early in vitro flowering was developed. Immature embryos of 4 week after pollination exhibited about 96% germination within 30 days of culture on MS medium containing sucrose (3%) (w/v), NAA and BA (6 and 9 μM) in combination. Protocorm-like bodies (PLBs) formed from the germinating seeds on the germination medium. Rooted plantlets were formed within 2-3 wk on MS medium containing sucrose (3%), NAA and BA (3 and 12 μM in combination) where about 29 shoot/buds produced per cycle of 4 wk interval. The well rooted plantlets produced 4-5 floral buds per spike when they were maintained on MS medium containing sucrose (3%), fresh apple juice (10%) (v/v) for four wk followed by on MS medium freed of apple juice but enriched with NAA and BA (3 and 12 μM respectively). The hardened plantlets were transferred to community potting mix where the about 80% transplants survived after two months of transfer.     

报春石斛的组织培养与快速繁殖

1植物名称报春石斛(Dendrobium primulinum Lindl)。2材料类别茎尖和茎段。3培养条件芽诱导培养基:(1)MS 6-BA2.0mg·L-1(单位下同) NAA0.1;(2)MS 6-BA2.0 NAA0.5。芽生长培养基:(3)MS 6-BA0.2。芽增殖培养     

Detection of Cymbidium mosaic virus in Protocorm-Like Bodies in Dendrobium Sonia using One-Step RT-PCR

Dendrobium orchids are clonally mass propagated by tissue culture techniques. However, Cymbidium mosaic virus is prevalent in vegetatively propagated Dendrobium spp. Therefore, a sensitive virus detection method is an important requirement for the production of CymMV-free orchid plants and control of virus spreading in the orchid industry. The presence of CymMV was screened in in vitro PLBs of Dendrobium Sonia using a one-step RT-PCR with specific primers to amplify a 858 bp fragment of a CymMV coat protein gene and flanking regions. Results were compared with those obtained from a conventional indirect ELISA. Only 22% samples showed the presence of CymMV in indirect-ELISA as compared to 78% in RT-PCR. Thus, the one-step RT-PCR appears to be a more sensitive method than indirect-ELISA for detecting CymMV in PLBs.     

In vitro Propagation and Conservation of Atropa acuminata Royle ex Lindl -An Indigenous Threatened Medicinal Plant

A micropropagation method has been developed for multiplication and conservation of Atropa acominata by induction of axillary shoot proliferation from shoot tips and nodal explants using Murashige and Skoog (MS) medium supplemented with BAP ( 1 mg I^-1) and IBA (1 mg I^-1). Revised tobacco (RT) medium with IAA (1 mg I^-1) was found most suitable for shoot elongation. Rooting was highest on full strength RT medium containing IBA (1 mg I^-1). In vitro raised plantlets were hardened and transferred to soil.     

超声处理对霍山石斛类原球茎悬浮培养细胞生理代谢的影响

为了了解超声处理对霍山石斛类原球茎产生的生理效应,研究了超声波功率和超声时间对霍山石斛类原球茎悬浮培养细胞生长以及多糖和蛋白质合成的影响;分析了培养基中碳、氮利用、细胞内活性氧水平以及蔗糖转化酶、硝酸还原酶、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和过氧化物酶（POD）活性的变化。结果表明,适当功率和时间的超声处理（300 W3 min）能显著促进霍山石斛类原球茎的增殖,最大细胞干重为34．6g·L^-1;明显促进培养基中碳和氮的利用;显著提高胞内可溶性多糖、可溶性蛋白质和H2O2的含量;细胞内蔗糖转化酶、硝酸还原酶以及SOD、CAT和POD的活性明显升高。适当的超声波处理能促进霍山石斛类原球茎的生长发育和提高细胞的生理活性。     

玫瑰石斛的组织培养及快速繁殖

1植物名称玫瑰石斛（Dendrobium crepidatum Lind1．ex Paxt．）。 2材料类别无菌萌发的种胚苗茎节或茎段。 3培养条件种胚诱导培养基：（1）MS＋NAA0．5mg．L^-1（单位下同）;（2）1／2MS＋马铃薯提取物20％。茎节或茎段分化培养基：（3）MS：（4）1／2MS。生根和壮苗培养基：（5）1／2MS＋IBA0．2-0．4＋活性炭0．1％;（6）MS＋NAA1＋6-BA0．5;     

In vitro propagation of Dendrobium macrostachyum Lindl.--a threatened orchid

In vitro propagation of Dendrobium macrostachyum, a threatened and endemic species was achieved through nodal explants. The nodal explants were cultured on Murashige and Skoog (MS) basal medium and MS medium supplemented with N6-benzyladenine (BA-2.22, 4.44 and 8.88 microM), Kinetin (KN-2.32, 4.65, and 9.29 microM) and Coconut water (CW, 5, 10 and 15%) individually or in combination with 2.69 microM alpha-naphthalene acetic acid (NAA). Axillary shoots were induced directly from nodal explants in medium containing BA, KN or CW. Optimal shoot induction (6 shoots/explant) was attained from nodal explants cultured on medium supplemented with 15% CW. Well developed shoots rooted at an average 5 roots per shoot in half strength MS medium devoid of any growth regulators.     

羚羊石斛的离体快速繁殖

1植物名称羚羊石斛(DendrobiumcochliodesSchltr.)。2材料类别种子。3培养条件萌发培养基:(1)VW;(2)VW+椰子乳;(3)1/2MS;(4)1/2MS+椰子乳;(5)N6;(6)N6+椰子乳;(7)N6+椰子乳+活性炭2g·L-1。继代增殖和分化成苗培养基:(8)N6+椰子乳+6-BA1.0mg·L-1(单位下同)+NAA0.2;(9)N6+椰子乳+6-BA0.2+NAA0.1;(10)N6+椰子乳+6-BA0.2+NAA0.1+活性炭2g·L-1;(11)花宝1号(美国Haponex公司产品,N∶P∶K=7∶6∶19)1.5g·L-1+花宝2号(美国Haponex公司产品,N∶P∶K=20∶20∶20)1.5g·L-1+椰子乳+6-BA0.2+NAA0.1。生根壮苗培养基:(12)N6+NA…     

鱼籽兰(Chloranthus errectus)的离体繁殖

本研究通过几种培养基对鱼籽兰外植体培养,选择出了有效的外植体和适合的培养基,并摸索出了有效的移载和栽培技术,培育出了外植体繁殖的成熟开花植株,为该植物提供了一条新的快速繁殖途径,对同属其它植物的快速繁殖具有参考意义。     

大苞鞘石斛的离体培养与快速繁殖

1植物名称大苞鞘石斛(Dendrobium wardianumWarner). 2 材料类别种子(授粉6个月). 3培养条件萌发培养基:(1)1/2MS 6-BA0.2mg·L-1(单位下同) NAA 0.2.继代增殖与分化培养基:(2)1/2MS;(3)1,2MS 6-BA 0.2 NAA 0.5:(4)1/2MS 6-BA 0.5;(5)CHB 6-BA 0.5;(6)1/2MS NAA 0.5.生根壮苗培养基:(7)CHB IBA 1.0;(8)花宝1号3g·L-1 蛋白胨2 g-L-1 IBA 1.0:(9)B5.以上萌发培养基和继代增殖与分化培养基均附加2.0%蔗糖,生根壮苗培养基附加3.0%蔗糖,椰乳均为100 mL·L-1,7.6 g·L-1琼脂固化,pH 5.4～5.6.培养温度为(23±1)℃,光照强度30～40μmol·m2·s-1,光照时间16h·d-1.     

铁皮石斛茎段离体快繁

1　植物名称　铁皮石斛 (Ｄｅｎｄｒｏｂｉｕｍｃａｎｄｉｃｕｍ)。2　材料类别　幼嫩茎段。3　培养条件　基本培养基为ＭＳ。芽增殖培养基 :(1 )ＭＳ +ＮＡＡ 0 .1～ 0 .5ｍｇ·Ｌ- 1 (单位下同 ) +6 ＢＡ 0 .5～ 2 .0 ;壮苗培养基 :(2 )ＭＳ无激素培养基 ;生根培养基 :(3 ) 1 /2ＭＳ +ＩＢＡ 0 .1。以上各种培养基蔗糖浓度均为 3 .0 % ,ｐＨ 5 .4～ 5 .8,培养温度 (2 5 +2 )℃ ,连续光照培养 ,光照强度 1 5 0 0ｌｘ。4　生长和分化情况4.1　丛生芽诱导　剪取幼嫩茎段 ,按常规表面灭菌后 ,切成约 2 .0ｃｍ左右的切段 ,每一切段应至少保留 …     

莲的离体快速繁殖技术

以莲(Nelumbo nucifera)授粉后18天的莲子胚芽为外植体,通过初代培养、继代培养和炼苗移栽,建立了莲离体快速繁殖体系。结果表明,将胚芽外植体诱导出无菌苗的最适初代培养基为MS固体培养基添加0.5 mg·L–1 6-BA、0.5 mg·L–1NAA、30 g·L–1蔗糖、0.5 g·L–1活性炭和0.8 g...     

柠檬香蜂草离体快速繁殖(简报)

以柠檬香蜂草带腋芽幼嫩茎段为外植体,MS为基本培养基,对其进行离体快繁研究。结果表明,在Ms＋6-BA1.0mgCL＋IBA0．5mg／L培养基上诱导产生不定芽的效果最好;在MS＋6．BA0．5mg/L＋IBAO．1mg／L分化培养基上分化率达3～4倍;在1／2MS＋IBA0．2mg／L生根培养基中正常发根,且根系粗壮。     

白檀离体快繁技术

以白檀(Symplocos paniculata)幼嫩茎段为实验材料, 通过对启动培养、增殖、生根培养及移栽的影响因子进行研究, 初步建立了白檀的组织培养体系。结果表明: 白檀外植体最适灭菌方案为0.1%升汞3分钟, 无菌苗获得率达81%; 最适初代启动培养基为1/2MS+30 g∙L^-1蔗糖+8 g∙L^-1琼脂, 出芽率达86.83%; 增殖最适培养基为1/2MS+1.0 mg∙L^-1 6-BA+0.02 mg∙L^-1 IBA+30 g∙L^-1蔗糖+8 g∙L^-1琼脂, 增殖系数达3.57; 最适生根培养基为WPM+0.5 mg∙L^-1 IBA+0.5 mg∙L^-1 NAA+20 g∙L^-1蔗糖+2 g∙L^-1 AC+8 g∙L^-1琼脂, 生根率达93%; 炼苗后, 移入园土:草炭土=1:1 (v/v)的基质中, 成活率达83%。     

In Vitro Propagation Of Four Watsonia Species

The genus Watsonia has a number of species with potential to be developed as new ornamental crop plants, but to date there are no reports on in vitro propagation of any member of this genus. Seeds from four Watsonia species, Watsonia gladioloides, Watsonia lepida, Watsonia laccata, and Watsonia vanderspuyiae were decontaminated and germinated on one-tenth strength MS media without hormones or sucrose. Shoots were induced from seedling hypocotyl segments when both an auxin [α-naphthaleneacetic acid (NAA)] and cytokinin [N⁶−benzylaminopurine (BA)] were present in the medium, while root and leaf explants failed to produce shoots. Multiplication of axillary shoots was greatest when only BA (0.5 mg l⁻¹) was added to the medium. Shoot explants propagated in a ‘liquid-shake’ culture exhibited greater growth rates than those on agar-solidified medium, but shoot production varied between species. Meristemoids were induced in all species, but no significant trend was found between growth index (GI) and meristemoid formation, suggesting that reduction in GI may not necessarily be a prerequisite for producing meristemoids. Corm formation was inconsistent and storage organs could only be induced in one of the four species, W. vanderspuyiae. This occurred best at 25°C with 3% sucrose and an agar level of 15%. Indole-3-acetic acid (IAA) and NAA at 1 mg l⁻¹ significantly increased mean number of roots per shoot explant on all four species, while indole-3-butyric acid (IBA) was more effective when applied at 0.1 mg l⁻¹. Plantlet survival ex vitro was negatively affected when NAA and 2,4-dichlorophenoxyacetic acid (2,4-D) were used to root shoot explants for all species. In W. laccata, all auxin treatments [IAA, IBA, NAA, phenylacetic acid (PAA), and 2,4-D] at a concentration of 1 mg l⁻¹ significantly reduced ex vitro survival of plantlets. Successful micropropagation of Watsonia is an important step in the further development of this genus as a horticultural crop.     

云南龙竹的快速繁殖和离体保存研究

本研究以干冷保存（-20 ℃,相对湿度15%）150 d 的云南龙竹（Dendrocalamus yunnanicus）颖果为实验材料,通过种子萌发建成丛芽无菌无性系进行快繁和离体保存。研究表明,丛芽增殖的最佳培养基为MS + 6-BA 2 mg/L + NAA 0.2 mg/L,蔗糖浓度以3%为佳;生根培养的最佳培养基为MS + IBA 1 mg/L + NAA 1 mg/L,以单芽诱导生根为好;离体保存丛芽的最适培养基为MS + 6-BA 0.5 mg/L + NAA 0.2 mg/L;当培养温度由室温 (25±3)℃降低至(20±3)℃和12 ℃时,其继代周期可由原来的2 个月分别延长至4 个月和8 个月。在组织培养过程中发现白化苗或叶色变异现象。     

In Vitro Clonal Propagation and Medium Term Conservation of Brahmi [Bacopa monnieri (L) Wettst]

A protocol has been developed for in vitro clonal propagation leading to conservation of Bacopa monnieri (L) Wettst, a medicinal plant of high commercial potential with legendary reputation as a memory vitalizer. Single node explants when cultured on Murashige and Skoog’s medium supplemented with BA (0.2 mg l^-1), showed active shoot proliferation (22.2 shoots/ explant in 8 weeks) without callus formation. Rooting was achieved on the same medium. The generated shoots could also be conserved for 12 months with high survival rate (up to 100%). The regenerants upon transfer to soil showed no morphological variation as compared with the donor plants. The medium optimized in the present study was applied for culture establishment and conservation of a total of 15 Brahmi accessions procured from different regions.     

铁核桃离体培养与快速繁殖

以优良单株‘纳雍-1’的单芽茎段为外植体,建立了铁核桃（Juglanssigillata）离体培养与快速繁殖的体系。结果表明,附加6-BA1．0mg·L-1 ＋活·IgK（AC）3．0g·L-1的DKw培养基适宜铁核桃腋芽诱导;适宜铁核桃芽增殖的培养基为DKW＋6-BA1．0mg·L-1 ＋IBA0．02mg·L-1,40d后增殖系数可达7．33;试管苗的茎尖和茎段均可用于增殖培养;一步生根法（低浓度的生长素IBA持续诱导）不利于铁核桃试管苗嫩茎生根;采用二步生根法,生根率最高可达71．73％,其中,不同IBA浓度、暗培养时间、蔗糖浓度和AC含量对试管苗嫩茎生根影响显著,铁核桃试管苗在附]sulBA5,0mg·L-1的1／4DKW培养基中暗培养12d,再转移到不含IBA的1／4DKW培养基（附加AC 3g-L-1和蔗糖20g·L-1）中生根效果最好;生根试管苗采用珍珠岩和营养土两步炼苗,60d后成活率达到87．50％。     

天女木兰的离体培养和快速繁殖

1植物名称天女木兰(Magnolia sieboldii),又名天女花、小花木兰、山牡丹. 2材料类别种子. 3培养条件种胚萌发培养基:(1)B5 6-BA 0.5 mg·L-1(单位下同) NAA 0.05.继代繁殖培养基:(2)B5 TDZ 0.3 NAA 0.03 VC(抗坏血酸)500.0或AC(活性炭)500.0;(3)B5 6-BA 2.0 NAA 0.2 VC 500.0或AC 500.0.继代苗生根培养基:(4)1/2B5 NAA0.5 AC 1000.0 VB(维生素B)20.0.愈伤组织诱导培养基:(5)B5 6-BA 2.0 2,4-D 0.5 VC 500.0.以上培养基均附加3.0%蔗糖、0.6%琼脂,pH 5.8～6.0,121～124℃下灭菌20 min.光照度1000 lx左右,光照14～16 h·d-1,培养温度白天(23±2)℃,夜间20℃.