苦荞胚性愈伤组织诱导与植株再生研究

以苦荞子叶和下胚轴为外植体,进行了不同浓度激素组合的MS和SH固体培养基对胚性愈伤组织诱导及植株再生的研究。结果发现,MS培养基比SH培养基更有利于胚性愈伤组织诱导;2,4-D是诱导愈伤组织的有效激素,KT能有效促进胚状体的形成;下胚轴和子叶都能有效诱导出胚性愈伤组织和再生植株。下胚轴在MS 1.5mg·L-12,4-D 1.5mg·L-1BA培养基,子叶在MS 2mg·L-12,4-D 0.5~1.5mg·L-1BA上能高效诱导出愈伤组织;愈伤组织在MS 2mg·L-12,4-D 0.1mg·L-1KT培养基中继代,能有效诱导胚性愈伤组织;来自下胚轴的胚性愈伤组织在1/2MS 2.0mg·L-1BA 0.5mg·L-1KT 0.1mg·L-1NAA培养基上能够高频再生出芽,来自子叶的胚性愈伤组织在1/2MS 1.0mg·L-1BA 0.1mg·L-1KT 0.1mg·L-1NAA培养基上芽诱导率较高;MS 1mg·L-1NAA是适宜的再生苗生根培养基。     

兰属"小金童"组织培养研究

兰属小金童(Cymbidium Golden elf. 'Sundust')茎尖组织培养研究表明:茎尖外植体在不添加植物生长调节剂的花宝1号培养基上诱导产生原球茎;原球茎在花宝1号3g/L + 6-BA 1.5mg/L的液体培养基上培养,能形成快速增殖的丛生原球茎;不同的培养方式对原球茎的增殖效果有显著差异;原球茎在不附加植物生长调节剂的1/2MS或花宝1号培养基上分化成无根小苗,然后在花宝1号+NAA 0.2mg/L+IBA 0.6mg/L培养基上诱导生根,进而再形成完整植株.     

兰属“小金童”组织培养研究

兰属小金童(Cymbidium Golden elf.‘Sundust’)茎尖组织培养研究表明:茎尖外植体在不添加植物生长调节剂的花宝1号培养基上诱导产生原球茎;原球茎在花宝1号3g/L 6-BA 1.5mg/L的液体培养基上培养,能形成快速增殖的丛生原球茎;不同的培养方式对原球茎的增殖效果有显著差异;原球茎在不附加植物生长调节剂的1/2MS或花宝1号培养基上分化成无根小苗,然后在花宝1号 NAA 0.2mg/L IBA 0.6mg/L培养基上诱导生根,进而再形成完整植株。     

几种植物生长调节物质对大花蕙兰组培原球茎增殖的影响

采用KC 培养基, 分别添加不同浓度的BA、KT、NAA 及KT+NAA 组合, 对大花蕙兰初代培养已分化出的原球茎进行了继代培养。结果发现:①BA 可加快原球茎的增殖速度, 但浓度超过0.5 mg/L 后增殖速度有所下降, 且原球茎变小呈暗绿色。②KT 的促进增殖效果好于BA, 但浓度超过1.0 mg/L 时, 增殖速度也有所下降, 原球茎变小。③NAA 促进原球茎增殖的效果明显好于BA、和KT, 但浓度大于1.0 mg/L 后增殖不再加快, 而且部分原球茎有变褐现象。④在KT 0.5~0.7 mg+NAA 0.7~1.0 mg/L 范围组合的KC 培养基上, 大花蕙兰原球茎增殖的速度和质量是比较理想的。考虑到较低的细胞分裂素和植物生长素有利于植物增殖过程中遗传性的稳定, 选择KT0.5 mg+NAA 0.7 mg/L 的KC 培养基作为大花蕙兰原球茎增殖培养基。     

外部因子对蝴蝶兰叶片原球茎状体发生的影响

对影响蝴蝶兰叶片原球茎状体 (PL B,Protocorm - like- body)发生和植株再生的外部因子进行了研究。结果表明 :BA是决定原球茎状体发生的主要因子 ;苹果汁、香蕉汁和椰子汁明显促进原球茎状体的形成 ;在 MS+BA5mg/L+椰子汁 15%的培养基中 ,蝴蝶兰叶片原球茎状体的诱导率可达 6 0 %以上 ;活性炭可有效防止外植体叶块变褐死亡 ;多效唑 1.5mg/L可促进再生植株根的形成 ,使叶片变厚变短变绿 ,使试管苗移栽成活率达 95%以上     

非洲紫罗兰叶片体细胞胚培养及快繁技术研究

以非洲紫罗兰叶片为材料进行胚状体诱导及快繁技术研究.结果表明:.在MS NAA0.1mg/L BA0.1 mg/L 2,4-D1.0 mg/L的培养基上培养15d利于诱导胚性细胞分化,起始黑暗培养5～10d可提高胚性细胞分化率;在MS BA0.05～1mg/L的培养基上可诱导胚状体大量发生;在MS NAA0.1mg/L十BA0.1 mg/L的培养基上能够获得茎芽快速增殖;在1/2MS NAA0.01mg/L的培养基上可以生根.     

不同培养基上白芨的种子萌发与幼苗形态发生

对无菌条件下白芨种子在不同培养基上的萌发及其幼苗形态发生进行了研究.结果表明:在白芨种子萌发过程中,种胚先转绿,然后在种胚远离残余胚柄的一端突破种皮,形成原球茎.在原球茎顶端分化出叶原基,并逐渐发育成叶片.在原球茎下部表面存在透明的毛状物,推测这些毛状物与幼根根毛是同功的.白芨种子萌发最适培养基为1/2 g·L-1 +1.0 mMS NAA.添加低浓度的外源细胞分裂素(6-BA或KT)与NAA组合均严重抑制种子的萌发,种子萌发率较低,形成的圆球茎较小,原球茎分化出的小苗也较细弱.白芨生根壮苗最适培养基为1/2 g·L-1 NAA+2.0 g·L-1活性炭+1.0 mg·L-1 +1.0 mMS GA3.生根壮苗培养基中加入活性炭和GA3有利于根的生长和壮苗,且植株长势健壮.炼苗基质为腐殖土和蛭石(1∶1),成活率可达90%以上.     

玉米幼胚高效再生系统的建立

建立了玉米幼胚高效再生系统。经研究发现,苏玉1号、农大3138、农大108的幼胚培养在含有2,4－D（2.5mg/L）的IM培养基上后,大多数幼胚能愈伤化并增大,形成基部相连、上部分开的微芽结构;微芽结构在转移到BM培养基上后,形成小植株;进一步转移到RM培养基上,它们长根并形成完整杆株。玉米幼胚高效再生植株与下列因素有关：玉米基因型、幼胚大小、幼胚长芽至分化时间、6－BA、IBA、Gelrite。不同品种玉米再生能力有显著差异,幼胚大小在1－2mm之间再生能力强,幼胚长芽至分化时间4－6d最好。激素6－BA浓度在0.5-0.6mg/L之间有利于微芽形成小植株,IBA浓度在0.6-1.0mg/L促进生根。Gelirte可代替琼脂粉用于玉米生根。     

红豆草体细胞胚胎发生的激素调控及L—脯氨酸对它的促进作用

在 LS 附加1mg/1 BA+1mg/l KT 的培养基上,红豆草(Onobrychis viciaefolia Scop.)无菌苗的下胚轴切段产生淡黄色的愈伤组织。愈伤组织转移到 LS 附加1mg/l BA 的培养基上,诱导体细胞胚胎发生,而在 LS 附加1mg/l KT 的培养基上抑制体细胞胚胎发生。同时,发现红豆草胚性愈伤组织中游离脯氨酸的含量仅为非胚性愈伤组织的2/5。向培养基中加入L-脯氨酸可以促进红豆草体细胞胚胎发生。最适浓度为1000mg/l。     

濒危植物紫斑牡丹胚离体培养

以MS为基本培养基 ,添加不同浓度的BA、NAA及其组合 ,对濒危植物紫斑牡丹的胚进行体外培养。实验结果显示在没有添加任何激素的培养基上 ,离体胚生长类似种子萌发时胚的生长模式 ,子叶扩大伸长 ,达到约种子大小时停顿 ,根伸长生长极其显著 ;BA促进子叶膨大生长 ,当BA浓度增大时根生长受到抑制 ;NAA促进愈伤组织形成 ,抑制根生长。 0 .1mg·L- 1BA促进紫斑牡丹胚的胚轴、根生长 ,有利于幼苗形成。     

从石刁柏原生质体培养再生植株

石刁柏,又名芦笋(Asparagus officinalisL.)是百合科天门冬属植物。其栽培品种含有丰富的维生素类及蛋白质。同时,石刁柏对于某些疾病有一定的药效,因此它已成为人们所喜爱的一种高级营养蔬菜。国外已有不少关于石刁柏试管苗繁殖的报告,但至今只有Bui Dang Ha等从石刁柏枝状叶分离的原生质体得到愈伤组织,并由此愈伤组织诱导获得了再生植株。此后,未见在石刁柏的原生质体培养方面再有新的工作。在本文中,我们利     

Plant Regeneration from Protoplasts of Ligusticurn wallichii Franch

The hypocotyls of the embryoid derived plantlets of Ligusticum wallichii Franch were used for protoplast preparation. Protoplasts were obtained with the enzyme mixture containing 1.5% Onozuka R-10, 0.3% Macerozyme R-10, 0.5% Snailase, 5 mmol/l CaCl2, 1 mmol/l KH2PO4, 0.6 mol/l manitol, at pH 5.6–5.8 and 27℃. Protoplasts were cultured in a modified MS liquid medium containing 1 mg/l 2,4-D + 0.5 mg/l 6- BA. The first divisions were found after twelve days, and the dividing cells formed cell colonies of 0.5–1 mm after about fourty days. When they were transferred to MS agar medium (with half quantity of macronutrients) supplemented with 2,4-D (0.5mg/l) and 6-BA(0.5mg/l), they grew into calli. At last, on the medium without any phytohormones, the growing calli differentiated embryoids which developed into plantlets with many green leaves and roots.     

墨兰的无菌播种和植株再生

墨兰的无菌播种结果发现,在不添加细胞分裂素的培养基上,种子可以发芽,但只有原球茎和根状茎产生;不可能进一步分化成苗,只有在含有不同激素成分的MS或KnudsonC培养基上,才有可能诱导芽的分化,其中以附加6-BA 0.5-1.0mg/L+NAA 0.1mg/L诱导效果最佳,在附加6-BA 2.0mg/L+NAA 0.4mg/L的MS培养基中,能加速芽的增殖,根状茎转入含有相同激素成分的液体增殖培养基中振荡培养,可大大提高芽的分化速率。添加0.5%活性炭对芽的分化有明显增效作用。在附加NAA 0.2mg/L的MS培养基中;幼苗的生根效果最佳。     

液体悬浮培养促进铁皮石斛原球茎高效诱导、增殖的研究

用正交设计方法对铁皮石斛原球茎具有高效增殖影响的激素（BA,NAA,KT,马铃薯汁）以及配比进行研究,结果表明：以茎段为外植体在培养基Ms＋BA2．0mg／L＋NAA2．0mg／L＋马铃薯汁10％＋糖3％,具有很好的原球茎诱导作用,50d后诱导率达95．20％;原球茎在1／2MS＋BA2．0mg／L＋NAA1．0mg／L＋KT0．5mg／L＋糖3％的培养基上,以液体悬浮培养,原球茎增殖达49．032g／50d。     

Plant regeneration from callus protoplasts of the forage legume Astragalus adsurgens Pall.

A reproducible release of viable protoplasts was obtained from friable calli of Astragalus adsurgens. Protoplasts underwent sustained divisions and formed cell colonies when cultured in either liquid or agarose-solidified Kao and Michayluk (1975) protoplast medium (KM8P) supplemented with 1.5 mg/l 2,4-D, 0.5 mg/l BA and 0.5 M glucose. Compared to liquid culture, agarose bead culture improved division frequency effectively, the two culture systems showing a plating efficiency of 0.8±0.5% and 6.5±0.7%, respectively. Upon transfer to Murashige and Skoog (1962) medium (MS) with 1–2 mg/l BA, alone or in combination with NAA or 2,4-D at 0.1 mg/l, the protoplast-derived calli produced complete plantlets through somatic embryogenesis. The maximum percentage of calli producing somatic embryos (52.5± 2.2%) occurred on MS medium containing 0.1 mg/l NAA and 1 mg/l BA, whereas the maximum number of calli regenerating plantlets (64.7±6.2) was obtained on MS medium with 0.1 mg/l NAA and 2 mg/l BA. Received: 25 April 1997 / Revision received: August 1997 / Accepted: 2 September 1997     

Plant Regeneration from Protoplasts of Peucedanum Terebinthaceum (Fisch.) Fisch. ex Turcz

Calll with many embryogenic cell colonies were produced from segments of seedlling of Peucedanum terebinthaceum (Fisch.) Fisch. ex Turcz. which were cultured on the 1/2MS agar medium (with half quantity of macronutrients) containing 1 mg/l 2,4-D. Cell suspension culture with high percentage of embryogenic cell colonies was established from the calli shaking in liquid medium. The cell suspension culture was used for protoplast preparation. Protoplasts were obtained with the enzyme mixture containing 1.5% Onozuka R-10, 0.3% Macerozyme R-10, 0.5% Snailase, 5 mmol/l CeCl2, 1 mmol/l KH2PO4, 0.6 mol/l mannital at pH 5.8 and 25℃. Cultured in a modified MS liquid medium containing 1 mg/l 2,4-D+ 0.5 mg/l zeatin, the protoplasts emered division after four days, and formed cell colonies of 0.5–1mm after about forty days. When transfered to 1/2 MS liquid medium supplemented with zeatin (0.5 mg/l), the cell colonies differentiated in to embryoids, then developed into plantlets with many green leaves and roots on the 1/2 MS agar medium devoid of phytohormones.     

Development of a plant regeneration system based on friable embryogenic callus in the ornamental Alstroemeria

 Stem segments of seedlings from two Alstroemeria breeding lines, cultured on media supplemented with 4 mg/l 2,4-dichlorophenoxyacetic acid and 0.5–1.0 mg/l 6-benzylaminopurine (BA), initiated soft callus, which became compact after subculture on a medium with only 0.5 mg/l BA. Friable embryogenic calli were initiated from compact callus on a medium supplemented with 10 mg/l picloram. Proembryos developed from friable embryogenic calli via embryos into plants after subculture on medium supplemented with 0.1 mg/l BA. The proembryos formed friable embryogenic calli again after culture on medium supplemented with 10 mg/l picloram. The total time needed to regenerate a complete plantlet from friable callus was approximately 6 months. This system for the production of embryogenic material is considered to have valuable applications for genetic transformation in Alstroemeria. Received: 22 April 1999 / Revision received: 16 July 1999 · Accepted: 20 July 1999     

Plant Regeneration from Protoplasts of Coriandrum sativum

Calli produced from stem segments of seedling of Coriandrum satwum which were cultured on MS agar medium containing NAA 1.0mg/L. The embryogenic cell colony suspension was estabilished on MS liquid medium containing NAA 1.0mg/L%2,4-D 0.2mg/L+BA 0.5 mg/L. The cell suspension culture was used for protoplast preparation. Protoplasts were obtained in the enzyme mixture containing 2.0% Onozuka R-10, 1.0% pectinase, 0.5% snailase, 0.5% dextran sulfate potassium Salt, 0.6mol/L mannital CPW solution at pH 5.8 and 25℃. Cultured in a KM8P liquid medium containing NAA 1.0mg/L+2,4-D 0.2mg/L+6-BA 0.5 mg/L, glucose 0.4mol/L and CM 20mi/L; the protoplasts entered the stage of derision after three days, cell clusters formed in 10 days and calli formed after about 50 days. When the calli were transferred to MS agar medium containing many growth substances, they differentiated into embryoids, and then developed into plantlet with many green leaves and roots on the 1/2 MS agar medium.     

Plant regeneration from protoplasts of peppermint (Mentha piperita L.)

Summary Enzymatically isolated leaf-derived protoplasts of peppermint (Mentha piperita L.) were cultured in modified B5 medium containing 1 mg/l NAA, 0.4 mg/l BA, 0.5% sucrose, 0.5 M mannitol and 0.1% Gelrite (first medium). After 30 d culture at 25°C in the dark, protoplasts formed colonies consisting of about 100 cells. Gelrite medium blocks were transferred into liquid medium to promote further growth. Colonies of 0.5 mm transferred to 0.2% Gelrite solidified medium (same components as first medium) formed green calli (1–2 mm) under incubation in the light. Green calli transferred to differentiation medium (B5, 0.1 mg/l NAA, 5 mg/l BA, 2% sucrose, 0.2 M mannitol, 0.2% Gelrite) developed shoot buds after 3–4 weeks. Whole plants were recovered following rooting of shoots in B5 medium without hormones.Abbreviations BA 6-benzylaminopurine - NAA -naphthaleneacetic acid - KIN kinetin - ZEA zeatin - CPW cell and protoplast wash solution - B5 Gamborg et al. (1968) mineral elements - MS Murashige and Skoog (1962) mineral elements     

不同激素对匙叶芋芽的诱导与增殖的影响

取日本引进的Spathiphyllum sp.根茎外植体接种在不同激素配比的MS培养基上,结果以MS BA 2mg/l KT 1mg/l诱导芽的效果最好;通过附加不同种类的细胞分裂素,不同浓度的BA,不同生长素的试验,证明MS BA 1mg/l IAA 0.1mg/l组成较有利于芽的增殖;将芽移入生根培养基,15天左右长出根,形成完整植株。