Callus initiation and subculture of Taxus chinensis

IntroductionTaxolisacompIexditerpenoidsecondaryproductofthegenushauSthatwasapprovedfortreatmentagainstovarianandbreastcancersandshowsprom-iseagainstothercancers.DuetotherelativescarcityofthefewnaturaIresourcesandthelowyieldoftaxol,thesupplyoftaxolisrestricted,limitingexpansionofcIinicaItrialsandtreatmentavailabiIity.lnterestinalternativemethodsfortaxoIproductionhasbeenintensifying.CellcuItureprovideaconvenientsystemforstudyingthebiosynthesisoftaxol.Itmaybeaviablealternativefortaxolproduct(…     

湿地松愈伤组织的诱导研究初报

取湿地松成熟胚作为外植体,在添加不同BA、NAA浓度配比的MS培养基上,诱导愈伤组织的最适培养基是MS+BA0.5mg/L+NAA0.5mg/L+0.6%琼脂+25g/L蔗糖。实验同时证明,NAA是胚诱导愈伤组织的必需因子。     

白芨的离体培养

为快速生产大量优质白芨种苗,以白芨种子为外植体进行离体快繁研究。结果表明:MS基本培养基可促进白芨种子的快速无菌萌发,当6-BA的浓度为1.0 mg/L时可以促进幼苗的快速分裂生长,添加了2 mg/L的2,4-D的培养基可有效促进愈伤组织的形成。种子苗原球茎经切割后在MS+2 mg/L 2,4-D+0.5 mg/L6-BA的培养基上愈伤组织诱导率为85%,1.0 mg/L的6-BA和0.15 mg/L的NAA配比可以诱导丛生芽的生长和增殖,添加了70 g/L香蕉泥和0.5 mg/L NAA的1/2MS培养基可有效促进根的生长。无菌发芽的种子苗移栽率可以达90%,而试管苗移栽的成活率较低,但随离体培养时间的延长有所提高。     

海南粗榧外植体灭菌与愈伤组织诱导

对海南粗榧茎段外植体灭菌和愈伤组织诱导实验研究表明,以海南粗榧温室扦插苗的嫩茎作为外植体,70%酒精1min,0.1%氯化汞溶液12min表面消毒处理效果最好;最适愈伤组织诱导培养基为MS+0.5mg/LNAA+0.5～1.0mg/L BA+2.5～4.0mg/L 2,4～D。     

北乌头组培快繁技术体系初步研究

本文对北乌头组培快繁技术体系进行了初步研究,同时比较了不同外源激素组合对北乌头外植体诱导的影响,其结果表明:北乌头愈伤组织诱导最佳外植体为叶片,丛生芽诱导最佳外植体为带芽茎段及茎尖,愈伤组织诱导最佳培养基为MS+1.0mg/L6-BA+1.0mg/L2,4-D,丛生芽诱导最适合培养基为MS+1.0mg/L2,4-D+0.2mg/LNAA,继代培养以WPM+1.0mg/L6-BA+0.5mg/LIBA效果最为显著,增殖率在4倍以上;北乌头生根最佳培养基为1/2MS+0.1mg/LNAA,生根率可到90%。     

优系欧李茎叶愈伤组织诱导与植株再生

以欧李春季萌发的幼茎、叶片为外植体诱导愈伤组织的产生,结果表明:叶片愈伤组织培养以改良MS为基本培养基,附加NAA 1.0 mg/L IBA 0.5 mg/L BA 0.2 mg/L效果较好;茎段愈伤组织培养以改良MS附加2,4-D0.5 mg/L NAA 0.3 mg/L BA 0.15 mg/L诱导愈伤组织效果好;愈伤组织再诱导不定芽以1/3 MS附加BA2.0 mg/L IBA 0.01 mg/L培养基效果佳。     

两种外源激素在白掌组织培养不同阶段的剂量研究

文章研究了不同浓度的细胞分裂素6-BA对白掌外植体诱导、愈伤组织增殖、不定芽增殖及生长素IBA对诱导生根的影响。结果表明：初代培养基以1／2MS CM100ml／L 蔗糖30g／L 琼脂6g／L 6-BA 1.0mg／L效果最好；在培养基MS NAA 0．1mg／L 蔗糖30g／L 琼脂6g／L中添加6-BA 1.0mg／L愈伤组织和不定芽的增殖倍数最高；在培养基1／2MS 活性炭2g／L 蔗糖30g／L 琼脂6g／L中添加2．0mg／L IBA可显著提高生根率。     

白刺花胚性愈伤组织诱导及体细胞胚发生

【目的】探讨不同植物生长调节剂对白刺花胚性愈伤组织诱导的作用,以及培养基中氮源和无机盐浓度对白刺花体细胞胚发生和植株再生的影响,以期建立白刺花体细胞胚发生、发育及调控技术体系,为白刺花种苗快速繁殖体系建立及遗传转化研究提供参考。【方法】以白刺花叶片为外植体,研究生长调节剂2,4-D(1.0、2.0、3.0、4.0 mg ·L -1 )、NAA(0、0.5、0.8、1.0 mg ·L -1 )、6-BA(0.2、0.5、1.0、2.0 mg ·L -1 )和TDZ(0、0.2、0.5、1.0 mg ·L -1 )组合对胚性愈伤组织诱导,及NAA(0、0.2、0.5 mg ·L -1 )、6-BA(0、0.5、1.0 mg ·L -1 )和TDZ(0、0.2、0.5 mg ·L -1 )组合对体细胞胚发生的调控作用,筛选最优生长调节剂组合;并研究培养基中KNO 3和NH 4NO 3比例对体细胞胚发生的作用,及MS培养基中无机盐浓度(1/5MS 、1/4MS、1/3MS、1/2MS)对体细胞胚萌发的影响,筛选最佳的体细胞胚发育及成熟萌发条件。【结果】白刺花叶片外植体胚性愈伤组织诱导适宜培养基为MS + 2,4-D 3.0 mg ·L -1 + NAA 0.5 mg ·L -1 + 6-BA 0.2 mg ·L -1 + TDZ 1.0 mg ·L -1 +蔗糖40 g ·L -1 +琼脂7.0 g ·L -1 ,诱导率为42.0%。采用MS基本培养基时,最佳的体细胞胚发生培养基为MS + NAA 0.5 mg ·L -1 + 6-BA 1.0 mg ·L -1 + TDZ 0.5 mg ·L -1 +蔗糖40 g ·L -1 +谷氨酰胺100 mg ·L -1 +琼脂7.0 g ·L -1 ,体细胞胚发生率为78.46%,总胚数为对照的3.6倍;MS培养基中,KNO 3浓度提高1倍、NH 4NO 3降至1/2时,体细胞胚发生率可提高至91.33%,总胚数为采用MS基本培养基时的1.4倍;1/3MS培养基有利于体细胞胚的萌发,萌发率为82.75%,幼苗长势良好,单株平均鲜质量为76 mg,幼苗驯化移栽1个月后成活率达90%以上。【结论】白刺花叶片接种于添加2,4-D、NAA、6-BA和TDZ不同组合的诱导培养基上,可脱分化形成愈伤组织或胚性愈伤组织,2,4-D浓度对愈伤组织形态和质地有较大影响。TDZ有利于体细胞胚的形成,适宜浓度的生长素与细胞分裂素组合及硝态氮和铵态氮的比例对体细胞胚的形成和发育具有调控作用,降低MS无机盐浓度可提高体细胞胚萌发率,本试验体系的再生植株移栽成活率达90%以上。     

驱蚊草组培快繁技术研究

以驱蚊草幼嫩茎段为外植体,将其接种在MS BA2.0mg/L 2,4-D0.2mg/L培养基上进行培养,15~20天嫩茎诱导形成愈伤组织,诱导率达90%,然后将愈伤组织接种于MS BA1.0mg/L 2,4-D0.5mg/L的培养基中进行培养,15天左右开始形成丛生芽,最后将成苗接种于1/2MS NAA0.5 mg/L培养基中,7~10天生根率可达100%。     

香蕉薄切片愈伤组织再生体系的建立研究

以巴西香蕉品种的薄切片为外植体高频地诱导出愈伤组织,并诱导出了不定芽,获得了再生植株。在实验过程中,利用正交实验中筛选出愈伤组织诱导的最佳培养基为MS+2,4-D 4.0 mg.L-1+KT 1.0 mg.L-1+NAA 1.0 mg.L-1+活性炭200 mg.L-1,诱导率可达100%,愈伤组织增殖的最佳培养基是MS+2,4-D 2.0 mg.L-1+6-BA 0.5 mg.L-1+NAA 0.5 mg.L-1+活性炭100 mg.L-1,继代3-4次的愈伤组织在6-BA浓度为5.0 mg.L-1的分化培养基上能分化出不定芽,继代10次之后愈伤组织在继代培养基上开始出现褐化现象,愈伤组织增殖明显受到抑制,褐化的愈伤组织逐渐死亡。     

透骨草组织培养的研究

以透骨草的嫩茎为材料,进行了愈伤组织诱导、愈伤组织分化、不定芽生根、试管苗的移栽、试管苗扦插、试管苗移植的研究,成功的建立起透骨草的嫩茎无性系。结果证明：1/2MS＋BA0.5mg/L＋2,4-D1.2 mg/L＋NAA0.2-0.8 mg/L为嫩茎愈伤组织的的理想培养基;1/2MS＋AgNO31.2 mg/L＋BA0.6 mg/L＋NAA0.1 mg/L为愈伤组织和不定芽分化的理想培养基;1/3MS＋IAA0.4 mg/L为不定芽生根培养和生根继代培养的理想培养基;炉灰渣是试管苗移栽和扦插的理想基质。移植试管苗后期出现生长旺盛、长势整齐的性状,翌年春季出现萌发早5d、根系相当于野生植株2倍的性状。     

芦苇植株再生体系优化

在前人工作基础上利用禾本科芦苇成熟种子为外植体建立了植株再生体系。芦苇成熟种子经2%洗涤灵溶液洗涤,70%乙醇处理30s,20%＂84＂消毒液处理25min后接种到MS＋4%蔗糖（pH 5.5）的种子萌发培养基中培养,种子萌发率为62%。萌发种子继代至MS＋1mg/L 6-BA＋0.1mg/L 2,4-D＋0.5mg/LNAA＋4%蔗糖（pH 5.5）培养基进行不定芽诱导,不定芽发生率为95%。无根丛生芽继代到不定根诱导培养基1/2MS＋0.5mg/L IBA＋0.5mg/L NAA＋4%蔗糖（pH 5.5）,不定根发生率为90%。     

白玉兰离体培养和快速繁殖

试验建立了白玉兰离体繁殖体系。种子为外植体 ,初代培养基为 :B5 +BA0 .5mg/L +NAA0 .0 5mg/L + 30g/L糖 +琼脂 6g/L ,萌发率 85 %。继代培养基为 :B5 +BA1.5mg/L +NAA0 .15mg/L + 30g/glucose/sucrose + 6g/Lagar+Vco0 .5g/L ,月增殖率可达 5 .0± 0 .1。生根培养为 :1/2B5+NAA0 .5mg/L +AC1.0mg/L +VB2 0mg/L。以试管苗茎段、根段做外植体 ,培养基B5 +BA2mg/L+ 2 .4 -D0 .5mg/L +VC0 .5g/L ,可获得少量愈伤组织 ,继代存活率仅为 30 %。没有再分化成苗     

红掌组织培养及快速繁殖

红掌组织培养用叶柄顶部作外植体 ,愈伤组织诱导培养基为 1 2MS +6 -BA1.0mg·L-1+2 ,4 -D0 .1mg·L-1+食用白沙糖 4 0g·L-1,继代增殖及完整植株形成培养基为改良MS +6 -BA0 .5～ 2 .5mg·L-1+NAA0 .5mg·L-1或MS中NH4 NO3 的 5 4+食用白沙糖 30g·L-1,每个过程均培养 4 5d ,每块组织增殖 2～ 3倍 ,形成完整植株 4～ 6株。移栽基质粗河沙∶珍珠岩 (2∶1) ,遮光率为 80 % ,保持较高湿度 ,每 7d喷施 1 2MS无机盐 ,2个月后成活率在 95%以上     

Callus induction from leaves of differentpaulownia species and its plantlet regeneration

The experiment was carried out on five different species ofPaulownia for callus induction from leaves. MS medium was adopted as basic medium, and from different combinations of NAA and BA the suitable media were determined for callus induction, bud differentiation, and root differentiation of five different species. MS+0.5NAA+4BA, MS+0.3NAA+2BA, MS+0.5NAA+4BA, MS+0.3NAA+6BA, and MS+0.3NAA+8BA were suitable media of callus inductions of leaves, respectively, forPaulownia tomentosa, Paulownia australis, Paulownia fortunei, Paulownia elongata andP. tmentosa x P. fortunei, and MS+0.3NAA+12BA, MS+0.3NAA+12BA, MS+0.5NAA+12BA, MS+0.5NAA+12BA, and MS+0.7NAA+12BA were suitable media for bud differentiation from leaf callus respectively for above five species. The rooting media was determined as 2MS+0.1NAA, 1/2MS+0.1NAA, 1/2MS, 1/2MS+0.3NAA, and 1/2MS+0.5NAA. These results provide reference data for breeding new fine varieties with different kinds ofPaulownia protoplasts fusions. Foundation Item: This paper was supported by National Nature Science Foundation of China (No. 39870631) and Nature Science Foundation of Henan Province (No. 994011100). Biography: Fan Guo-qiang (1964), male Ph. Doctor, Professor in Forestry Department of Henan Agriculture University. Zhengzhou 450002, P. R. China. E-mail: guoqiangf@263.net. Responsible editor: Song Funan     

刚果12号桉花药愈伤组织诱导初报

由不同浓度的2,4-D,BA和GA3组成的16种培养基上对刚果12号桉花药愈伤组织的诱导,试验表明:在MS基本培养基中附加2,4-D1.0mg/L+BA1.0mg/L+GA30.5mg/L+蔗糖8%的诱导率最高,达58.06%。     

虎杖愈伤组织的诱导及其白藜芦醇形成初探

将虎杖P olygonum cusp id a tum不同外植体经不同消毒时间处理后,接种在添加不同激素种类和水平的相同基本培养基上或相同激素种类和水平基本培养基上进行诱导实验,同时对根和根茎芽、叶、韧皮诱导的愈伤组织进行白藜芦醇含量的测定.结果表明:基本培养基以M S较好,外植体叶对激素种类较为敏感,其中适当浓度的NAA诱导愈伤组织比2,4-D的效果好,KT比BA好,添加KT的培养基上诱导愈伤组织比较紧密,有利于分化,在M S+NAA 2 m g/L+KT 0.1 m g/L培养基上诱导愈伤组织较好,根茎芽的诱导率最高,为73%;愈伤组织的生长趋势从接种的第3天开始生长,到21 d时生长达到最高峰,干质量为0.461 2 g,以后生长速度减慢;对不同材料诱导的愈伤组织进行白藜芦醇含量的测定,其中根茎部芽的诱导的愈伤组织中白藜芦醇含量最高,其次是叶和根,最低的为韧皮.     

山地杨快速繁殖研究

本文以日本王子造纸公司培育的美洲黑杨(Populus deltoides,简称DD)与辽杨(P.maximowiczii,简称MM)的杂交品种MD110品系的2a生穗条为材料,以水培萌条的顶芽、叶片、叶柄为外植体,在进行愈伤组织的诱导培养时,以芽的生长点为外植体诱导愈伤组织的效果最好。MS+1.15 mg/L 6-BA+2.5％蔗糖为最佳诱导培养基。在愈伤组织的芽分化培养中,在琼脂固体培养基1/2(N)MS+0.30 mg/L6-BA+0.10 mg/L NAA+2.5％蔗糖上,分化率可达94％。在MS+0.20mg/L6-BA+2.5％蔗糖的液体培养基上,芽的分化率也可以达98％。在生根培养中,与萘乙酸相比,吲哚丁酸更有利于生根,且根生长粗壮。生根效果较好的培养基为1/2(N)MS+0.05 mg/L NAA+2.5％蔗糖。     

Effects of plant growth regulators,carbon sources and pH values on callus induction in Aquilaria malaccensis leaf explants and characteristics of the resultant calli

The endangered tropical tree, Aquilaria malaccensis, produces agarwood for use in fragrance and medicines. Efforts are currently underway to produce valuable agarwood compoundsn tissue culture. The purpose of this study was to develop an optimal growth medium, specifically, the best hormone combination for callus suspension culture. Using nursery-grown A. malaccensis, sterilized leaf explants were first incubated on basic Murashige and Skoog(MS) gel medium containing 15g/L sucrose and at pH 5.7. Different auxin types including 1-naphthaleneacetic acid(NAA), 2,4-dichlorophenoxyacetic acid(2,4-D), and indole-3-butyric acid(IBA), were tested at various concentrations(0.55, 1.1 and 1.65 μM) using the basic medium. Leaf explants were incubated for 30 days in the dark. Callus induced by 1.1 μM NAA had the highest biomass dry weight(DW) of 17.3 mg; however the callus was of a compact type. This auxin concentration was then combined with either 6-benzylaminopurine(BAP) or kinetin at 0.55, 1.1, 2.2 or 3.3 μM to induce growth of friable callus. The 1.1μM NAA + 2.2μM BAP combination produced friable callus with the highest biomass(93.3mg DW). When testing the different carbon sources and pHs, sucrose at 15g/L and pH at 5.7 yielded highest biomasses at 87.7mg and 83 mg DW, respectively. Microscopic observations revealed the arrangement of the friable cells as loosely packed with relatively large cells, while for the compact callus, the cells were small and densely packed. We concluded that MS medium containing 15 g/L sucrose, 1.1 μM NAA + 2.2 μM BAP hormone combination, and a pH of 5.7 was highly effective for inducing friable callus from leaf explants of A. malaccensis for the purpose of establishing cell suspension culture.     

Shoot proliferation and callus regeneration from nodular buds of Drepanostachyum luodianense

This research reports on an efficient shoot proliferation and callus regeneration system for bamboo.Young, semi-lignified branches with one lateral bud from Drepanostachyum luodianense(Yi et R. S. Wang) Keng f.were used as explants. Disinfection with 0.1% HgCl_2 for 8 min was the optimum treatment and the best medium for bud initiation was Murashige and Skoog(MS) medium containing 3.0 mg L~(-1)6-benzyladenine(BA). Multiple shoots were induced from nodal shoot segments on MS medium containing 5.0 mg L~(-1) BA, 0.5 mg L~(-1) kinetin(Kin), and 1.0 mg L~(-1) naphthaleneacetic acid(NAA). The highest frequency of callus formation(65.6%) was on MS medium containing 4.0 mg L~(-1)2,4-dichlorophenoxyacetic acid(2, 4-D), 0.5 mg L~(-1) NAA, and 0.1 mg L~(-1) thidiazuron(TDZ). The optimum medium for callus proliferation was MS medium with 4 mg L~(-1)2,4-D, 0.5 mg L~(-1) TDZ and 0.5 mg L~(-1) NAA, and the optimum hormone combination was 4 mg L~(-1) BA ? 0.5 mg L~(-1) NAA for callus redifferentiation(up to 85.6%). A 100% rooting was achieved on MS medium supplemented with 2.0 mg L~(-1) NAA and 0.5 mg L~(-1)3-indole butyric acid(IBA). Rooted plantlets were acclimatized in a greenhouse in humus soil ? perlite(1:1) substrate. These micropropagated callus induction and regeneration systems for bamboo will be useful for genetic engineering and multiplication.