Micropropagation of a mature colchicine-polyploid and irradiation-mutant of Betula pendula Roth

Tetraploid plantlets were regenerated from cultured apical and axillary buds of a 23-year-old colchicine-polyploid and irradiation-mutant Betula pendula Roth tree. Bud explants were grown on modified Murashige and Skoog medium supplemented with 2.0 mg l(-1) benzylaminopurine (BAP) and 0.01 mg l(-1) 1-naphthaleneacetic acid (NAA). The medium allowed both induction of adventitious buds and development of shoots. The cut ends of new shoots produced new buds and shoots during a 4-week culture period. The micropropagated shoots were rooted on modified Murashige and Skoog medium containing 0.1 mg l(-1) NAA as the sole growth regulator. Plantlets were transferred to a peat/soil mixture (1:1) in the greenhouse, acclimated and then transplanted to a cold frame. The regenerated plantlets had a tetraploid chromosome set (4n = 56) and an altered leaf morphology typical of colchicine-polyploid birches. The leaves were hypertrophied and asymmetrical, with curly leaf margins. The mutant nature of the parent tree was also evident in the light-green color of the leaves of the plantlets.     

Rapid Micropropagation of Edible Bamboo Dendrocalamus asper

Abstract

Micropropagation protocols for Dendrocalamus asper using nodal shoots and seeds culture are described. Multiple shoots were induced through forced axillary branching. Ninety-five percent of the nodal shoot explants taken from juvenile primary and lateral branches, produced multiple shoots through axillary buds activation within 2 to 8 weeks on Murashige & Skoog's (MS) medium supplemented with 0.1-15 mg/l benzyladenine (BA). The cultured seeds also produced multiple shoots (1-20) within 6 weeks on this medium. The multiple shoot differentiation was influenced by the concentration of BA in the medium. The in vitro generated shoots were excised and subculture on MS + 3.0 mg/l BAP for further shoot multiplication. Fifteen to 20 fold rate of shoot multiplication was achieved by regular subculturing. These shoots were multiplied for more than 3 years without loss of vigor. Ninety-five percent of the shoots were rooted, when propagules (each consisting of cluster of 3 shoots) were transferred on to MS medium with 3.0 mg/l NAA or 10 mg/l indole-3-butyric acid (IBA).

To date, 18,000 plants (through axillary bud initiated from nodal ex-plants) and 6,000 plants from seed culture have been hardened and acclimatized. 12,000 plants have been field transferred.     

火炬松丛生芽诱导及植株再生

采用火炬松带子叶项芽为外植体建立了组织培养再生体系。结果表明,火炬松丛生芽的形成受诱导培养基中激素浓度及外植体年龄的影响较大。适合火炬松丛生芽诱导的培养基为改良GD 6-BA 4 ms／L NAA 0．02 mg/L(诱导率86．7％),截取外植体的最佳苗龄为14-21 d。已分化的丛生芽继代培养在无6-BA但附加0．5- 1．0 g/L活性炭或0．02-0．05 mg/L NAA的培养基上伸长很快。伸长的丛生芽接种在附加0．05 ms／L NAA的1／2改良GD培养基中,1个月后有12．5％产生不定根。     

龙脑樟愈伤组织诱导及增殖研究

以龙脑樟的茎段作为外植体,进行腋芽的诱导获得无菌苗,继而腋芽小叶被用于愈伤组织的诱导并进行增殖培养。结果表明外植体适宜的灭菌方法为：0.1%升汞灭菌10 min,污染率为21.6%;腋芽小叶愈伤组织诱导最佳培养基配方为：MS＋BA 4.0 mg/L＋NAA 0.1 mg/L,诱导率可达72.7%,较佳增殖培养基组合为：MS＋BA 5.0 mg/L＋NAA 0.5 mg/L。     

色木槭芽直接增殖途径的培养条件

为建立色木槭芽直接增殖的诱导培养体系,以色木槭野生树木休眠芽萌发枝条为材料,进行芽增殖的培养条件研究。结果表明,4月份是适宜休眠芽培养的取材时期。嫩茎在2%次氯酸钠浸泡20 min是最佳消毒方案。NAA浓度对休眠芽萌发和嫩茎生长的影响具有显著差异。MS+0.1 mg/L NAA+20 g/L蔗糖(pH5.8)是适合休眠芽萌发和嫩茎伸长的培养基。培养30天时,腋芽萌发率可达63.3%,嫩茎平均高度可达15.9 mm。6-BA对芽直接增殖的促进效果好于KT。不同激素组合中,IBA与6-BA组合对芽增殖和丛生芽生长的效果好于NAA与6-BA组合、NAA与KT组合、IBA与KT组合。MS+0.1 mg/L IBA+1 mg/L 6-BA+20 g/L蔗糖是适合芽增殖和丛生芽生长的培养基。培养30天时,芽增殖率可达90%,增殖倍数可达3.19,且茎芽生长正常。     

杉木无性系规模化组培繁育技术研究

为了建立高效的杉木规模化组培快繁体系,以杉木未木质化的春梢茎段为外植体,MS为基本培养基,研究不同生长调节剂及其浓度组合对茎段萌发和丛生芽增殖的影响.结果表明：采用未木质化的春梢茎段,剖开后匍匐放置于培养基上,腋芽容易萌发,最佳初代培方式为MS＋6-BA1.0mg/L＋ NAA0.5 mg/L培养基上培养10天后,转到MS＋ 6-BA0.8 mg/L＋NAA0.3 mg/L继续培养,最佳增殖培养基为MS＋ 6-BA0.8 mg/L＋ NAA0.3 mg/L,无根无菌植株采用常规扦插方法生根,可成功培育杉木优良无性系苗木.     

Rapid clonal multiplication of a woody tree, <Emphasis Type="Italic">Vitex negundo</Emphasis> L. through axillary shoots proliferation

An efficient and improved shoot regeneration technique for the micropropagation of Vitex negundo, an aromatic and medicinal shrub through in vitro culture of nodal segments with axillary buds, is described. Thidiazuron (TDZ) used at 1.0 μM was the most effective in inducing bud break and growth, and also in initiating multiple shoot proliferation at the rate of 25 microshoots per nodal explant with axillary buds, after 4 weeks of culture. By repeated subculturing of nodal explants, a high-frequency multiplication rate was established. Optimum shoot multiplication and elongation was achieved when TDZ exposed explants were subcultured on Murashige and Skoog (MS) media containing a combination of 1.0 μM 6-benzyladenine (BA) and 0.5 μM α-naphthalene acetic acid (NAA). Efficient rooting was achieved directly in soilrite when basal portion of the shoots were treated with 500 μM indole-3-butyric acid for 10 min which was the most effective in inducing roots, as 97% of the microshoots produced roots. Plantlets went through a hardening phase in a controlled plant growth chamber, prior to ex-vitro transfer. Micropropagated plants grew well, attained maturity and flowered. No phenotypical differences for morphogenesis were observed among the regenerants.     

野百合组织培养的研究

以野百合鳞茎、叶片为外植体通过继代培养和生根培养,诱导不定芽的发生,进而获到再生植株。试验结果表明:用MS培养基培养野百合鳞茎、叶片组织,均能够诱导不定芽产生;在培养基中附加BA2.5mg/L和NAA0.2mg/L有利于鳞茎不定芽的诱导且芽苗长势好,而叶片则在培养基中附加BA1.5mg/L、NAA0.2mg/L的培养效果最佳;从不定芽诱导出根以1/2MS效果最好。     

日本樱花茎段再生体系的建立

以成年日本樱花茎段为外植体,系统研究了取材时间、冷藏时间、消毒方法、激素等诸多因素对茎段再生的影响,建立了日本樱花茎段丛生芽再生体系。研究结果表明,一般中午取材、4℃冷藏2 d、75%酒精浸泡3 min后用0.1%HgCl2浸泡7 min,可获得日本樱花无菌材料。将无菌材料接种至MS+NAA0.05 mg.L-1+6-BA2.0 mg.L-1的分化培养基上,腋芽明显萌动并伸长;在MS+6-BA4.0 mg.L-1+NAA0.3 mg.L-1+GA30.05 mg.L-1的增殖培养基上,腋芽不但能较快增殖形成大量丛生芽,而且形成的小芽能继续长大。待小芽长至3 cm~3.5 cm时,将其切割下来转移至1/2MS+NAA0.6 mg.L-1+IBA0.2 mg.L-1生根培养基中,最终获得日本樱花的完整植株。     

西蒙得木组织培养繁殖技术的研究

通过对西蒙得木〔Simmondsia chinensis(Link)〕组织培养大批量繁殖技术的研究,探索基本培养基、生长调节物质及蔗糖含量等对嫩梢增殖及生根的影响,结果表明:采用春季生长的1年生实生苗以及6年生高产的优株茎节作外植体,培养于含有ZT 1～3 mg/L或BA 1～3mg/L和NAA 0.01～0.1 mg/L的MS培养基中,经30～40天,几乎所有腋芽均能长成嫩梢,切取长达4～6 cm的嫩梢,培养于含有NAA的1/2MS培养基中,经1周暗培养,1个月后生根率达93％,小植株移栽于含砂土的基质中,保持湿润,获得成功。用幼苗及优株作外植体的培养效果无显著差异,说明此法不受遗传型的限制。     

仙人球的组织培养与快速繁殖

以全盛球种仙人球的茎切段和整个子球为外殖体进行试管培养，结果表明，在外殖体的切口处可诱导形成愈伤组织，在其刺座上的疣突处可直接产生小球体。经试验筛选出最适宜的培养基为：不定芽(小球体)诱导，MS+6-BA 8mg/L+KT 1mg/L+NAA 0.1mg/L；从生芽分化和继代，MS+6-BA 3.5 mg/L+KT 0.5mg/L+NAA 0.1 mg/L；生根，1/2MS+NAA 0.1mg/L+IBA 0.1mg/L+活性炭0.5%。     

杂交马褂木组织培养技术研究

外植体采自杂交马褂木的幼树或扦插苗,采集部位为其顶芽或腋芽,最佳培养基为MS+BA2.0 mg/L+NAA0.1 mg/L。采用DCR为基本培养基,加入抗褐化试剂柠檬酸50.0mg/L,Vc 50.0 mg/L,继代增殖率可以达到2.8倍,最高达到4倍。生根培养基为1/2MS+NAA1.5 mg/L+活性炭0.5 g/L,瓶苗生根率高达61%。组培苗直接移栽到红心土,成活率达到93%以上。     

In vitro regeneration of plantlets from mature embryos of Pinus ayacahuite

A plantlet regeneration protocol was developed for Pinus ayacahuite var. ayacahuite (Ehrenb.). Embryos from mature seeds from ten provenances were cultured in a 16-h photoperiod for 3 days on a medium containing 30 mM sucrose and 0.7% agar. Cotyledons from these embryos were subcultured onto MCM medium (Bornman 1983) supplemented with 50 micro M N(6)-benzyladenine and 90 mM sucrose for 2 weeks. Bud development and shoot elongation were maximized by subculturing the explants on half strength AE medium (von Arnold and Ericksson 1981), supplemented with 60 mM sucrose and 0.05% activated charcoal every 30 days. Seed source had a significant effect on the responses of the embryos to the bud induction protocol. For the provenance with the best response to bud induction, about 79% of the cultured cotyledons formed buds, and each cotyledon formed a mean of 9.1 buds, so that about 70 shoots could be induced from each seed. The best rooting response (40% rooting) was obtained by treating the shoots for 8 h with 100 micro M naphthalene acetic acid.     

水栒子组织培养快繁体系研究

以水栒子（Cotoneaster multiflorus Bunge.）的新梢带侧芽茎段为外植体,研究了不同激素组合对其外植体诱导、丛生芽分化增殖以及试管苗生根的影响。结果表明：芽诱导最适培养基为MS＋BA0.8＋NAA0.04,诱导率为68%;分化增殖最佳培养基是MS＋6-BA1.2＋NAA0.04,增殖倍数为4.53;最佳生根培养基是1/2MS＋IBA0.4＋NAA0.04,生根率为91%;试管苗移栽的最佳基质组合及配比为沙子∶熟表土∶草炭土∶果渣=3∶3∶2∶2,成活率达91%。     

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。     

花叶络石组织培养研究

文章以花叶络石（Trachelospermum jasminoides ‘Variegatum’）幼嫩茎段为外植体,研究了不同消毒时间的外植体消毒效果、激素质量浓度配比对腋芽增殖及生根的影响和移栽基质配比,建立花叶络石快速繁殖体系。结果表明：0．1％升汞消毒10 min 效果最好,成活率达72％;花叶络石腋芽增殖较好的培养基组合为 MS ＋BA2．0 mg／L ＋NAA0．05 mg／L,增殖倍数4．23;最佳的生根培养基为1／2MS ＋IBA0．3 mg／L,生根率达到97．2％;花叶络石移栽较优的基质配比为蛭石∶珍珠岩∶泥炭＝6∶1∶3,成活率达93％。     

樟脑型樟树芽器官离体培养技术

以樟脑含量＞70％的樟脑型樟树优株的嫩枝为外植体,进行芽器官离体培养技术研究.结果表明：每年的1～4月外植体灭菌消毒接种,无菌活体获取率比较高;适宜樟脑型樟树组培的基本培养基：M2 （MS＋Ca （NO3）2.4H2O 300 mg/L）;芽诱导培养激素组合：6-BA4 mg/L＋NAA1.5 mg/L＋KT 0.5 mg/L;芽增殖培养激素组合：6-BA3.0 mg/L ＋NAA0.5 mg/L ＋KT0.2 mg/L;在培养基中添加L-半光氨酸10 mg/L抑制芽褐化效果良好.     

树莓组织培养技术

以未萌发的腋芽和带芽茎段为外植体,对树莓进行了组织培养和快速繁殖研究.结果表明,未萌发的腋芽是最佳的外植体,MS+6BA1.0 mg/L+NAA0.2 me/L为最合适的增殖培养基;将产生的不定芽转到1/2MS+IBA1.0 mg/L的生根培养基中生根良好,移栽存活率达80%以上.     

现代郁金香离体培养与快速繁殖

以现代郁金香的内层鳞片、幼茎和茎尖为外植体进行了离体培养与快速繁殖试验,筛选出最佳培养基;（1）诱导鳞片产生丛芽为MS＋0．4～1．0mg／LBA＋0．4mg／LNAA;（2）茎尖诱导培养为MS＋2mg／LBA＋0．1mg／LNAA;（3）茎段诱导培养为MS＋1．0mg／LBA＋0．2mg／LNAA）（4）丛芽增殖培养为MS＋0．4mg／LBA＋0．2mg／LNAA和MS＋0．4mg／LBA＋0．2mg／LIAA;（5）生根诱导培养为1／2MS＋0．4mg／LKT和0．1～1．0mg／LNAA。