铁棍山药组织培养快繁及试管珠芽离体再生体系研究

以河南温县铁棍山药带腋芽的茎段为外植体进行铁棍山药种苗快繁及珠芽离体再生体系的研究。结果表明:（1）75%乙醇浸泡30 s和5% NaClO消毒15 min配合使用灭菌效果最好;腋芽诱导最适培养基为MS+0.5 mg·L^-1 6-BA+0.1 mg·L^-1 NAA,培养20 d后诱导的多芽体倍数最高,为2.22,高度最高为3.3 cm;继代增殖最适培养基为MS+1.5 mg·L^-1 6-BA,增殖倍数可达4.1;生根培养最适培养基为1/2MS+0.2 mg·L^-1 6-BA+1.0 mg·L^-1 NAA+0.02%活性炭,平均生根天数为12 d,生根率达100%,根系最长为1.04 cm。（2）用单芽带外植体的接种方式,其珠芽诱导率及诱导的珠芽数显著高于只接单芽的接种方式,珠芽诱导率达88.9%,平均珠芽数为1.50,大小为0.38 cm×0.54 cm;蔗糖浓度为1%～3%有利于珠芽诱导,珠芽整齐度好,形状规则,试管苗叶色浓绿;离体珠芽芽诱导的最适培养基为MS+1.5 mg·L^-1 6-BA+0.2 mg·L^-1 NAA,珠芽在18～22 d发芽,30 d后诱导率最高为83.3%。该实验结果为铁棍山药试管苗的工厂化生产奠定了技术基础。     

茄子野生近缘种托鲁巴姆试管微扦插繁殖技术研究

嫁接栽培是茄果类蔬菜防治土传病害和提高产量的重要措施之一。茄子野生近缘种托鲁巴姆(Solanums torvum)因综合抗性强,成为茄子和番茄嫁接的常用优良砧木。但是,由于托鲁巴姆种子的发芽率、发芽势和发芽指数较低,苗龄较长,限制了其在工厂化育苗中的大规模应用,因此迫切需要开发其他方法及相应技术体系提高托鲁巴姆的育苗效率,降低育苗成本。为优化托鲁巴姆微扦插技术,该研究探索并优化了试管内微扦插繁殖托鲁巴姆技术,以无菌播种获得初代无菌苗的茎段为外植体,通过在培养基中添加植物生长调节剂,对比不同浓度植物生长调节剂对托鲁巴姆微扦插繁殖过程中的影响。结果表明:(1)托鲁巴姆在不同培养基中,腋芽诱导、继代增殖和生根培养的效果存在显著差异,初代芽诱导的最佳培养基为MS+KT 0.5 mg·L^-1+IBA 0.1 mg·L^-1,出芽率达90%。(2)继代扦插最佳培养基为MS+IBA 0.4 mg·L^-1,培养30 d的增殖系数达6.11,植株长势健壮。(3)最佳生根培养基为1/2MS+IBA 0.2 mg·L^-1,生根培养30 d,单株一级根数4.56条,最长根长125.80 mm、根粗0.50 mm,根系发达。采用试管内微扦插技术繁殖托鲁巴姆种苗,操作简单,增殖系数较高,可满足快速繁育种苗的要求。该研究结果为托鲁巴姆的工厂化规模育苗提供了新途径。     

豇豆子叶节再生体系建立及Pti4基因遗传转化

为获得豇豆(Vigna unguiculata)遗传转化体系,以‘成豇七号’带1 片子叶的子叶节作为外植体,对其高效再生体系和农杆菌介导抗病基因Pti4 的遗传转化进行了研究。结果表明,豇豆无菌苗、不定芽诱导和不定芽伸长培养的最适培养基分别为MSB₅ + 6-BA 3.0 mg L^-1、MSB₅ + 6-BA 1.0 mg L^-1 + KT 0.06 mg L^-1 和MSB₅ + 6-BA 0.5 mg L^-1 + IBA 0.2 mg L^-1。不定芽在MS 培养基上能迅速诱导生根,获得完整植株。以豇豆子叶节为受体,通过农杆菌介导成功将 Pti4 整合到‘成豇七号’抗性芽基因组中。因此,豇豆高效再生体系的建立为遗传育种研究奠定了基础。     

黑木相思愈伤组织诱导及植株再生

以黑木相思(Acacia melanoxylon)优良单株(AMY12004)的当年新生枝条带腋芽茎段为外植体, 灭菌后接入MS培养基上培养, 以其无菌萌芽的叶片、茎段和叶柄为实验材料, 通过间接器官发生途径建立黑木相思愈伤组织诱导及高频植株再生体系。研究结果表明, 诱导愈伤组织的最佳外植体为茎段; 愈伤组织诱导的最佳培养基为MS+1.5 mg·L–16-BA+0.2 mg·L–1NAA+3%蔗糖, 诱导率为93.33%; 愈伤组织再分化的最佳培养基为MS+2.0 mg·L^–16-BA+0.5 mg·L^–1NAA+3%蔗糖, 分化率为79.17%, 再生系数为9.58; 再生芽生根的最佳培养基为MS+0.5 mg·L^–1IBA+0.5 mg·L^–1NAA+4%蔗糖, 生根率为96.05%, 移栽存活率为81.40%。     

裸果木再生体系建立与不定芽发生研究

为探究裸果木再生体系建立的影响因素,确定其不定芽发生的起源,该研究以裸果木健壮植株的茎段为外植体,采用6 BA和IBA不同浓度组合,筛选愈伤增殖及不定芽再生的最佳浓度组合,确定生根诱导的关键影响因素,建立再生体系,并对其不定芽分化进程进行解剖结构分析,以确认其起源。结果表明：(1)裸果木茎段的最佳愈伤增殖培养基为MS+1 mg·L^-1 IBA+1 mg·L^-1 6 BA+30 g·L^-1蔗糖+7 g·L^-1琼脂,主体间效应分析表明IBA为关键影响因素;愈伤大小随IBA浓度增加呈现先升高后下降的趋势。(2)最佳不定芽诱导培养基为MS+0.5 mg·L^-1 6 BA+30 g·L^-1蔗糖+7 g·L^-1琼脂,诱导不定芽数量为4.9个/块,生芽率达92.3%。(3)生根诱导中,SH基本培养基和蔗糖浓度为关键因素,最佳生根培养基为SH+0~10 g·L^-1蔗糖+7 g·L^-1琼脂,生根率达91.3%。(4)解剖结构观察发现,不定芽起源于愈伤表层的分生细胞,为外起源。该研究通过器官发生途径建立了裸果木的再生体系,确定了不定芽为外起源,为裸果木这一珍稀濒危的林木种质资源保护及可持续利用奠定了研究基础,并为其未来的发展利用提供了有效途径。     

濒危药用植物桃儿七的离体培养研究

以桃儿七种子诱导的无菌苗为材料,研究了外源激素对芽诱导、增殖和生根的影响,建立了桃儿七离体培养再生体系。结果表明：芽诱导阶段,采用3.0 mg·L^-1 6-BA+0.5 mg·L^-1 GA₃激素组合,出芽率最高可达85-71%,缩短出芽时间30~40 d,在添加3.0 mg·L^-1 6-BA+0.2 mg·L^-1 IAA的MS培养基上利于增殖,增殖速度快,增殖系数1.63。以WPM+1.5 mg·L^-1 IAA+0.5 mg·L^-1 NAA的培养基培养30 d 后,生根率可达60.1%。     

佛手山药组织培养的研究

以佛手山药块茎、叶片、茎段为外植体, 探讨了其组织培养技术。结果表明：块茎培养以暗培养MS+6-BA1.0 mg·L^-1+NAA0 1 mg·L^-1效果较好;叶片诱导的适宜培养基为MS+ 6-BA0.5～1.0 mg·L^-1+NAA2.0 mg·L^-1, 暗培养;茎段培养都是光培养,无节茎段以MS+6-BA1.0 mg·L^-1+NAA0.5 mg·L^-1较好;带节茎段的初代培养则以MS+6-BA0.5～1.0 mg·L^-1+NAA0.1 mg·L^-1效果较好,继代增殖培养基为MS+6-BA0.5 mg·L^-1+NAA0.1 mg·L^-1,生根培养基为1/2MS +NAA0.5 mg·L^-1。     

大花蕙兰'红酒'× 莲瓣兰'边草素花' 杂交种组培快繁技术

该研究以大花蕙兰‘红酒''(Cymbidium hybridum ‘hongjiu'')×莲瓣兰‘边草素花''(C. tortisepalum ‘biancaosuhua'')F1代杂交种原球茎和根状茎为材料,比较了不同激素配比增殖分化、生根的培养基,建立了适用杂交兰组培快繁体系。结果表明:1/2MS+6-BA 1.0 mg·L^-1+NAA 1.0 mg·L^-1+AC 0.05%+香蕉80 g·L^-1对原球茎增殖效果最佳,增殖率达到307%; 1/2MS+6-BA 1.5 mg·L^-1+NAA 1.0 mg·L^-1 +AC 0.05%+香蕉80 g·L^-1有利于原球茎分化,分化率为82%; 1/2MS+TDZ 2.0 mg·L^-1+NAA 0.1 mg·L^-1+AC 0.05%+香蕉80 g·L^-1对根状茎增殖分化效果最佳,增殖率为293%,分化率为79%; 1/2MS+IBA 0.5 mg·L^-1+NAA 0.3 mg·L^-1+AC 0.05%+香蕉80 g·L^-1为最佳生根培养基,生根率达到84.7%,且根粗苗壮,叶色浓绿。此体系为杂交兰种苗的规模化生产提供了技术支持。     

加拿大金露梅离体培养与快繁体系的建立

以加拿大金露梅嫩芽、叶片为外植体进行试验,通过观察确定嫩芽为初代培养的外植体。运用L₉(3⁴)正交试验筛选出最适合的腋芽初代培养、芽苗继代增殖及组培苗生根的最佳培养基,从而为金雨点建立了一套“腋芽诱导—继代增殖—生根培养”的快速繁殖体系。结果表明,诱导腋芽的分化培养基为MS+6-BA 2 mg·L^-1+NAA 0.1 mg·L^-1,其中MS、6-BA和NAA均为诱导的主要因子,影响顺序为MS＞6-BA＞NAA,诱导率达96.33%;继代增殖培养中激素6-BA是影响加拿大金露梅芽增殖生长的主要激素,差异极显著（p=0.000 1）,并且以浓度为2.0 mg·L^-1的增殖效果最好,确定最佳培养基为MS+6-BA 2.0 mg·L^-1+NAA 0.6 mg·L^-1+KT 0.2 mg·L^-1,半月增殖倍数达6.12;对生根培养3种激素（KT、NAA、IBA）进行方差分析, KT和NAA的影响均达到了显著水平(p=0.000 1; 0.001 0),是影响生根的主导因子,而IBA的p值为0.424 5,因此可以忽略其影响,在诱导时确定用NAA和KT两种激素。之后对生根数量、生根率进行多重比较表明最终确定生根培养基为MS+NAA 0.1 mg·L^-1+KT 1.0 mg·L^-1,生根数为8.63条,生根率为95.33%。     

南高丛蓝莓快繁技术体系研究简

以南高丛蓝莓试管无菌丛生芽为材料,对南高丛蓝莓丛生芽的诱导与增殖、继代次数对丛生芽诱导增殖的影响、瓶内生根、瓶外生根、不同生根方式试管苗移栽成活率的大小进行了研究。南高丛蓝莓丛生芽诱导与增殖培养基以WPM+ZT 2.0 mg·L^-1较佳,增殖倍数可达3.50;继代6次丛生芽增殖倍数可达24.00;瓶内生根生根培养基以WPM+ZT 0.5 mg·L^-1+IBA 0.1 mg·L^-1为佳,生根率可达80.73%±3.17%,生根周期为100 d;试管芽用25 mg·L^-1 IBA溶液浸蘸10 s,以1/6 WPM为营养液加珍珠岩作基质,生根率可达到80.00%±5.00%,生根周期为40 d;瓶外生根试管苗移栽成活率是瓶内生根试管苗的2倍。基本建立了南高丛蓝莓的试管快繁技术体系,为南高丛蓝莓的工业化育苗奠定了技术基础。     

In vitro propagation of the nitrogen-fixing tree-legume Acacia mangium Willd.

In vitro propagation was initiated from 2-week-old and 7-month-old explants of Acacia mangium. Juvenile explants (2 week-old) of 5- to 10-mm lengths composed of two leaves were cultured on Murashige and Skoog (MS) medium containing 1.0 or 2.0 mg L^-1 6-benzyladenine (BAP). After 6 weeks, most explants had formed a large cluster of 14–18 axillary shoots produced by prolific branching of the primary axillary shoot after elongation. The maximum multiplication rate (40) was obtained in the first subculture; the rate decreased to 10–20 in the second one. The mean length of shoots was not significantly affected by BAP concentrations during the subsequent cultures. Rooting ability of juvenile explants was greatly affected by BAP concentrations used in the multiplication medium. When both types of explants were multiplied on a MS medium containing 1.0 mg L^-1 BAP and transferred to a half-strength MS medium containing 0.05 mg L^-1 IBA, only 10% of the juvenile explants were rooted versus 70% of the 7-month-old explants. Rooted plants transferred onto artificial substrate were all nodulated, when inoculated with a specific Bradyrhizobium sp. strain.     

Micropropagation ofFagus sylvatica L.

Summary An in vitro shoot multiplication system was established from juvenileFagus sylvatica L. tissues, and plantlets were regenerated. Embryonic axes were excised from beech seeds and germinated in vitro on media supplemented with 6-benzyladenine (BA) to obtain plantlets with axillary shoots. Shoot multiplication was maintained by sequential subculture of axillary shoot tips and basal segments on Woody Plant Medium supplemented with 0.5 mg/liter BA+2 mg/liter zeatin+0.2 mg/liter naphthaleneacetic acid (NAA). The effeciency of shoot multiplication clearly depended on the kind of explant used. Transfer to fresh medium every 2 wk during the 6-wk multiplication cycle improved multiplication rates. In the rooting stage, an initial 7-day dark period significantly improved rooting capacity and accelerated the emergence of roots on auxin-treated shoots. Adventitious buds were induced on the intact hypocotyls of the whole plantlets derived from the initial embryonic axis explants, especially on those cultured on medium with 1 mg/liter BA. Cotyledon and hypocotyl segments isolated from seedlings grown in vitro from embryos also exhibited capacity for adventitious bud formation, especially when cultured on media supplemented with 0.5 mg/liter BA + 0.1 mg/liter NAA.     

Effect of plant growth regulators on regeneration of plantlets from bud cultures ofCymbopogon nardus L. and the detection of essential oils from thein vitro plantlets

Cymbopogon nardus L. could be propagated via tissue culture using axillary buds as explants. The aseptic bud explants obtained using double sterilization methods produced stunted abnormal multiple shoots when they were cultured on Murashige and Skoog medium (MS) supplemented with 1.0 mg L^-1 or 2.0 mg L^-1 benzyladenine (BA). Stunted shoots that cultured on MS + 1.0 mg L^-1 BA + 1.0 mg L^-1 N⁶-isopentenyl-adenine (2iP) could induce elongation of shoots from about 60% of the stunted shoots. Normal multiple shoots could be induced at the highest (19.7 shoots per bud) from the bud explants within six weeks when cultured on proliferation medium consisted of MS supplemented with 0.3 mg L^-1 BA and 0.1 mg L^-1 indole-3-butyric acid (IBA). The separated individual shoot produced roots when transferred to basic MS solid medium. The essential oils that were contained in the mature plants namely citronellal, geraniol and citronellol were also found in thein vitro C. nardus plantlets. Citronellal was the main essential oil component in the matured plants while geraniol was the main component in thein vitro plantlets.     

In vitro micropropagation of the macarronesian evergreen treePersea indica (L.) K. Spreng

Summary Shoot propagation ofPersea indica (L.) K. Spreng was achieved using seedling axillary buds cultured on MS (Murashige and Skoog, 1962) medium with 1 mg/l (2.8 μM) N⁶-benzyladenine (BA). Forty percent of the obtained shoots did not elongate, but showed bud proliferation, which was maximal (three axillary buds per shoot) at the end of the seventh subculture. Sixty percent of the shoots elongated, did not show bud proliferation, and formed calluses at their base. Successful rooting (84.6%) was achieved dipping the base of each elongated shoot in 3 g/l (16.11 mM) indolebutyric acid (IBA) for 1–2 s, and transferring to half strength MS medium without growth regulators. These shoots presented an acclimatization success of 100%. Results suggest that micropropagated elongated shoots ofP. indica can be adequately used in reforestation programs.     

褐纹报春苣苔组织培养与快速繁殖

褐纹报春苣苔(Primulina glandaceistriata)是一种极具观赏价值的喀斯特地区野生花卉,目前尚未有褐纹报春苣苔组培快繁的研究报道。该研究以褐纹报春苣苔的叶片为外植体,通过两种途径建立其组培快繁体系。结果表明:适宜的不定芽诱导培养基为MS+6-BA 2.0 mg·L~(-1)+NAA 0.10 mg·L~(-1),适宜的不定芽增殖培养基为MS+ZT 2.0 mg·L~(-1)+NAA 0.10 mg·L~(-1)+活性炭0.05 g·L~(-1),增殖系数为11.09;适宜的愈伤组织诱导培养基为MS+TDZ 2.0 mg·L~(-1)+NAA 0.10 mg·L~(-1),适宜的愈伤组织分化培养基为MS+ZT 1.0 mg·L~(-1)+NAA 0.10mg·L~(-1),分化系数为12.46;适宜的生根培养基为1/2MS+IBA 0.1 mg·L~(-1)+活性炭0.05 g·L~(-1)或1/2MS+IBA0.5 mg·L~(-1)+活性炭0.05 g·L~(-1),生根率为100%。该研究结果成功建立了褐纹报春苣苔的组培快繁体系,为今后褐纹报春苣苔的种苗繁殖和遗传转化提供了技术支持。     

Micropropagation of<Emphasis Type="Italic">Schizandra chinensis</Emphasis> BAILLON using glucose from cotyledonary nodes

A protocol for the micropropagation ofSchizandra chinensis has been developed using regenerated shoots from axillary bud explants. In preparing to do so, we found that seed type (i.e., mature vs. pre-mature) significantly influenced the rate of germination. The Woody Plant (WP) medium proved to be superior to the Murashige and Skoog (MS) medium for germination purposes. Multiple shoots were induced from cotyledonary nodes of axenic seedlings on WP media containing 6-benzylaminopurine (BA) alone or in combination with 1-napthaleneacetic acid (NAA). High frequencies of shoot proliferation and the greatest number of shoots per explant (11.6) were observed with the use of 1.0 mg L^-1 BA. We also established a culture method for proliferating shoots by repeatedly subculturing the original cotyledonary nodes on a shoot multiplication medium each time newly formed shoots were harvested. To induce root formation, glucose was supplied as a carbon-source substitution for sucrose. The best rooting rate was obtained from a WP medium supplemented with 3% glucose and 0.5 mg L^-1 NAA. Following transplantation in the field, 82% of the plantlets survived.     

<i>In vitro</i> Germination and Micropropagation of <i>Aconitum vilmorinianum</i>: An Important Medicinal Plant in China

Aconitum vilmorinianum, a well-known traditional Chinese herb, is recently being threatened by overexploitation and environment disturbance. This study was conducted to provide propagation methods through in vitro germination and explant cultivation. Germination was stimulated up to 66.00% on Murashige and Skoog (MS) medium containing 2.0 mg L⁻¹ 6-benzylaminopurine (BAP), 0.1 mg L⁻¹ 1-napthaleneacetic acid (NAA), and 30 g L⁻¹ sucrose. Three bacteria (Pantoea agglomerans, Erwinia persicina, and Pseudomonas tolaasii) would be responsible for consistent contamination during germination. The latter two were effectively eradicated after disinfected. The influence of explant types and hormone combinations on direct and indirect organogenesis was evaluated in the present work. The frequency of shoot induction from axillary bud explants was 100% on the MS fortified with 2.0 mg L⁻¹ BAP and 0.3 mg L⁻¹ NAA. Shoots multiplication was optimized on MS medium supplemented with 0.1 mg L⁻¹ thidiazuron (TDZ) and 0.1 mg L⁻¹ NAA. High callus induction percentage (96.67%) was obtained from stem segments on MS medium with 2.0 mg L⁻¹ 2,4-D, then successfully regenerated into shoots on MS medium in the presence of 0.1 mg L⁻¹ TDZ and 0.2 mg L⁻¹ NAA. The present work could be useful for the utilization and conservation of this valuable species.     

In vitro differentiation and plant regeneration of Albizia chinesis (Osb.) Merr

Summary Petiolar and distal cotyledonary segments (PCS and DCS) of Albizia chinensis were cultured on Murashige and Skoog's (MS; 1962) medium and induced to form adventitious shoot buds in the presence of either cytokinins 6-benzylamino purine (BAP), kinetin (KN) or thidiazuron (TDZ). Superiority of BAP in inducing shoot bud and differentiation was observed. PCS was more morphogenic to shoot bud differentiation than DCS. TDZ was highly effective in inducing shoot buds, but arrested shoot growth, while KN produced more callus during differentiation of shoots. Rapid and high rate of shoot multiplication per explant was achieved through subculture in MS medium containing BAP (1.0 mg l⁻¹) and indole-3-acetic acid (IAA) (0.5 mg l⁻¹). BAP at low concentration was required to enhance shoot multiplication and elongation. Successful rooting of regenerated shoots was carried out in a two-step culture procedure in MS media with indole-3-butyric acid (IBA) (2.0 mg l⁻¹) and subsequent subculture in IBA-free medium.     

黄花蒿组培快繁与种质离体保存的研究

以带侧芽的黄花蒿(Artemisia annua L.)茎段为外植体,以MS为基本培养基,进行组织培养和种质保存研究.结果表明,培养基MS 6-BA 1.0 mg L-1 IBA 0.1 mg L-1、MS 6-BA 0.5 mg L-1 IBA 0.1 mg L-1和MS NAA0.1 nag L-1 IBA 0.5 rng L-1可分别用于黄花蒿的芽诱导、增殖和生根培养,培养20 d的增殖倍数为5.5倍,生根率98.3%.培养基MS CCC 1.0 mg L-1、MS CCC 2.0 nag L-1、MS PP3334.0 mg L-1可用作离体保存,连续保存200 d的存活率分别达72.3%、77.0%、69.2%.活力检测表明,黄花蒿种质经保存后的增殖、生根能力没有下降.因此,可通过诱导腋芽增殖建立黄花蒿快繁体系,及在培养基中添加CCC或PP333拼能使材料长期保存.