药用植物金荞麦愈伤组织诱导与植株再生

目前转基因技术已成为植物定向遗传改良的重要手段,而建立稳定高频的离体再生系统是实现遗传转化的基础和前提.本试验以25 ～30 d苗龄的金养麦(Fagopyrum dibotrys)无菌苗叶片、茎节间、叶柄为外植体进行愈伤组织诱导与植株再生研究.结果表明:叶片在MS +2,4-D 4.0 mg/L +6-BA 1.0 mg/L培养基上愈伤组织诱导率达到89％.茎节间在MS +2,4-D 2.0 mg/L +6-BA 2.0 mg/L培养基上愈伤组织诱导率为87％.叶柄在MS +2,4-D 4.0 mg/L +6-BA 2.0 mg/L+ IBA 0.2 mg/L培养基上的最高诱导率仅为54％.愈伤组织分化不定芽的适宜培养基为MS +6- BA2.0 mg/L +TDZ0.2 mg/L +NAA0.2 mg/L;金荞麦不定芽在1/2 MS +NAA 0.5 mg/L的培养基上生根效果最好.组培再生植株经炼苗后移栽到田间成活率达80％以上,且生长表现正常.高频完整再生体系的建立,为金荞麦进一步遗传操作和扩大药材资源奠定了基础.     

矮生龙船花的组织培养和快速繁殖

矮生龙船花(IxoracoccineaL.)的带节茎段在MS 2,4D2.0mg/L培养基上产生大量的愈伤组织;在MS 6BA1.0mg/L NAA0.2mg/L培养基上芽的增殖系数达413,并产生少量的愈伤组织;在MS NAA0.2～2.0mg/L培养基上只产生芽而无愈伤组织形成。愈伤组织在MS 6BA0.5mg/L NAA0.5mg/L培养基上产生大量的不定芽,丛生芽在MS 6BA0.5mg/L NAA0.5mg/L培养基上生长较快并产生较多分枝,将分枝节下或切成段后在MS 6BA0.5mg/L NAA0.5mg/L培养基上能迅速生长并产生新的分枝。试管内小苗在1/2MS NAA0.5mg/L培养基上的生根壮苗效果较好。矮生龙船花试管苗成活率为935%。     

黄花补血草愈伤组织的诱导和植株再生

以黄花补血草(Limonium aureum(L.)Hill.)无菌苗为材料,研究了不同激素配比条件下不同外植体愈伤组织的诱导及植株再生.结果表明,黄花补血草无菌苗叶片、叶柄和幼根均可作为离体培养的外植体,但叶片和叶柄的诱导率明显高于幼根.将外植体接种于分别添加0.5 mg/L NAA或1.0 mg/L 2,4-D或0.3-0.5 mg/L 6-BA 0.5-2.0 mg/L NAA的MS培养基上,经过14-28 d培养后,可脱分化产生乳白色、红色或浅绿色颗粒状或致密愈伤组织,频率达到70%以上.在MS 0.3 mg/L 6-BA 1.0 mg/L NAA培养基上,红色和绿色颗粒状愈伤组织经过1-2次继代后,均可分化产生不定芽,进而形成丛生芽,分化率达到100%.将高约3 cm的丛生芽切下,接种于分别添加0.5 mg/L IAA或0.5 mg/L IBA的1/2 MS培养基上可产生不定根,获得完整的再生植株.     

骆驼蓬的组织培养及植株再生

以骆驼蓬(Peganum harmala L)无菌苗下胚轴切段为材料,在不同的培养基上进行愈伤组织的诱导,发现在MS基本培养基附加2．0mg/L 2,4—D、0．5mg／L 6—BA和3％蔗糖时,可100％的诱导出愈伤组织。愈伤组织在附加2．0mg／L 6—BA、0．5mg／L NAA、500mg／L CH和3％蔗糖的MS培养基上诱导出丛生芽,进而发育成苗,苗的分化频率在30％左右。分化苗或其茎切断在附加0．2mg／L IBA、0．2mg/L NAA和3％蔗糖的l／2MS培养基上出现根的分化,分化频率在90％以上。再生植株经炼苗后移栽成活,成活率在80％以上。     

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。     

罗布麻子叶和下胚轴再生植株的培养

本文报道了用罗布麻（Apocynum venetam)幼苗的子叶和下胚轴切段诱导出愈伤组织和再生植株。结果表明,愈伤组织的诱导在附加0.5mg/L6－BA和0.1-1mg/L NAA的MS培养基上为最好。在附加0.5mg/L NAA的MS培养基上培养愈伤组织能促进芽的分化,当NAA浓度增加到1mg/L时则能抑制芽的分化。随后在附加0.4mg/L IBA的MS培养基上诱导生根,获得完整再生植株。     

茎用芥菜细胞质雄性不育系子叶原生质体培养和高效植株再生

利用茎用芥菜细胞质雄性不育系原生质体培养获得了再生植株,并研究了影响原生质体培养的因素.结果表明,子叶是茎用芥菜原生质体培养最佳的外植体,10 d苗龄的子叶原生质体在改良MS培养基上培养3 d后发生第1次细胞分裂,6 d后发生第2次分裂,3周后形成细胞团,5周后形成肉眼可见的小愈伤.培养基中缺少NAA或2,4-D都会降低愈伤组织的再生能力.在含一定浓度的NAA(0.25 mg/L)和2,4-D(0.25 mg/L)培养基上诱导的愈伤组织质地致密且有光泽,芽的分化能力高;在MS+BA l mg/L+NAA 0.2 mg/L的培养基上芽的分化频率高达近29%,再生芽在1/2MS+NAA0.1 mg/L培养基上生根,形成完整植株.     

茎用芥菜细胞质雄性不育系子叶原生质体培养和高效植株再生（英文）

利用茎用芥菜细胞质雄性不育系原生质体培养获得了再生植株,并研究了影响原生质体培养的因素.结果表明,子叶是茎用芥菜原生质体培养最佳的外植体,10 d苗龄的子叶原生质体在改良MS培养基上培养3 d后发生第1次细胞分裂,6 d后发生第2次分裂,3周后形成细胞团,5周后形成肉眼可见的小愈伤.培养基中缺少NAA或2,4-D都会降低愈伤组织的再生能力.在含一定浓度的NAA(0.25 mg/L)和2,4-D(0.25 mg/L)培养基上诱导的愈伤组织质地致密且有光泽,芽的分化能力高;在MS+BA l mg/L+NAA 0.2 mg/L的培养基上芽的分化频率高达近29%,再生芽在1/2MS+NAA0.1 mg/L培养基上生根,形成完整植株.     

皂质芦荟的组织培养

将皂质芦荟的茎段及幼苗作为外植体进行组织培养,试验结果表明：皂质芦荟的茎段经40d左右可诱导形成愈伤组织,再经20d萌生再生芽：幼苗培养需要30d左右基部直接分化再生芽。同时经试验筛选出愈伤组织形成、再生芽分化和生根的最适培养基为：MS＋6－BA 2.0mg/L NAA 0.1mg/L、MS 6-BA 3.0mg/L NAA 0.1mg/L和MS NAA 0.3mg/L。     

冬凌草离体培养体系的建立及主要次生代谢产物的测定

以冬凌草叶片为外植体,研究不同浓度激素组合对冬凌草愈伤组织诱导及植株再生的影响,并对不同外植体(茎、叶)诱导愈伤、芽的分化能力及再生植株内主要次生代谢产物的含量进行了比较研究。结果表明:在MS 2.0 mg/L 6-BA 1.0 mg/L NAA培养基上诱导愈伤组织效果较好;在MS 2.0 mg/L 6-BA的培养基上诱导芽的效果较好;叶片和茎段在愈伤诱导培养基上均能产生大量的愈伤组织,但其再分化能力以茎段最好;再生苗生根培养基以0.3 mg/L IBA最好;以叶为外植体诱导的再生植株中冬凌草甲素、迷迭香酸的含量均高于以茎为外植体诱导的再生植株。     

川芎高频再生体系的建立

为建立川芎(Ligusticum chuanxiong Hort.)高频再生体系,优化了诱导和分化培养基及培养条件.以叶柄为外植体,以MS为基本培养基,KT2.0 mg/L+ IAA0.5mg/L的激素组合对不定芽分化最有利.在此基础上,针对外植体来源、培养条件和愈伤组织继代时间3个因素进行优化.结果表明:采用川芎无菌苗叶柄作为外植体,黑暗条件下诱导出愈伤组织,再在光照下继代培养15d后转入分化培养基中对不定芽诱导最为有利,分化率为44.4％.分化后得到的不定芽在含NAA0.5 mg/L和IBA 0.5 mg/L的1/2MS培养基上生根率达90％,移栽存活率为95％.     

细裂银叶菊叶的组织培养繁殖研究

以细裂银叶菊叶片为材料,进行愈伤组织的诱导、分化培养及生根诱导培养。结果表明：叶片愈伤组织的诱导以MS 2,4-D2mgL^-1 BA1mgL^-1 NAA0．1mgL^-1。培养基较好：分化培养以MStBA0．5mgL^-1 NAA0．1mgL^-1为好：生根诱导以1／2MS NAA0．01mgL^-4效果最好。     

川芎高频再生体系的建立

为建立川芎(Ligusticum chuanxiong Hort.)高频再生体系,优化了诱导和分化培养基及培养条件。以叶柄为外植体,以MS为基本培养基,KT 2.0 mg/L+IAA 0.5 mg/L的激素组合对不定芽分化最有利。在此基础上,针对外植体来源、培养条件和愈伤组织继代时间3个因素进行优化。结果表明:采用川芎无菌苗叶柄作为外植体,黑暗条件下诱导出愈伤组织,再在光照下继代培养15 d后转入分化培养基中对不定芽诱导最为有利,分化率为44.4%。分化后得到的不定芽在含NAA 0.5 mg/L和IBA 0.5 mg/L的 1/2MS培养基上生根率达90%,移栽存活率为95%。     

长柄双花木愈伤组织诱导及植株再生

以长柄双花木当年生嫩梢上的叶柄、嫩茎、嫩叶为外植体,对影响长柄双花木愈伤组织诱导和继代、分化主要因素进行研究。结果表明:在培养基MS+NAA0.5mg/L+2,4-D2.0mg/L上,三种外植体均可诱导出愈伤组织,其中叶片愈伤组织诱导率最高。该培养基还可作为愈伤组织继代培养基,但继代培养周期不超过2周。愈伤组织接种在MS+BA2mg/L上分化不定芽,根的诱导在1/2MS+IBA0.5mg/L培养基上进行。     

驱蚊草组织培养及其愈伤组织诱导研究

运用正交设计方法,利用植物组织、细胞培养技术,成功地建立了驱蚊草组织培养快繁技术体系。研究出:不定芽诱导的最适培养基为MS 6-BA0.2mg/L NAA0.3mg/L,增殖倍数为6.73;生根培养基为1/2MS NAA0.4mg/L,平均每株生根数为11.2条,生根率为90.0%。通过对其离体茎段的培养研究实验,得出驱蚊香草的最适愈伤组织诱导培养基为MS 2,4-D0.5mg/L 6-BA2.0mg/L NAA0.3mg/L,发愈率为93.3%,愈伤组织多为淡黄色,质地疏松。     

In Vitro Method for Propagation of Centella asiatica (L) Urban by Shoot Tip Culture

A rapid clonal propagation system has been developed for the medicinally important herb Centella asiatica (L) Urban by shoot tip (2–3 cm long) culture. The shoot tips isolated from mature plants were inoculated on MS medium incorporated with BA alone or in combination with NAA and Kn. The optimum number of shoots (3.38) with optimum number of leaves per shoot (4.25) were attained on MS medium supplemented with 4.0 mg l^?1 BA and 0.1 mg l^?1 NAA. On transferring the microshoots on full strength MS medium supplemented with various concentrations of IBA (1.0-3.0 mg l^?1) and NAA (0.5-2.0 mg l^?1), profuse rooting (46.8 per shoot) was obtained in MS basal medium with 2.0 mg l^?1 IBA with root length of 19.7 cm. Well rooted plantlets were acclimatized successfully by adjusting the temperature and humidity for 3–4 weeks after transfer to pots filled with sterilized vermiculite soil: sand (1:1)mixture. This micropropgation protocol could be useful for raising a stock of genetically homogenous material for field cultivation within a very short period.     

In vitro studies on Pea (Pisum sativum L.): I. Callus formation,regeneration and rooting

Abstract

Callus production, shoot formation via organogenesis and rooting of the regenerated shoots are reported in an Egyptian variety of Pisum sativum L. Calli were initiated from hypocotyl, leaf, root and mature embryo explants when cultured on MS medium containing B5 vitamins and supplemented with 2 mg/l 2,4-D+1 mg/l kin. Among the different types of explants, hypocotyl showed best potential for callus proliferation. Hypocotyl, leaf and immature cotyledon explants were used for shoot organogenesis. The best results of shoot formation were achieved when hypocotyl explants were cultured on MS-medium supplemented with 2 mg/l BA+1 mg/l NAA. However, immature cotyledon explants showed the highest frequency of shoot formation with 1 mg/l BA. Data of in vitro rooting showed that maximum root frequency occurred on culture medium containing half strength of MS salts, 40 g/l sucrose and 2 mg/l NAA.     

在1/3海水培养基上筛选豆瓣菜耐盐变异体

The responses of stem segments of watercress ( Nasturtium offtcinale R. Br. ) to 6-BA, NAA and 2,4-D were studied. MS medium supplemented with 2.0 mg/L 6-BA, 0.2 mg/L 2,4-D was used for callus initiation and maintainance. MS medium supplemented with 4.0 mg/L 6-BA was suitable for plant regeneration and MS medium without plant hormone supplement was used for rooting and plant propagation. For screening of salt. tolerant calli, stem segments of watercress were plated onto callus initiation medium containing 1/3 natural seawater. Seventeen out of the 325 plated explants produced calli. The growth curves demonstrated that the growth rate of salt-tolerant calli on saline medium almost matched that of the control calli on normal medium. Some of the salt-tolerant calli were transferred to the normal regeneration medium or saline regeneration medium to induce plant regeneration. In the first case, buds and shoots were regenerated in the same way as those of control calli on normal regeneration medium. More than 1 000 regenerated shoots were obtained of which 83 regenerated shoots were cut and transferred to saline MS base medium. At first, all shoot growth was inhibited, but 40 days after the transfer, rapid-growing axillary shoots were observed on 16 of the original shoots but none on the control shoots on saline MS base medium. Moreover, green spots appeared on most calli 10 days after they were transferred to saline medium, however buds appeared only on 5 calli from the 30 transferred calli and at the end only 2 rapid-growing shoots were obtained from two calli. In total, 18 variant lines were obtained through propagation of the salt-tolerant shoots on saline MS base medium. RAPD analysis was performed in 10 of the 18 salt-tolerant variant lines and DNA variation was detected in all the tested variant lines.     

Organogenesis and Plantlet Regeneration in Vitro of Populus euphratica

The research of organogenesis and in vitro plantlet regeneration of Populus euphratica Oliver was carried out using the tender shoots from mature tree as initial explants and MS medium as the basic medium. The effects of plant growth regulators (PGR) on the regeneration were compared. The results showed that the concentration of PGR was not strictly required for the organogenesis of the excised organs and callus, but the ratio of BA to NAA was important. Calli could be induced from the excised leaves and stems cultured on the medium with 0.5 mg/L BA and 0.5 mg/L NAA. The embryonic callus could be multiplied in dark on the medium supplemented with 0.25 mg/L BA and 0.5 mg/L NAA. For the adventitious bud regeneration of the leaf and callus, supplement with 0.5 mg/L BA and 0.1 mg/L NAA was appropriate, giving a regeneration frequency of 82.9% and 100%, respectively. The suitable level of BA and NAA for the excised stem's was 0.1 mg/L and 0.01 mg/L respectively, yielding a regeneration frequency of 83 %. Rooting occurred on the MS medium with half strength of macronutrient and addition of 0.015 mg/L NAA, and the rooting rate could reach up to 86.2%. The techniques of somatic cell cloning of P. euphratica was established in vitro. The problems of deterioration of the subcultured shoots were also discussed.     

Induction of root and shoot formation from root meristems ofPetunia hybrida

Summary A procedure has been developed for the induction of root or shoot formation from root meristems of germinated seeds ofPetunia hybrida. Root formation was obtained on Murashige and Skoog (MS) medium supplemented with a combination of 6-benzylaminopurine (BA) (0–0.5 mg/l) and naphtaleneacetic acid (NAA) (0.05–2.0 mg/l). Induction of predominantly shoot formation was obtained on MS medium containing the following combinations of hormones (in mg/l): 0.05–0.5 NAA and 0.25–2.0 BA. Complete plant formation was obtained after rooting of the shoots on MS medium supplemented with IAA (0–2.0 mg/l) or NAA (0-0.5 mg/l).