盾叶薯蓣类原球茎的离体诱导及快繁体系的建立

为解决盾叶薯蓣离体培养中试管苗移栽困难的难题,以盾叶薯蓣带腋芽的茎段为外植体,借助正交试验设计方法,离体诱导出类原球茎并建立了类原球茎微繁殖技术体系。结果表明:以带腋芽茎段为外植体诱导致密愈伤组织的适宜培养基为MS+6-BA2.0mg/L+NAA0.4mg/L+KT0.6mg/L+蔗糖3%;类原球茎诱导和增殖培养基为:MS+6-BA4.0mg/L+KT1.0mg/L+蔗糖6%;类原球茎生根培养基:1/2MS+NAA 0.3mg/L+IAA 0.8mg/L+活性炭0.3%+蔗糖1.5%。经该途径诱导得到的生根类原球茎植株经炼苗后移栽的成活率可达到90%以上。     

‘SK4—316’胡萝卜体胚的诱导和培养

以'SK4-316'胡萝卜无菌苗的下胚轴为外植体,研究不同培养基配方和培养条件对愈伤组织诱导、体细胞胚间接发生及其同步化培养的影响,以及不同脱分化时间、脱分化培养基及外植体续存时间对体细胞胚直接发生的诱导及其培养的影响.结果表明:含3%蔗糖、0.8%琼脂的1/2MS + 2,4-D 2.5 mg/L + 6-BA(或KT)0.5 mg/L + CH 300 mg/L是诱导愈伤组织的良好培养基;1/2MS + 2,4-D 1.25 mg/L + KT 0.25 mg/L + 6-BA 0.25 mg/L(含3%蔗糖)适于愈伤组织分化并诱导体胚发生,0.02% ABA对体胚的诱导有促进作用,0.06% ABA或15% PEG能促进体胚成熟;外植体在MS + 2,4-D 1.0 mg/L固体培养基上脱分化培养48 h,再转入MS + CH 300 g/L液体培养基中可诱导体胚直接发生,但随着外植体续存于诱导培养基中时间的延长,体胚发生变异的几率也渐增.     

药用植物灯盏花的组织培养

以灯盏花花葶、花盘及叶柄为外植体,MS为基本培养基,通过不同的激素种类和浓度配比,建立灯盏花组培快繁体系。结果如下:在所有实验方案中,花葶的出愈率最高,是理想的快速繁殖材料。较适宜的诱导愈伤组织的培养基为MS+BA1.0mg/L+IBA0.05mg/L+蔗糖3.0%,诱导不定芽的培养基为MS+BA2.0mg/L+IAA1.0mg/L+蔗糖3.0%或MS+Kt3.0+IAA0.5mg/L+蔗糖3.0%,而根的诱导则是在1/2MS+NAA1.0Mg/L+蔗糖3.0%的培养基上进行。同时对组织培养过程中灯盏花植株再生的方式进行了讨论。     

剑叶龙血树组织培养与快速繁殖的研究

用剑叶龙血树的腋芽、顶芽或茎段作外植体,构建了剑叶龙血树组织培养再生体系。试验结果如下:剑叶龙血树腋芽、顶芽或茎段诱导愈伤组织的最佳培养基为MS+6-BA 2.0mg/L+NAA 0.5mg/L,诱导率达95%;选择MS+6-BA 2.0mg/L+NAA0.2mg/L+30g/L的蔗糖+8g/L琼脂培养基有利于芽的分化和增殖,在1/2MS+NAA 0.2mg/L+IBA0.4mg/L+活性炭的培养基中诱导生根效果最好。     

连钱草组培快繁技术研究

用连钱草无菌茎尖为外植体进行快速繁殖,分别诱导、分化、生根形成再生植株进行快速繁殖,并移栽成活。结果表明在MS+6-BA1.5mg/L+NAA0.1mg/L培养基上诱导丛生芽效果最佳。在MS+IBA1.0mg/l+KT1.0mg/L培养基中根的诱导率为100%。     

菜心组织培养技术初探

为建立菜心(Brassica campestris ssp.chinensis var.utilis)的快繁技术体系,以花药和子叶-子叶柄为外植体进行组织培养研究。结果表明,花药培养以选取未开放的花蕾为宜,且花柱略高于花瓣,此时小孢子多数处于单核靠边期。菜心花粉的萌发率不高,且秋冬季的花粉比夏季的萌发率高。菜心花药愈伤组织诱导培养基为:MS+1.0 mg L–1 KT+1.0 mg L–1 2,4-D+3%糖+6 g L–1琼脂+8%椰乳,不定芽诱导培养基为:MS+2.0 mg L–1 6-BA+0.5 mg L–1 NAA+1.0 g L–1活性炭+2%糖+6 g L–1琼脂或MS+2.0 mg L–1 ZT+0.5 mg L–1 IAA+0.5 g L–1 AgNO3+1.0 g L–1活性炭+2%糖+6 g L–1琼脂。花药培养的不定芽诱导率为36.7%,不定芽培养出现褐化现象,不能形成再生植株;而以子叶-子叶柄为外植体培养获得的植株再生率可达80%。     

匙叶芋芽的诱导和植株再生

植物名称:匙叶芋Spathiphyllum sp.材料类别:根茎。培养条件:基本培养基为MS。诱导芽培养基,附加激素:(1) BA 2.0 mg/L(单位下同)+KT1.0,(2) KT0.5+NAA 0.2,(3) BA 0.5+IAA 1.0。芽增殖培养基,附加激素BA 1.0+IAA 0.1。生根培养基,MS减半,蔗糖2％,琼脂0.7％。培养温度25±2℃,每天光照10～12小时,光照强度2000 lx。     

甘肃贝母离体小鳞茎直接发生途径的研究

目的：建立甘肃贝母组织培养小鳞茎的直接发生技术体系。方法：采用单因子试验设计，筛选适宜甘肃贝母组织培养的外植体及灭菌方法，探究外植体种类、茎段大小、培养温度、激素组合、培养方式等对小鳞茎诱导和增殖的影响。结果：休眠芽的最佳灭菌方式为75%乙醇灭菌30 s,0.1%升汞灭菌1 min，无菌水冲洗3～5次。休眠芽为外植体时培养效果最好，其次为茎段和小鳞叶，叶片效果较差。茎段大小对甘肃贝母组培小鳞茎诱导影响不大。固体培养基更适宜直接诱导小鳞茎，培养基配方为：1/2 MS+6-BA 0.5 mg/L+NAA 0.1 mg/L或1/2 MS+6-BA 1.0 mg/L+NAA 1.0 mg/L，蔗糖3%、琼脂0.7%,pH 5.8。一定的变温培养有利于其鳞茎增殖。适宜的生根培养基为1/2 MS+NAA 0.1 mg/L+IAA 0.5 mg/L+1.5%蔗糖+0.7%琼脂，pH 5.8。结论：建立了甘肃贝母离体培养小鳞茎直接发生技术体系，可为甘肃贝母种质保存及工厂化育苗提供技术支撑。     

巴戟天组织培养和快速繁殖研究

以巴戟天顶芽及嫩茎节段为外植体,以MS为基本培养基,通过不同的激素种类和浓度配比,建立巴戟天组培快繁体系。结果表明,外植体表面消毒以70%酒精预处理60s,再用0.1%HgCl2浸泡10min,效果较好,茎节为外植体优于顶芽。培养基MS+BA1.0mg/L+IBA0.05mg/L利于诱导出芽,可用于初代培养;MS+BA1.0mg/L+IBA0.2mg/L利于形成丛生芽,用于继代增殖,繁殖系数6.0/50d;1/2MS+IBA0.4~0.8mg/L适宜诱导生根获得再生植株,生根率100%;生根苗移栽于排水良好的火土或砂土中,成活率90%。     

铁皮石斛愈伤组织诱导研究

研究不同培养基和光照条件对铁皮石斛愈伤组织诱导的影响。结果表明,外植体直接接种于培养基上,最适宜培养条件是MS 5.92 g·L^-1＋2,4-D 5 mg·L^-1＋IAA 1.5 mg·L^-1＋KT 0.62 mg·L^-1＋蔗糖37.5 g·L^-1＋琼脂0.8% (pH 5.9～6.0),暗培养15 d后再光培养;外植体捣碎后平铺于培养基上,最适宜培养条件是MS 4.74 g·L^-1＋2,4-D 1 mg·L^-1＋IAA 1.5 mg·L^-1＋KT 0.25 mg·L^-1＋蔗糖30 g·L^-1＋琼脂0.8% (pH 5.9～6.0),25 ℃持续光培养。     

Micropropagation of <Emphasis Type="Italic">Vitex agnus-castus</Emphasis>, (Verbenaceae)—a valuable medicinal plant

This study describes in vitro shoot induction and plant regeneration from a mature apical meristem and nodal explants of the endangered medicinal shrub Vitex agnus-castus. Multiple shoots were induced directly from the axis of nodal and apical meristem explants on Murashige and Skoog (MS) medium containing 3% sucrose and different concentrations (1.0, 1.5, 2.0, and 2.5 mg/l) of 6-benzyl aminopurine (BAP) in combination with Kinetin (Kin) and α-naphthalene acetic acid (NAA), both at 0.1 mg/l. BAP and Kinetin were used as supplements to MS basal medium, either individually or in combination with auxins. The optimal concentration of BAP for inducing bud break was found to be 2.0 mg/l when Kinetin was at 0.1 mg/l. Regeneration frequency was highest for both apical meristem and nodal explants (94.5% and 90.3%, respectively) when explants were cultured on MS medium supplemented with BAP (2.0 mg/l) and Kin (0.1 mg/l). A maximum of 7.7 ± 0.4 and 6.7 ± 0.2 shoots were obtained per explant for apical meristem and nodal explants, respectively. Regenerated shoots, transferred to MS medium supplemented with either 1.0 or 1.5 mg/l BAP combined with 0.1 mg/l GA₃, showed maximum elongation of 6.7 ± 0.4 and 6.0 ± 1.3 cm in apical meristem and nodal explants, respectively. In vitro regenerated shoots transferred to half-strength MS medium supplemented with 0.1 mg/l IBA induced 90.4% of the shoots to form roots after 30–35 d of culture. Up to 80% of the regenerated shoots were successfully established in soil in the greenhouse.     

海滨锦葵胚轴愈伤组织诱导及植株再生

以海滨锦葵(Kosteletzkya virginica)胚轴为外植体, 在9种不同激素配比的培养基上进行愈伤组织诱导、继代培养、不定芽分化及生根培养, 确定了植株再生的最适培养条件: (1)愈伤组织诱导最适培养基为MS + IAA 1.0 mg.L-1 + KT 0.3 mg.L-1 + sucrose 30 g.L-1 + agar 8 g.L-1, 愈伤组织诱导率为93.94%; (2)不定芽诱导最适培养基为MS + IAA 0.1 mg.L-1 + ZT 0.5 mg.L-1 + sucrose 30 g.L-1 + agar 8 g.L-1, 不定芽诱导率为65.83%; (3)生根最适培养基为MS + sucrose 30 g.L-1 +agar 8 g.L-1, 生根率为96.67%。炼苗移栽后, 成活率可达85%。     

Establishment of a highly efficient transformation system for pepper (Capsicum annuum L.)

Application of modern genetic manipulation has been limited in pepper ( Capsicum annuum L.) due to the lack of an efficient transformation system. Following the development of an efficient protocol for in vitro regeneration of pepper cotyledons, we investigated the key factors affecting transformation and established a highly efficient genetic transformation system using the pepper cotyledon as starting material. In this system, cotyledon explants are preconditioned for 2 days on kanamycin (km)-free DM1 medium [Murashige and Skoog (MS) salts/Gamborg B5 vitamins basal medium supplemented with 20 g/l sucrose, 5,000 mg/l DJ nutrients and a hormone combination of 1.0 mg/l indoleacetic acid (IAA) and 5.0 mg/l 6-benzyladenine (BA) solidified with 0.7% agar, pH 5.8], followed by co-cultivation with Agrobacterium tumefaciens on DM1 for 2 days and delay selection on DM1 with 500 mg/l carbenicillin (carb) for 2 days. The explants are then placed on DM1 containing 10 mg/l AgNO(3), 50 mg/l km-sulfate and 500 mg/l carb. After 4-5 weeks, the explants with buds are transferred to EM1 medium (MS salts/Gamborg B5 vitamins basal medium supplemented with 20 g/l sucrose, 5,000 mg/l DJ nutrients, 10 mg/l AgNO(3) and a hormone combination of 1.0 mg/l IAA, 3.0 mg/l BA and 2.0 mg/l gibberellic acid, solidified with 0.7% agar, pH 5.8) with 50 mg/l kanamycin and 500 mg/l carbenicillin for the elongation of buds. After 3-6 weeks, 1- to 2-cm-long elongated shoots are excised and planted on RM1 medium (MS basal medium supplemented with a hormone combination of 0.2 mg/l NAA and 0.1 mg/l IAA, solidified with 0.8% agar, pH 5.8) with 25 mg/l km and 200 mg/l carb for rooting. We tested four genotypes of pepper, and all presented a high differentiation efficiency (81.3% on average), elongation rate (61.5%) and rooting efficiency (89.5%). Polymerase chain reaction analysis results showed that 40.8% of the regenerated plantlets were transgenic plants.     

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。     

Plants obtained from the Khair tree (Acacia catechu Willd.) using mature nodal segments

An in vitro method for obtaining plants of Acacia catechu has been developed using nodal explants from mature `elite' trees growing in the field. Maximum shoot bud development (eight to ten) from a single explant was achieved on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP) (4.0 mg/l) and α-naphthaleneacetic acid (0.5 mg/l). Addition of adenine sulphate (25.0 mg/l), ascorbic acid (20.0 mg/l) and glutamine (150.0 mg/l) to the medium was found beneficial for maximum shoot bud induction. The shoot buds developed into healthy and sturdy shoots on MS medium containing BAP and kinetin at 1.0 mg/l. Excised shoots were rooted on 1/4-strength MS medium with indole-3-acetic acid at 3.0 mg/l and 1.5% sucrose to obtain complete plants. Received: 17 June 1997 / Revision received: 11 September 1997 / Accepted: 27 September 1997     

金线莲快速繁殖及促根壮苗试验

以金线莲(Anoectochilus roxburghii)试管苗带节茎段和顶芽为外植体,用正交试验筛选芽快速增殖的培养基配方,并研究生根粉(ABT)对促根壮苗的作用。结果表明,丛生芽增殖最佳配方为MS+BA 3.5mg/L+KT 1.5mg/L+NAA 0.6mg/L,两个月增殖倍数达3.5;3mg/L 3号生根粉对金线莲生根壮苗效果最佳,使植株生根条数、最长根、植株增高和增叶数分别比对照增加12.8%、34.1%、13.2%和22.4%,从而提高移栽成活率,3种不同基质均达到94%以上。     

毛白杨悬浮细胞系的建立及再生植株的获得

以毛白杨基因型TC152无菌苗为材料,研究毛白杨悬浮细胞系建立与植株再生,结果表明,通过悬浮培养和固体培养两种方法诱导毛白杨悬浮细胞分化不定芽,最终获得无菌生根苗。愈伤组织在MS＋1.5mg·L-12,4-D＋30g·L-1蔗糖的液体培养基中振荡培养,12d可建立悬浮细胞系;悬浮细胞系继代培养基为MS＋0.8mg·L-12,4-D＋30g·L-1蔗糖,继代周期为7d,悬浮细胞在MS＋1.0mg·L-16-BA＋0.1mg·L-1NAA＋0.5～1.0mg·L-1ZT＋30g·L-1蔗糖培养基中悬浮培养,可分化大量不定芽,每个培养瓶中可得到40～50个芽,个别不定芽玻璃化;不定芽在1/2MS＋0.6mg·L-1IBA＋20g·L-1蔗糖＋5.5g·L-1琼脂培养基上可分化不定根。悬浮细胞通过固体平板培养增殖为愈伤组织块后,在MS＋1.0mg·L-16-BA＋0.1mg·L-1NAA＋1.0mg·L-1ZT＋30g·L-1蔗糖＋5g·L-1琼脂的固体培养基上,不定芽分化率可达到70.00%。     

Effect of ABA on the in Vitro Induction of Floral Buds of Dendrobium candidu Wall.ex Lindl.

An in vitro flowering system of Dendrobium candidurn Wall. ex Lindl., a wild species of orchid, was established. Callus was induced from seeds and protocorms formed on MS agar medium supplemented with 0.3 mg/L NAA under diffused light. Floral buds were induced when protocorms were cultured on MS agar medium supplemented with 2 mg/L 6-BA and 0.5 mg/L NAA, the frequency of floral bud induction being 27.0%. When protocorms were precultured on MS medium supplemented with 0. 5 mg/L ABA for 15 days and then transfered onto MS medium supplemented with 2 mg/L 6-BA, the frequency of floral bud induction increased greatly reach 84.0 % during a period of 5 months. Meanwhile the number of branches and floral buds also increased and in some instances inflorescence appeared. Never the less, no floral bud was formed if protocorms continued to be cultured on the ABA-containing MS medium.     

石斛离体培养中ABA对诱导花芽形成的影响

由兰科植物铁皮石斛（Ｄｅｎｄｒｏｂｉｕｍ ｃａｎｄｉｄｕｍ Ｗａｌｌ．ｅｘ Ｌｉｎｄｌ．）种子诱导形成的愈伤组织,在光照下置于ＭＳ附加０．３ ｍ ｇ／ＬＮＡＡ 的培养基上繁殖,可以形成原球茎。将原球茎转入ＭＳ含２ ｍ ｇ／Ｌ ６－ＢＡ 和０．５ ｍ ｇ／ＬＮＡＡ 的培养基上,花芽形成频率为２７．０％ 。原球茎先在０．５ ｍ ｇ／ＬＡＢＡ的培养基上预培养１５ ｄ,再转入含２ ｍ ｇ／Ｌ６－ＢＡ 的ＭＳ培养基上培养,花芽形成频率明显提高,可达８４．４％ ,而且每株植株花的数目增加;但是在仅有ＡＢＡ 的ＭＳ培养基上培养的原球茎再生的植株未见花芽形成     

正交实验法在香果树种质离体保存中的应用

以香果树带芽茎段为外植体,采用正交实验法研究活性炭、琼脂、封口材料和培养容器对香果树试管苗离体保存的影响。结果表明,香果树带芽茎段用200 ml三角瓶,无菌培养容器封口膜封口,保存于MS+KT 2.0 mg/L+6-BA 2.0 mg/L+NAA 0.1 mg/L+30 g/L蔗糖+0.5 g/L活性炭+7.5 g/L琼脂培养基上6个月后,成活率可达86.3%。离体保存的试管苗没有发生遗传变异。