生姜离体叶片愈伤组织悬浮细胞系建立与植株再生

以‘莱芜大姜’为试材,研究了生姜离体叶片愈伤组织的诱导以及细胞悬浮系建立与植株再生。结果表明,以生姜试管苗叶片为外植体,接种到MS+1.0 mg/L 2,4-D+0.5 mg/L 6-BA+30 g/L蔗糖的培养基上,可有效诱导出生长迅速、质地疏松的愈伤组织。将获得的愈伤组织接种到MS+0.15 mg/L 2,4-D+6.0 mg/L 6-BA+30 g/L蔗糖的液体培养基上,25℃黑暗条件下震荡培养25-30 d,可建立分散性好、生长迅速的悬浮细胞系,细胞悬浮系培养的适宜参数为:初始接种量为1.0-1.5 g,继代培养的适宜间隔期为15 d,继代培养液体培养基更新比例为3/4。将悬浮细胞接种到固体培养基MS+0.2 mg/L NAA+10.0 mg/L 6-BA+30 g/L蔗糖上可获得再生植株。     

盐生植物海马齿离体再生

建立盐生植物海马齿(Sesuvium portulacastrum)的离体再生体系,为其生物技术改良奠定基础。以海马齿叶片、茎和腋芽为外植体, 在不同激素配比的培养基上进行愈伤组织诱导、继代培养以及不定芽的分化和生根培养。结果表明: 最适愈伤组织诱导的外植体为叶片, 其次为幼嫩的茎段和腋芽。以叶片为外植体, 愈伤组织诱导率最高的培养基为MS+2.0mg·L^–12, 4-D + 0.5 mg·L^–16-BA + 3%sucrose; 芽分化最适培养基为MS + 1.0 mg·L^–1 2, 4-D + 0.2 mg·L^–1 6-BA + 3% sucrose;生根最适培养基为MS + 3%sucrose + 0.1%AC。炼苗移栽后, 成活率可达80%。     

抗茎枯病芦笋品种离体培养的研究北大核心CSCD

选用芦笋(Asparagus officinalis)茎枯病高抗品种格兰蒂为材料,研究各种生长素及细胞分裂素对其茎尖诱导的愈伤组织、不定芽增殖和生根的效果。实验结果表明,最适芦笋茎尖诱导愈伤组织的培养基为:MS+6-BA0.3 mg/L+2,4-D 1.5 mg/L+蔗糖25 g/L+琼脂7 g/L,pH 5.8;最适愈伤组织增殖诱导、分化不定芽的培养基为:MS+NAA 0.3 mg/L+6-BA 0.5 mg/L+蔗糖30 g/L+琼脂7 g/L,pH 5.8;最适生根培养基为:1/2 MS+0.5 mg/L IBA+1 mg/L PP333+0.3 mg/L 6-BA+蔗糖30 g/L+琼脂7 g/L,pH 5.8。     

麻疯树悬浮细胞系的建立与优化

以麻疯树无菌幼苗为外植体,研究疏松愈伤组织诱导方案及不同培养条件对麻疯树悬浮细胞生长的影响,旨在建立麻疯树悬浮细胞体系.结果表明,麻疯树疏松愈伤组织诱导的最适培养基及激素组合为:MS+2,4-D0.6mg/L+BA 1.0 mg/L+蔗糖30 g/L,此培养基上诱导出的愈伤组织湿润松散,颜色鲜艳.接种愈伤组织进行悬浮培养的液体培养基最适激素组合为:NAA 0.2mg/L+2,4-D 1.0 mg/L+BA 0.5 mg/L.初代愈伤组织适宜用于悬浮培养,摇床转速应低于120 r/min为宜,这样培养的悬浮细胞分散度最高.培养基中添加500 mg/L水解酪蛋白能有效地促进悬浮细胞的生长.悬浮细胞振荡培养过程中悬浮细胞生长的时间进程为起始培养的第5天前,细胞增殖十分缓慢;第5-11天期间生长迅速;第13天后基本停止生长.在上述优化培养条件下,麻疯树悬浮细胞系增长速率最快,细胞生长状态最佳.     

冬凌草组织培养及其愈伤组织诱导研究

实验采用L9（34）正交试验方法优化冬凌草组织培养快繁培养基,筛选冬凌草增殖、生根和愈伤组织诱导的最佳培养基配方。结果表明：不定芽诱导的最适培养基为MS＋0.2 mg/L IBA＋0.1 mg/L6-BA＋0.2 mg/L NAA;生根培养基为1/2 MS＋1 mg/L IBA＋1 g/L活性炭;以冬凌草茎段为外植体诱导愈伤最佳培养基为MS＋1.5 mg/L6-BA＋2 mg/L2,4-D。     

曼陀罗茎段愈伤组织诱导和再生植株的研究

本试验以曼陀罗茎段为外植体,在附加不同植物激素组合的培养基中对愈伤组织的诱导和植株再生进行研究。结果表明:采用修改的MS培养基(除去甘氨酸,维生素B1含量增加至0.5mg/L,pH5.5)附加2mg/L2,4-D可由曼陀罗茎段诱导大量胚性愈伤组织;愈伤组织继代选用0.5mg/L2,4-D为宜;不定芽的诱导采用MS培养基(20g蔗糖,8g琼脂,0.1g水解干酪素) 6-BA(0.5mg/L);幼苗进一步转接至1/2MS IBA(0.2mg/L)生根培养基中,可完成曼陀罗茎段愈伤组织诱导和再生植株的组织培养过程。     

白刺花胚性愈伤组织诱导及体细胞胚发生和萌发

探讨不同因素对白刺花下胚轴、子叶2种外植体胚性愈伤组织诱导及体细胞胚发生和萌发的影响。以B5和MS为基本培养基,研究2,4-D、6-BA和TDZ对白刺花下胚轴和子叶胚性愈伤组织的诱导;在MS培养基上添加不同浓度2,4-D,研究胚性愈伤组织增殖情况;采用ABA,探究对体细胞胚发生的影响。结果表明:下胚轴比子叶更易诱导胚性愈伤组织,筛选出2种外植最佳的胚性愈伤组织诱导培养基均为MS+2.0 mg/L 2,4-D+0.5 mg/L TDZ+0.5 mg/L 6-BA,胚性愈伤组织诱导率分别为77.3%和41.0%。15.0 mg/L ABA、0.2 mg/L 2,4-D和2.0 mg/L 6-BA有利于体细胞胚发生,1/3MS+0.2 mg/L NAA+0.1 mg/L 6-BA+2.0 g/L活性炭+25 g/L蔗糖+7 g/L琼脂的培养基可使体细胞胚萌发率达80%以上,再生植株移栽成活率高达90%。白刺花外植体种类及培养基类型均会影响胚性愈伤组织的诱导,其中下胚轴诱导效果优于子叶;MS培养基较适合启动细胞脱分化形成愈伤组织,2,4-D对胚性愈伤组织的增殖保持有调控作用,ABA有利于体细胞胚的发生。     

小桐子愈伤组织的诱导

本文以小桐子的叶片、叶柄、茎段及下胚轴和子叶作为外植体,研究不同外植体类型对愈伤组织诱导的影响,结果表明叶柄的诱导率最高,其次为茎段的诱导率。同时以小桐子的下胚轴作为外植体,研究植物生长调节剂种类及浓度配比对愈伤组织诱导的影响,结果显示6-BA与2,4-D的组合更适宜小桐子愈伤组织的诱导,MS+6-BA0.5mg/L+2,4-D0.1mg/L为小桐子愈伤组织诱导的最佳培养基,其愈伤组织诱导率高达96.7%。本研究为小桐子愈伤组织的分化、植株再生及相关的遗传转化研究提供了参考。     

红叶石楠离体植株再生频率的影响因素研究

以红叶石楠带芽茎段及叶片为外植体,分析激素和培养条件等因子对愈伤组织诱导及植株再生的影响。结果表明,MS+0.10mg/L 2,4-D+0.50mg/L NAA+0.50mg/L 6-BA+0.50mg/L KT为最佳愈伤组织诱导培养基,暗培养的愈伤组织诱导率高于光培养,其愈伤组织诱导率可达100%(带芽茎段)和98%(叶片)。MS+0.50mg/L IBA+2.00mg/L 6-BA+2.00mg/L KT为最佳分化增殖培养基,分化率91%以上,增殖倍数6.8以上,均达到最高。1/2MS+0.50mg/L IBA+0.01mg/L NAA为最佳生根培养基,生根率92%,生根量4.4根/株,均达到最高。     

睡菜离体快繁技术的研究

以睡菜的幼嫩茎段为外植体,接种到附加不同浓度激素配比(6-BA/NAA)的MS培养基,诱导睡菜愈伤组织、芽及根的生长。研究发现,外植体在1.0mg/L 6-BA+0.1mg/L NAA+MS的培养基上培养10d,可观察到浅绿色的愈伤组织。愈伤组织转接到4.0mg/L 6-BA+0.3mg/L NAA+MS培养基上2周左右可生成芽。对带芽的愈伤组织再进行诱导生根进而形成完整再生植株,最适根诱导培养基为0.3mg/L 6-BA+1.0mg/L NAA+MS培养基。该实验采用植物离体快繁技术成功建立了睡菜再生体系,为睡菜种苗规模化奠定了技术基础。     

The initiation of callus and regeneration from callus culture ofTulipa gesneriana

Intergeneric Fragaria vesca x Potentilla fruticosa hybrids were produced using in vitro culture. Hybrid plants were not obtained by direct embryo rescue, but were regenerated from cotyledon-derived callus. Experiments with F. vesca indicated that using cotyledon halves was not more productive than using entire cotyledons. A polarity was observed in cotyledons and in cotyledon halves, with callus and regenerated shoots produced more frequently from proximal ends. Cotyledons from 17% of hybrid embryos produced callus and regenerated mature plants. The technique enabled rapid multiplication of some embryos, with the production of more than one hybrid plant. In some cases more than 100 shoots were obtained from one embryo, demonstrating the potential usefulness of this technique for the production of intergeneric hybrids.Abbreviations BA 6-benzylaminopurine - IAA indole-3-acetic acid - NAA -naphthaleneacetic acid     

Influence of benzimidazole on callus growth and shoot formation on callus cultures

The influence of benzimidazole on callus growth and shoot formationin vitro was examined. At concentrations of 8.5 x 10^–4mMto 8.5 x 10^–1mM benzimidazole failed to stimulate callusgrowth or shoot formation. Concentrations of 0.85 mM and 1.7mM inhibited kinetin-dependent callus growth while lower concentrationswere without effect. It may be inferred that benzimidazole hasno cell division properties such as is associated with the cytokinins,though it mimics, in appearance, cytokinins actions in severalphysiological processes. (Received August 12, 1975; )     

The initiation of callus and regeneration from callus culture ofTulipa gesneriana

Cold treatment of seeds, obtained from crosses between cultivars ofT. gesneriana L., affects the developmental stage of embryos, which in turn influences the frequency of callus induction and the development of different callus types. Cold-treated, mature embryos and basal segments ofin vitro-derived bulblets, were suitable explants for the initiation of regenerative callus on medium with 2,4-dichlorophenoxyacetic acid. The bulblets were initiated on flower-stalk segments from cold-stored bulbs ofT. gesneriana ‘Christmas Marvel.’ Histological analyses of regenerative callus revealed the regeneration of bulb-like structures. The influences of culture medium, culture conditions, growth regulators and acetylsalicylic acid, an inhibitor of ethylene, on the initiation and establishment of regenerative callus cultures are discussed.     

Agrobacterium-mediated transformation of seedling-derived maize callus

Efficient production of seedling-derived Type I callus was demonstrated for several corn genotypes including commercial inbred lines. Seeds were germinated on MS-based medium containing 10 mg l(-1) picloram and 3 mg l(-1) 6-benzylaminopurine, which induced the development of axillary buds in the area of coleoptilar node. Nodal sections of 7-10-day old seedlings were isolated, split longitudinally, and placed on callus induction medium supplemented with 2.2 mg l(-1) picloram and 0.5 mg l(-1) 2,4-dichlorophenoxyacetic acid. For lines L4 and L9 the frequency of embryogenic callus induction was 38-42% based on calli per split nodal section. Frequency of callus induction from split nodal sections of seeds germinated on media without growth regulators was 0-3%. Seedling-derived callus of five genotypes was used for Agrobacterium-mediated transformation. Two constructs containing the green fluorescence protein gene and genes for either neomycin phosphotransferase II or glyphosate selection were used in transformation experiments. Transformation frequency varied from 2 to 11% and about 60% of the T(0) plants had 1-2 copies of transgenes.     

Breakdown of phosphatidylinositol in soybean callus

We have investigated the breakdown of membrane-bound phosphatidylinositol (PI) in homogenates of soybean (Glycine max) callus. The breakdown of PI was stimulated by the detergent deoxycholate. At pH 7.0 and 1·gl^-1 of deoxycholate the loss of PI was rapid and extensive: more than 80% was broken down within 10 min. The breakdown of PI was also stimulated by millimolar concentrations of Ca²⁺. The products of breakdown of added PI (purified from soybean callus) in this system were identified from their chromatographic mobilities as 1,2-diacylglycerol, myo-inositol 1-phosphate and myo-inositol 1:2-cyclic monophosphate.Abbreviations DOC deoxycholate - EDTA ethylenedi-aminetetraacetic acid,-acetate - Pi Inorganic phosphate - PI phosphatidylinositol - PS phosphatidylserine - TLC thinlayer chromatography     

Metabolism of kinetin by soybean callus

Soybean callus metabolised applied 6-furfurylamino (8^-14C) purine very rapidly to polar compounds, some of which were retained on a Dowex 50 cation exchange resin, and were unaffected by alkaline phosphatase; while others were apparently phosphorylated and were detected in the aqueous fraction. Adenine was detected as an intermediate and it can be concluded that it was formed as a result of the rapid and efficient removal of the furfuryl side chain. The formed adenine was rapidly incorporated into the polar metabolites. Exactly how the presence of this cytokinin stimulates cell division is not apparent from the results.The financial support of the C.S.I.R., Pretoria, and The Natal University Development Fund is gratefully acknowledged.