青阳参组织培养及愈伤组织的成分分析

用青阳参（Cynanchum otophyllum）的嫩枝和芽在MS + 2.0 mg/L 2,4-D + 0.1 mg/L KIN的培养基上诱导愈伤组织。通过不同的培养基和激素配比实验,发现 6,7-V + 2.0 mg/L 2,4-D + 0.3 mg/L KIN 最适合愈伤组织的生长。但在6,7-V + 1.0 mg/L 2,4-D + 0.1 mg/L KIN 培养基中的愈伤组织次生代谢物含量最高。愈伤组织的生长周期为27 d,但在33 d时次生代谢产物的含量最高。从愈伤组织中分离到7个化合物：⑴9,10,11-三羟基-十八碳-12(Z)-烯酸甲酯 (methyl 9,10,11-trihydroxy-12-octadecenoate),(2) 胡萝卜甙 (daucosterol),(3)β 谷甾醇 (β sitosterol),（4） 华木酸 (betulinic acid),（5）齐端果酸 (oleanolic acid),（6）棕榈酸 (hexadecanoic acid),（7）十八碳-9-烯酸 (9-octadecenoic acid)。首次报道从植物愈伤组织中分离到多羟基十八碳烯酸,并讨论了化合物（1）对植物细胞生长的可能影响。     

中华结缕草(Zoysia sinica Hance)组织培养和再生植株研究

以中华结缕草(Zoysia sinica Hance)成熟种子为外植体在附加2．5mg／L2,4-D、0．25mg／L 6-BA和1～2mg／L VB1的改良MS培养基(MSm)上愈伤组织的诱导率最高为43．0％。愈伤组织的最佳继代培养基为MSm附加0．1mg／L 6-BA和2．0mg／L 2,4-D。在无生长调节物质的MS培养基(MS0)上,外观呈白色到淡黄色、含有密实颗粒的愈伤组织再生率为30％～60％。     

荞麦组织培养及高频植株再生体系的建立

通过对荞麦(Fagopyrum esculentum Moench)不同外植体、不同激素配比的比较研究,建立了荞麦离体培养高效植株再生体系。荞麦子叶切段在含2．0 mg/L 2,4-D和1．0 mg/L 6-BA的MS培养基上愈伤组织诱导率为89．6％,而下胚轴切段在含2．0 mg/L 2,4-D和1．0-2．0 mg/L 6-BA MS培养基上愈伤组织诱导率高达100％。在2．0 mg／L 6-BA、0．1 mg,L IAA和1 mg／L KT的MS培养基上通过愈伤组织间接分化或外植体直接分化形成不定芽。来自子叶和下胚轴的愈伤组织的分化率分别为42．5％和73．6％,下胚轴的分化率明显高于子叶。将生长状态良好的不定芽转至含1．0 mg/L IBA和0．5mg/L NAA的1/2 MS培养基上生根,生根率达到100％。再生植株移栽到盆土中,成活率达91．6％,并且生长状态和特征均表现正常。     

野牛草幼穗愈伤组织的诱导及植株再生

以野牛草[Buchloe dactyloides(Nutt．)Engelm．]幼穗为外植体,建立了愈伤组织诱导、继代培养和植株再生体系。结果表明,雌穗比雄穗难以脱分化形成愈伤组织;小于8mm雄幼穗在2mg／L2,4-D培养基上的愈伤组织诱导率为80．0％～86．8％;添加10mg／L AgNO3对愈伤组织诱导率影响不明显,但可改善愈伤组织质量。2mg／L 2,4-D结合0．1mg／L 6-BA的培养基有利于愈伤组织的继代培养;继代超过3次、继代间隔超过3周,愈伤组织分化能力明显下降。雄穗愈伤组织在含1．0mg／L 6-BA培养基上,弱光条件下分化出芽的频率较高,达31．8％～35．0％;附加3％麦芽糖既可减轻褐化程度,又利于丛生芽的分化。分化苗在1/2MS 0．3mg／L IBA培养基上的生根率为62．5％。     

费尔干猪毛菜愈伤组织的诱导、分化及植株再生体系的建立

探讨了植物生长调节剂、外植体等因子对费尔干猪毛菜（Salsola fergartica Drob．）愈伤组织诱导、分化与植株再生的影响。结果表明：2,4-D与6-BA组合使用时,在一定浓度范围内均有愈伤组织产生,最佳的诱导培养基为MS＋2,4-D2．0mg／L＋6-BA0．2mg／L,下胚轴为诱导愈伤组织的最佳材料;愈伤组织继代培养较好的培养基为MS＋NAA0．1mg／L＋6-BA1．0mg／L;愈伤组织不定芽分化培养基为MS＋6-BA0．5mg／L＋NAA0．05mg／L;根分化的培养基为1／2MS＋IBA0．2mg／L,且生根率可达72％。以带柄子叶为外植体,诱导丛生芽的最佳分化培养基为MS＋6-BA1．0mg／L,再生频率可达90．74％。     

地黄组织培养及植株再生的研究

以地黄根茎所获无菌苗为材料,对其愈伤组织诱导、分化和再生植株的获取进行了初步研究。结果表明：取叶片、茎段、叶柄进行愈伤组织诱导,筛选出最适培养基为MS附加2,4-D0．5mg／L、BA1．0mg／L,愈伤组织诱导率可达100％。将叶片接种在分化培养基中,诱导不定芽,其最适分化培养基为MS附加BA 3mg／L、NAA 0．1mg／L,分化率为77．5％。试管苗在改良的MS(大量与微量元素、铁盐和有机物质各1／2)附加NAA 0．05mg／L的培养基上,经过15～20d培养,生根率可达100％。     

桃儿七成熟胚的离体培养研究

以MS为基本培养基,通过添加不同激素和附加物等措施对桃儿七种子成熟胚进行了无菌苗和愈伤组织诱导初步研究。研究结果表明：0．5mg／L IAA＋0．2mg／L KT＋0．3mg／L GA3处理可获得生长良好的无菌苗,诱导率达41．67％。1．0mg／L 2,4-D＋0．4mg／L KT＋0．3mg／L GA3处理愈伤组织诱导率可达36．67％,添加500mg／L水解酪蛋白（CH）或10mg／L D-生物素有益于愈伤组织的诱导。光照和变温培养有利于提高无菌苗和愈伤组织的诱导率,有效降低褐化率。     

火龙果愈伤组织诱导与植株再生

为了建立火龙果愈伤组织诱导与植株再生体系,以火龙果茎段、幼苗和子叶为外植体进行离体培养试验。结果表明：茎段诱导愈伤组织的最优培养基为1／2MS＋2,4-D2．0mg·L^-1＋6-BAO．5mg·L^-1,诱导子叶愈伤组织的最适培养基是1／2MS＋2,4-D2．0mg·L^-1＋6-BA1．0mg·L^-1,诱导愈伤组织分化的最优培养基为1／2MS＋6-BA4．0mg·L^-1＋NAA0．5mg·L^-1,最佳生根培养基为1／2MS＋6．BA1mg·L^-1＋NAA0-3mg·L^-1。     

盾叶薯蓣组织培养技术的优化

以盾叶薯蓣的根状茎、茎段、叶柄、幼叶为材料,进行愈伤组织诱导、分化及再生植株形成的研究。结果表明：盾叶薯蓣不同外植体均能诱导出愈伤组织,其中茎段愈伤组织的诱导率最高;不同激素配比的培养基对愈伤组织的形成有很大的影响：以LS为基本培养基,2,4-D浓度为4．0mg／L、6-BA浓度为1．0mg／L的激素配比诱导率最高,达62．5％;以改良MS为基本培养基,2,4-D浓度为2．0mg／L、6-BA浓度为0．5mg／L的激素配比诱导率最高,达71．4％。筛选到优化的分化培养基为改良MS附加2．0mg／L的6-BA和0．5mg／L的Vc,且能直接诱导出根,并形成完整植株。     

植物生长调节剂对白头翁叶片培养的影响

以白头翁试管苗叶片为外植体,以MS为基础培养基,探讨不同细胞分裂素和生长素对叶片不定芽诱导、愈伤组织的诱导、增殖、再分化的影响。实验结果表明：细胞分裂素TDZ适合于白头翁叶片培养,与生长素搭配使用使叶片发生分化;2,4-D对愈伤组织的诱导能力相对较强。叶片直接诱导不定芽的激素组合为TDZ0．3mg／L＋NAA0．1mg／L;愈伤组织增殖的激素组合为TDZ0．2mg／L＋2,4-D0．2mg／L,将愈伤组织转接到TDZ和NAA组合的培养基上时会再分化。     

Determination of phenolic antioxidant compounds produced by calli and cell suspensions of sage (Salvia officinalis L.)

Sage (Salvia officinalis L.) calli were established by culturing internodal segments, excised from aseptic seedlings, on MS basal medium gellied with agar and supplemented with 0.05 mg/L dichlorophenoxyacetic acid (2,4-D) in presence of benzyladenine (BA) or zeatin (ZEA) or kinetin (KIN), at 1.5 mg/L. Suspended cells were established by transferring one callus to 50 mL of liquid MS basal medium devoid of agar and containing the same type of hormonal supplementation used in respective calli growth. The highest growth of calli and suspensions occurred with 1.5 mg/L ZEA. However, with this cytokinin supplementation, as well as with 1.5 mg/L KIN, both in presence of 0.05 mg/L 2,4-D, suspensions differentiated small root shaped structures. Well shaped, majority single cell suspensions were formed under the effect of 0.05 mg/L 2,4-D and 0.5 mg/L KIN. Calli grown with 0.05 mg/L 2,4-D and 1.5 mg/L BA and suspended cells grown with 0.05 mg/L 2,4-D and ZEA or KIN at 1.5 mg/L, or KIN at 0.5 mg/L, were searched for phenolics production. Twelve phenolic compounds were identified in calli: gallic acid, 3-O-caffeoylquinic acid, 5-O-caffeoylquinic acid, caffeic acid, rosmarinic acid, hesperetin, epirosmanol, hispidulin, genkwanin, carnosol, carnosic acid, and methyl carnosate. With the exception for genkwanin and epirosmanol all of these phenolic compounds were also produced by the sage suspension cultures grown in the presence of 1.5 or 0.5 mg/L KIN. Genkwanin was the only phenolic absent in the suspensions grown with 1.5 ZEA. Suspended cells, grown with 0.5 mg/L KIN, and calli cultures showed the highest specific accumulation of the total phenolics, with rosmarinic acid representing 94-97 percnt;.     

Callus Induction in Small Flowered Willow Herb (<Emphasis Type="Italic">Epilobium parviflorum</Emphasis> Schreb)

In order to determine the most suitable in vitro tissue culture and plant regeneration conditions for the small flowered willow herb (Epilobium parviflorum Schreb), various explants were cultured on semi-solid MS media containing factorial combinations of plant growth regulators. Callus induction from hypocotyl, cotyledon, petiole and leaf explants was achieved on media containing 2,4-dichlorophenoxy acetic acid (2,4-D) and kinetin (KIN). All other growth regulator combinations [□-naphtaleneacetic acid (NAA) ± benzylaminopurine (BAP), NAA ± thidiazuron (TDZ), indol acetic acid (IAA) ± Zeatin (ZEA)] tested failed to respond. The best results with cotyledon- and petiole- derived callus were obtained from MS medium supplemented with 1.0 mg l^?1 2,4-D + 0.1 mg l^?1 KIN and 2.0 mg l^?1 2,4-D + 0.2 mg l^?1 KIN. It was observed that B5 basal medium was more effective than MS basal medium for producing seedling and the most effective seed sterilizing solution was 25 % (v/v) sodium hypochlorite (NaOCl). No plant regeneration was observed in either callus induction or during the subculturing stage. This is the first report on in vitro tissue culture study within the genus Epilobium.     

Somatic embryogenesis and plant regeneration from seeds of wild<Emphasis Type="Italic"> Dicentra spectabilis</Emphasis> (L.) LEM

Regeneration via somatic embryogenesis from callus was studied in Dicentra spectabilis. To obtain somatic embryogenic callus, we cultured D. spectabilis seeds on MS basal media supplemented with various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D). The highest percentage of embryogenic callus formation was observed on media containing 1.0 mg/l 2,4-D under dark conditions. Somatic embryogenesis was studied by transferring the callus onto MS basal medium containing different concentrations (0.0, 0.1, 0.5, 1.0, 2.0 mg/l) of KIN (kinetin) and/or BAP. Somatic embryogenesis on MS basal media with 1.0 mg/l of KIN was excellent under light conditions. Somatic embryos were rooted by transferring them to half-strength MS basal media containing 2 g/l Phytagel. About 64.2% of the somatic embryos converted to rooted plantlets, 4% showed secondary embryogenesis and 31.8% did not develop and died. Rooted plantlets showed a 46% survival rate when acclimatized ex vitro.Abbreviations BAP 6-Benzylaminopurine - 2.4-D 2,4-Dichlorophenoxyacetic acid - KIN Kinetin - SEM Scanning electron microscopyCommunicated by H. Lörz     

Morphogenetic effects of 2,4-dichlorophenoxyacetic acid on pinto bean (Phaseolus vulgaris L.) leaf explants in vitro

Roots, callus and/or globular structures were produced on primary leaf and distal cotyledon explants of pinto bean (Phaseolus vulgaris L. cv. UI 114) cultured on semisolid MS medium with a wide range of 2,4-D concentrations (0.01 to 80 mg/L) with either 0 or 1.0 mg/L kinetin. Explants rooted at lower 2,4-D concentrations than at those favoring globule formation on callus, although roots, callus and globules often developed from the same explant. Isolated opaque green globular structures developed when callus initiated on media with 3 or more mg/L 2,4-D was subcultured in liquid MS + 30 mg/L 2,4-D. These structures multiplied with a fresh weight doubling time of 8–9 days in MS + 30 mg/L 2,4-D. Although this multiplicative behavior and opaque color were reminiscent of embryoids reported for other species, no cotyledons or roots were seen.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - KIN kinetin - MS Murashige-Skoog medium Cooperative investigations of the Agricultural Research Service, U.S. Department of Agriculture and the Michigan Agricultural Experiment Station, East Lansing, Michigan 48824. Michigan Agricultural Experiment Station Journal article No. 11923     

红豆杉的愈伤组织诱导及培养研究（初报）

以红豆杉雄球花、茎切段等为外植体诱导产生了愈伤组织。适宜愈伤组织诱导及其生长的培养基为１／２ＭＳ培养基附加２,４—Ｄ２．０ｍｇ／Ｌ和ＮＡＡ０．１ｍｇ／Ｌ。在不同浓度激素配比的培养基上均未观察到愈伤组织发生再分化。     

Study on Tissue and Protoplast Culture of Wild Cotton (Gossypium davidsonii)

This paper deals with the study on the condition of callus formation, embryogenesis, organogenesis, plant regeneration and protoplast culture of wild cotton (G. davidsonii) Callus cultures derived from several organs such as root, stem, leaf, cotyledon and hypocotyl. The results obtained in these cultures showed that the modified MS medium containing 2,4-D 1.0+KT 0.1; 2,4-D 0.1+KT 0.01; NAA (IAA) 2.0+KT 0.1 and NAA (IAA) 1.0+KT 0.1 mg/L were favorable to callus formation. Modified MS medium containing 2,4-D was suitable for initiated callus of G. davidsonii Besides, suspension cultures from callus of G. davidsonii were saccessfully initiated. Optimum concentration of 6BA (or ZT, or 2ip) and NAA (IAA) was for shooting, somatic embryo or leaf formation. Plantlets regenerated from somatic embryo at lower concentration of 6BA, or ZT, or 2ip. As to protoplast culture of this species, the age and physiological condition of callus or suspension cells and concentration of enzymes used for protoplast isolation affected the yield and survival of protoplasts. Protoplast of this species cultured in modified MS medium containing 2,4-D 0.5+NAA 0.5+ZT 0.1–0.2 mg/L. and divied after 3–4 days. The rate of division was 3--4% and cell cluster formed after 14 days, then these cells died.     

中华结缕草(Zoysia sinica Hance)组织培养和再生植株研究

以中华结缕草(Zoysiasinica Hance)成熟种子为外植体在附加2.5mg/L2,4-D、0.25mg/L6-BA和1～2mg/LVB₁的改良MS培养基(MS_m)上愈伤组织的诱导率最高为43.0%。愈伤组织的最佳继代培养基为MSm附加0.1mg/L6-BA和2.0mg/L2,4-D。在无生长调节物质的MS培养基(MS₀)上,外观呈白色到淡黄色、含有密实颗粒的愈伤组织再生率为30%～60%。     

Somatic embryogenesis and plant regeneration from leaf derived callus of winged bean [Psophocarpus tetragonolobus (L.) DC.]

Somatic embryos were obtained from callus cultures derived from leaf explants of the winged bean, Psophocarpus tetragonolobus (L.) DC. Initiation and development of the somatic embryos occurred with a two-step culture method. Callus cultures initiated on MS medium with NAA and BAP, upon transfer to a new medium with IAA and BAP, produced somatic embryos. Maximum embryogenesis of 60% was obtained on induction medium with 0.5 mg/l NAA plus 1.0 mg/l BAP followed by transfer to a secondary medium with 0.1 mg/l IAA and 2.0 mg/l BAP. Optimal embryo germination and plantlet development was achieved on MS medium with 0.2 mg/l BAP plus 0.1 mg/l IBA. The regenerated plants were successfully transferred to glasshouse conditions.Abbreviations MS Murashige and Skoog (1962) medium - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA 1-naphthaleneacetic acid - IAA Indole-3-acetic acid - IBA Indole-3-butyric acid - BAP 6-benzylaminopurine - KN Kinetin