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。     

2个甜瓜品种高效再生体系的建立

以甜瓜厚皮品种绿宝石和薄皮品种甘甜一号为材料,研究了5d龄无菌苗的子叶外植体在附加6-BA和IAA的不同浓度组合培养基上的分化情况.结果表明:2个品种在不定芽和愈伤组织的分化上存在着较大的差异,绿宝石在含有6-BA2.0mg·L-1的分化培养基上的不定芽诱导率最高,达93.75%;而甘甜一号品种在含有6-BA1.0mg·L-1和6-BA2.0mg·L-1的分化培养基上的不定芽分化频率均为100%.另外,随着IAA浓度的增大,疏松愈伤组织的分化均有加重的趋势;而提高6-BA的浓度,虽然分化进程有所加快,但玻璃化-褐化程度加重.本项研究为应用转基因技术改良甜瓜品种的重要育种性状奠定了组织培养方面的基础.     

拟南芥胚愈伤组织诱导和植株再生

CullusInductionandPlantletRegenerationinEmbryoExplantsofAra－bidopsisthalianaLIXiU-Ru(BeijingAgriculturalCollege,Beijing102206)##D1植物名称拟南芥（Arabidopsisthaliana）。2材料类别胚。3培养条件（1）愈伤组织诱导培养基：B5或1／2MS＋2,4－D0．5mg·L－1（单位下同）＋BA0．05十2％蔗糖或葡萄糖;（2）诱导生芽培养基：B5、MS或1／2MS＋2,4-D0．05＋BA0．5＋3％蔗糖;（3）诱导生根培养基：1／2MS＋2,4-D1．0。将种子表面消毒后,接种在B5培养基上,1天后剥胚,将胚接种在培养基（1）上进行愈伤组织诱导。…     

百脉根愈伤组织原生质体再生植株

百脉根无菌苗幼茎在含２．０ｍｇ／Ｌ－,２,４－Ｄ,０．１ｍｇ／Ｌ２－ｉｐ的ＭＳ培养基上诱导和继代培养愈伤组织。选取绿色松散颗粒愈伤组织分离原生质体。原生质体培养在调整珠ＫＭ８Ｐ,Ｖ－ＫＭ,ＭＳ和ＳＨ培养基上「含３００ｍｇ／Ｌ,ＣＨ,２％ＣＷ,２％蔗糖,６％葡萄糖,２．０ｍｇ／Ｌ,２,４－Ｄ,０．５ｍｇｇ／Ｌ,ＢＡ,５ｍｍｏｌ／Ｌ ＭＥＳ」,原生质体再生细胞均能分裂,并形成小愈伤组织,但以ＫＭ８０为     

何首乌愈伤组织诱导和植株再生

1　植物名称　何首乌 (Ｐｏｌｙｇｏｎｕｍｍｕｌｔｉｆｌｏｒｕｍ)。2　材料类别　蒙山阳坡路旁野生植株上的嫩芽。3　培养条件　诱导愈伤组织培养基 :(1 )ＭＳ 6 ＢＡ 1ｍｇ·Ｌ-1(单位下同 ) ＮＡＡ 1 ;(2 )ＭＳ 6 ＢＡ 2 ＮＡＡ 1 ;(3) 1 /2ＭＳ ＮＡＡ 0 .3;(4) 1 /2ＭＳ 2 ,4 Ｄ 0 .1 ;(5 ) 1 /2ＭＳ 2 ,4 Ｄ0 .3;(6 ) 1 /2ＭＳ 2 ,4 Ｄ 1 ;(7)ＭＳ 6 ＢＡ 3 2 ,4 Ｄ 1。芽分化培养基 :(8)ＭＳ 6 ＢＡ 1 ;(9)ＭＳ 6 ＢＡ 2 ;(1 0 )ＭＳ 6 ＢＡ 5 ＮＡＡ 0 .1 ;(1 1 )ＭＳ 6 ＢＡ 2 ＮＡＡ 0 .4;(1 2 …     

怀牛膝愈伤组织的诱导和植株再生

1植物名称怀牛膝(Achyranthes bidentata)种子由河南省温县农业科学研究所提供. 2材料类别子叶. 3培养条件以MS为基本培养基.诱导愈伤组织及再分化的培养基:(1)MS 2,4-D 1.0 mg·L-1(单位下同) 6-BA 0.1;(2)MS 2,4-D 1.0 6-BA 1.0.生根培养基:(3)MS NAA 0.05.以上培养基均添加3%蔗糖、0.60%琼脂,pH 5.8～6.2,培养温度为(25±2)℃,光照度2 000 lx,光照时间14 h·d-1.     

油桐叶片愈伤组织诱导及植株再生

以油桐无菌苗叶片为试材,研究不同种类及浓度的植物生长调节剂对愈伤组织诱导、分化、增殖及生根的影响。结果表明：叶片愈伤组织的最佳诱导培养基为1／2MS＋2．omg·L-16-BA＋1．0mg·L～2,4-D,诱导率达100％;最佳分化培养基为1／2MS＋3．0mg·L～6-BA＋0．1mg·L-1。IBA＋0．05mg·L—IAA,分化率为86．36％;最佳继代增殖培养基MS＋3．0mg·L～6-BA＋0．05mg·L。IBA;最佳生根培养基为1／2MS＋O．1mg·L-1。IBA,生根率93．83％。炼苗后移栽到泥炭土：珍珠岩：蛭石=2：1：1的基质中,成活率达92％以上。     

花生成熟胚胚轴的植株再生和快繁

利用正交试验筛选出了以花生成熟胚胚轴为外植体获得较高频率的不定芽分化和植株再生的条件,并在此基础上对花生试管苗的快速繁殖进行了研究,研究表明,4d龄实生苗的花生胚轴,接种于MS NAA0.4mg/L TDZ0.2mg/L诱导培养基上培养28d后,转移到MSo可获得丛生的再生苗。将丛生苗分离,接种到P1培养基（MS NAA0.4mg/L BAP0.5mg/L）,小苗长大,把较大的试管苗切成带叶节的茎段,接种于含不同浓度的TDZ和BAP的快繁培养基,结果表明,TDZ0.2mg/L最适宜试管苗的快繁,繁殖系数可达4.8/月,试管苗经IBA诱导生根后,移栽田间,开花结果。     

转抗虫基因欧美黑杨离体快繁技术研究

以抗虫欧美黑杨的叶,带腋芽茎段为外植体进行离体快繁技术研究。最佳接种时间为8月份,新芽生长迅速。基本培养基为MS,较适初培养基为MS＋6－BA0.5mg/L（以下单位同）＋NAA0.01mg/,附加30g/L,蔗糖,7g/L琼脂。愈伤组织诱导并同时分化出新芽培养基为MS＋6－BA1.5 NAA0.3,附加40g/L蔗糖,6g/L琼脂。继代增殖培养基为MS 6-BA1.0 NAA0.1 GA2.0,附加30g/L蔗糖,5g/L琼脂。生根培养基为MS＋IBA2．0。     

向日葵离体再生体系的建立

为了建立高效的向日葵离体再生体系,从基因差异、外植体取材、生长素和细胞分裂素浓度、附加物的添加等方面出发,对向日葵愈伤诱导、分化、生根等过程进行了系统优化。结果表明：杂交材料相对于自交材料更容易实现再生;最佳外植体是生长4 d的子叶;最佳愈伤诱导培养基是MS培养基 (MS) +2.0 mg/L 6-苄基腺嘌呤 (6-BA)+0.5 mg/L奈乙酸 (NAA)+1.0 mg/L激动素 (KT),诱导率最高可达100％;最佳分化培养基是MS+0.2 mg/L 6-BA+0.5 mg/L NAA+0.3 mg/L KT+0.3 mg/L硝酸银 (AgNO3)+0.2 g/L活性炭 (AC),芽分化率可达71％;最佳生根培养基是1/2 MS+0.6 mg/L吲哆丁酸 (IBA),生根率最高为77％。方差分析表明,材料基因型、外植体生长时间、激素、AgNO3、AC对向日葵再生呈现显著性影响。     

Genetic transformation of Populus deltoides and P.x euramericana clones using Agrobacterium tumefaciens

The susceptibility of different Populus euramericana (Neva, PE68-022 x P. nigra, 71-060 x P. nigra) and P. deltoides (PE68-022 x P. deltoides) clones to wild-type Agrobacterium tumefaciens strains (A281 and 82.139) was evaluated in an inoculation experiment, and differences in the frequency of tumor formation (0-48) were found. Co-cultivation experiments demonstrated high transformation ability of oncogenic binary A. tumefaciens strains as compared to disarmed strains. Using oncogenic binary strains, transgenic calluses were obtained from all tested clones. The presence of acetosyringone did not influence the transformation frequency of the disarmed strains. Co-inoculation experiments were performed using leaf discs and a bacterial suspension containing both wild-type and disarmed strains. No positive effects on transformation efficiency were noticed in these conditions either. The transformation of tumors and kanamycin resistant calluses was confirmed by DNA analysis.     

欧美黑杨离体再生途径及影响因子的研究

先期完成了以腋芽发育为再生方式的研究后 [1 ] ,又对不定芽的发育进行了探索 .分别以欧美黑杨特选品系的不同部位外植体进行愈伤组织诱导及植株再生研究 ,易诱导并易分化出新芽的外植体为幼嫩茎段 ;通过不同激素浓度合理配比 ,进行适宜的诱导及分化培养基筛选 ,培养基为 MS+6 - BA 1.2 mg/ L +NAA 0 .5 mg/ l,附加 6g/ L琼脂 ,诱导的愈伤组织分化出密集的再生芽 ;糖分在诱导分化过程中作用突出 ,其合适浓度为 4 0 g/ L ;愈伤组织的形态及培养时间对分化频率影响较大 ,最佳的形态为致密的绿色 ,白色及粉红色的愈伤组织较疏松 ,分化率很低 .不同部位外植体、激素含量、蔗糖浓度、愈伤组织形态及培养时间是明显影响欧美黑杨愈伤组织诱导分化的因子 .     

Efficient plantlet regeneration from protoplasts isolated from suspension cultures of poplar (Populus alba L.)

We developed an efficient plant regeneration system from protoplasts for poplar (Populus alba L.). Protoplasts were isolated from 4-day-old suspension cultures derived from seed-induced calli with a yield of 6.96× 10⁶ cells/g fresh weight cells and then cultured at a concentration of 2.5×10⁵ cells/ml in NH₄NO₃-free Murashige and Skoog (MS) medium supplemented with 5 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 0.05 μM thidiazuron (TDZ) and 0.5 M glucose as a osmoticum. The plating efficiency of the cultured protoplasts was calculated at 26.5% at day 7 and 31.7% at day 14. Cell colonies were observed after culturing for 4 weeks. Regenerated colonies were propagated through subculture in liquid MS medium supplemented with 5 μM 2,4-D. Buds were induced from regenerated calli on MS medium containing 10 μM kinetin or 1 μM TDZ. Regenerated shoots were rooted on half-strength MS medium, and the plantlets were transplanted in soil. Randomly amplified polymorphic DNA analysis did not detect any DNA polymorphism among the regenerated plants. Received: 7 March 1997 / Revision received: 16 June 1997 / Accepted: 5 July 1997     

小黑杨快繁与再生体系的优化

利用已获得小黑杨无菌苗叶片,研究不同培养基和植物激素对不定芽诱导、丛生苗抽茎及幼苗生根的影响。结果表明：培养基和激素对小黑杨叶片不定芽的诱导及幼苗生根有一定影响。小黑杨叶片不定芽诱导的最佳培养基：MS+6 BA 0.5 mg·L^-1+NAA 0.05 mg·L^-1,诱导率为100%。丛生苗诱导的最佳培养基：MS+6-BA 0.2 mg·L^-1+NAA 0.05 mg·L^-1。幼苗生根的最佳培养基有两种：MS+NAA 0.25 mg·L^-1或MH+IBA 0.2 mg·L^-1,生根率大于99%。     

Plant regeneration from hypocotyl segments of Lupinus luteus cv. L. Aurea

Complete plants of Lupinus luteus L. cv. Aurea that were regenerated from hypocotyl segments, bloomed, produced seeds and were efficiently nodulated by Bradyrhizobium sp. strains. The highest rates of shoot formation were obtained on A medium plus 1.3% agar with 10.0 M 2-isopentenyladenine (2iP) and 0.11 M naphthaleneacetic acid (NAA); the best rooting was achieved on a medium with 0.5 M NAA plus 0.05 M 2iP. Afterward, plantlets were transferred to either perlite or peat-containing pots and irrigated with a N-free nutrient solution until maturity. Direct rooting of hypocotyls could also be obtained on A medium with 1% agar.     

A rapid and efficient method for regeneration of plantlets from embryo explants of cumin (Cuminum cyminum)

A new, simple and efficient method was developed for multiple shoot regeneration of cumin from imbibed embryo cultures. This method yielded a large number of shoots within short period of time (30–50 days) without any subculturing. The effects of different media, different embryo explants and various combinations of plant growth regulators (PGRs) on callus formation and shoot regeneration in cumin were investigated. Simultaneous callus formation and shoot regeneration was obtained. The best response for multiple shoot regeneration was observed on B5 medium containing 1.0 mg l^–1 BAP, 0.2 mg l^–1 NAA and 0.4 mg l^–1 IAA, with an average of 140 shoots per explant.     

Transgenic Populus tremula: a step-by-step protocol for its Agrobacterium-mediated transformation

In recent years, Populus species have acquired an important place in basic and applied research of woody plants. The practical role of Populus species in world forestry and their importance to research as a woody-plant model have led to increasing interest in tissue-culture and molecular techniques, as well as the development of transformation procedures for this genus. A simple technical procedure is described here step-by-step, for the first time, as a routine method for transforming Populus tremula using a disarmed Agrobacterium tumefaciens hypervirulent strain. The procedure begins with the inoculation of stem explants with bacterial suspension, followed by a short period of co-cultivation on a highly regenerative medium. Transformed shoots are selected on regeneration medium containing antibiotics and the presence of the inserted target genes is checked using a rapid and efficient PCR test. Selected shoots are transferred to a rooting medium, under the same selection pressure, and propagated via stem cuttings. Selected plants can be hardened and transferred to the green-house within 4 months of inoculation. The method has proven efficient for several gene constructs, selection on Kan or Hyg, and three different Agrobacterium strains.     

Improved micropropagation of Populus spp. by Pluronic F-68

Stem explants and leaves (without petioles) excised from axenic shoots of Populus tremula cv. Ahle or P. tremula × tremuloides cv. Münden were cultured in the presence of the non-ionic, co-polymer surfactant, Pluronic F-68. Stem explants developed shoots within 10 d of culture and significant (p<0.05), but genotype-dependent, increases in total shoot fresh weight (maximum 2 × control) occurred in cultures supplemented with 0.001–0.1% Pluronic F-68 over a 72 d period. Similarly, increases in both fresh weight (up to 10-fold) and number of shoots per P. tremula × tremuloides leaf explant (5-fold maximum) over 60 d occurred with Pluronic F-68 at 0.001%.Abbreviations BAP 6-benzylamino purine - IBA indolebutyric acid - MS0 Murashige and Skoog medium [14] lacking growth regulators - NAA -naphthaleneacetic acid - WPM woody plant medium