不同杂种优势类群玉米幼胚愈伤组织的诱导及植株再生特性的评价

以Reid、唐四平头和其他种质等3个杂种优势类群共30份玉米自交系为实验材料,以玉米幼胚作为外植体,研究基因型、培养基、激素、继代培养次数对玉米幼胚愈伤组织诱导及植株再生的影响.研究结果表明供试材料均能进行愈伤组织诱导,但是只有部分自交系能再生植株.通过对不同杂种优势类群自交系玉米幼胚培养特性的分析,从Reid、唐四平头和其他种质类群的供试自交系中,筛选出了胚性愈伤组织诱导频率和绿苗分化率均较高、适合于遗传转化的受体材料,如黄野四/京24//C108/黄野四选系、黄早四/先早17//吉853选系、R43//黄野四/711选系、京7、京7黄、3189/4380选系、4380/陕综5选系、8103、先早17、18-599红、501、178和冀53.     

紫果猕猴桃幼胚愈伤组织诱导及植株再生

以紫果猕猴桃(Actinidia arguta var．purpurea)幼胚为外植体,诱导愈伤组织并进行植株再生。结果表明：不同的培养基和不同的培养条件对幼胚愈伤组织的诱导率及分化率不同;0．2mg／L ZT与0．5mg／L GA。配合使用有利于促进愈伤组织的诱导;7％蔗糖、600mg／L CH与400mg／L Gln都有利于促进愈伤组织的形成;在添加0．5mg／L 6-BA、0．05mg／L NAA与0．5mg／L GA3的MS培养基中植株的再生率达93．3％。     

不同基因型玉米愈伤组织诱导与植株再生研究

以5个玉米品系幼胚为外植体,研究了基因型、2,4-D浓度以及胚龄对愈伤组织诱导的影响;6-BA对愈伤组织分化的影响;以及IBA对再生芽生根的影响。结果表明：除。31外,其他基因型的外植体在相同条件下均可诱导出愈伤组织,但是不同基因型间存在显著差异;2,4-D浓度和胚龄显著影响愈伤组织的诱导,且2,4-D浓度为2．0mg／L,胚龄在11—13d之间时,玉米愈伤组织诱导率较高且质量较好。将愈伤组织转入分化培养基后,6-BA促进了愈伤组织的再分化;在生根培养基中,IBA促进了再生芽生根,经过炼苗后移栽获得再生植株。     

莲胚愈伤组织诱导及植株再生的研究

关于莲胚组织培养国内外有过研究,但迄今利用组织培养技术从莲胚诱导愈伤组织形成再生植株的成功例子尚未见有报道。     

玉龙草胚性愈伤组织的诱导及植株再生

1　植物名称　玉龙草 (Ｏｐｈｉｏｐｏｇｏｎｊａｐｏｎｉｃｕｓｃｖ .“Ｎａｎｕｓ”)。2　材料类别　嫩芽。3　培养条件　胚性愈伤组织诱导培养基为 :(1)ＭＳ 2 ,4 Ｄ 1.0ｍｇ·Ｌ- 1(单位下同 ) 6 ＢＡ 0 .0 1;(2 )ＭＳ 2 ,4 Ｄ 2 .0 6 ＢＡ 0 .0 1;(3)ＭＳ 2     

诱发玉米幼胚愈伤组织及其再生植株耐盐性变异的研究

本文以玉米获白×Mo17单交种的幼胚为材料。以10g/1NaCl为起始浓度,利用逐步提高NaCl浓度的方法,筛选出耐20g/1NaCl的细胞变异体,并获得再生植株。同时对影响耐盐变异系筛选、分化的 因素进行了研究。讨论了愈伤组织变异体及其再生植株的稳定性。     

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％。     

玉米叶片愈伤组织的诱导及其植株再生

植物名称:玉米(Zea mays)品种小金黄。材料类别:幼叶。培养条件:以MS培养基为基本培养基,生长调节物质配比为:(1)2,4-D 4mg╱L(单位下同) NAA2 IAA2;(2)2,4-D8 NAA4 IAA4;(3)2,4—D 8 NAA4 IAA4 ZT0.5 BA0.5 区T 0.5:(4)2,4-DS NAA4 IAA4 ZTl     

野罂粟胚性愈伤组织诱导和植株再生

1 植物名称 野罂粟（Papaver nudicaule L．）。     

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。…     

玉米幼胚胚性愈伤组织再生能力相关性状遗传研究

本文对来源于美国、墨西哥和中国的144份不同玉米自交系幼胚胚性愈伤组织的再生能力相关性状进行了研究,发现其再生能力受到环境、基因型及环境与基因型互作三方面的影响。其中各性状之间的相关性表现为：绿点率(green embryonic callus rate, GCR)、分化率(embryonic callus differentiating rate, CDR)及再生绿苗数(the plantlet number of embryonic callus regeneration, CPN)之间呈极显著正相关,且这三者与褐化率(embryonic callus browning rate, CBR)呈极显著负相关; 两次继代的克隆指数(embryonic callus cloning index for the first subculture, CCI1; embryonic callus cloning index for the second subculture, CCI2)呈显著正相关,且CCI2与GCR有一定的正相关关系,与CBR呈负相关关系;生根率(embryonic callus rooting rate, CRR)则与GCR、CDR及CPN呈一定正相关。经过广义遗传力计算发现：胚性愈伤组织的两次继代克隆指数CCI1、CCI2和CRR的遗传力较低,其他性状的遗传力较高。此外,经Ward法双向聚类分析,共发现了11个具有高再生能力的自交系材料,且通过生根培养发现其再生绿苗的生根情况良好,因而可将它们作为玉米转基因受体的骨干自交系。     

Screening wheat genotypes for high callus induction and regeneration capability from anther and immature embryo cultures

Plant regeneration via tissue culture varies with the genotype and is an important factor in establishing cell selection and genetic transformation systems. To select genotypes – especially Japanese ones – with a high regeneration capability, we screened 107 wheat genotypes (78 domestic, 29 foreign) for callus induction and regeneration capability from anther and immature embryo cultures. For anther culture, 83 of 107 genotypes tested induced calli and 45 regenerated plants. Only 9 genotypes, however, produced green plants, 25 produced only albino plants, and 11 produced both green and albino plants. Glennson 81 was the highest in callus induction, followed by Orofen, Danchi–komugi and Chris. The genotypes with a relatively high regeneration capability were Framala 80 at 24% and Glennson 81 at 19%, these two genotypes produced only green plants. For immature embryo culture, 97 genotypes showed a 90% callus induction rate and 74 genotypes regenerated plants. Very few genotypes produced albino plants. The genotypes with a high regeneration capability were Genaro 81 at 90%, Chinese Spring at 80%, and Norin 75 at 75%. This revised version was published online in June 2006 with corrections to the Cover Date.     

渗透胁迫条件下小麦成熟胚愈伤组织的诱导及植株再生的研究

用不同浓度的PEG6000及NaCl对5个小麦品种的成熟胚组织培养物进行处理,研究在渗透胁迫条件下基因型和激素对成熟胚愈伤组织的诱导及分化的影响。结果表明,小麦整株水平与细胞水平的抗性存在一定相关,不同基因型对干旱与盐胁迫的敏感程度不同,成熟胚愈伤组织的诱导率和植株再生率表现出明显的差异。初步得到了晋麦47、长武134、红芒麦的耐旱愈伤组织以及晋麦47、长武134的耐盐愈伤组织,并获得了晋麦47和长武134具有一定抗性的再生芽。     

An efficient and rapid regeneration via multiple shoot induction from mature seed derived embryogenic and organogenic callus of Indian maize (Zea mays L.)

An efficient method for in vitro micro propagation and genetic transformation of plants are crucial for both basic and applied research. Maize is one of the most important cereal crops around the world. Regeneration from immature embryo is hampered due to its unavailability round the year. On the contrary mature embryo especially tropical maize is recalcitrant toward tissue culture. Here we report a highly efficient regeneration (90%) system for maize by using 2 different approaches i.e., embryogenic and organogenic callus cultures. Seeds were germinated on MS medium supplemented with 5 mg/l 2,4-D and 3 mg/l BAP. Nodal regions of 2 wks old seedlings were longitudinally split upon isolation and subsequently placed on callus initiation medium. The maximum frequency of embryogenic callus formation (90%) was obtained on MS medium supplemented with 2 mg/l 2,4-D and 1 mg/l BAP in the dark conditions. The compact granular organogenic callus formation (85% frequency) was obtained on MS medium supplemented with 2.5 mg/l 2,4-D and 1.5 mg/l BAP at light conditions. MS medium supplemented with 2 mg/l BAP, 1 mg/l Kinetin and 0.5 mg/l NAA promoted the highest frequency of shoot induction. The highest frequency of root formation was observed when shoots were grown on MS medium. The regenerated plants were successfully hardened in earthen pots after adequate acclimatization. The important advantage of this improved method is shortening of regeneration time by providing an efficient and rapid regeneration tool for obtaining more stable transformants from mature seeds of Indian tropical maize cultivar (HQPM-1).     

Regeneration and transformation of Egyptian maize inbred lines via immature embryo culture and a biolistic particle delivery system

Summary A regeneration system was developed for elite Egyptain maize inbred lines using immature embryos as explants. This system proved to be highly genotype-dependent. Line Gz 643 was identified as the best line, revealing the highest regeneration frequency (42.2%). Addition of l-proline and silver nitrate to culture media greatly enhanced the formation of embryogenic type II callus and the regenerability of some of the tested lines. Transformation of the scutellar tissue of immature embryos from inbred line Gz643 was performed with the particle delivery system using a single plasmid carrying both the GUS and Bar genes (pAB-6) or by co-transformation with two plasmids, pAct1-F (GUS) and pTW-a(Bar). Different transformation parameters were evaluated, i.e. ostomic treatment, acceleration pressure, and number of shots. Osmotic treatment (0.25 M sorbitol + 0.25 M mannitol) along with the use of either acceleration pressure 1300 psi and one shot per plate (for co-transformation with pAB-6) or 1100 psi and two shots per plate (for transformation with pAct1-F and pTW-a) gave the best results, as expressed by the number of blue spots in the β-glucuronidase (GUS) assay. Stable transformation was confirmed in Ro transformed plants by means of histochemical GUS assay and herbicide application. PCR and Southern blot analysis proved the integration of the full-length genes in some of the transgenics.     

不同培养基对水稻胚性愈伤组织诱导及分化的影响

以水稻成熟胚为材料诱导愈伤组织,统计在不同基本培养基上的愈伤诱导率以及绿苗分化率,分析不同基本培养基及外源激素的含量和比例对愈伤组织生长及分化的影响。结果表明,试验材料对基本培养基具有选择性,MS培养基对籼稻种胚愈伤的诱导培养效果较好,NB培养基则更适合粳稻种胚愈伤的诱导培养;诱导继代培养基中加入多种氨基酸组合可有效提高出愈率和分化率,特别是粳稻的愈伤组织的诱导和分化需要多种氨基酸的共同作用;不同基因型水稻材料对激素和氨基酸组合的需求不同。     

陆地棉中棉所24胚性愈伤组织的诱导及植株再生

以陆地棉“中棉所24”为材料进行了全固体体细胞培养,获得了愈伤组织和再生植株。愈伤组织诱导阶段采用0．01IAA 0．01KT 0．012,4-D的培养基效果好,继代时间多为30～50d;激素由高到低的继代可明显提高胚性愈伤分化率,IAA和KT含量均较低,IAA／KT比例为1：1～1：6,胚性愈伤最高分化率为50．22％。     

Tissue culture and plant regeneration from immature embryo explants of Calotropis gigantea (Linn.) R. Br.

Callus cultures were established from immature embryos of Calotropis gigantea (Linn.) R. Br. on a modified basal medium of Murashige & Skoog supplemented with 1 mgl^-1 2,4-D. In addition to 0.1 mgl^-1 of NAA the optimal BAP concentration for promoting shoot bud formation and growth was 2 mgl^-1. Rooting was induced when shoots were transferred to auxin-supplemented Bonner's solution or half-strength MS basal salt solutions.Abbreviations NAA -naphthaleneacetic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - IBA indole-butyric acid - BAP 6-benzylaminopurine - Kin kinetin     

Application of activated charcoal and nanocarbon to callus induction and plant regeneration in aromatic rice (Oryza sativa L.)

The investigations of nanotechnology with the application on agricultural products also have been few reported, especially the plant regeneration. The effects of activated charcoal and nanocarbon on the callus induction and plant regeneration of aromatic rice were studied. Activated charcoal was added into the callus induction and regeneration medium. The presence of activated charcoal in the callus induction medium (100–500 mg L^?1), activated charcoal significantly reduced the percentage of the callus induction and biomass accumulation (fresh weight, dry weight and size). Whereas, the regeneration medium supplemented with 100 mg L^?1 of activated charcoal showed the highest percentage of plant regeneration (61.90%) and the ratio of the number of seedlings to the number of regenerated calli (RSR; 3.06) that derived from the callus induction medium (without activated charcoal). Moreover, the induced calli derived from the callus induction medium supplemented with nanocarbon at 5 mg L^?1 showed the highest percentage of callus induction (94.70%), the percentage of green spots (95.83%), the percentage of plant regeneration (60.42%) and the RSR (3.12) when transferred the calli into the regeneration medium (without nanocarbon). After that, nanocarbon was also added into the regeneration medium. The percentage of green spots (96.08%), the percentage of plant regeneration (62.75%) and the RSR (3.16) obtained from the regeneration medium supplemented with 20 mg L^?1 of nanocarbon showed the highest values. This experiment showed that the optimum concentration of activated charcoal and nanocarbon had potential to enhance the callus induction and plant regeneration frequencies in tissue culture medium of aromatic rice.     

Callus induction and plant regeneration from immature embryos of rye and triticale

Callus cultures were established from the scutellum, scutellar node and radicle region of immature embryos of rye and octoploid triticale on modified Murashige-Skoog basal medium supplemented with various growth regulators. 2, 4-D, 2, 4, 5-T and 2, 4, 5-Cl, POP were found suitable for initiation and maintenance of callus cultures. Cytokinins had no or inhibitory effect on callus induction and growth. On basal medium containing 5 mg/l of 2,4,5-Cl₃ POP, 16% of triticale and 17% of rye primary cultures exhibited shoot bud regeneration after 3–4 weeks. Transfer of such cultures to basal medium supplemented with zeatin or zeatin in combination with IAA further promoted shoot elongation and plantlet formation. Plantlets were rooted on basal medium containing 1 mg/l NAA and were eventually transferred to soil. Chlorophyll variants were observed in about 6% of triticale cultures.