In vitro regeneration of Irvingia gabonensis by somatic embryogenesis

A productive genotype of Irvingia gabonensis were cultured in vitro for induction embryogenic calli, somatic embryogenesis and regeneration of plantlets. Fragments of young leaves were used as primary explants. Callogenesis was initiated by culture of explants during 30 days on Murashige and Skoog medium half strength (MS/2) supplemented with 1-6 mg L(-1) of 2,4-dichlorophenoxyacetic acid (2,4-D). The highest percentage of explants forming calli is 85.1% at 3 mg L(-1) of 2,4-D. Somatic embryos were obtained after a subculture of embryogenic calli during 60 days on MS/2 supplemented with 1-3 mg L(-1) of BAP. The highest percentage of embryogenic calli which differentiates somatic embryos is 63.8 +/- 2.3% at 1 mg L(-1) of 6-benzylaminopurine (BAP). The highest number of somatic embryos per callus which is 43.6 is obtained with 2 mg L(-1) of this phytohormone. When isolated from calli and sub-cultured during 30 days on MS/2 supplemented with 2 mg L(-1) of BAP, somatic embryos germinate with a highest percentage of 83%. The subculture of germinated somatic embryos on the same Basal Medium (BM) supplemented with 4 mg L(-1) of BAP and 2 mg L(-1) of Naphthalene Acetic Acid (NAA) during 80 days gives rise to the plantlets with 82.7 +/- 4.8% of success. With this combination, each plantlet has average length of 5.6 cm, bears 3.3 leaves and 7.2 roots with 1 or 2 pivoting roots. Plantlets acclimatized on a mixture sterilized soil/vermiculite at equal volume survive at 93%. Results of this study constitute a new way for a production of Irvingia gabonensis seedlings with pivoting root and they permit to arrest the difficulties of natural and horticultural reproduction.     

Growth optimization and organogenesis of Gerbera jamesonii Bolus ex. Hook f. in vitro

Regeneration potentials in Gerbera jamesonii Bolus ex. Hook f. from tissues culture system was studied using leaf, petiole and root explants. In vitro regeneration, callus induction and root formation were optimized by manipulation of growth regulators during organogenesis. Various kinds of plant growth regulators such as 6-Benzylaminopurine (BAP), alpha-Naphthalene acetic acid (NAA), 2, 4-Dichlorophenoxyacetic acid (2,4-D), Indole-3-acetic acid (IAA), Indole-3-Butyric acid (IBA), N6-[2-Isopentenyl]adenine (2iP), Kinetin and Zeatin were used to initiate cultures. These plant growth regulators were added to Murashige and Skoog medium in different combinations and concentrations. Adventitious shoots were obtained from petiole explants cultured on Murashige and Skoog (MS) medium supplemented with 2.0 mg L(-1) BAP and 0.5 mg L(-1) NAA. Effectiveness of shoot regeneration medium, type of growth regulator used and duration of induction period were investigated. Leaf explants cultured on MS medium supplemented with 1.0 mg L(-1) BAP and 2.0 mg L(-1) 2, 4-D showed the best results for callus induction. Root explants were found to be non-regenerative in all experiments conducted. Petiole segment was identified as the best explant for regeneration of this species. Regenerated plants were rooted on Murashige and Skoog basal medium. Plantlets were then transferred to field with 75% survival rate.     

几个籼稻品种的成熟胚愈伤组织植株再生体系的建立

组织培养技术在作物改良上的成功应用需要合适的植株再生体系。本试验以7个籼稻成熟胚为材料，通过优化激素配比，建立适合于水稻遗传转化的高频植株再生体系。研究结果表明，NB 2mg／L2，4-D适合于供试品种的成熟胚愈伤组织的诱导，诱导率达90％以上；在分化培养基中添加3．0-3．5mg／L KT，0．5～1．0mg／L NAA和5．0mg／L ABA的激素配比，明恢81、N175、航1号的最高分化率分别达72．7％、80．0％和78．0％；移栽成活95％以上。     

芦荟库拉索(Aloe vera L)再生体系的建立

对芦荟(AloeveraL.)组织培养和再生体系进行了研究,初步获得芦荟的初代和继代培养基为MS 6-BA2.5mg/L NAA0.2mg/L 蔗糖30mg/L,生根培养基成分为1/2MS IBA0.2mg/L NAA0.1mg/L 蔗糖30mg/L,愈伤组织培养采用粗壮的芦荟基部的白色部分,诱导愈伤组织的培养基为B5 6-BA2.0mg/L 2,4-D2.0mg/L 蔗糖30mg/L,采用Vc5.0mg/L抑制愈伤组织诱导过程中褐变的产生。诱导愈伤组织和外植体直接出芽2种方式并存。试管苗移栽后存活率为100%。采用潮霉素为抗性筛选标记,选择浓度为20￣30mg/L。     

In vitro study on regeneration of Gladiolus grandiflorus corm calli as affected by plant growth regulators

In this study, in vitro organogenesis of Gladiolus grandiflorus cultivar pink corm segments were evaluated by culturing corm calli in modified MS medium supplemented with 3% sucrose and 0.7% agar with different concentration of BAP (0, 1, 2 and 4 mg L(-1) medium) and NAA (0, 0.5, 1 and 2 mg L(-1) medium) in factorial experiment of Completely Randomized Design (CRD). In order to obtain Gladiolus calli, corm segments (Aprox. 5 x 5 x 1 mm in size) were kept in modified MS medium (Murashige and Skoog, 1962) that was supplemented with 1 mg L(-1) 2, 4-D, 3% sucrose and 0.7% agar. The results showed that increasing the concentration of BAP from 0 to 2 mg L(-1) medium simulated plantlet regeneration but no significantly effect was obtained on shoot and cormel organogenesis between 2 and 4 mg L(-1) BAP concentration in medium. Increasing of NAA content in media without BAP developed rootlet significantly. Interaction results showed that increasing BAP content against decreasing of NAA concentration stimulates the shoot and cormel proliferation.     

短光低温不育水稻宜D S成熟胚培养的研究

对短光低温不育水稻宜D S成熟胚在组织培养中的愈伤组织诱导和分化进行了研究,结果表明:愈伤组织诱导率与绿苗分化率之间无对应关系;诱导培养基的成分对转分化后的愈伤组织有后效作用,2,4-D与6-BA和NAA搭配使用比单独使用2,4-D诱导的愈伤组织更易于分化;同时还发现,不同诱导培养基诱导的愈伤组织在转分化后对分化培养基的要求不同.     

小麦不同外植体的组织培养效果研究

良好的组织培养效果是提高植物基因转化效率的基础。以山东省近年来育成并大面积推广的10个优良品种（系）为材料,对这些品种的花药、幼穗、幼胚和成熟胚的组织培养效果进行研究,旨在筛选每一基因型最适合于组织培养的外植体类型,为应用基因工程技术进行小麦遗传改良提供基础材料。结果表明,禽伤诱导率和再生成苗率与基因型和外植体类型（花药、幼穗、幼胚和成熟胚）密切相关。8802在花药和成熟胚培养中表现突出,花药出愈率达119．5％,愈伤分化成苗率19．9％;烟农19的幼穗愈伤直接分化成苗率最高,这43．5％;8802、烟农19和潍麦8号三个基因型的幼胚培养效果差异不显著,其愈伤分化成苗率分别为26．3％、24．5％和24．8％。蔗糖浓度在3％～9％之间,对成熟胚的愈伤组织诱导率影响很小。高浓度蔗糖降低愈伤生长速度。在一定蔗糖浓度下,愈伤诱导率与2,4-D浓度密切相关,高浓度的2,4-D对愈伤组织再生不利。MS＋4mg／L2,4-D对于成熟胚的脱分化相对较好,MS和MS＋0．4mg／L NAA＋0．6mg／L KT作为成熟胚的愈伤组织再生培养基,对不同基因型具有一定的适用性。     

Callogenic studies of Achyranthes aspera leaf explant at different hormonal combinations

With the objective to promote in vitro callus induction, leaf segments of Achyranthes aspera were inoculated on basal MS medium supplemented with 3.0% sucrose and 0.8% agar with different concentrations of 2,4-D alone and in combination with NAA, BAP, IAA, IBA and Zeatin. The explants were maintained in growth room at 25 +/- 1 degrees C and 16 h light cycle. The best callus induction was obtained with 2,4-D (1.0 and 2.0 mg L(-l)) in combination with NAA (0.5 mg L(-1)). Callus induction and good texture from leaf explant was also observed at 2,4-D with BAP. On these combinations morphologically, light green, soft, compact and non-embryogenic callus (Type III callus) was observed. While morphology of callus and callogenic response was poor at 2,4-D alone or in combination with other hormones at different concentrations.     

低浓度2,4-D提高水稻体细胞成苗研究

 以较低浓度(0.2、0.4、0.8 mg/L)的2,4-D作为水稻(Oryza sativa L.)的外植体(种子)诱导愈伤组织培养基中的生长素类激素，发现0.8 mg/L的2,4-D能较早地使胚的盾片产生大量的愈伤组织，但这些愈伤组织在原培养基上不能分化出绿芽。含0.2 mg/L的2,4-D能在胚芽鞘节等部位产生一些较光滑、致密、白色颗粒状的结构或愈伤组织，虽然它们出现较迟，但这些结构物能在原诱导培养基上逐渐转化成芽簇，并产生一些纤细的根。将芽簇分割成小块，转移到加有KT等的成苗培养基上，能形成大量的幼苗，绿苗频率达80%以上，明显高于一般报道的水平。试管苗经大田种植后生长整齐一致，能产生出正常的种子。     

An evaluation of callus induction and plant regeneration in twenty-five turf-type tall fescue (Festuca arundinacea Schreb.) cultivars

In order to investigate the genetic variation in tissue culture response and to find the cultivars with high regeneration ability for genetic transformation, twenty-five turf-type tall fescue ( Festuca arundinacea Schreb.) cultivars, including many elite ones released recently, were evaluated for their callus induction and plant regeneration responses. Callus induction was initiated from mature seeds on a Murashige and Skoog (MS) medium containing 9·0 mg l^–1 2,4-dichlorophenoxyacetic acid (2,4-D). Induced calli were subcultured on the same medium with 2·0 mg l^–1 2,4-D and then transferred to a MS medium supplemented with 2·5 mg l^–1 6-benzylaminopurine (BAP) for plant regeneration. Significant differences were observed among the twenty-five cultivars in both callus induction and plant regeneration ( P  < 0·001). Callus induction rate of viable seeds varied from 4·4% to 51·9%. Callus regeneration rates ranged from 16·7% to 58·8%. Overall regeneration rates (number of regenerated calli over number of cultured viable seeds) ranged from 1% to 22%. Approximately 94% of the regenerants were green plantlets.     

单倍体籼稻无性系微芽的离体调控

 以籼稻单倍体无性系微芽为材料,研究了激素及秋水仙碱等处理对籼稻单倍体微芽的诱导、分化、扩增及加倍的影响。2 mg/L的2,4-D可提高籼稻单倍体微芽的培养力。NAA对籼稻单倍体微芽的扩增有明显影响,0.5 mg/L的NAA可成倍扩增微芽,培养35 d后,其芽数比原始芽数增加了46.21倍,比不加NAA的对照增加了2.8倍,且单芽重仅0.079 mg。籼稻单倍体微芽扩增的适宜培养基为：N6附加MET 2.5 mg/L、NAA 0.5 mg/L、6-BA 2 mg/L。500 mg/L秋水仙碱溶液处理籼稻单倍体微芽48 h,其二倍体得率较高,绿苗率及二倍体率分别为42.9%和60.0%;秋水仙碱处理愈伤组织的效果不佳,虽然提高秋水仙碱处理浓度可提高二倍体率,最高可达100%,但由于绿苗分化率下降,使总的二倍体得苗率比不处理的对照低。     

低浓度2，4－D提高水稻体细胞成苗研究

以较低浓度（０．２、０．４、０．８ｍｇ／Ｌ）的２，４－Ｄ作为水稻（ＯｒｙｚａｓａｔｉｖａＬ．）的外植体（种子）诱导愈伤组织培养基中的生长素类激素，发现０．８ｍｇ／Ｌ的２，４－Ｄ能较早地使胚的盾片产生大量的愈伤组织，但这些愈伤组织在原培养基上不能分化出绿芽。含０．２ｍｇ／Ｌ的２，４－Ｄ能在胚芽鞘节等部位产生一些较光滑、致密、白色颗粒状的结构或愈伤组织，虽然它们出现较迟，但这些结构物能在原诱导培养基上逐渐转化成芽簇，并产生一些纤细的根。将芽簇分割成小块，转移到加有ＫＴ等的成苗培养基上，能形成大量的幼苗，绿苗频率达８０％以上，明显高于一般报道的水平。试管苗经大田种植后生长整齐一致，能产生出正常的种子。     

云蔗03-194甘蔗组培快繁技术

以甘蔗新品种云蔗03-194的幼嫩叶片作为外植体,采用不同2,4-D浓度对幼嫩叶片的切片进行愈伤诱导,以不同6-BA和KT激素配比对甘蔗愈伤进行分化诱导和增殖培养,以不同NAA浓度及香蕉汁添加量诱导甘蔗组培苗生根。结果表明:适宜甘蔗幼嫩叶片愈伤诱导的培养基为MS+2,4-D 1.5mg/L,诱导分化培养以MS+6-BA1mg/L+KT 0.5 mg/L为宜,增殖培养以6-BA 2.0 mg/L+KT 1.0 mg/L为宜,适宜的生根培养基为MS+NAA 1.5mg/L+30mL香蕉汁。以河沙:红壤土(2∶1)为假植基质,假植成活率达92%～96%,植株生长健壮,长势良好。     

红杨桃胚乳愈伤组织的诱导和三倍体植株再生

以红杨桃成熟干种子为材料,萌发后剥离胚乳进行培养。结果表明,最佳诱导愈伤组织的培养基为MS+2,4-D　2mg/L+BA 0.2mg/L,诱导频率可达93.5％:将淡绿色致密的愈伤组织转至MS+ZT　3mg/L+NAA0.2mg/L的培养基上,经连续继代后,形成芽原基并发育成小植株,壮苗后取茎尖染色体显微观察,证明再生植株多为三倍体,其发生频率可达73.7％。      

甘蔗近缘属斑茅组织培养高频再生体系的建立

以甘蔗近缘属植物斑茅的幼嫩叶片为材料,研究MS、SH、B5三种基本培养基和不同激素(2,4-D、NAA、6-BA)配比对斑茅愈伤组织诱导及继代培养的影响,并筛选出斑茅细胞高频再生体系的最优培养基配方。研究结果表明,斑茅幼嫩叶片愈伤组织诱导培养基以MS+2,4-D 3.0 mg/L+6-BA 0.4 mg/L为最适,愈伤组织继代培养基以MS+2,4-D 3.0 mg/L为最适。在此条件下诱导的愈伤组织质地疏松,具有较强的分化能力。目的是建立斑茅组织培养的高频再生体系,为斑茅材料的细胞悬浮培养、染色体加倍和细胞融合等研究提供一定参考。     

“热研11号”黑籽雀稗植株再生体系的建立

以热带牧草"热研11号"黑籽雀稗(Paspalum atratum cv.Reyan No.11)种子为材料,对其外植体植株的再生过程进行系统研究.结果表明,以MS无机盐 9.0mg/L维生素B1 9.5mg/L维生素B6 4.5mg/L尼克酸 1.0mg/L水解酪蛋白 30.0g/L蔗糖 8.0g/L琼脂为基本成分(MSM),附加植物激素类物质2.0mg/L2,4-D时,适合种子的愈伤组织形成,愈伤诱导率可达65%:继代培养基附加1.0mg/L2,4-D和0.1mg/LKT:分化的培养基附加6.0mg/L6-BA,分化率可达50%;生根培养基附加0.5mg/L激素类物质NAA,生根率100%.完成植株再生约需13周.     

贵港报春苣苔的叶片组培与植株再生

以贵港报春苣苔的叶片为外植体,研究不同培养基对其不定芽诱导和增殖、愈伤组织诱导与分化以及生根的影响。结果表明,外植体叶片以纵切为宜,不定芽诱导最适培养基为MS+6-BA 4.0 mg/L+IAA 1.5 mg/L,愈伤组织诱导最适培养基为MS+6-BA3.0～5.0 mg/L+2,4-D0.5～1.0 mg/L,不定芽诱导率以及愈伤组织诱导率均为100.00%;愈伤在MS+KT 1.0 mg/L+NAA 0.2 mg/L+potato 30 g/L+banana 30 g/L+apple 20 g/L+coconut juice 100 mL/L培养基上分化系数达12.64;不定芽在MS+ZT 1.0 mg/L+NAA 0.10 mg/L+potato 30 g/L+banana 30 g/L+apple 20 g/L+coconut juice 100 mL/L培养基的增殖系数为8.55;不定芽在3/4 MS+NAA 0.01～0.05 mg/L+活性炭1.0～3.0 g/L培养基上的生根率为100.00%。综上所述,叶片纵切后能通过不定芽途径以及愈伤组织途径建立贵港报春苣苔的组织培养技术体系,在该体系下不定芽增殖系数高、愈伤分化高,组培苗生根好。     

海马齿再生体系的优化及GUS基因的转化

以海马齿无菌实生小苗的叶片为外植体,对不同激素浓度和组合以及不同pH值等培养条件进行实验比较.结果表明:适于愈伤组织诱导的培养基为MS+2,4-D 2.0 mg/L+6-BA 0.5 mg/L+蔗糖3%,诱导率达100%;适于芽分化的培养基为MS+NAA 3.0 mg/L+6-BA 0.5 mg/L,分化率达到77.4%;诱导愈伤组织形成培养基的最适pH值为4.5,而诱导愈伤组织分化培养基的最适pH值为5.0.以小苗叶片为外植体材料和农杆菌介导的方法进行GUS基因的转化,组织化学染色检测发现共培养2 d后的外植体中有蓝色斑点产生,在转化2个月后的外植体上形成的愈伤组织也存在点状的蓝色斑点.说明GUS基因已转入海马齿外植体中.     

The influence of different hormone concentration and combination on callus induction and regeneration of Rauwolfia serpentina L. Benth

The influence of media composition on callus induction and subsequent regeneration of Rauwolfia serpentina L. Benth has been studied. High frequency (96.43%) callus induction was obtained when nodal segments from in vitro raised shoots were cultured on MS medium supplemented with 0.5 mg L(-1) BA and 2.0 mg L(-1) NAA. The callus differentiated into adventitious shoots when it was subcultured on MS medium supplemented with 2.0 mg L(-1) BA with 0.2 mg L(-1) NAA. Regenerated shoots were best rooted on half-strength MS medium with 1.0 mg L(-1) each of IBA and IAA.     

水稻成熟胚盾片诱导愈伤组织再生体系的建立

采用水稻成熟胚盾片诱导愈伤组织作为分化再生的外植体，通过优化激素组合和调节培养基渗透压，建立了适合水稻遗传转化的高效再生体系，将水稻成熟胚诱导愈伤组织分化再生的过程划分为3个不同时期，即成熟胚盾片愈伤组织诱导，胚性细胞诱导保持，胚性愈伤组织分化再生，以CultureI(LS 2,4-D2.0mg/L)诱导成熟胚盾片愈伤组织，愈伤组织剥离后接种于Culture II-3(LS 2,4-D 2.5mg/L 山梨醇3g/L)上诱导胚性愈伤组织，继代后接种于Culture Ⅲ-2(MS NAA 0.5mg/L 6-BA 1.0mg/L Kinetin 2.0mg/L 山梨醇8g／L）上分化成苗，培养结果表明，该再生体系所获得的胚性愈伤组织分化再生率均在60％－80％之间，利用本研究建立的高效再生体系建立水稻外源基因遗传转化系统，使转化过程中所得转化愈伤组织高效再生成苗，从而顺利获得转化植株，可以提高水稻外源基因转化的效率，为水稻品种的定向遗传改良提供优良的遗传转化技术体系。