玉米自交系愈伤组织诱导及植株再生研究

以玉米自交系7922、H99、P2、Mo17、吉846、吉63等的幼胚为外植体诱导愈伤组织，研究了不同培养基、不同激素种类和浓度及不同碳源的种类和浓度对愈伤组织诱导的影响。结果表明：NB培养基的胚性愈伤组织诱导率最高；对于不同基因型玉米自交系2,4-D浓度为2 mg/L时胚性愈伤组织诱导率均最高；在以20 g/L蔗糖和20 g/L葡萄糖及30 g/L蔗糖和10 g/L葡萄糖作碳源的诱导培养基上愈伤组织诱导率最高，但不能长期继代使用。对不同激素组合对玉米愈伤组织再生分化的影响进行了探讨。     

铁兰液体振荡培养再生与快速繁殖

以铁兰叶片为外植体,诱导获得脆性愈伤组织,液体振荡进行胚性愈伤组织培养及快速增殖,固体培养诱导胚状体萌发、不定芽生根,最后移栽成活,实现了铁兰的快速繁殖。在优化的培养条件下,铁兰无菌苗叶片愈伤组织诱导率90%以上,振荡后的愈伤组织100%成为胚状体,体积增殖系数可达5.0;90%以上的胚状体可分化为不定芽,95%以上的不定芽可诱导生根,移栽成活率95%以上。研究结果表明成功进行了铁兰的液体振荡培养及快速繁殖。     

玉米幼胚愈伤组织诱导及再生

利用综3、87-1、郑58、昌7-2、137和K12等6个骨干自交系玉米幼胚进行愈伤组织诱导并建立再生体系。结果显示,2,4-D对玉米幼胚愈伤组织的诱导形成是必需的,浓度在2.0～2.5 mg/L;6-BA不利于愈伤组织的形成,但能促进幼胚直接长芽、长根,尤其以郑58最明显。材料综3、郑58、K12诱导愈伤组织比较容易,诱导率均≥85%;材料137和87-1表现中等,诱导率约在50%～60%;材料昌7-2表现最差,愈伤组织诱导率≤25%。     

玉米自交系郑58和丹340幼胚再生体系的建立

研究选取9个优良玉米品种的自交系,以幼胚为外植体,以N6、MS培养基为基本培养基,诱导愈伤组织,探讨基因型、培养基类型、外植体大小、激素浓度等因素对愈伤组织诱导率的影响,分析不同基因型愈伤组织生长状态的差异,对诱导成功的愈伤组织进行分化和再生。研究结果表明,基因型对愈伤组织诱导率起决定作用,且基因型和培养基之间存在相互作用;外植体大小在1.5～2.0 mm时的幼胚较适合诱导胚性愈伤组织;2,4-D对玉米幼胚愈伤组织的诱导形成起关键作用,浓度在2.0～2.5 mg/L时大多能诱导出胚性愈伤组织。     

玉米胚性愈伤组织转基因受体系统建立的研究初报

通过对几个常用玉米自交系的幼胚培养,探讨不同基因型、不同培养条件对幼胚离体培养的影响。结果表明,相同自交系在MS和N6不同培养基上胚性愈伤的诱导率存在显著或极显著的差异;不同自交系材料在相同培养条件下其愈伤诱导率存在显著或极显著差异。并发现N6A培养基为供试材料的最适培养基,18-599(红)为胚性愈伤诱导率最高的基因型。研究的结果还表明,不同基因型要求的最适培养基存在差异,2,4-D对愈伤组织的诱导率影响显著,NAA的添加不能显著提高愈伤组织的诱导率。     

柑桔组织培养中的次生胚胎发生研究

以MT为基本培养基并附加各种植物激素和生理活性物质的一系列培养基中,6～8周龄的柑桔胚珠诱导产生愈伤组织和胚状体后进一步诱导形成次生胚状体,通过萌发或诱导丛芽再生出完整植株。外源激素的种类和组合对愈伤组织的诱导,胚状体的形成和次生胚状体的发生以及成苗都有显著的影响。染色体计数分析表明,通过次生胚胎发生途径的再生植株,其遗传性是稳定的。     

胚龄和2,4-D浓度对玉米自交系幼胚愈伤组织诱导率的影响

玉米幼胚愈伤组织的诱导由于受多种因素的影响使其相对于其它禾本科植物而言较为困难,其中胚龄和2,4-D浓度是影响玉米幼胚愈伤组织诱导的两个较为主要的因素.本文分析了胚龄和2,4-D浓度对玉米幼胚愈伤组织诱导率的影响,提出了玉米幼胚组织培养的最适的胚龄因自交系而异,对于吉853,吉842两种自交系胚龄以11～13d为宜,而4112自交系则以12～14d为宜;3种自交系诱导的适宜2,4-D浓度都为1mg/L,自交系间差异不大.为进一步研究能长期继代的玉米幼胚胚性愈伤组织的诱导,建立胚性愈伤组织的培养体系提供了参考依据。     

玉米幼胚组织培养体系优化

以5个骨干玉米自交系幼胚为材料, 研究幼胚大小、2, 4-D浓度、KT浓度对幼胚愈伤组织诱导及分化的影响。结果表明, 5个骨干玉米自交系均可诱导出愈伤组织, 但不同条件下诱导率存在显著差异。最适幼胚大小为2.0～3.0 mm, 因基因型不同而有所变化;2, 4-D浓度在1.0～3.0 mg/L有利于愈伤组织诱导, 同一材料不同2, 4-D浓度对胚性愈伤组织诱导率的影响具有显著差异, 不能以初级愈伤诱导率高低作为选择胚性愈伤诱导率的标准;KT可显著提高愈伤组织的分化率, 随着KT浓度的增加分化率呈先上升后下降的趋势, 最适KT浓度为1.0～2.5 mg/L。     

农杆菌介导优良玉米自交系Ⅱ型胚性愈伤组织遗传转化体系的建立

以玉米自交系8902、340、4112未成熟幼胚为材料,诱导、继代筛选出良好的Ⅱ型胚性愈伤组织,用农杆菌(EHA105和LBA4404)介导法转化其愈伤组织.利用本室构建的植物双元表达载体中携带的GUS报告基因对农杆菌转化系统的条件进行了研究,如农杆菌的侵染浓度、菌种类型、侵染时间、A8(乙酰丁香酮)浓度及基因型等,建立了农杆菌介导优良玉米自交系Ⅱ型胚性愈伤组织高效遗传转化体系。     

小麦不同外植体离体培养与再生研究

为了研究小麦组织培养的影响因素,对6个小麦品种的幼胚、幼穗以及成熟胚进行组织培养,研究不同基因型、外植体以及培养基对愈伤组织诱导及分化再生的影响.结果表明,不同小麦品种幼胚、幼穗以及成熟胚愈伤组织诱导率均在90%以上,且差异不大,而分化率和再生率却表现出很大差异.总体来说,供试外植体幼胚再生率最高,幼穗次之,成熟胚最低.绵阳19幼胚再生率最高,为54%,其最佳激素培养基为MS KT 1.0 mg/L IAA 1.0 mg/L;其次为小偃107和小偃22的幼胚愈伤组织.幼穗愈伤组织中绵阳19愈伤组织再生率最高,为27.5%,其次为洛阳8716.成熟胚愈伤组织培养中陕354再生率最高,为25.1%,优于其幼穗愈伤组织,其次为绵阳19的成熟胚愈伤组织.     

苯乙酸促进水稻花药愈伤组织的再分化和直接成苗

 研究了苯乙酸（phenylacetic acid，PAA）应用于水稻花药培养的效果。以 2,4-D 为对照，发现PAA不影响愈伤组织诱导率，但显著提高了愈伤组织分化率（p<0. 05），从而显著提高了植株再生率（p<0. 05）。其促进效果与品种、培养基无机盐成分有关，对籼稻品种效果较好。PAA还能有效地促进花药愈伤组织不经转移到分化培养基上而直接在诱导培养基上分化成苗。直接再生的绿苗大多具有旺盛的不定根，可直接移栽到土壤中去。当PAA浓度为10 mg/L 时，特青、02428和亚优2号的直接植株再生率分别达0. 56%, 1. 64%和2. 69%。提高PAA浓度可进一步提高直接植株再生率。利用基于PAA的一步成苗培养法已构建成了3个水稻杂种花培后代加倍单倍体（DH）群体。     

In vitro shoot regeneration ofSolanum tuberosum cultivars: interactions of medium composition and leaf, leaflet, and explant position

Summary Shoot regeneration was investigated on explants from different leaves and leaflets of three potato cultivars Posmo, Folva and Oleva. Explants were excised from glasshouse grown plants and grown for 6 days on callus induction medium with indole-3-acetic acid or 2,4-dichlorophenoxyacetic acid. Explants were then transferred to auxin free shoot regeneration medium with gibberellic acid and 6-benzyladenine or zeatin. By using the optimum combinations and concentrations of plant growth regulators and by excision of explants from particular regions of proximal leaflets from newly unfolded leaves, shoot regeneration frequencies of 97.0% were obtained for cv. Posmo and 32.1% for cv. Folva. Shoot regeneration frequency of cv. Oleva was very low and could not be improved by the different treatments.     

基因型对油菜子叶外植体再生植株的影响

为了了解细胞核和细胞质基因在油菜再生中的作用，采用MS培养基附加不同的激素，对6个基因型相关材料的子叶外格体进行了愈伤组织诱导及植株再生研究。愈伤组织诱导频率、生长速度以及不定芽的生根频率，主要受细胞核基因的影响，细胞质施加的影响不明显。细胞核基因极显著地影响着不定芽的再生，在此过程中，细胞质也起了显著的作用。陕2A与PolimaA的胞质效应表现一致，两种不育材料可能属同种类型来源的细胞质。     

Callus induction and in vitro plant regeneration of rice (Oryza sativa L.) under various conditions

The objective of the present study was to develop an effective protocol for optimum callus induction and complete plant regeneration for four varieties of rice (Oryza sativa L.) i.e., Super Basmati, Basmati-370, Basmati-371 and Fakhre Malakand. Calli were induced from mature seed scutelum. The Murashige and Skoog (MS) and Chu's N6 media containing hormone 2, 4-D (2, 4-Dichlorophenoxy acetic acid) in different concentrations were used for callus induction. Fakhre Malakand produced maximum calli on N6 media containing 3 mg L(-1) 2,4-D. while other three varieties showed maximum callus induction on N6 media containing 2.5 mg L(-1) 2,4-D. N6 media was found better than MS media for callus induction. For complete plant regeneration the calli of two varieties i.e., Basmati-370 and Basmati-371 were plated on N6 media containing different concentrations of NAA (1-Naphthalene acetic acid) and BAP (6-benzyl aminopurine). The maximum regeneration frequency (%) was observed on N6 media containing NAA 1 mg L(-1) and BAP 2.5 mg L(-1). It took 27-30 days for the callus to regenerate into a complete plant. Basmati-370 produced 4-7 plantlets per callus whereas Basmati-371 produced 4-8 plantlets per callus with regeneration frequencies of 61 and 69%, respectively.     

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。     

玉米花粉诱导小黑麦单倍体的诱导率遗传规律分析

为揭示玉米花粉诱导小黑麦产生单倍体中诱导率的遗传规律,以六个诱导率不同的春性小黑麦品系为材料,采用完全双列杂交方法,对与小黑麦单倍体诱导率相关的颖果率、得胚率、颖果得胚率及成苗率的配合力、遗传力及杂种优势等进行了分析。结果表明,小黑麦亲本的颖果率、颖果得胚率、得胚率及成苗率一般配合力(GCA)差异较大,品系08春H区61和08春H鉴25颖果率的GCA效应值较高,而品系08春H鉴10得胚率的GCA效应值最高。四个性状的特殊配合力(SCA)效应值也有较大差异,同一性状在不同组合间SCA效应值的变幅也较大,颖果率的SCA效应值变幅为-14.23~36.21,得胚率的SCA效应值变幅为-20.32~25.65,颖黑得胚率的SCA效应值变幅为-9.58~18.11,成苗率的SCA效应值变幅为-0.99~5.31。四个性状的母体遗传效应所占遗传分量均最大,非遗传效应次之。得胚率、颖果得胚率和成苗率的超亲优势均达到了极显著水平,且均表现为负向优势,颖果率的超亲优势和中亲优势在参试杂交组合中的变幅最大。四个性状的狭义遗传力均超过40%,其中成苗率的狭义遗传力最高(67%);得胚率、颖果得胚率以及成苗率的广义遗传力分别为41.46%、59.52%和66.80%,都达到了极显著水平。可见,根据母本的诱导率,参考配合力高低选择优良F1代进行诱导有利于提高玉米花粉诱导小黑麦单倍体的效率。     

不同菜用大豆品种的胚胎发生与再生

以8个菜用大豆品种的未成熟子叶为外植体,在MS(附加高浓度2,4-D)培养基诱导愈伤组织形成、体细胞胚胎发生与再生植株.结果表明:八个菜用大豆品种的未成熟子叶均可诱导产生愈伤组织与体细胞胚胎发生,出愈率为14.71%～100%.体细胞胚胎发生率变化在0～36.76%之间,绿子叶菜用大豆没有形成体细胞胚.体细胞胚可正常萌发产生小植株,移栽成活结实.菜用大豆的出愈率和体细胞胚胎发生率与基因型有关.     

In vitro direct plant regeneration and Agrobacterium‐mediated transformation of lucerne (Medicago sativa L.)

In vitro direct plant regeneration of lucerne was achieved by simultaneous application of thidiazuron (TDZ) and 6‐benzyladenine (BA) in Murashige and Skoog (MS) medium. Seedlings were germinated and grown for 6 d on growth regulator–containing MS medium. The shoot tip, consisting of the apical meristem along with parts of the cotyledonary leaves and hypocotyl, was then cultured on a medium containing the growth regulator(s). Adventitious budding of the shoot tip was promoted synergistically by treatment with TDZ and BA, and a maximum of thirty‐five shoots per explant was obtained on a medium supplemented with 2 mg L^?1 TDZ and 1 mg L^?1 BA. Plant regeneration frequency varied from 67 to 93%, and five Indian lucerne cultivars responded well to the regeneration protocol. The Agrobacterium‐mediated transformation frequency from co‐cultivated explants was 13% following multiple shoot induction. Southern analysis of the T₀ plants and T₁ progenies confirmed stable inheritance of the hpt marker gene. Agrobacterium infection of the explant caused a significant reduction in the plant regeneration frequency (23%) and the number of shoots induced (11) when compared with uninfected explants. A single shoot tip provided sufficient material to regenerate and establish twenty‐seven lucerne plants, whereas only nine plants could be regenerated from an Agrobacterium co‐cultivated explant. This transformation protocol could represent a valuable improvement over existing ones for lucerne.     

Somatic embryogenesis and regeneration of five multipurpose cassava landraces extensively integrated in African cropping system

Successful development and adoption of transgenic cassava (Manihot esculenta Crantz) varieties in Africa depend on in vitro regeneration of landraces because of their agronomic value and amenability to genetic modification. The study investigated somatic embryogenesis from axillary bud and immature leaf-lobe explant and regeneration via shoot organogenesis in five cassava landraces. Landraces exhibited significant (P < 0.05) differences in frequencies of primary somatic embryo production, number of embryos per explant, and frequency of secondary embryos. The frequency of primary somatic embryogenesis ranged from 7.8 to 14.5%, whereas the number of embryos per explant varied from 5.3 to 10.6. However, only frequency of primary somatic embryos and number of embryos per explant showed significant (P < 0.05) differences when immature leaf lobe was used as explant. The frequency of primary somatic embryogenesis ranged from 42.7 to 49.2%, whereas number of embryos per immature leaf-lobe explant varied from 9.5 to 15.2 per explant. Secondary embryogenesis was cultivar-independent in the case of immature leaf-lobe explant. Cyclic and green (mature) embryogenesis showed no significant (P < 0.05) landrace differences in both axillary bud and immature leaf-lobe explants. Shoot regeneration from cotyledon of somatic embryos had significant (P < 0.05) landrace differences. The mean shoot-bud formation frequency of axillary bud and immature leaf-lobe explants was 51.4% and 37.2%, respectively. Root formation was efficient, with greater than 70% of shoots forming root in all landraces. Similarly, landrace differences were detected for the survival of acclimatized regenerated plants, with a mean of 94.7% for both explants. In conclusion, somatic embryos were produced from immature leaf and axillary bud explants of the landraces and the embryos were converted to plantlets via shoot organogenesis.     

大豆子叶节高频愈伤组织诱导及其植株再生

以萌发7d的大豆子叶节切断及其切片为外植体,研究了萌发培养基、不同激素及其配比对大豆子叶节愈伤组织诱导与分化的影响.结果表明:当不添加BA时,在子叶节切断处几乎无愈伤组织的产生,只有添加BA时,才能产生愈伤组织.外植体切取方式、愈伤组织诱导培养基中BA和TDZ的浓度配比对愈伤组织诱导率产生很大影响.BA浓度为0.05～...