甘蓝型油菜高效离体再生体系的建立

以甘蓝型油菜(Brassica napusL.)HC8为材料,从无菌苗苗龄、预培养基激素浓度、预培养天数、6-BA及NAA的浓度等方面对影响油菜组织培养的因素进行了分析研究,建立了甘蓝型油菜品系HC8的离体再生技术体系。结果表明,该油菜组织培养的最佳苗龄为5 d;最佳预培养时间为5 d,最佳2,4-D浓度为1.0 mg/L;子叶最佳诱导培养基为MS+2.0 mg/L 6-BA+0.05 mg/LNAA+3.5 mg/L AgNO3或MS+3.0 mg/L 6-BA+0.1 mg/L NAA+3.5 mg/L AgNO3,该条件下子叶愈伤组织诱导率最高可达100%,再生频率及分化频率分别可达88.0%和108.33%;下胚轴最佳诱导培养基为MS+4.0 mg/L 6-BA+0.05 mg/L NAA+3.5 mg/L AgNO3,子叶愈伤组织诱导率最高可达95.24%,再生频率及分化频率分别可达81.82%和104.55%;最佳生根培养基为MS+0.5 mg/L NAA,生根率最高为90.0%。     

中国菟丝子离体再生体系的建立

选取萌发3～5d、长度3～5cm的中国菟丝子（Cuscuta Chinensis Lam）幼苗,将其分为上、中、下3部分并作为外植体进行离体培养与植株再生研究。结果表明,其上、中部片段更适宜愈伤组织诱导;诱导培养基以添加1mg L^-1 NAA和1mg L^-1 BA的MMS培养基效果最好,此培养基也可用于愈伤组织的继代培养,愈伤组织在上述培养基中已生长一年之久。分化培养基为添加1mg L^-1 BA的MMS培养基,平均每块愈伤组织可以产生2．8株植株。     

黄斑橡胶榕离体再生体系的建立

以黄斑橡胶榕(Ficus robusta“Yellow spot”)的顶芽为外植体进行离体培养与植株再生研究。结果表明,黄斑橡胶榕的诱导培养基以MS BA 2.0 mg L-1 NAA 0.1 mg L-1为宜;继代增殖以MS BA 1.0 mg L-1 NAA 0.05 mg L-1为宜,每个外植体可产生3个芽;生根培养基以MS IBA 0.1 mg L-1 AC 100 mg L-1为宜,生根率96.67%以上。试管苗移栽成活率达到98%以上。     

樱桃砧木Colt离体叶片再生

以樱桃砧木Colt试管苗的叶片为外植体,通过先诱导愈伤组织分不定芽以及叶片直接分化不定芽两种途径诱导再生,结果表明,在MS附NAA1．0mg/L、KT3．0mg/L、ZT0．25mg/L培养基中,愈伤诱导率可达100％;诱导的愈伤MS附加NAA0．2mg/L、IAA0．5mg/L、6－BA0．5mg/L、KT1．0mg/L,GA0．5mg/L培养基中,不定芽分化率为21．3％,在MS附加6-BA6．0mg/L、NAA1．0mg/L、GA0．5mg/L中,叶片一叶柄不定芽诱导率可达48．3％。     

荞麦高频离体再生及发根农杆菌转化体系的建立

荞麦无菌苗下胚轴切段在不同激素配比的MS培养基上诱导愈伤组织,出愈率均为100％。在2．0mg/L2,4-D和1．5mg/L 6-BA组合下诱导产生的愈伤组织;转入2．0mg/L 6-BA和1．0mg/L KT的MS培养基,再生苗分化率在80％以上。根尖色体分析表明再生植株具一定的遗传稳定性。发根农杆菌A4转化荞麦下胚轴和子叶获得发状根,纸电泳检测所有随机取样测定的发状根均有相应冠瘿碱的存在。     

黑莓品种‘Arapaho’离体叶片不定芽诱导影响因素分析及再生体系建立

以黑莓(Rubus spp.)品种‘Arapaho’无菌苗叶片为外植体,通过正交和单因素实验分别研究了基本培养基类型、6-BA和1BA质量浓度以及暗培养时间、外植体的叶位和接种方式对不定芽诱导的影响,并研究了IBA质量浓度对不定芽生根的影响;在此基础上,初步建立了黑莓品种‘Arapaho’离体叶片的再生体系.正交实验结果表明:基本培养基类型对叶片不定芽诱导率及平均不定芽数的影响最大,而IBA质量浓度对叶片不定芽诱导率及6-BA质量浓度对平均不定芽数的影响较小;适宜‘Arapaho’叶片不定芽诱导的最佳培养基为含有2.0mg·L-16-BA和1.0 mg·L-1IBA的MS培养基.单因素实验结果表明:暗培养时间、外植体的叶位及接种方式对不定芽诱导率有显著影响;最适宜的暗培养时间为21 d;植株中、上部叶片的再生能力较强,其中第3和第4位叶的不定芽诱导效果最佳;叶面朝上接种更有利于不定芽的诱导.在含0.2 mg·L-1 IBA的MS培养基中,不定芽生根率达100.0％,且根数多、长势良好.黑莓品种‘Arapaho’离体叶片的再生体系为:以无菌苗的第3和第4位叶为外植体,经过适当修剪后叶面朝上接种于含有2.0 mg·L-16-BA和1.0 mg·L-1IBA的MS培养基上,暗培养21 d后置于光照条件下培养30 d;将不定芽转接到含有0.5 mg·L-16-BA和0.3mg·L-1 NAA的MS培养基上进行继代培养;当不定芽高约2 cm时转接到含有0.2 mg·L-1IBA的MS培养基上进行生根培养,最终获得完整植株.     

胡杨雄花和花序轴离体培养的适宜培养基

本文以胡杨（Populus euphratica Oliver）雄花和花序轴为外植体,在11种培养基（表1）上诱导愈伤组织,在8种培养基（表2）上诱导芽分化,在5种培养基（基本培养基为1／2MS,表3）上诱导根分化,得到如下结果：     

野牛草成熟胚离体培养及植株再生

1植物名称野牛草[Buchloe dactyloides(Nutt.)texoka]. 2材料类别成熟胚. 3培养条件(1)愈伤组织诱导培养基:MS 2,4-D1.5～6.0 mg·L-1(单位下同) 6-BA 0.1 脯氨酸1 000 水解酪蛋白(CH)500 谷氨酰胺500 α-酮戊二酸100 硫代硫酸银(STS)5;(2)愈伤组织继代培养基:MS 3/2MS(有机) 2,4-D 2.5 6-BA 0.1 CH1 000 聚乙烯吡咯烷酮(PVP)200或维生素C(vC)200;(3)再生培养基:不附加任何植物生长调节物质的MS基本培养基(MS0).所有培养基中均添加3%蔗糖、0.56%琼脂,pH 5.8.愈伤组织诱导及继代培养为暗培养,不定芽分化及植株再生过程中光照12 h·d-1,光照度为1 500lx,培养温度为(25±1)℃.     

蚂蝗七的离体快速繁殖

1植物名称蚂蝗七(Chirita fimbrisepala Hand.- Mazz.)。2材料类别花梗和带苞片的花蕾。3培养条件(1)愈伤组织诱导和不定芽分化培养基;MS 6-BA 0.1mg·L~(-1)(单位下同) NAA 0.1;(2)继代增殖培养基:MS 6-BA 0.05 NAA 0.1;(3)壮苗生根培养基:1/2MS 1.0%蔗糖 0.3%活性炭。培养基(1)和(2)均加入3.0%蔗糖和0.6%琼脂,pH 6.2,培养温度(25±3)℃;在愈伤组织     

樱桃砧木Colt离体叶片再生

以樱桃砧木Colt试管苗的叶片为外植体 ,通过先诱导愈伤组织分化不定芽以及叶片直接分化不定芽两种途径诱导再生。结果表明 :在MS附加NAA 1 0mg/L、KT3 0mg/L、ZT0 2 5mg/L培养基中 ,愈伤诱导率可达 1 0 0 % ;诱导的愈伤在MS附加NAA 0 2mg/L、IAA0 5mg/L、6 BA 0 5mg/L、KT 1 0mg/L、GA 0 5mg/L培养基中 ,不定芽分化率为 2 1 3% ;在MS附加 6 BA 6 0mg/L、NAA 1 0mg/L、GA 0 5mg/L中 ,叶片 -叶柄不定芽诱导率可达 48 3%。     

提高小麦愈伤组织分化频率的因素

研究了影响小麦愈伤组织诱导、芽分化及其植株再生的一些因素。结果表明：在愈伤组织诱导和继代过程中添加ＡＢＡ（１ ．０ｍｇ／Ｌ） 有利于小麦中晚期幼胚致密愈伤组织的诱导及再生能力的保持;外植体来源尤其是基因型对长期培养的愈伤组织再生能力有很大影响; 不同的外源激素（ ＫＴ、６ＢＡ、ＩＡＡ、ＴＤＺ和玉米素等） 也影响芽分化频率,其中ＴＤＺ可明显提高芽分化频率;在转入分化培养前对愈伤组织进行干燥处理可有效地提高其芽分化频率;在生根培养基中添加适量的ＩＡＡ 或ＮＡＡ 可有效促进生根。     

Direct organogenesis and plantlet regeneration from mature zygotic embryos of masson pine (Pinus massoniana L.)

Mature zygotic embryos of masson pine were cultured as initial explants to investigate the process of direct organogenesis. Adventitious buds were initiated on DCR medium (Douglas-fir cotyledon revised medium) supplemented with 0.5 mg l⁻¹ N⁶-benzyladenine (BA) and 0.05 mg l⁻¹ indolebutyric acid (IBA) or α-naphthaleneacetic acid (NAA). The highest induction frequency of adventitious buds was 99.3%. Subsequent transfer of buds to medium with lower concentrations of plant growth regulators in time was necessary for differentation of high quality adventitious buds. After culturing on elongating medium, in which the proportion of cytokinins to auxins was reduced, shoots higher than 2 cm were transferred for root induction to GD medium with half of the concentration of macro-salts (½ GD) and with 2 mg l⁻¹ IBA and 0.05 mg l⁻¹ BA. The average root frequency was over 70%. After adventitious roots had appeared, the shoots were transferred to ½ GD medium with a lower concentration of IBA (0.2 mg l⁻¹) for further root development.     

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。     

A system for routine plantlet regeneration of sunflower (Helianthus annuus L.) from immature embryo-derived callus

Immature embryos from inbred commercial cultivars of sunflower (Helianthus annuus L.) were used as donor material for induction of regenerable tissue in vitro. Optimum regeneration frequencies were obtained by transferring the tissue through a sequence of defined media using a specified timetable. The first medium was characterized by a high sucrose content (12%), 10 M 2,4-dichlorophenoxyacetic acid and 5 M abscisic acid. Within 7 days, proliferation of smooth, white, dedifferentiated tissue from the cotyledons was evident. After 3 weeks, the tissues were transferred through a series of 3 media, designed to promote shoot formation, shoot elongation and rooting. Regenerates were obtained with all 7 genotypes tested. From 1983 to 1984, approximately 500 primary R₀ plantlets were regenerated, grown to maturity in a greenhouse and self-pollinated. The resultant R₁ seeds were subsequently field-grown and the plants were evaluated for variation.     

骨干玉米自交系丹598遗传再生体系的建立

目的:以玉米骨干自交系丹598的幼胚为外植体,诱导愈伤组织建立遗传再生体系。方法:探讨胚龄、培养基种类、2,4-D浓度对愈伤组织诱导的影响。结果:在授粉后16～18 d,2,4-D浓度为2.0 mg/L时诱导最佳;设置N6、NB、改良NB、MS、MB等5种培养基,筛选出改良NB培养基为最佳诱导培养基;分化培养基中添加1 mg/L激动素、0.5 mg/L 6-卞基嘌呤和0.5 mg/L萘乙酸能促进绿苗分化和根系生长。结论:建立了玉米自交系丹598的优良再生体系,为以后的基因转化工作打下了良好基础。     

香石竹叶片离体再生体系的建立

以香石竹(Dianthus caryophyllus Linn.)无菌苗叶片为外植体,从不同细胞分裂素及其他激素配合使用等方面进行筛选,建立香石竹叶片离体再生体系.结果表明,不同的细胞分裂素影响叶片不定芽分化频率,其中较低浓度的6-BA(0.5 mg·L-1)和TDZ(0.001 mg·L-1)配合使用能有效诱导香石竹叶片不定芽分化;添加一定浓度的PP333(4 mg·L-1)可提高叶片不定芽分化频率和平均芽数.香石竹叶片不定芽分化的适宜培养基为:MS 0.002mg·L-1TDZ 0.5 mg·L-16-BA 0.2 mg·L-1IAA 4 mg·L-1PP333;壮苗培养基为:MS 0.2 mg·L-1 6-BA 0.2 mg·L-1IAA;生根培养基为:1/2 MS.不定芽诱导频率达到42.61%,平均芽数为4.53个.     

Micropropagation of sugarcane (Saccharum spp.)

A method for callus induction, adventitious bud regeneration, shoot multiplication and rooting of in vitro formed shoots of Helianthus annuus L. var. Argentario is described. Hypocotyl and cotyledon explants formed callus on medium containing 2 mgl^–1 naphthalene acetic acid and 0.5 mgl^–1 benzyladenine. Adventitious buds were formed on hypocotyl segments on medium containing 0.5–2 mgl^–1 benzyladenine. The optimal level of sucrose concentration for shoot regeneration from hypocotyls was 1.5%. Multiplication from shoot apices was promoted by kinetin (2 mgl^–1) plus gibberellic acid (5 mgl^–1), benzyladenine (2 mgl^–1) plus gibberellic acid (10 mgl^–1) or at lower frequency by benzyladenine (1 mgl^–1). A general feature of the plantlets formed in vitro was the precocious flowering.     

狭叶黄芩组织培养再生体系的建立

以狭叶黄芩的茎段为外植体,研究不同消毒剂处理、不同植物生长激素配比对狭叶黄芩茎段腋芽诱导、愈伤组织诱导、丛生芽分化、增殖、生根及移栽的影响。结果表明：最佳消毒方式为0.1% HgCl₂消毒5 min,污染率最低为8.25%;诱导腋芽最佳培养基为MS+1 mg·L^-1 6-BA+1 mg·L^-1 NAA,诱导率可达73.66%;诱导愈伤最佳培养基为MS+1 mg·L^-1 6-BA+1 mg·L^-1 2-4D,诱导率为91.33%;愈伤组织分化的最佳培养基为MS+1 mg·L^-1 6-BA+0.5 mg·L^-1 NAA,分化率为44.71%。芽增殖的最佳培养基为MS+1 mg·L^-1 6-BA+0.5 mg·L^-1 NAA,其芽增殖倍数为5.85;最佳生根培养基为1/2MS+0.2 mg·L^-1 IBA,生根率可达到74.07%;试管苗移栽时蛭石：珍珠岩：园土比例按1：1：3的体积比搭配使用,移栽成活率最高达到79.24%,并且植株生长旺盛。本研究建立狭叶黄芩再生体系,为狭叶黄芩野生资源在妥善的保护基础上开发应用提供一定的理论支持。     

Somatic embryogenesis and plant regeneration from hypocotyl protoplasts of sunflower (Helianthus annum L.)

A sunflower genotype (Helianthus annuus L. cv. Florom-328) able to regenerate plants from in vitro cultures was identified by screening hybrids and inbred lines. Protoplasts of this genotype were isolated from dark grown hypocotyls and were cultured in droplets of agarose-solidified V-KM medium covered by liquid V-KM supplemented with naphthaleneacetic acid (NAA) and benzylaminopurine (BAP). One week later colonies were subjected to 2,4-dichlorophenoxyaceticacid for a one week period. Further culture in V-KM with reduced concentrations of NAA and BAP resulted in the appearence of somatic embryos. Maturation of embryos was achieved by culture on MS medium supplemented with NAA, BAP, gibberellic acid A₃ and the ethylene inhibitor AgNO₃. Embryos were then transferred onto hormone free MS medium for germination. The frequency of shoot formation in the best case was 9.6 percent of viable colonies (1.3 percent of protoplasts plated). Some of the shoots with roots could be transplanted into soil, others were grafted on hypocotyls of in vivo germinated seedlings. Eighty percent of grafted shoots and over 95 percent of rooted shoots survived. The plants flowered and produced 5 to 10 seeds each. Factors affecting the frequency of embryo formation and plant regeneration are discussed.Abbreviations BAP 6-benzylaminopurine - GA3 gibberellic acid - MES morpholinoethanesulfonic acid - MS Murashige and Skoog medium - NAA naphthaleneacetic acid - V-KM protoplast culture medium of Binding and Nehls - 2,4D 2,4-dichlorophenoxyacetic acid     

A shoot regeneration protocol effective on diverse genotypes of sunflower (Helianthus annuus L.)

Summary We describe a protocol, and several experiments that helped lead to its development, for sunflower regeneration. Important factors for sunflower regeneration were explant age, cytokinin type and concentration, basal medium, and explant source. We could not induce shoot regeneration from the explants derived from mature tissues including leaf, petiole, and stem. However, use of juvenile explants such as embryo meristem and primordial leaf tissues allowed routine regeneration of 17 different sunflower genotypes. High frequency of shoot regeneration was achieved with these explants taken from seedlings up to 5 d after germination. Explant age was less critical for embryo meristem explants than for primordial leaf tissues. Of the four basal media tested, MS and B5 media produced higher shoot-regeneration frequencies than did Anderson and woody plant media. The highest shoot-regeneration frequency was obtained with MS medium supplemented with 2 μM BA and without auxin. Addition of 1 μM naphthalene-acetic acid to the medium significantly reduced both the percentage of explants producing shoots and average number of shoots per explant. Regenerated shoots were grown to maturity in a greenhouse.