红江橙胚发生及影响因素的研究

红江橙花后４周的幼果,直径约１ｃｍ左右,这时期的胚珠最适于胚性愈伤组织的诱导。以ＭＴ为基本培养基,加上适当浓度的ＩＡＡ、ＢＡ和ＭＦ可显著提高胚性愈伤组织诱导率。体胚发生,以ＭＴ＋ＩＡＡ０．１ｍｇ１＾－１＋ＢＡ（ＺＴ）１ｍｇ１＾－１培养基较好,涌诱导产生正常体胚,且频率较高。胚性愈伤组织继代次数对体胚发生也有一定的影响,继代６次以内,胚性愈伤组织具有旺盛产生体胚能力;继代至第９次时,产生体胚的能力明     

一品红离体培养诱导体细胞胚的研究（简报）

一品红的嫩茎离体培养在附加不同浓度的２,４－Ｄ和６－ＢＡ的ＭＳ培养基上,２．４Ｄ浓度为１．０～２．０ ｍｇ· Ｌ~（－１）的愈伤组织生长量最大。这些愈伤组织转移到ＭＳ＋６－ＢＡ１．０ ｍｇ·Ｌ~（－１）＋ＮＡＡ０．１ ｍｇ·Ｌ~（－１）的分化培养基上后,产生了胚性愈伤组织,观察到大量不同时期体细胞胚。带有体细胞胚的愈伤组织进一步转移到无激素 ＭＳ培养基上,便发育成小植株。     

栀子组织和细胞培养生产天然食用色素的研究：Ⅰ.愈务组织…

在诱导出愈伤组织的基础上,对各种不同的培养条件进行分析研究,考察它们对愈伤组织生长和栀子黄色素产生的影响,筛选出适宜的生长培养基：Ｂ５＋ＴＢＡ １ｍｇ／１＋ｋＴ０．２３ｍｇ／、ＭＧ－５＋ＩＢＡ １ｍｇ／＋ＫＴ０．２３ｍｇ／、Ｂ５＋ＩＡＡ１．５ｍｇ／ｌ和生产培养基;Ｍ－９＋ＩＡＡ １ｍｇ／ｌ、Ｍ－９＋ＩＡＡ １．５ｍｇ／ｌ。并且,获得了几个色素含量较高的愈伤组织系。另外,还研究了含色素和不含色素的愈     

蛇床幼茎离体培养中体细胞胚胎形成的观察

蛇床幼茎外植体经诱导产生了愈伤组织。在ＭＳ＋２,４－Ｄ,０．２ｍｇ／Ｌ＋ＺＴ０．４ｍｇ／Ｌ培养基中,愈伤组织转变成胚性愈伤组织。转入ＭＳ＋ＮＡＡ０．２ｍｇ／Ｌ＋ＺＴ０．８ｍｇ／Ｌ培养基以后,胚性愈伤组织分化出体细胞胚胎。体细胞胚胎在ＭＳ＋ＮＡＡ０．５ｍｇ／Ｌ培养基中可直接发育成为完整植析。显微观察表明,体细胞胚胎产生于愈伤组织的表层细胞或内部细胞。在鱼雷胚期已有螺纹导管的分化。子叶期的维管组织从两     

茶（Camellia sinensis Kuntze）离体培养体细胞胚胎发生的组织细胞学研究

以茶树叶片为外植体．以ＭＳ培养基附加４ｍｇ·Ｌ－１６－ＢＡ,２ｍｇ·Ｌ－１ＩＡＡ．３ｍｇ·Ｌ－１ＧＡ３和０．２—０．３％活性碳诱导出愈伤组织和胚状体,进一步形成小植株．切片观察表明．茶叶愈伤组织胚状体的发生,起源于愈伤组织表层及其内部的单个细胞和细胞团,胚状体发育顺序与合子胚大致相似．经球形胚、心形胚、鱼雷胚和子叶胚阶段．但发育过程中常有畸形胚出现．     

在离体条件下拐芹（Angelica polymorpha）形成胚性愈伤组织和非…

以拐芹（Ａｎｇｅｌｉｃａ ｐｌｏｙｍｏｒｐｈａ）的叶柄为外植体,在附加２,４－Ｄ（１ｍｇ／Ｌ）＋ＫＴ（０．５ｍｇ／Ｌ）ＭＳ培养基上诱导脱分化,４０天内可形成愈伤组织。在相同的培养基上经过继代培养,可在２０天内形成胚性愈伤组织（Ｅｍｂｒｙｏｇｅｎｉｃ Ｃａｌｌｕｓ,ＥＣ）和非胚性愈伤组织（Ｎｏｎ－Ｅｍｂｒｙ－ｏｇｅｎｉｃ Ｃａｌｌｕｓ,ＮＥＣ）。将不同天数的培养物作为样品,提取其内源激素;气液相色谱     

大叶紫花苜蓿愈伤组织原生质体再生植株

大叶紫花苜蓿下胚轴诱导的愈伤组织在继代培养基上生长快速,易于分散。继代第１２ｄ的愈伤组织原生质体的得率为６．５×１０７／ｇ鲜重。原生质体培养基为ＳＨ基本培养基,含有１．０ｍｇ／Ｌ２,４－０、０．５ｍｇ／ＬＢＡ、２．０ｇ／ＬＣＨ、２％蔗糖、６％葡萄糖、５ｍｍｏｌ／ＬＭＥＳ,培养密度为１．０×１０５／ｍＬ。培养至第１２ｄ时的原生质体再生细胞植板率为３．７％。由原生质体形成的小愈伤组织在含２．０ｍｇ／Ｌ２,４－Ｄ的ＭＳ固体培养基上大量增殖。增殖的愈伤组织转移至２．０ｍｇ／Ｌ２－ｉｐ＋０．１ｍｇ／ＬＮＡＡ的Ｂ５培养基上,形成体细胞胚并发育成完整植株。     

决明组织培养的研究

以草决明无菌苗子叶为外植体,接种于７类诱导愈伤组织的培养基上：Ａ．ＭＳ＋２．０２,４－Ｄｍｇ／Ｌ（以下单位省略）＋０．３ＢＡ＋０．２ＮＡＡ．Ｂ．ＭＳ＋０．２２,４－Ｄ十０．２ＢＡ＋２．０ＮＡＡ．Ｃ．ＭＳ＋１．０２,４－Ｄ＋０．５ＢＡ＋０．２ＫＴ．Ｄ．ＭＳ＋０．７ＢＡ＋１．５ＮＡＡ＋０．１ＫＴ．Ｅ．ＭＳ＋１．５２,４－Ｄ＋０．７ＢＡ十０．２ＭＡＡ．Ｆ．ＭＳ＋０．４２．４－Ｄ＋１．０ＮＡＡ＋０．１ＫＴ．Ｇ．ＭＳＢ（ＭＳ的无机成份和Ｂ５有机成分）＋０．１５ＮＡＡ＋ＢＡ,ＫＴ和ＺＴ各０．５。８～１５天后分别有９０～９９．６％的子叶片被诱导出愈伤组织,并且Ｆ．与Ｃ．类培养基对诱导愈伤组织比较理想,放于Ｇ培养基上的愈伤组织有９．２～３０．２％的芽分化率。芽在生根培养基１／２ＭＳ＋０．２ＩＢＡ中、９８％生根,形成完整的再生植株。     

深山含笑的组织培养和快速繁殖

组织培养深山含笑的实验结果表明：利用休眠芽和种子萌发时实生苗的上胚轴及下胚轴为外植体均能诱导出愈伤组织和不定芽,其中无菌实生苗的上胚轴最易诱导出不定芽,无菌实生苗的下胚轴最易诱导出愈伤组织。外体植体在ＭＳ＋１．０－２．０ｍｇ Ｌ＾－１ ２,４－Ｄ培养基上只产生愈伤组织;在ＭＳ＋３．０ｍｇ Ｌ＾－１ ＢＡ＋０．２ｍｇ Ｌ＾－１ ＮＡＡ培养基上产生较多的不定芽和较多的愈伤组织;在ＭＳ＋２．０ｍｇ Ｌ＾     

二棱大麦体细胞胚性无性素的建立

对１２个品种的二棱大麦幼穗和６个品种的幼胚在附加５００ｍｇ／Ｌ酪蛋白,１５０ｍｇ／Ｌ天门冬素和２ｍｇ／Ｌ ２,４－Ｄ的ＭＳ培养基（Ｍ３２０）中进行了离体培养,并诱导产生出愈伤组织。把由Ｄ１幼胚所获得的愈伤组织转移到含２,４－Ｄ０．５ｍｇ／Ｌ（Ｍ３０５）培养基中后诱导了产生出了体细胞胚性愈伤组织,幼苗和根。筛选出了体细胞胚性无性系。在长达二年的继代培养中,该细胞系仍保持旺盛的再生和分化成苗能力。     

Somatic embryogenesis from leaf derived callus of Tylophora indica (Burm. f.) Merrill

Mature leaf explant derived callus of Tylophora indica (Burm. f.) Merrill yielded somatic embryos on MS medium supplied with BA(1-2 mg/L) or kinetin(1-5 mg/L) or kinetin/BA (1-2 mg/L) used along with IAA(0.1-1 mg/L). Maximum somatic embryos (30) could be recovered from 100 mg of embryogenic callus within 60 days at an optimum concentration of 2 mg/L of BA which was also best suited for providing the maximum conversion rate (90%) of embryoids to plantlets. Kinetin (1-5 mg/L), used as the sole growth hormone, induced the development of embryoids showing either shoot or root primordia in 30% of the cultures. However, embryoids with shoot primordia developed roots upon transfer to medium containing IAA(0.1 mg/L) and kinetin(2 mg/L). Embryoids from all cultures germinated in the initiation medium and were transplanted to sterile vermiculite for hardening. After two weeks of hardening, the plantlets were transferred to the green house where they grew and established well showing a high rate of survival (90%).     

Plant regeneration by somatic embryogenesis from callus cultures of sweet sorghum

Tissue culture methods were developed for the induction, maintenance, and regeneration of embryogenic callus in sweet sorghum (Sorghum bicolor) cultivars Keller, Rio, and Wray. No significant differences were observed in production of embryogenic callus in cultures established from developmentally immature or mature embryo explants cultured on LS medium with 2 mg/1 2,4-D plus 0.5 mg/1 kinetin. Prolific callus production did not occur until the third four-week culture period. Long-term maintenance of embryogenic callus was dependent upon the selective transfer of embryogenic callus, with other callus types discarded. High-frequency plant regeneration was achieved and quantified on a fresh weight basis of embryogenic callus.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - BA benzyladenine - IAA indoleacetic acid - IBA indolebutyric acid - LS Linsmaier and Skoog basal medium (Linsmaier and Skoog, 1965)     

Somatic embryogenesis and plant regeneration in Gloriosa L

Friable callus was initiated from shoot apices of Gloriosa superba L. on basal MS medium supplemented with 2, 4-D (4mg L(-1)) + Kn(5 mg L(-1)) + CH(10 mg L(-1)) + CW(20%). Subculture of callus on the same medium after 4-5 weeks showed induction of large number of somatic embryos, which was confirmed with histological studies. Development of embryoids in plantlet took place when the embryogenic callus was transferred to basal MS medium supplemented with BAP (5 mg L(-1)), CH(50 mg L(-1)) +CW(20%). Roots were developed by subculturing them on to the medium containing Kn or BAP (5 mg L(-1)) and IBA (4 mg L(-1)). Plantlets were successfully transferred to pots containing mixture of soil, sand and farmyard manure (2:1:1).     

Pollen embryogenesis in anther cultures of Solanum carolinense L.

The direct differentiation of bicellular pollen grains of Solanum carolinense L. (Horse-nettle; Solanaceae) into embryoids and plantlets was induced by culturing whole anthers on Murashige and Skoog's medium supplemented with IAA. The highest frequency of embryogenic induction occurred at 10 mg/l IAA. Developmentally, both the generative and vegetative cells of the pollen grain contributed to embryoid formation whose pattern of development was similar to that of zygotic embryos. In a previous study, it was show that 2,4-D promoted callus formation by pollen grains in cultured anthers of S. carolinense. It appears then that there are two distinct pathways of androgenesis in this species that are determined by the type of auxin present in the medium.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - BA benzyladenine - KIN kinetin - MS Murashige and Skoog     

Somatic embryogenesis in elm

We show that isolated zygotic embryos of Ulmus minor and U. glabra can produce embryogenic cultures provided they are isolated from immature seeds before storage proteins begin to accumulate. Rates of somatic embryogenesis were highest among zygotic embryos collected 6 weeks post-anthesis when they were at the midcotyledonary stage, were about 5 mm long and had a fresh weight of approx. 10 mg. At this time, induction was even possible in Murashige and Skoog basal medium with no plant growth regulators, but addition of 2,4-dichlorophenoxyacetic acid was necessary at earlier stages of zygotic development. In medium supplemented with benzyladenine (BA) only, no embryogenic induction was observed. The formation of callus was an essential step not only for the induction of embryogenic masses, but also for the maintenance of embryogenic competence through successive subculture of callus on induction media supplemented with 0.1 mg l(-1) BA. Nine embryogenic U. minor lines and 24 U. glabra lines have been maintained in this way for 3 years. However, conversion into plantlets has occurred only rarely.     

Induction of embryogenic callus of loose skin mandarins]

Young ovules from 3 cultivars and undeveloped ovules in mature fruits from 8 cultivars of loose skin mandarin of Citrus were cultured on 4 different media respectively to induce embryogenic calli. Results showed that the combination of EME(MT + 500 mg/L malt extract) and MKT (EME + 10 mg/L KT) media performed well in the induction of embryogenic calli from young ovules; MGS(EME + 1 mg/L GA3 + 40 mg/L sulfate adenine) medium was better than MDB (MT + 0.01 mg/L 2,4-D + 0.1 mg/L BA) medium in inducing calli from the undeveloped ovules, and the darkness was conducive to the induction of embryogenic calli. There was no chromosome number variation in the induced calli. All of the examined cells were diploid with 2n = 2x = 18 chromosomes.     

Carbon source dependent somatic embryogenesis and plant regeneration in cotton, Gossypium hirsutum L. cv. SVPR2 through suspension cultures

Highly reproducible and simple protocol for cotton somatic embryogenesis is described here by using different concentrations of maltose, glucose, sucrose and fructose. Maltose (30 g/l) is the best carbon source for embryogenic callus induction and glucose (30 g/l) was suitable for induction, maturation of embryoids and plant regeneration. Creamy white embryogenic calli of hypocotyl explants were formed on medium containing MS basal salts, myo-inositol (100 mg/l), thiamine HCI (0.3 mg/l), picloram (0.3 mg/l), Kin (0.1 mg/l) and maltose (30 g/l). During embryo induction and maturation, accelerated growth was observed in liquid medium containing NH3NO4 (1 g/l), picloram (2.0 mg/l), 2 ip (0.2 mg/l), Kin (0.1 mg/l) and glucose (30 g/l). Before embryoid induction, large clumps of embryogenic tissue were formed. These tissues only produced viable embryoids. Completely matured somatic embryos were germinated successfully on the medium fortified with MS salts, myo-inositol (50 mg/l), thiamine HCl (0.2 mg/l), GA3 (0.2 mg/l), BA (1.0 mg/l) and glucose (30 g/l). Compared with earlier reports, 65% of somatic embryo germination was observed. The abnormal embryo formation was highly reduced by using glucose (30 g/l) compared to other carbon sources. The regenerated plantlets were fertile but smaller in height than the seed derived control plants.     

Plant regeneration from isolated cells ofLavandula latifolia medicus

Summary Single cells were obtained from hypocotyl-derived callus ofLavandula latifolia Medicus. Cells were plated in Murashige and Skoog medium supplemented with indoleacetic acid (IAA), benzyladenine (BA), and several IAA-BA combinations. Cell division required the simultaneous presence of IAA and BA in the culture medium, but callus formation was only achieved with 0.1 or 1 mg/liter IAA and 2 mg/liter BA. To induce organogenesis, calli were transferred to various regeneration media. Shoot-bud differentiation efficiency depended on the composition of both the callus induction and the shoot regeneration media, best results being obtained when calli grown in 1 mg/liter IAA and 2 mg/liter BA were subcultured to media containing 2 mg/liter BA and 15% coconut milk. Under these conditions, up to 75% of calli formed shoots that subsequently were rooted and established in soil.     

Plant Regeneration Through Somatic Embryogenesis in Taxus wallichiana

We describe an efficient process for regeneration of Taxus wallichiana (Zucc) plants from callus cultures derived from zygotic embryos. Zygotic embryos cultured on half strength Lloyd and McCown’s basal medium supplemented with SH vitamin (1/2 WPMSH), 0.5 mg I^?1 6-benzyladenine (BA) and 1.0–2.0 mg I^?1 á-Napthaleneacetic acid (NAA) produced compact yellow (CY) callus after 4 weeks of culture. The 8-week-old CY call! (lines CY-A and CY-B) were initially slow growing but proliferated on transfer to WPM basal medium supplemented with 8.0 mg I^?1 2,4-D, 0.1–0.9 mg^?1 NAA and 0.3–1.0 mg^?1 BA after 4 weeks. Four morphologically distinct calli lines were obtained, of which only two call! lines, CY-B-FW and CY-B-FY were embryogenic. The 12-week-old callus line CY-B-FW developed globular somatic embryos on transfer to secondary medium after 8 weeks and matured in maturation medium after 4 weeks. Only 10% of the mature somatic embryos regenerated into complete plantlets after 4 weeks on conversion medium. Although the frequency of conversion was low, complete regenerated plantlets via somatic embryogenesis were obtained after 7–8 months of initiation of culture. Taxane analysis showed that the paclitaxel accumulation was higher in embryogenic callus than in non-embryogenic callus.     

Direct somatic embryogenesis and plantlet regeneration from seedling leaves of winged bean,Psophocarpus tetragonolobus (L.) DC

Excised seedling leaf segments of winged bean [Psophocarpus tetragonolobus (L.) DC.] underwent direct somatic embryogenesis under appropriate incubation conditions. Initiation and development of the somatic embryos occurred using a two-step culture method. The culture procedure involved incubation for 28 days on MS basal medium supplemented with 0.1–0.5 mg/l NAA and 1.0–2.0 mg/l BA (induction medium) before transfer to MS medium supplemented with 0.1 mg/l IAA and 2.0 mg/l BA (embryo development medium). The initial exposure to low levels of NAA coincident with high levels of BA in the induction medium was essential for embryogenic induction. Maximum embryogenesis (43.3%) was obtained with 0.2 mg/l NAA and 2.0 mg/l BA, and at least 14 days on induction medium were required prior to transfer to the embryo development medium. The conversion frequency of cotyledonary embryos was 53.3% upon culture on MS medium containing 0.1 mg/l ABA for 7 days followed by transfer to MS medium supplemented with 0.1 mg/l IBA and 0.2 mg/l BA. Following conversion, the regenerated plantlets were transferred to soil and showed normal morphological characteristics.Abbreviations MS Murashige and Skoog (1962) medium - 2,4-D 2,4-dichlorophenoxyacetic acid - NAA 1-naphthaleneacetic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid - BA 6-benzylaminopurine - ABA abscisic acid