李砧木Marianna离体叶片再生体系优化研究

以李砧木‘Marianna’试管苗新梢顶端第1片叶为外植体,研究激素组合、基本培养基种类及外植体类型等对不定芽再生的影响。结果表明:1/2 MS基本培养基和WPM培养基再生率显著高于MS和SH培养基;叶片附带叶柄的外植体再生率和再生不定芽数显著高于叶柄和切除叶柄的叶片外植体;最佳再生培养基为1/2MS+2.0mg/L TDZ+0.1 mg/L IBA+0.25%琼脂+3.0%蔗糖,最高再生率和再生不定芽数分别为81.7%和7.46±1.38个;最佳生根培养基为1/2MS+0.5~1.0 mg/L IBA,能获得96.7%生根率、较高的生根数和根长。     

Successful propagation in vitro of apple rootstock MM106 and influence of phloroglucinol.

Successful in vitro propagation of clonal apple rootstock MM106 was achieved by culturing axillary buds on MS basal medium with BAP (1 mg/L), GA3 (0.5 mg/L) and IBA (0.1 mg/L). Use of liquid medium (LM) in initial cultures reduced phenol exudation to a greater extent and gave maximum sprouting percentage when transferred to solid MS medium. Phloroglucinol (PG) did not enhance sprouting of buds but increased the rate of multiplication when added in the medium. Maximum number of shoots were obtained when MS medium was supplemented with BAP (0.5 mg/L), GA3 (1 mg/L), IBA (0.1 mg/L) and PG (100 mg/L). For rooting, in vitro regenerated shoots were placed in IBA (30 mg/L) for 3 hr and transferred to solidified auxin free medium. Rooting was recorded in about 80% of shoots. Inclusion of PG in rooting medium was not beneficial but shoot cultures grown in its presence gave higher rooting percentage. Rooted plantlets showed about 70% survival rate in potting mixture of sand:soil:perlite (1:1:1).     

Tissue culture and plant regeneration of Helianthemum songaricum Schrenk]

The cotyledonary segments of sterile seedlings of Helianthemum Songaricum Schrenk were cultured on different media containing different phytohormons. It was found that the calli could be induced efficiently on MS basal medium supplemented with 10.0 mg/L NAA and 0.2 mg/L 6-BA. When calli were transferred on MS medium with 2.0 mg/L 6-BA and 0.2 mg/L NAA, shoots were produced. The frequency of shoot differentiation reached about 85%. The regenerated shoots were rooted on 1/2MS medium added with 0.5 mg/L NAA. The rooting rate was about 76%. Regenerated plantlets were successfully transplanted in soil, with a success rate of 67%.     

Micropropagation of sweet orange, Citrus sinensis Osbeck. for the development of nucellar seedlings

Protocol for micropropagation of elite plants of sweet orange (Citrus sinensis) through nucellar embryo culture has been standardized. Three to four nucellar embryos and a zygotic embryo could be excised from a single mature seed and successfully generated as healthy plants in basal MS medium. MS medium supplemented with NAA (1 mg/L) or 2, 4.D (1 mg/L) promoted callus development in both nucellar and zygotic embryos. GA3 (1 mg/L) enriched medium induced plantlets initiation but their growth was very poor. No significant differences were observed between initial growth patterns of nucellar and zygotic seedlings developing from the same ovule. Five to six shoots were obtained from collar region of both category of embryos in MS medium supplemented with BAP (1 mg/L) within 60 days of inoculation. The number of plantlets were almost doubled after their transfer in the same medium and culture for another 30 days. Higher doses of BAP resulted in initiation of callus directly from the embryos. The regenerated shoots (2-3 cm) could be rooted in MS medium supplemented with either only NAA (0.75 mg/L) or NAA (0.50 mg/L) and IBA (2.0 mg/L). A number of plantlets could be obtained from a nucellar embryo grown shoot within a limited time period.     

白花蛇舌草叶片离体培养及试管无性系的建立

以白花蛇舌草叶片为材料,建立了白花蛇舌草叶片高效不定芽发生、植株再生体系。研究了不同激素及其组合对外植体不定芽发生的效应。结果显示,在MS基本培养基中单纯添加6BA,当6BA浓度为0.1mg/L和0.5mg/L时,不能诱导离体叶片发生不定芽,6BA浓度在1.0～5.0mg/L范围内,诱导率随BA浓度升高而增加,适宜浓度为3.0mg/L;当6BA与NAA配合使用时,其诱导率随着培养基中6BA与NAA的相对比值的提高而提高;以MS+BA3mg/L+NAA0.01mg/L作为继代培养基,建立起白花蛇舌草高效、稳定的试管无性系,为白花蛇舌草遗传转化研究奠定了基础。     

半日花的组织培养及植株再生(简报)

半日花(Helianthemum Songaricum Schrenk)是半日花科半日花属落叶小灌木,分布于新疆、甘肃和内蒙古等荒漠地区海拔1000米至1300米山地。为亚洲中部荒漠特有种和古地中海植物区系孑遗种,是生长了7000万年的远古植物。比具有“活化石”之称的水杉和银杏还要早几千万年。植物学家将它及与之同龄的四合木、沙冬青和革苞菊等十几种植物统称为“古老残遗濒危植物”。故而在研究亚洲中部荒漠,特别是研究我国荒漠植物区系的起源以及与地中海     

In vitro adventitious shoot formation from leaf cultures of Clerodendrum inerme (L) Gaertn.

In vitro adventitious shoots (about 28) of Clerodendrum inerme were regenerated from leaf segments on MS medium containing BA (4 mg/L). These shoots developed directly from the leaf explants without callusing after 5 weeks. Leaf explant when cultured in MS medium containing BA (2 mg/L) and NAA (0.5 mg/L) developed compact callus that became nodular and regenerated shoots (about 50) after 5 weeks. The in vitro developed shoots were rooted in MS medium supplemented with IAA (2 mg/L). The hardened plantlets were successfully established in the field with 90% survival.     

盾叶薯蓣类原球茎的离体诱导及快繁体系的建立

为解决盾叶薯蓣离体培养中试管苗移栽困难的难题,以盾叶薯蓣带腋芽的茎段为外植体,借助正交试验设计方法,离体诱导出类原球茎并建立了类原球茎微繁殖技术体系。结果表明:以带腋芽茎段为外植体诱导致密愈伤组织的适宜培养基为MS+6-BA2.0mg/L+NAA0.4mg/L+KT0.6mg/L+蔗糖3%;类原球茎诱导和增殖培养基为:MS+6-BA4.0mg/L+KT1.0mg/L+蔗糖6%;类原球茎生根培养基:1/2MS+NAA 0.3mg/L+IAA 0.8mg/L+活性炭0.3%+蔗糖1.5%。经该途径诱导得到的生根类原球茎植株经炼苗后移栽的成活率可达到90%以上。     

Regeneration of grape (Vitis labruscana cv. Kyoho) by shoot-tip culture

We investigated the optimal levels of growth regulators, culture media, and pH on callus growth and organogenesis of in-vitro cultured ‘Kyoho’ grapes. Calli were induced by culturing leaf blades on an MS basal medium supplemented with 1 mg/IL BA and 0.01 mg/L 2,4-D. In addition, calli originating from the exocarp and mesocarp of grape fruits devel-oped on MS media supplemented with 0.1 mg/L IAA, NAA, or 2,4-D, or with 0.2 mg/L BA. In testing the potential for plant regeneration from shoot tips on various media, we found that the Nitsch medium, with I mg/L BA, was optimal for caulogenesis. The type of shoot development depended on the pH of the medium, with vigorous multiple-shoot devel-opment occurring at pH 6.0, and single shoots forming at pH 5.0. Finally, we were able to obtain rooted seedlings from the regenerated shoots that had been cultured on 1/4-strength Nitsch medium supplemented with 0.03 mg/L NAA.     

An efficient in vitro regeneration system of fieldpea (Pisum sativum L.) via. shoot organogenesis

In-vitro regeneration in fieldpea was achieved from immature embryonic axes and cotyledonary node explants of six genotypes on modified MS media supplemented with different concentration of plant growth regulators, 6-Benzylamino purine (BAP) and Naphthalene acetic acid (NAA). The best regeneration response, leading to multiple shoot formation efficiency (22.34 shoots/explant) was observed in the medium supplemented with 1.0 mg/L BAP and 0.2 mg/L NAA and best frequency (67.55?±?4.74) was achieved on medium containing 2.0 mg/L BAP and 0.4 mg/L NAA. The shoots were subcultured on a medium supplemented with a combination of 1.0 mg/L GA₃, 2.0 mg/L BAP and 0.4 mg/L NAA, which resulted in elongation of 85 % of shoots. Rooting attempted from the elongated shoots, on half strength MS medium and supplemented with three different auxins IBA, IAA and NAA separately, exhibited similar results. Alternatively, micro-grafting of in vitro regenerated shoots onto pre-germinated root stocks raised in green house facility was attempted with high success rate (75 %). The grafted plants could be successfully hardened, fertigated with Hoagland solution and distilled water in a ratio of (1:10) for acclimatization and further development. All the genotypes tested, produced multiple shoots that could be established to mature fertile plant, hence, the medium combinations used were found to be genotype neutral.     

In vitro induction of multiple shoots and plant regeneration in cotton (Gossypium hirsutum L.)

Induction of multiple shoots in cotton (Gossypium hirsutum L. cv. Anjali-LRK 516) has been achieved with cotyledonary nodes devoid of cotyledons and apical meristems. Explants from 35-day-old seedlings yielded the maximum number of shoots (4.7 shoots/explant) using Murashige and Skoog (MS) basal medium supplemented with 6-benzylaminopurine and kinetin (2.5 mg/1 each). Explants from 35-day-old seedlings raised in glass bottles produced a higher number of multiple shoots (8.3 shoots/explant) than those grown in glass tubes and cultured on the same shoot induction medium. Elongation of multiple shoots was obtained on liquid or agar MS basal medium without phytohormones. In vitro shoots were rooted on half-strength agar-solidified MS basal medium or with 0.05 or 0.1 mg/1 naphthaleneacetic acid. Hardening and survival of tissue culture plantlets was 95% under greenhouse conditions.Abbreviations BAP 6-Benzylaminopurine - GA₃ Gibberellic acid - MS Murashige and Skoog medium - NAA -Napthaleneacetic acid     

Micropropagation of Ulmus pumila L. from mature trees

Summary Multiple shoots were induced from nodal segments of mature trees of Ulmus pumila L. on Murashige and Skoog (MS) medium supplemented with benzyladenine (BA). Further multiple shoots were obtained from nodal segments taken from in vitro proliferated shoots when cultured in MS medium containing 0.5 mg.l^–1 BA. Rooting of the shoots was achieved on half or full strength MS medium or in MS medium supplemented with 0.1 mg.l^–1 naphtaleneacetic acid (NAA). Rooted plantlets were able to resume independent growth after a short period of acclimatization.     

Effect of TDZ on plant regeneration from mature seeds in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis>)

Pea (Pisum sativum L. cv. Espace) seeds directly cultured on thidiazuron (TDZ)-containing medium formed high numbers of shoots. The number of shoots per seedling depended on the concentration and duration of the TDZ treatment. The best treatment was 12-wk incubation on MS medium supplemented with 4 mg/l TDZ followed by 4-wk culture on MS medium supplemented with 0.5 mg/l benzylaminopurine (BA) and produced more than 400 shoots/seedling. Isolated shoots rooted at a high frequency on MS medium containing 2–3 mg/l indole-3-butyric acid and 2 mg/l α-naphthalene acetic acid. In addition to the formation of shoots, bud-containing tissues (BCT) were formed at the cotyledonary nodes, shoot nodes, tendrils, stipules, and internodes. The BCT from the cotyledonary nodes and the shoot nodes was maintained in its pure state on MS medium supplemented with 4 mg/l TDZ by repeated culture. Shoot development was accomplished when the BCT were left on MS medium supplemented with 4 mg/l TDZ without subculture prior to transfer onto MS medium supplemented with 0.5 mg/l BA.     

提高西瓜离体培养植株再生效率的研究

本文以“京欣1号”母本和“伊选”西瓜4天苗龄子叶为外植体,研究离体培养植株高频率再生体系。结果表明：“伊选”子叶远轴端外植体的再生频率仅为10％,子叶近轴端外植体在5mg／LBA 0．1mg／L IAA的激素组合下植株再生频率为100％,平均每个外植体的丛生芽数在所有组合中最多,为10.3个;“京欣1号”母本子叶近轴端外植体在2mg／LBA 0．5mg／L IAA激素组合下植株再生频率为100％,平均每个外植体的丛生芽数在所有组合中最多,达6．9个。本试验条件下,子叶近轴端外植体接种4天即分化出不定芽,至再生苗的移栽仅需40天,在MS 0．1mg／L NAA的生根培养基上的生根率为97．3％,移栽成活率达98．5％。     

Plant regeneration from axillary buds of a mature tree of Ficus religiosa

Dormant axillary buds of nodal explants collected from a mature (35-year-old) tree of Ficus religiosa L. sprouted on MS medium supplemented with 6-benzyladenine (BA, 5 mg l^–1) and indole-3-butyric acid (IBA, 0.2 mg l^–1 ) within 4 days. Multiple shoots were obtained when these explants were transferred to MS medium containing 1.5 mg l^–1 BA and 1.5 mg l^–1 adenine sulphate (ADS). These multiple shoots (1 cm) were allowed to elongate to a height of 4–5 cm by maintaining them on MS medium containing 2 mg l^–1 BA, 0.5 mg l^–1 1-naphthalene acetic acid (NAA), and activated charcoal (0.3% w/v). Nodal segments taken from these in vitro-proliferated shoots yielded multiple shoots when cultured on the multiple shoot-inducing MS medium mentioned above. Root induction in these shoots (4–5 cm in height) was achieved by transferring them onto MS medium supplemented with 2 mg l^–1 IBA and 0.1 mg l^–1 NAA for 1 week; upon transfer to half-strength MS basal medium these shoots exhibited root proliferation. These rooted plantlets were successfully established in soil after a short period of acclimatization. Received: 17 April 1997 / Revision received: 2 September 1997 / Accepted: 20 September 1997     

An improved method for micropropagation and regeneration of date palm (Phoenix dactylifera L.)

We developed a novel large-scale micropropagation pathway for date palm (Phoenix dactylifera L.) based on organogenesis. We obtained organogenic stems from shoot tip explants of the Moroccan date palm cultivar Najda, and investigated shoot proliferation from these organogenic stems in vitro on various media; Beauchesne medium (BM) and Murashige and Skoog medium (MS) at full-strength, half-strength, and one-third-strength, containing various concentrations (0, 0.25, 0.5, and 1 mg/L) of 2-naphthoxyacetic acid (NOAA) and kinetin. The optimal medium during the multiplication phase was half-strength Murashige and Skoog medium (MS/2) supplemented with 0.5 mg/L NOAA and 0.5 mg/L kinetin (23.5 morphologically superior shoots per explant, with low vitrification rates). For the shoot elongation phase, shoots were transferred to the same proliferation medium, or to MS or MS/2 media without plant growth regulators (PGRs). Shoots elongated rapidly and showed a high rate of root formation on media supplemented with PGRs. For example, on MS/2 medium containing 1 mg/L NOAA and 1 mg/L kinetin, the average shoot length was 15.1 cm, the average number of roots per shoot was 6.2, and their average length was 3.4 cm. On PGR-free media, shoots were shorter with wider and greener leaves, and had fewer roots. The plantlets were transferred to a greenhouse for acclimation. The survival rate after 2 months was related to the medium used during the elongation phase; >90 % of shoots that were cultured on PGR-free media survived, while there was a poor survival rate of shoots that had been cultured on media containing PGRs.     

Production of biomass and flavonoid of Gynura procumbens (Lour.) Merr shoots culture in temporary immersion system

Gynura procumbens (Lour.) Merris one of medicinal plant which was carried out used as antioxidant, anticancer, anti-inflammatory, hepatoprotective, and antimicrobial. Many strategies were used to increase the production of biomass and valuable compounds. This study was to investigate the variation effect of growth regulators and immersion frequency on production of biomass and flavonoid contained of G. procumbens shoots culture in temporary immersion bioreactor. Stem nodes were used as an explants and induction of shoots were done in solid MS medium supplemented with many kinds of growth regulator. The best treatments were used to produce biomass and flavonoid compounds in temporary immersion bioreactor; there are combination of IAA 2?mg/L and BA 4, 6, 8?mg/L and immersion frequency (5 min each 3?h; 15?min each 12?h). Results showed that the growths of G. procumbens shoots in solid MS medium were influenced by supplementation of growth regulators. MS medium supplemented with single cytokinine (6 mg/L kinetin) and combination of auxin (IAA) and cytokinine (BA) caused increasing of shoots growth. Production of biomass of G. procumbens in temporary immersion bioreactor was achieved in long immersion interval (12?h) and highest flavonoid production was obtained in combination treatment of immersion frequency 15?min each 12?h and MS medium supplemented with IAA 2?mg/L, BA 8?mg/L.     

中国木薯栽培种通过器官发生再生植株研究

本文研究了中国木薯栽培种四种外植体通过器官发生再生植株的条件。结果表明:在MS附加0.05mg/L TIBA,1mg/L BA的培养基上“NZ 188”初步的萌发胚状体“切头”后切口处可直接产生丛芽,出芽率为43%。“SC201”胚状体子叶块在MS附加0.5 mg/L NAA,0.5mg/L BA的培养基上可直接出芽,出芽率为42%,在MS附加0.5mg/L IBA,1.5mg/L BA培养基上·出芽率为31%,AgNO_3和ABA单独使用或配合使用均不利于芽的再生。“NZ188”胚状体下胚轴在MS附加0.5mg/LNAA,0.5mg/L BA的培养基上形成的愈伤组织转入MS附加1mg/L NAA,2mg/L BA的培养基上,3周后大多数愈伤组织有绿点出现、仅4.4%外植体分化出芽。“HZ188”无菌苗茎段接种在MS附加0.05mg/L TIBA,2mg/LBA的固体培养基上,2周后形成大量愈伤组织,4周后仅见一块愈伤组织分化出芽。     

In vitro culture of Cuminum cyminum regeneration of flowering shoots from calli of hypocotyl and leaf explants

Calli from hypocotyl explant of Cuminum cyminum L. (Cumin) grew rapidly on Gamborg's B₅ basal medium with the following supplements, (i) 0.5 mg/l — 2,4-D (ii) 4 mg/l — NAA plus 2 mg/l — Kinetin and (iii) 0.2 mg/l — NAA plus 0.2 mg/l — BAP, whereas calli from leaf explant in these media grew slowly. Hypocotyl and leaf calli produced roots when transferred to basal medium only and shoots in basal medium with 0.5 mg/l NAA and 0.1 mg/l BAP. Ninety percent of the shoots produced roots when they were transferred to half strength MS inorganic salts supplemented with 0.5 mg/l each of IBA and NAA.Fifty to sixty percent of rootless as well as rooted shoots produced terminal umbellate flowers on this medium.     

草木樨状黄芪高频离体再生体系的建立

以草木樨状黄芪无菌苗茎切段为材料,在含１－２mg/L2,4-D和０－０．５mg/L６ＢＡ的MS培养基上培养获得大量愈伤组织,愈伤组织诱导率在９５％以上,愈伤组织在附加０．２mg/LＫＴ,１mg/L６ＢＡ,０或０．５mg/LＮＡＡ,５００mg/LＣＨ 和２００mg/L ＹＥ的ＭＳ培养基上诱导丛生芽,并进而发育成苗。苗的分化频率为１００％。分化苗或其茎的切段在不国源植物激素的１／２ＭＳ培养基上可出现根的分化,分化频率达９０％以上,再生植株经炼苗后移栽成活率达８０％以上。