植物激素对空心泡叶片不定芽形成的影响

空心泡是蔷薇科悬钩子属植物,属小果类果树,其聚合果肉质多浆,芳香,果味酸甜。空心泡目前尚处于野生状态,是一种很有利用前景的植物。有关悬钩子属植物组织培养已有许多报道,所有这些工作都     

In vitro shoot regeneration from cotyledons of immature ovules ofImpatiens platypetala Lindl.

Summary An efficient and reproducible protocol has been developed for in vitro shoot regeneration from cotyledonary explants derived by germinating immature ovules ofImpatiens platypetala Lindl. ‘TR6-27-2’. Cotyledonary explants were cultured on a modified Murashige and Skoog (MS) agar-solidified medium containing 7.5g · liter^−1 sucrose, 22.2µ M N⁶-benzyladenine (BA), and 0.54µM α-naphthaleneacetic acid (NAA). The induction of organogenic tissues occurred after 6 to 8 wk in culture. Exogenous auxin and cytokinin were essential for the induction of organogenic tissues and survival of explants, and BA was most effective for the induction of organogenic tissues, compared with other cytokinins tested. The addition of glutamine (500 mg · liter^−1) was also important for growth of organogenic tissues after induction and for reducing explant death during culture. The induction of organogenic tissue was also influenced by the type of cotyledon cultured and the age of the donor seedlings. On average, eight shoots per explant were induced from organogenic tissues larger than 0.5 cm in diameter 6 to 8 wk after transfer to a modified MS agar-solidified medium without NAA and BA reduced to 4.44µM. Shoots longer than 0.5 cm in length were successfully rooted 2 to 4 wk after transfer to a basal MS medium containing 30g · liter^−1 sucrose.     

In vitro shoot bud differentiation and plantlet regeneration in Celastrus paniculatus Willd

Multiple shoot buds could be induced directly from internode explants of Celastrus paniculatus inoculated on Murashige and Skoog’s (MS) medium containing different growth regulators. The best response was obtained when 4.44 μM 6-benzylaminopurine (BAP) was incorporated in the medium. Incorporation of indole-3-acetic acid (IAA) and α-naphthalene acetic acid (NAA) did not improve response, rather promoted callusing. Adventitious shoot buds could be multiplied and elongated on MS medium containing 2.22 μM BAP. Rooting of shoots (80 %) was obtained when their bases were dipped in pre-autoclaved indole-3-butyric acid (IBA) solution (2.45 mM) for 10 min followed by their implantation on medium containing 1/4 MS salts, 1.0 % sucrose and 0.6 % agar. Out of 500 plantlets subjected to hardening, 410 were successfully hardened under greenhouse conditions. Twenty plants were established in field while remaining of them were transferred to nursery conditions without any mortality.     

In vitro adventitious shoot regeneration from leaves of Prunus spp.

A high level of adventitious shoot regeneration was obtained from proliferating shoots in vitro for a range of Prunus spp. There was a significant variability in clone response to a range of adventitious shoot regeneration treatments. Treatment of apricot clone H.152 with Quoirin macroelements (C.R. Rech., Stu. Cult. Fruit. Maraîchères Gemblaux (1977) 93–117), and both apricot clone H.146 and hybrid plum clone P.1869 with half-strength Murashige and Skoog medium, consistently induced regeneration. Thidiazuron (TDZ) alone, or in combination with naptthaleneacetic acid (NAA), was most effective in stimulating adventitious shoot production, the optimum concentration being clone-dependent. Addition of silver nitrate (AgNO₃) to regeneration media enhanced regeneration by 10–40% and reduced the variability between experiments. Regeneration with AgNO₃ was obtained also for three other plum clones belonging to the P. marianna, P. domestica and P. insititia species.     

Adventitious shoot formation from leaf explants of Rhododendron

A protocol for high frequency adventitious shoot regeneration adventitious shoot regeneration from leaf explants of Rhododendron spp. has been developed. The highest percentage of regeneration and the greatest number of shoots were obtained when leaf explants were cultured on Anderson's medium containing 4.9 M IBA and 73.8 M 2iP. Genotypic variation was observed for adventitious shoot regeneration potential among the seven cultivars tested. Regeneration frequencies ranged from 0 to 96%. Lodestar had the highest rate of regeneration after 3 months of culture with 96% shoot regeneration and an average of 14 shoots per explant. Regenerated shoots were rooted in soil in about 2 months. This protocol should be useful in applying gene transfer techniques to Rhododendron improvement.Abbreviations IAA 1-H-indole-3-acetic acid - NAA 1-naphthaleneacetic acid - IBA 1-H-indole-3-butyric acid - 2,4-d 2,4-dichlorophenoxyacetic acid - BA 6-benzyladenine - 2iP N-(3-methyl-2-butenyl)-1-H-purine-6-amine     

In vitro plant regeneration of Aristolochia indica through axillary shoot multiplication and organogenesis

Protocols for in vitro plant regeneration via axillary and adventitious shoot regeneration were established in an important medicinal plant, Aristolochia indica L. (Aristolochiaceae). Basal Murashige and Skoog's (MS) medium supplemented with 0.54 μM α-naphthaleneacetic acid (NAA) and 13.31 μM benzyladenine (BA) induced the maximum number of shoots (45-50) from shoot tip and nodal segment cultures. Phenolic accumulation in leaf and internodal stem derived callus cultured in MS medium containing NAA or 2,4-dichlorophenoxyacetic acid and BA or kinetin was controlled by the addition of 1.0 mg l^-1 phloroglucinol (PG) to the callus induction medium. Basal medium supplemented with 2.69 μM NAA, 13.31 μM BA and 1.0 mg l^-1 PG induced the best results in terms of shoot bud regeneration from leaf derived callus. Direct de novo development of shoots from leaf segments was achieved using 13.31 μM BA along with 50 mg l^-1 activated charcoal. The microshoots were rooted in White's medium supplemented with 2.46 μM indolebutyric acid. More than 85% of rooted plants survived in the soil. This revised version was published online in June 2006 with corrections to the Cover Date.     

美国黄松成熟胚的离体培养与不定芽的形成

以美国黄松成熟胚为外植体进行离体培养诱导不定芽。正交试验和统计分析结果表明基本培养基的种类对外植体不定芽的诱导起主要作用。在GD培养基上附加0.5mg/L的6－BA,外植体不定芽的诱导率达55％,平均增值率为6,最大增值率达10。NAA不利于外植体不定芽的诱导,培养基中加入适量的添生炭有利于不定芽的形成和生长。     

A liquid cytokinin pulse induces adventitious shoot formation from Douglas-fir cotyledons

Summary The effects of high-concentration, 2-h liquid pulses of N⁶-benzylaminopurine (BA) and thidiazuron (TD) on adventitious bud and shoot formation were tested in cotyledons of Douglas-fir (Pseudotsuga menziesii). Seedling age proved important; on average, cotyledons from the youngest seedlings formed 10-fold more buds than cotyledons from the oldest seedlings. Optimal cytokinin concentrations for the youngest cotyledons were 400 and 800 M BA, and 100 and 200 M TD. Shoots developed best from buds induced with 300, 400, and 800 M BA. Four gelling agents were tested; BRL agarose yielded more than three times the number of buds, and Gelrite nearly twice the number of buds, as either Sigma agar or Difco Bacto-Agar. One of the best treatments (400 M BA, agarose) yielded more cotyledons with buds, and more buds per cotyledon, than when cytokinins were incorporated into the growth medium.Abbreviations BA N⁶-benzylaminopurine - TD thidiazuron - SH Schenk and Hildebrandt (1972) - NAA 1-naphthaleneacetic acid Paper 2691 of the Forest Research Laboratory, Oregon State University, Corvallis. Mention of trade names or commercial products does not constitute endorsement or recommendation for use by the authors or Oregon State University.     

Adventitious shoot formation from embryonic explants of red pine (Pinus resinosa)

Adventitious shoots were induced from excised embryos of Pinus resinosa Ait, on half-strength Le-Poivre (LP) medium containing 1–70 μ M N⁶-benzyladenine (BA). At lower concentrations of BA, only 2–3 shoot primordia (from as many as 22 formed per embryo) developed into shoots when subcultured onto medium containing 0.5% activated charcoal. Concentrations of 10 to 70 μ M of BA produced significantly higher numbers of shoot primordia and most of them developed into shoots. Ten to 17 day culture on medium containing 10–25 μ M BA proved optimal for maximum adventitious shoot production. Less than three days of incubation on the cytokinin medium did not stimulate the formation of adventitious shoots. Twenty-four day culture on the same medium produced several shoots, but most of them failed to develop normally and formed callus. Coconut milk (0.1–5% v/v) inhibited adventitious shoot formation. Using optimal conditions, seeds from 11 open-pollinated selected trees were compared to test for genetic differences in the potential to produce adventitious shoots from embryos. No significant differences were observed with regard to the shoots produced per embryo among the different seed collections. More than 200 plants produced through this technique were tested for variation in several isozymes by electrophoresis. No variations were observed.     

Plantlet multiplication from white pine (Pinus strobus L.) embryos in vitro: bud induction and rooting

White pine embryos were grown on 4 different media with 6 different benzyladenine (BA) concentrations. Maximum adventitious shoot initiation and growth were obtained on a modified Lepoivre medium with 20 M BA. Modified Schenk and Hildebrandt, Murashige and Skoog, and Gresshoff and Doy media were also tested. Shoot elongation was achieved on half-strength basal medium lacking growth regulators. Three rooting experiments involving indole-3-butyric acid (IBA), sucrose concentration, shoot orientation, IBA pulse length, and light or dark were carried out. Treatment of shoots in an upright position with 50 M IBA for eight days followed by culture in a medium with 3% sucrose in the light produced the most rooting (50% at 3 months). Rooted shoots were transplanted to the greenhouse for further growth.     

Somatic embryogenesis in <Emphasis Type="Italic">Pinus nigra</Emphasis>: embryogenic tissue initiation,maturation and regeneration ability of established cell lines

The effect of plant growth regulators (PGR), 6-benzyladenine (BA), 2,4-dichlorophenoxyacetic acid (2,4-D), α-naphthaleneacetic acid (NAA) and sugars (sucrose, maltose, glucose, fructose) on the initiation of somatic embryogenesis of Pinus nigra Arn. was investigated. Megagametophytes containing immature zygotic embryos have been used as explants. The experiments were done in the years 2000 and 2001. Higher initiation frequencies were obtained in 2001 when the zygotic embryos showed uniformity, being in the precotyledonary stage of development. Embryogenic tissue initiation occurred on all the media tested, including PGR-free medium. Relatively high initiation frequencies were obtained on media containing either NAA (9.09 %) or 2,4-D (7.14 %) alone. Somatic embryos were present as bipolar structures and showed differences in morphological features among cell lines. Plantlet regeneration occurred in cell lines containing bipolar somatic embryos composed of compact meristematic embryo “head” and suspensor organized into bundles.We highly appreciate the financial support from VEGA, Slovak Grant Agency, project No. 2/2089/22.     

In vitro adventitious root induction in Antirrhinum majus L.

A broadly applicable method for the successful induction of root systems in a number of cultivars of A. majus has been determined. This involves a double filter-paper bridge with a liquid medium for root induction and allows the transfer of culture-grown plantlets to a glasshouse environment with minimal disturbance to the plant as a whole. 100% survival of transferred plantlets has been achieved with the inclusion of a few simple precautions upon shoot transfer and during the initial stages of plant establishment in vivo.     

Optimized somatic embryogenesis in Pinus strobus L.

Summary Somatic embryogenesis (SE) initiation in Pinus strobus was optimized by the manipulation of plant growth regulator (PGR) concentrations in the culture medium. Modified Litvay medium (MLV) of Litvay et al. (1985) supplemented with lower than routinely used PGR concentration increased initiation of established embryogenic cultures from approximately 20 to 53%. The original developmental stage of zygotic embryos had a pronounced effect on the SE response. The optimum stage was the pre- to shortly post-cleavage stage. A substantial genetic influence on initiation of SE was indicated by a significant variance component due to families. Genotype X collection date and genotype X media interactions had large effects on initiation of SE. The PGR levels in the culture medium prior to maturation had a significant effect on subsequent production of mature somatic embryos. Embryogenic tissue initiated and proliferated on medium with a low level of PGR consistently produced a high number of somatic embryos, indicating that optimized initiation protocol also enhanced somatic embryo production. Somatic embryos of 93 embryogenic lines (representing five families) that were initiated on media with different PGR concentrations were converted to plants at an overall frequency of 76%, and grown in the greenhouse. With these improved protocols, application of P. strobus SE in commercial clonal forestry is feasible as an alternative to traditional breeding and reforestation.     

In vitro shoot organogenesis from excised immature cotyledons and microcuttings production in Stone Pine

Adventitious buds were induced on isolated immature cotyledons of Pinus pinea L. in the presence of benzyladenine (BA). The response to different BA concentrations also depended upon the culture medium used (modified MS, SH and GD). A wide range of BA concentrations (5, 25 or 50 M) can be applied to the GD and SH media, which are the media with the lower nitrogen content, without damaging effects. In the MS medium, which has the highest nitrogen concentration, the range of BA that can be applied was narrower and the highest BA concentration was lethal. The addition of indolebutyric acid (0.05, 0.25 or 0.5 M) to the induction medium, decreased the response of cotyledons. The increase in the concentration of sucrose from 3% to 5% did not increase the number of responding cotyledons. The addition of activated charcoal (0.5 and 3 g l^-1) or indolebutyric acid (1.5 or 3 M) did not speed up the elongation of explants. Elongation of the buds produced shoots with two different phenotypes, each phenotype having a different multiplication rate.Abbreviations BA benzyladenine - GD Gresshoff & Doy medium - IBA indolebutyric acid - MS Murashige & Skoog medium - SH Schenk & Hildebrandt medium     

Comparative adventitious shoot induction in kentucky coffeetree root and petiole explants treated with thidiazuron and benzylaminopurine

Summary Adventitious shoot induction and elongation was compared between root and petiole explants of Kentucky coffeetree (Gymnocladus dioicus L.) explants treated with a factorial combination of benzylaminopurine (BA) and thidiazuron (TDZ). Petiole explants initiated more adventitious shoots compared to root explants. Up to 83% of petiole explants initiated shoots compared to 67% of root explants. Maximal shoot induction was approximately 12 or five shoots per responding explant for petiole and root explants, respectively. For both explant types, TDZ was more effective than BA for shoot induction. There was an interaction between BA and TDZ on shoot induction in petiole explants, with the greatest percentage of explants forming shoots and the highest number of shoots initiated on the combination of 0.5 μM TDZ plus 10μM BA and 1.0μM TDZ plus 5 or 10 μM BA. In contrast, increasing concentrations of BA inhibited shoot initiation in root explants with and without TDZ. While BA inhibited shoot initiation in root explants, it promoted shoot initiation in petiole explants. In contrast, TDZ was equally effective at inducing shoots in root and petiole explants. This suggests that root and petiole explants of Kentucky coffeetree could be a useful model system for studying the differences, in apparent mode of action between TDZ and BA on adventitious shoot initiation.     

Gelling agents, silver nitrate and sequestrene iron influence adventitious shoot and callus formation from Rubus leaves

Summary Blackberry (Rubus spp.) cultivars Chester Thornless, Kotata, Marion, and Navaho were evaluated for morphogenic potential using the top one to four leaves from in vitro-grown shoots. In each experiment adventitious shoots were regenerated via organogenesis without an intervening callus phase. Rooting was spontaneous when shoots were transferred to a medium without growth regulators or with indole-3-butyric acid (IBA). Three gelling treatments (agar, Gelrite, and a combination) calibrated for equal gel firmness [110 g/(1.1 cm)² π] did not affect explant regeneration or the number of shoots per explant, but did affect callus production. Significantly more callus (P≤0.05) was induced on regeneration medium (RM) with 10 μM N⁶-benzyladenine (BA), 0.5 μM IBA, and 0.19% Gelrite, than on medium containing either 0.71% agar, or the 0.35% agar/0.11% Gelrite combination. Explants on RM with 5 μM IBA produced significantly more callus, but fewer shoots, compared to zero or 0.5 μM IBA treatments for all gel treatments. Adding 200 mg l⁻¹ sequestrene iron [sodium ferric ethylene diamine di-(O-hydroxyphenylacetate)] at the first transfer onto RM induced more shoots per explant than the control, but did not influence the amount of callus produced. Sequestrene iron in the second transfer on RM significantly increased the regeneration (caulogenesis) frequency from 30 to 40% for ‘Marion’ and from 23 to 43% for ‘Kotata’. Silver nitrate significantly reduced callus production for all treatments, but did not affect the frequency of caulogenesis or the number of shoots per explant. Part of a thesis submitted by C. W. V. Tsao in partial fulfillment of the requirements for the MS degree. The use of trade, names in this publication does not imply endorsement by the US Department of Agriculture or Oregon State University.     

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.     

In vitro adventitious shoot formation on mature-phase leaves and petioles of Liquidambar styraciflua L.

Adventitious shoots were induced to form on leaves and petiole segments of mature-phase Liquidambar styraciflua L. Shoot organogenesis occurred directly, without the formation of a distinct callus stage, and well-defined shoots were visible in 6–9 weeks. Prolific shoot production was supported by Woody Plant Medium supplemented with relatively high levels of benzyladenine (2.5 mg/l). Changes in benzyladenine concentration and the addition of 0.1 mg/l naphthaleneacetic acid to the medium altered the relative abundance of shoots on various parts of a leaf. Shoot formation occurred most frequently at or near breaks in major vasculature. Wounding of the leaves by slashing across the lamina and vasculature significantly increased the total number of shoots formed per explant and also altered the pattern of organogenesis. Differences in organogenic response were seen between the cultivars ‘Moraine’ and ‘Variegata’. Shoots derived from leaves were easily rooted and acclimated to greenhouse conditions. Three new variegation types arose in vitro as a result of adventitious shoot formation on ‘Variegata’ leaves.     

Thidiazuron promotes adventitious shoot regeneration from pothos (Epipremnum aureum) leaf and petiole explants

Summary Regeneration of adventitious shoots of pothos (Epipremnum aureum Linden and Andre) ‘Jade’ was obtained using leaf and petiole explants preprated from shoot tips of 3-yr-old greenhouse-grown plants. Explants were cultured on Murashige and Skoog (MS) basal medium supplemented with thidiazuron (TDZ), 6-(4-hydroxy-3-methy-trans-2-butenyl-amino)purine (zeatin) or N-isopentenylaminopurine (2iP) individually with α-naphthaleneacetic acid (NAA) in 18 combinations. Callus was initiated from cut surface and along the midrib or major vein of leaf sections. Shoot regeneration from leaf and petoole explants occurred in 30d on medium containing 1, 5 or 10μM TDZ with 0.5 or 1.0μM NAA except petioles on medium with 10 μM TDZ and 1.0 μM NAA where regeneration failed. More time (50d) was needed for shoot regeneration when explants were cultured on medium containing either 2iP or zeatin with NAA. Regeneration frequencies were up to 20% and 50% for leaf and petiole explants, respectively. Shoot numbers per responding explant attained 30 for leaf and petiole explants on medium containing TDZ but only one to four on medium containing either 2iP or zeatin. These results indicate that TDZ is a more effective cytokinin for in vitro regeneration of pothos than either zeatin or 2iP.. Shoots elongated readily and rooted well on MS basal medium, without plant growth regulators. Plantlets acclimatized rapidly and grew vigorously in the greenhouse after transfer to pots containing a commerecial potting medium.     

Effect of various bud induction treatments on elongation and rooting of adventitious shoots of Canary Island pine (Pinus canariensis)

Three-day-old cotyledonary explants of Pinus canariensis were subjected to 30 induction treatments using half-strength Bornman's medium containing various combinations of N⁶- benzyladenine, zeatin, kinetin and 2-isopentenyl-adenine. The highest numbers of buds were obtained with 10 M 6-benzyladenine, but both kinetin and zeatin influenced shoot elongation. Shoots were maintained on half-strength Schenk and Hildebrandt medium with 2% sucrose and 0.05% activated charcoal. For rooting, shoots were pulsed for 4 h in a 100 M indole-3-butyric acid aqueous solution (pH 4.2–4.5), and planted in peat:vermiculite:perlite (1:1:1). After 8 weeks, the numbers of rooted shoots were similar for most treatments. Therefore, the bud induction treatments did not significantly influence rooting of adventitious shoots of Canary Island pine.