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1.
Randall P. Niedz Terence J. Evens 《In vitro cellular & developmental biology. Plant》2011,47(6):682-694
The objective of this study was to improve shoot regeneration from grapefruit. Because many commercially grown citrus types
are apomictic, important in vitro applications such as Agrobacterium-mediated transformation commonly use epicotyl explants from in vitro seedlings; thus, adequate adventitious shoot production is an important prerequisite for efficient use of these applications.
Eight plant growth regulators were studied—six cytokinins (6-benzylaminopurine, kinetin, zeatin trans-isomer, 6-[γ,γ-dimethylallylamino] purine, zeatin riboside trans-isomer and meta-topolin) and two auxins (α-naphthalene acetic acid and indole-3-acetic acid). An iterative design strategy
was followed that included mixture and mixture-amount experimental designs suitable for resolving proportional and concentration
effects; in vitro effects of cytokinins and auxins are affected by both proportion and concentration. One-centimeter-long explants were excised
from the epicotyl of etiolated, in vitro-grown seedlings. Explants were placed onto experimental formulations and cultured in growth cabinets at 27°C over 6 wk, which
included 2 wk in the dark followed by 4 wk in the light. The results indicated that (1) 6-benzylaminopurine or zeatin riboside
were the most effective cytokinins for inducing shoot regeneration in citrus; (2) zeatin riboside singly or in combination
with indole-3-acetic acid resulted in the highest quality, the greatest number of explants with buds/shoots, and the greatest
shoot number; and (3) 6-benzylaminopurine and indole-3-acetic acid improved shoot regeneration vs. 6-benzylaminopurine at a considerably lesser cost than zeatin riboside and indole-3-acetic acid. 相似文献
2.
Root, hypocotyl and cotyledonary explants of niger (Guizotia abyssinica Cass) CV. Sahyadri were aseptically cultured on Murashige and Skoog's basal medium (MS) containing BAP and kinetin. Multiple shoot regeneration was induced from hypocotyl and cotyledonary explants while root explants produced only callus on MS medium supplemented with BAP. BAP (1 mg l-1) was optimum for shoot regeneration. Regenerated shoots were transferred to MS medium without auxins, with auxins and with increasing concentrations of sucrose for rooting. Complete plantlets were obtained in all cases; however, 0.5 mg l-1 NAA was the best for induction of roots. Ninety-seven per cent of the plantlets survived and completed their life cycle when transferred to natural conditions.Abbreviations BAP
6-benzylamino purine
- NAA
-naphthaleneacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid 相似文献
3.
For centuries Hypericum perforatum has been used in natural medicine. In the last decades, it has also attracted the attention of pharmaceutical industry due
to its promising anti-depressant properties. The important factor in pharmaceutical application of plant material is its stable
content of active compounds. Such stability requires standardized conditions of growth, e.g. an in vitro culture. Our aim was to establish a medium allowing for an effective regeneration of shoots from the standardized leaf explants in
in vitro conditions. Cultures of the leaf explants carried out in darkness, on Murashige and Skoog agar medium, supplemented
with auxins (2,4-dichlorophenoxyacetic acid, 2-metoxy-3,6-dichlorobenzoic acid, α-naphtaleneacetic acid, indole-3-acetic acid)
and cytokinins (kinetin, N6-(benzyl)adenine, thidiazuron) resulted in callus formation. The callus produced roots on media containing indole-3-acetic
acid or α-naphtaleneacetic acid alone. On media supplemented with auxins and cytokinins, indirect shoot organogenesis was
also observed. The most efficient shoot formation was observed with 2.85 μM of indole-3-acetic acid and 4.44 μM of benzyladenine.
Regenerated shoots were rooted on Murashige and Skoog without plant growth regulators medium or on a medium supplemented with
indole-3-acetic acid. From a single leaf explant (one fifth of the leaf) after a month of the culture, 35 regenerated shoots
were obtained (allowing for the formation of about 180 vegetative shoots per leaf). Successful multiplication of shoots from
a standardized explant makes it possible to obtain a great quantity of uniform plant material for biotechnological purposes. 相似文献
4.
The plant hormone auxin has been shown to be involved in lateral root development and application of auxins, indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA), increases the number of lateral roots in several plants. We found that the effects of two auxins on lateral root development in the indica rice (Oryza sativa L. cv. IR8) were totally different from each other depending on the application method. When the roots were incubated with an auxin solution, IAA inhibited lateral root development, while IBA was stimulatory. In contrast, when auxin was applied to the shoot, IAA promoted lateral root formation, while IBA did not. The transport of [3H]IAA from shoot to root occurred efficiently (% transported compared to supplied) but that of [3H]IBA did not, which is consistent with the stimulatory effect of IAA on lateral root production when applied to the shoot. The auxin action of IBA has been suggested to be due to its conversion to IAA. However, in rice IAA competitively inhibited the stimulatory effect of IBA on lateral root formation when they were applied to the incubation solution, suggesting that the stimulatory effect of IBA on lateral root development is not through its conversion to IAA. 相似文献
5.
David G. Davis Prudence A. Olson 《In vitro cellular & developmental biology. Plant》1993,29(3):97-101
Summary All parts of leafy spurge seedlings can be regenerated when isolated and placed onto B5 medium. One-centimeter isolated hypocotyl
segments were tested successfully for their usefulness as a bioassay system by comparing the response of auxins, herbicides,
and cytokinins. Indole-3-acetic acid (IAA) was the most effective auxin to stimulate root formation. IAA was effective whether
the hypocotyl segments remained on the same medium up to 60 days, or the segments were transferred to basal media after 2
or 5 days (pulse treatment). Pulse treatments with the other auxins resulted in stimulation of root formation; continuous
or 5-day pulses of higher concentrations of indole-3-butyric acid,α-naphthaleneacetic acid and especially 2,4-dichlorophenoxyacetic acid and picloram formed excessive callus instead of roots.
Picloram did not stimulate root formation, whether the treatment was continuous or pulse-treated. No roots formed with continuous
picloram at 0.1 mg/liter or greater, but transfer to basal media did result in root and shoot formation at about 50% of the
number formed on the controls. Lesser picloram concentrations had no effect. Shoots formed readily on untreated (control)
segments, but continuous treatment with all three cytokinins, kinetin, zeatin, and zeatin riboside, increased the numbers
of shoots about equally. Root formation was inhibited by the cytokinins at the higher concentrations (0.1 to 0.2 mg/liter).
With the exception of a 5-day pulse of 0.04 mg/liter IAA, the auxins did not stimulate shoot formation, but generally inhibited
shoot formation, even in pulse-treated cultures. 相似文献
6.
Giovanni Iapichino Marcello Airò 《In vitro cellular & developmental biology. Plant》2008,44(4):330-337
Multiple shoots were induced on stem segments of an 8-y-old plant of Metrosideros excelsa Sol ex Gaertn. “Parnel”. Axillary shoots produced on uncontaminated explants were excised, segmented, and recultured in the
same medium to increase the stock of shoot cultures. The Murashige and Skoog (MS) medium, augmented with different concentrations
of 2- isopenthenyladenine (2iP) and indole-3-acetic acid (IAA), either singly or in combinations, as potential medium for
shoot multiplication by nodal segments was tested. In the following experiment, equal molar concentrations of four cytokinins
[2iP, kinetin, zeatin, and N
6-benzyladenine (BA)] in combination with equal molar concentrations of three auxins [IAA, α-naphthaleneacetic acid (NAA),
and indole-3-butyric acid (IBA)] were tested for ability to induce axillary shoot development from single-node stem segments.
The highest rate of axillary shoot proliferation was induced on MS agar medium supplemented with 1.96μM 2iP and 1.14μM IAA
after 6 wk in culture. Different auxins (IAA, IBA, and NAA) were tested to determine the optimum conditions for in vitro rooting of microshoots. The best results were accomplished with IAA at 5.71μM (89% rooting) and with IBA at 2.85 or 5.71μM
(86% and 86% rooting, respectively). Seventy and 90 percent of the microshoots were rooted ex vitro in bottom-heated bench (22 ± 2°C) after 2 and 4 wk, respectively. In vitro and ex vitro rooted plantlets were successfully established in soil. 相似文献
7.
Aerobic Methylobacteria Are Capable of Synthesizing Auxins 总被引:1,自引:0,他引:1
Obligately and facultatively methylotrophic bacteria with different pathways of C1 metabolism were found to be able to produce auxins, particularly indole-3-acetic acid (IAA), in amounts of 3–100 g/ml. Indole-3-pyruvic acid and indole-3-acetamide were detected only in methylobacteria with the serine pathway of C1 metabolism (Methylobacterium mesophilicumand Aminobacter aminovorans).The production of auxins by methylobacteria was stimulated by the addition of L-tryptophan to the growth medium and was inhibited by ammonium ions. The methylobacteria under study lacked tryptophan decarboxylase and tryptophan side-chain oxidase. At the same time, they were found to contain several aminotransferases. IAA is presumably synthesized by methylobacteria through indole-3-pyruvic acid. 相似文献
8.
Summary Stem segments from apical shoot tips of Polygala myrtifolia were used as primary explants to establish in vitro cultures. Axillary shoots produced on non-contaminated explants were excised and recultured in the same medium to increase
the stock of shoot cultures. Equal molar concentrations of five cytokinins [2-isopentenyladenine, kinetin, zeatin, N
6-benzyladenine (BA), and adenine] were tested for ability to induce axillary shoot development from double-node stem segments.
The highest rate of axillary shoot proliferation was induced on Murashige and Skoog agar medium supplemented with 1.8 μM BA. Seven indole-3-acetic acid (IAA) concentrations (0, 2.9, 5.7, 8.6, 11.4, 14.3, 17.1 μM) were tested to determine the optimum conditions for in vitro rooting of microshoots. Up to 72% of the microshoots rooted with 14.3 μM IAA. Other auxins tested, α-naphthaleneacetic acid and indole-3-butyric acid, were less effective than IAA in inducing adventitious
root formation. All rooted plantlets having more than three roots were successfully established in soil. 相似文献
9.
Micropropagation of Pinus caribaea Morelet 总被引:1,自引:0,他引:1
Adventitious shoot formation was induced in excised mature embryos of Pinus caribaea using a modified Murashige and Skoog medium (MSM) supplemented with 6-benzyladenine. The highest frequency (96%) of adventitious bud production was observed when embryos were exposed to 8.9 M BA for one week prior to transfer to a growth regulator-free medium. Increased BA concentration and longer exposure to BA significantly reduced survival rates of explants. Dilution of the basal medium to 1/4× and 1/8× decreased shoot formation but 1/2× was just as effective as full-strength. Addition of auxins, glyphosate and coconut water to the rooting medium did not improve rooting success beyond that of spontaneous rooting. Sucrose at 1.5% significantly increased rooting of shoots. Plantlets were successfully transferred to the soil after preincubation in liquid medium.Abbreviations BA
6-benzyladenine
- NAA
naphthaleneacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- MSM
modified Murashige and Skoog medium
- CBM
Cupressus basal medium
- GDM
modified Gresshoff and Doy medium
- SH
Schenk and Hildebrandt medium 相似文献
10.
Micropropagation of Salvia brachyodon through nodal explants 总被引:1,自引:0,他引:1
A protocol for in vitro propagation of Balkan endemic plant Salvia brachyodon Vandas from nodal segments was developed. 6-benzylaminopurine was more effective in axillary buds promotion when compared
to thidiazuron. The rooting of regenerated shoots was induced by transferring them to the media supplemented with auxins.
All tested auxins (indole-3-acetic acid, indole-3-butyric acid, and α-naphthaleneacetic acid) stimulated the rooting of S. brachyodon shoots. The acclimatization of in vitro rooted shoots was successful. 相似文献
11.
This report describes an efficient plant regeneration system for the medicinal plant Lythrum salicaria via direct adventitious shoot development from leaf and stem explants. Leaf explants were much more responsive to regeneration
than stem segments. Of the hormonal combinations tested, those involving thidiazuron (TDZ; 0.1, 0.3 or 0.5 mg dm−3) were more effective than the combinations of other hormones and 0.1 mg dm-3 TDZ combined with either indole-3-acetic acid
(IAA) or indole-3-butyric acid (IBA) was the most productive. Rooting was readily achieved when multiple shoots were singled
out and cultured on medium containing different auxins. IAA was the most effective on root development in terms of both the
number of roots per shoot and the frequency of rooted shoots. More than 90 % of the regenerants survived after hardening for
four weeks at gradually decreased air humidity. 相似文献
12.
Summary Callus derived from Symphytum officinale L. regenerants was cultured in the presence of various phytohormones. The growth rate of callus was stimulated by all phytohormones at various concentrations. With 1-naphthaleneacetic acid no organ differentiation could be observed. With indole-3-butyric acid at low concentrations only roots were formed, whereas 6-benzylaminopurine, kinetin and zeatin at various concentrations induced either root or shoot formation or the simultaneous regeneration of both. Minor amounts of fructans were formed at high 6-benzylaminopurine-, zeatin- and at all indole-3-acetic acid-concentrations. The concentration of 1-naphthaleneacetic acid had no influence on the fructan content. Highest rates of fructan synthesis occurred at low zeatin-concentrations up to 1.5 mg/l. Only zeatin at all concentrations induced the synthesis of polyfructans, whereas appreciable amounts of oligofructans were formed under the influence of all other phytohormones.Abbreviations NAA
1-naphthaleneacetic acid
- IBA
indole-3-butyric acid
- BAP
6-benzylaminopurine
- IAA
indole-3-acetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- DP
degree of polymerisation
- GLC
gas liquid chromatography
- TLC
thin layer chromatography 相似文献
13.
María Teresa Herrera Margarita Cacho M. Purificación Corchete Jorge Fernandez-Tarrago 《Plant Cell, Tissue and Organ Culture》1990,22(3):179-182
Shoot tips from seedlings of Digitalis thapsi L. were cultured on Murashige and Skoog's medium and the effect of various auxins (2,4-D, NAA and IAA) were analyzed alone or in combination with cytokinis (BA and kinetin). Shoot multiplication and direct rooting of the new shoots were obtained after four weeks of culture in MS medium without hormones, but callus formation and the appearance of abnormal phenotypes were frequent. The addition of auxins to the cultures prevented the formation of callus but not the appearance of variant phenotypes. Both drawbacks could be avoided by combination of NAA or IAA with BA or kinetin. The best results for shoot multiplication and direct rooting were obtained with 0.5 mg l-1 NAA and 0.1 or 0.5 mg l-1 kinetin.Abbreviations BA
6-benciladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- Kin
kinetin
- NAA
naphtalene acetic acid
- MS
Murashige and Skoog 相似文献
14.
Camillo Branca Gabriele Bucci Paolo Domiano Ada Ricci Anna Torelli Maria Bassi 《Plant Cell, Tissue and Organ Culture》1991,24(2):105-114
In order to understand better the relationship between auxin structure and activity on morphogenesis and cell elongation, six different auxins were tested on the regeneration of tomato (Lycopersicon esculentum Miller var. Alice) from cotyledons and on pea (Pisum sativum L. var. Alaska) stem elongation. The auxins were: indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 1, 2-benzisoxazole-3-acetic acid (BOA), 1,2-benzisothiazole-3-acetic acid (BIA), 1-naphthalenacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D). All these compounds obey the minimum requirement rules for auxin activity and all were effective on cell elongation. At the dose of 10 M and in the absence of cytokinin, they all, except 2,4-D, induced roots, while in the presence of cytokinin they induced shoots, roots, hairy root-like filaments (HRLF) or callus depending on their concentration. The morphogenetic pattern did not change by varying cytokinin concentration. We conclude that auxin structure plays a minor role in morphogenesis or cell elongation, because it is only responsible for variations in the level of auxin activity. 相似文献
15.
Summary A method of plant regeneration from hypocotyl segments of Platanus acerifolia Willd, has been developed. Hypocotyl slices were cultured on Murashige and Skoog (MS) basal medium supplemented with a range
of combinations of cytokinins [6-benzyladenine (BA) or kinetin] and auxins [indole-3-butyric acid (IBA), indole-3-acetic acid,
α-naphthaleneacetic acid or 2,4-dichlorophenoxyacetic acid] for adventitious shoot induetion. The highest regeneration frequency
was obtained with MS medium containing 2.0 mg l−1 (8.88 μM) BA and 0.5 mg l−1 (2.46 μM) IBA. Adventitious buds and shoots were differentiated from hypocotyl-derived cellus or directly from the wounded sites within
4–8 wk. The regenerated shoots were elongated and proliferated efficiently on multiplication medium. Complete plantlets were
transplanted to the soil and grew normally in the greenhouse after root formation on rooting medium for 4–6 wk. 相似文献
16.
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. 相似文献
17.
A. R. Pathak 《Plant biosystems》2017,151(1):1-5
Hemidesmus indicus (Asclepiadaceae) leaf explants were utilized for establishing culture in MS medium fortified with individual cytokinins, auxins, and their combinations. Optimum response (80%) was observed in N6-benzyladenine (BA, 20 μM) + indole-3-acetic acid (IAA, 1 μM) with 19.67 ± 0.81 shoots per explant. Roots were induced in ¼MS + indole-3-butyric acid (IBA, 20 μM). 相似文献
18.
Soil microorganisms are capable of producing auxins in the presence of the physiological precursor, L-tryptophan (L-TRP). This study was designed to assess the influence of L-TRP on radish (Raphanus sativus) yield when applied to soil. The amount of L-TRP added to soil to give optimum radish growth in glasshouse studies was 3.0 mg kg-1 soil which enhanced the root yield by 1.31-fold over the control. The root/shoot ratio was increased by 1.10-fold upon this amendment. One L-TRP application was sufficient to promote growth. The best time to apply L-TRP was at the onset of seedling emergence. The application of L-TRP promoted radish yield comparable to those plants treated with indole-3-acetic acid, indole-3-acetamide and indole-3-lactic acid. Foliar application of L-TRP had no effect on the root and shoot dry weight. A field study was conducted in which L-TRP applications at a rate of 20.4 and 204 mg m-2 significantly enhanced the radish yield in fertilized plots receiving fertilization. The shoot dry weight was increased by 1.29-fold and the root dry weight by 1.15-fold over the control in response to 20.4 mg L-TRP m-2. These findings indicate that L-TRP, applied at the appropriate times and concentrations, can increase radish yield. The effect of L-TRP on radish growth could be attributed to i) substrate-dependent auxin production in soil by the indigenous microflora, ii) uptake directly by plant roots followed by metabolism within their tissues, and/or iii) a change in the balance of rhizosphere microflora affecting plant growth. 相似文献
19.
C. A. Martínez A. M. Giulietti J. Rodríguez Talou 《Plant Cell, Tissue and Organ Culture》2011,104(1):91-100
Hypericum perforatum L. (St. John’s wort) produces a number of phytochemicals having medicinal, anti-microbial, anti-viral and anti-oxidative
properties. Plant extracts are generally used for treatment of mild to medium cases of depression. Plant regeneration can
be achieved in this species by in vitro culture of a variety of explants. However, there are no reports of regeneration from
petal explants. In this report plant regeneration from petal explants of St. John’s wort was evaluated. Petals of various
ages were cultured on agarized Murashige and Skoog 1962 (MS) medium supplemented with auxin and cytokinin (kinetin), maintained in the dark and callus and shoot regeneration determined
after 28 days. At an auxin to cytokinin ratio of 10:1, callus and shoot formation were induced by all levels of indole-3-acetic
acid (IAA), indole-3-butyric acid (IBA) and 1-naphthaleneacetic acid (NAA), while 2,4-dichlorophenoxyacetic acid (2,4-D) induced
only callus formation. The optimum level of auxin for shoot regeneration was 1.0 and 0.1 mg/l kinetin, where the regeneration
frequency was 100 percent for all three auxins. The highest number of shoots per explant (57.4 and 53.4) was obtained with
IAA and IBA, respectively. In the absence of auxin, kinetin levels of 0.1 and 0.25 mg/l induce callus and shoot formation
at low frequency but not at lower levels. Callus and shoot formation did not occur in the absence of growth regulators. Petal-derived
shoots were successfully rooted on half-strength MS medium without a requirement for exogenous auxin and flowering plants
were established under greenhouse conditions. From these results it can be concluded that auxin type is a critical factor
for plant regeneration from petal explants of Hypericum perforatum and there is no absolute requirement for high levels of cytokinin. 相似文献
20.
High frequency bud break and multiple shoots were induced in nodal explants collected between November to February from a 5 year old tree of Morus australis Poir syn. M. acidosa Griff. on Murashige and Skoog's medium supplemented with 6-benzylaminopurine (1.0 mg/1). Incorporation of gibberellic acid (0.3 mg/l) along with BAP (1.0 mg/l) not only induced faster bud break from nodal explants as well as from apical shoot buds, but it also enhanced the frequency of bud break. Nodal explants were more responsive than apical shoot buds. The shoots formed in vitro were multiplied further as nodal segments, and an average multiplication rate of 6-fold per subculture was established within 4–5 months. The shoots were successfully rooted on half-strength MS containing a combination of indole-3-acetic acid, indole-3-butyric acid and indole-3-propionic acid, each at 1.0 mg/1. The plantlets were successfully hardened off and established in natural soil.Abbreviations BAP
6-benzylaminopurine
- GA3
gibberellic acid
- KN
kinetin
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- IPA
indole-3-propionic acid
- MS
Murashige and Skoog (1962) medium
- NAA
1-naphthalene acetic acid 相似文献