共查询到20条相似文献,搜索用时 31 毫秒
1.
Summary Growth characteristics of tobacco protoplasts containing rolA linked to its own promoter, or the rolB, or rolC genes of Agrobacterium rhizogenes linked to the Cauliflower Mosaic Virus 35S RNA promoter were compared with those from untransformed plants. RolA protoplasts require auxin and cytokinin for callus formation. Protoplasts overexpressing rolB and C form callus in the absence of exogenously applied auxin and cytokinin, respectively. Long term callus growth requires auxin, but the requirement for cytokinin is not critical. Optimal transient expression of an auxin responsive promoter element occurred at lower external levels of auxin in rolB and rolC protoplasts compared with untransformed protoplasts. Addition of putrescine was required for auxin responsive transient gene expression in rolA protoplasts suggesting that polyamines, or their products affect gene expression in rolA plants.Abbreviations T-DNA
transferred DNA
- TL-DNA
left transferred DNA
- NAA
naphthalene acetic acid
- PEG
polyethylene glycol
- GUS
glucuronidase
- CaMV
cauliflower mosaic virus 相似文献
2.
Joni Esrom Lima Vagner Augusto Benedito Antonio Figueira Lázaro Eustáquio Pereira Peres 《Plant cell reports》2009,28(8):1169-1177
We analyzed the impact of ethylene and auxin disturbances on callus, shoots and Agrobacterium rhizogenes-induced hairy root formation in tomato (Solanum lycopersicum L.). The auxin low-sensitivity dgt mutation showed little hairy root initiation, whereas the ethylene low-sensitivity Nr mutation did not differ from the control Micro-Tom cultivar. Micro-Tom and dgt hairy roots containing auxin sensitivity/biosynthesis rol and aux genes formed prominent callus onto media supplemented with cytokinin. Under the same conditions, Nr hairy roots did not form callus. Double mutants combining Rg1, a mutation conferring elevated shoot formation capacity, with either dgt or Nr produced explants that formed shoots with little callus proliferation. The presence of rol + aux genes in Rg1 hairy roots prevented shoot formation. Taken together, the results suggest that although ethylene does not affect hairy root
induction, as auxin does, it may be necessary for auxin-induced callus formation in tomato. Moreover, excess auxin prevents
shoot formation in Rg1. 相似文献
3.
Isabelle Roussy Frédéric Dubois Rajbir S. Sangwan Brigitte S. Sangwan-Norreel 《Plant cell reports》1996,16(3-4):142-146
Summary The effect ofin planta treatments with auxin inhibitors such as 2,3,5 triiodobenzoic acid (TIBA) on regeneration of plantsin vitro is not known. Here, we show the beneficial effect of preconditioning sugarbeet plants in the greenhouse with TIBA (3 mg/1) for efficientin vitro plant regeneration via a callus phase from cultured leaf explants. Without this treatment, no shoot developed on the control leaf-calluses. Several hundred plants were routinely regenerated using this protocol. More importantly, the number of shoots per explantcallus increased drastically over the subsequent subculture period. The most favorable media for callus induction contained a combination of an auxin and a cytokinin (0.1 mg/1 2,4-dichlorophenoxyacetic acid and 1 mg/1 N-6 benzylaminopurine) or a cytokinin alone (2.2 mg/1 thidiazuron). However, only the callus derived from leaves of TIBA-treated genotypes and induced on thidiazuron-medium produced numerous shoots. Histological studies showed the formation of meristematic zones only in the organogenic callus developed on thidiazuron-coutaining medium. The analysis of peroxidase activity showed that the activity was higher for the TIBA-treated plants than for the untreated control plants. 相似文献
4.
Summary Methods of plant regeneration from callus and protoplasts of Helianthus giganteus L. are described. Embryogenic callus was obtained from leaf explants and plants were regenerated from these calli on MS media with different combinations of benzyladenine and naphtaleneacetic acid. Leaf protoplasts isolated from in vitro grown plants formed somatic embryos when cultured in agarose solidified droplets of V-KM medium containing benzyladenine and naphtaleneacetic acid. Embryos developed into plantlets on media with reduced auxin contents. Regenerated plants were successfully planted in soil.Abbreviations BA
benzyladenine
- IAA
indoleacetic acid
- MS
Murashige and Skoog medium
- NAA
naphtaleneacetic acid
- V-KM
protoplast culture medium of Binding and Nehls 相似文献
5.
R. V. Sairam C. Wilber J. Franklin B. Smith J. Bazil R. Hassel D. Whaling K. Frutiger C. A. Blakey R. Vierling S. L. Goldman 《In vitro cellular & developmental biology. Plant》2002,38(5):435-440
Summary A protocol for high-frequency callus, somatic embryogenesis, and plant regeneration for Tripsacum is described. Plants were regenerated from complete shoot meristems (3–4 mm) via organogenesis and embryogenesis. In organogenesis,
the shoot meristems were cultured directly on a high cytokinin medium comprising 5–10 mgl−1 (22.2–44.4 μM) 6-benzyladenine (BA). The number of multiple shoots varied from six to eight from each meristem. The time required for production
of plants from organogenesis was rapid (4–6 wk). In contrast, callus was induced on an auxin medium and continuously cultured
on an auxin medium for production of somatic embryos. Prolific callus with numerous somatic embryos developed within 3–4 wk
when cultured on an auxin medium containing 5 mgl−1 (22.6μM), 2,4-dichlorophenoxyacetic acid (2,4-D). The number of shoots induced varied from two to five per callus. Regardless of
the cultivars used, the frequency of callus induction and plant regeneration was between 48% and 94%. The seed germination
procedures also were modified and resulted in a maximum of 60–80% seed germination. Finally, the rate of T-DNA transfer to
complete shoot meristems of Tripsacum was high on the auxin medium and was independent of whether super-virulent strains of Agrobacterium were used or not. 相似文献
6.
A method to obtain plants from embryogenic callus of Brassica nigra and protoplasts of hypocotyl expiants is described. Callus was initiated on Murashige and Skoog medium containing kinetin (kn) and 2,4-dichlorophenoxy acetic acid (2,4-D). Lowering of auxin induced embryo formation. Supplementation with gibberellic acid (GA3) enhanced embryogenic response tenfold. Passage through liquid medium devoid of growth regulators was essential for the growth of embryos. Secondary embryos were produced on transfer to solid basal medium. Embryogenic callus retained its morphogenic ability even after 12 subcultures. Both primary and secondary embryos produced fertile plants. Hypocotyl-derived protoplasts were also regenerated to plants following the same protocol. The survival of plants on transfer to soil was about 80%. The seeds from plants derived from callus and protoplasts were viable.Abbreviations 2,4-D
2,4-dichlorophenoxy acetic acid
- NAA
naphthalene acetic acid
- IAA
indole acetic acid
- kn
kinetin
- GA3
gibberellic acid 相似文献
7.
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
8.
G. E. Hanning B. V. Conger 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1982,63(2):155-159
Summary The purpose of this investigation was to demonstrate callus induction and plantlet formation from cultured leaf segments of 12–15 week-old Dactylis glomerata L. (orchardgrass) plants. Flat half-leaf sections, approximately 2–3 mm square, from the three innermost (youngest) leaves were isolated and individually plated serially beginning at the leaf base on a solid SH medium containing 30 M of 3,6-dichloro-oanisic acid (dicamba). Callus formed on leaf sections from all 50 plants used in the study. After transfer to SH medium with 1 M dicamba, plantlets formed from leaf sections of 9 of the 50 plants. In most cases plantlets formed from embryogenic callus but in a few cases embryoids formed directly on the leaf surface without an intervening callus state. These developed into plantlets when transferred to low auxin medium. The response for both callus and plantlet formation decreased with increasing distance both spatially and temporally from the shoot apex. Histological examination of embryogenic callus revealed the presence of non-zygotic embryos in various stages of development. The results provide further support for compentency (if not totipotency) of Gramineae leaf cells. 相似文献
9.
The experimental results are presented on production of plants-regenerants of the sugar beet (Beta vulgaris L.) via callus formation or direct organogenesis from the leaf tissues. The method of aseptic treatment was developed for the seeds with strong bacterial and fungal invasion. The regenerant plants were obtained in the presence of various concentrations of synthetic hormones, such as cytokinin (6-benzaminopurine) and auxin (naphthylacetic acid), and inhibitor of auxin transport in plants (2,3,5-triiodobenzoic acid). This combination of growth regulators made it possible to avoid callus formation. The genotype of initial plants affected the capacity for callus formation and regeneration. The temperature influenced rhizogenesis in regenerants. 相似文献
10.
Hormone-regulated inflorescence induction and TFL1 expression in Arabidopsis callus in vitro 总被引:4,自引:0,他引:4
To study hormone-regulated inflorescence development, we established the in vitro regeneration system of Arabidopsis inflorescences in the presence of cytokinin and auxin. Media containing a combination of thidiazuron (TDZ) and 2,4-dichlorophenoxyacetic acid (2,4-D) were used to induce callus formation. Higher frequencies of calli were obtained by using the inflorescence stems as explants. After transferring the calli to media containing a combination of zeatin and indole-3-acetic acid (IAA), the inflorescences were induced from the calli. The morphology of regenerated inflorescences was similar to that of inflorescences in plants; however, flowers of regenerated inflorescences often lacked a few floral organs. Furthermore, TFL1, a gene involved in floral transition in Arabidopsis, was activated during the inflorescence induction. Our results suggest that the TFL1 gene plays an important role in hormone-regulated inflorescence formation. 相似文献
11.
Dominick V. Basile Nasrin Akhtari Yolanda Durand M. S. R. Nair 《In vitro cellular & developmental biology. Plant》1993,29(3):143-147
Summary
Artemisia annua L. is the source of a potent antimalarial, artemisinin. As part of a program to produce artemisinin through tissue culture,
a series of 14 multifactorial experiments were conducted to determine suitable conditions for initiating and maintaining friable
callus fromA. annua. In the first six experiments, three different nutrient formulations [Gamborg B5 (B5), Murashige and Skoog (MS), and Whetmore
and Rier (WR)], each with 32 combinations of auxins and cytokinins [2,4-dichlorophenoxyacetic acid (2,4-D) with benzyladenine
(BA), or 1-naphthaleneacetic acid (NAA) with 6-furfurylaminopurine (kinetin)], were tested. Both B5 and WR nutrients supported
friable callus formation from leaf explants with some combinations of auxin and cytokinin. Inasmuch as friable callus seemed
to be produced over a wider range of auxin and cytokinin concentrations in combination with B5, the remaining experiments
were conducted solely with this nutrient formulation. In the remaining eight experiments, it was determined that friable callus
formed when combinations of NAA with kinetin or 2,4-D and BA were used with B5 medium. Lighter colored, more friable callus
formed in response to 2,4-D and BA than with NAA and kinetin. No single combination of concentrations of auxin and cytokinin
seemed to be “ideal” for producing friable callus. Ranges of 2,4-D from 0.5 to 2.0 with BA between 0.025 and
0.1, or NAA between 0.5 and 2.0 with kinetin between 0.5 and 1.0 mg/liter, produced acceptable results. 相似文献
12.
Summary Simple one step micropropagation system for Cephaelis ipecacuanha A. Richard was developed using root cultures grown in vitro. Adventitious shoots were directly formed on the cut end of root segments without callus formation, on phytohormone-free B5 solid medium in the dark. When the shoots attached with root segments were further cultured under 16 h light / 8 h dark, they developed into plantlets, which could be transplanted to soil. The regenerated plants grew well in a greenhouse with showing normal appearance and accumulated alkaloids. The influence of auxin on adventitious shoot formation was also investigated.Abbreviations MS
Murashige-Skoog (Murashige and Skoog 1962)
- 1/2 MS
half strength MS
- B5
Gamborg B5(Gamborg et al. 1968)
- WP
woody plant (Lloyd and McCowm 1980)
- RC
root culture (Thomas and Davey 1982)
- HF
phytohormone free
- IAA
indole-3-acetic acid
- NAA
1-naphtaleneacetic acid
- TIBA
2,3,5-triiodobenzoic acid
- 2,4,6-T
2,4,6-trichlorophenoxyacetic acid
- SEM
scanning electron microscopy
- C.V.
coefficient of variation 相似文献
13.
Somatic embryos of Cyclamen persicum Mill. could be produced through a callus phase from juvenile explant material including anthers, ovaries and zygotic embryos. The auxin 2,4-D (1.0–1.5 mg l-1) and coconut milk (10% v/v) in MS medium were important factors for the induction of somatic embryogenesis. Somatic embryos germinated into plantlets in MS medium without growth regulators. The plants grew well in the greenhouse and flowered normally. The plants were phenotypically identical to the mother plants with a few exceptions.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
1-naphthylacetic acid
- IAA
3-indoleacetic acid
- BA
6-benzyladenine
- ABA
abscisic acid
- CM
coconut milk 相似文献
14.
Summary Root hair formation and the development of transfer cells in the rhizodermis was investigated in various existing auxinrelated mutants ofArabidopsis thaliana and in the tomato mutantdiageotropica. Wild-type Arabidopsis plants showed increased formation of root hairs when the seedlings were cultivated in Fe- or P-free medium. These extranumerary hairs were located in normal positions and in positions normally occupied by nonhair cells, e.g., over periclinal walls of underlying cortical cells. Defects in auxin transport or reduced auxin sensitivity inhibited the formation of root hairs in response to Fe deficiency completely but did only partly affect initiation and elongation of hairs in P-deficient roots. Application of the ethylene precursor 1-aminocyclopropane-1-carboxylic acid or the auxin analog 2,4-dichlorophenoxyacetic acid did not rescue the phenotype of the auxin-resistantaxr2 mutant under control and Fe-deficient conditions, indicating that functionalAXR2 product is required for translating the Fe deficiency signal into the formation of extra hairs. The development of extra hairs inaxr2 roots under P-replete conditions was not affected by auxin antagonists, suggesting that this process is independent of auxin signaling. In roots of tomato, growth under Fe-deficient conditions induced the formation of transfer cells in the root epidermis. Transfer cell frequency was enhanced by application of 2,4-dichlorophenoxyacetic acid but was not inhibited by the auxin transport inhibitor N-1-naphthylphthalamic acid. In thediageotropica mutant, which displays reduced sensitivity to auxin, transfer cells appeared to develop in both Fe-sufficient and Fe-deficient roots. Similar to the wild type, no reduction in transfer cell frequency was observed after application of the above auxin transport inhibitor. These data suggest that auxin has no primary function in inducing transfer cell development; the formation of transfer cells, however, appears to be affected by the hormonal balance of the plants.Abbreviations ACC
1-aminocyclopropane-1-carboxylic acid
- TIBA
triiodobenzoic acid
- NPA
N-1-naphthylphthalamic acid
- STS
silver thiosulfate 相似文献
15.
H. J. Newbury 《Plant Cell, Tissue and Organ Culture》1986,7(1):39-42
Nine varieties of Antirrhinum majus L. have been used in a study of in vitro multiplication of plants using shoot-tip culture. Acceptable multiplication rates were obtained in several media with only variety Victory showing significantly lower rates of shoot production. Wounded shoots of this variety produced callus in the absence of added auxin and some of this callus produced prolific roots. 相似文献
16.
Effects of cytokinin on adventitious root formation in callus cultures ofVigna unguiculata (L.) walp
Woong-Young Soh Pil-Son Choi Duck-Yee Cho 《In vitro cellular & developmental biology. Plant》1998,34(3):189-195
Summary Yellowish compact callus, induced from cowpea hypocotyls on Murashige and Skoog(MS) medium (1962) containing 0.2 mg/l(0.93
μM) kinetin and 0.4 mg/l (1.81 μM) 2,4-dichlorophenoxyacetic acid (2,4-D), was subcultured on MS medium containing cytokinin alone, auxin alone, or auxins
plus cytokinins in order to determine the effect of cytokinins on root organogenesis in callus cultures. The callus actively
proliferated on the same medium but did not show any organogenic activity macroscopically as well as microscopically. On medium
with N6-benzyladenine (BA) and 1-naphthaleneacetic acid (NAA), the yellowish compact callus first changed to pale green compact callus
and then many green spots appeared on its surface under light culture. But the yellowsih compact callus remained yellowish
and white spots appeared on its surface in dark culture. These spots gradually became white nodular structures. Adventitious
root formation from the nodular structures occurred not only on the same medium, but also on medium with either auxin or cytokinin
but not both. Yellowish compact callus on medium with auxin alone was transformed to yellowish friable callus, which did not
develop adventitious roots. The yellowish friable callus could gain rhizogenic activity only after morphological modification
to pale green compact callus on medium with auxin plus cytokinin. The modified callus did not form adventitious roots on medium
with auxins but only with cytokinins. Therefore, it is suggested that cytokinins have stimulating effects on root formation
from callus that previously did not show rhizogenic activity on medium with auxins alone. In addition, the rhizogenic potential
of cowpea callus was discriminated from that of leaf explants, which formed adventitious roots directly on medium with auxin
alone. 相似文献
17.
Analogues of auxin modifying growth and development of some monocot and dicot plants 总被引:1,自引:0,他引:1
The dependence of morphogenetic processes in the formation of vegetative and generative organs in spring oilseed rape and
barley on exogenously applied physiological analogues of auxin: 2,4-D (2,4-dichlorphenoxyacetic acid), NAA (naphthalene-1-acetic
acid), TA-12 (1-[2-chloroethoxycarbonyl-methyl]-4-naphthalenesulfonic acid calcium salt) and TA-14 (1-[2-dimethylaminoethoxicarbonylmethyl]naphtalene
chlormethylate) were investigated. The experiments were performed with hypocotyl tissue cultures of oilseed rape and barley
microspores in vitro. The auxin analogues applied revealed differences of morphogenetic competence in dedifferentiation-redifferentiation processes
that occurred in oilseed rape cultures. TA-12 and TA-14 applied together with NAA and BA (6-benzylaminopurine) caused more
intensive callus growth in comparison with 2,4-D. Rhizogenesis was induced when 2,4-D was substituted by TA-12. Compound TA-14,
unlike TA-12, facilitated the appearance and development of cotyledons in callus tissues. Hower the compounds TA-12 and TA-14
have no positive effect in monocot plant — barly anther culture for callogenesis and regeneration in comparison to indole-3-acetic
acid (IAA). TA-14 and TA-12 showed similar but not identical auxin properties and demonstrated high efficiency as modifiers
of rape-dicot plant growth and morphogenesis. 相似文献
18.
Peter F. Straub Debra M. Decker John L. Gallagher 《Plant Cell, Tissue and Organ Culture》1988,15(1):73-78
Phragmites australis tissue cultures were initiated from mature seeds on MS medium supplemented with 1 mgl-1 each of 2,4-D and IAA. Cultures displayed typical embryogenic callus that was compact and bright yellow. Selection for embryogenic callus established long-term regenerable cultures. Removal of auxin from the basal medium allowed numerous complete plants to be recovered from the cultures. Histological study indicated both the presence of embryogenic-type cells and the bipolar development of regenerated plants. 相似文献
19.
The frequency of embryo formation was determined in normal and 5-methyltryptophan-resistant (5-MTr) cell lines of wild carrot (Daucus carota L.) grown in the presence or absence of 2-isopentenyladenine (2-ip) and 2,4-dichlorophenoxyacetic acid (2,4-D). 2-ip stimulated the intitation of embryo formation and also accelerated embryo development. 2.4-D inhibited embryo differentiation at several stages: at 0.1 mg/l, it stopped regeneration at the earliest stage, resulting in callus growth instead of embryo formation; at 0.04 mg/l 2,4-D, some globular embryos were produced, but they did not develop into more advanced embryos. Variant cell lines with higher levels of auxin (indole-3-acetic acid, IAA) were used to study the effect of an elevated endogenous concentration of auxin on embryogenesis. IAA at these concentrations suppressed regeneration in the same manner as the exogenous auxin, 2,4-D, did. This result confirms the hypothesis that high levels of IAA are responsible for the suppression of regeneration in the 5-MTr cell lines. 相似文献
20.
High frequency somatic embryogenesis and plant regeneration from zygotic embryo-derived callus cultures of three Allium species 总被引:1,自引:0,他引:1
P. van der Valk O. E. Scholten F. Verstappen R. C. Jansen J. J. M. Dons 《Plant Cell, Tissue and Organ Culture》1992,30(3):181-191
The plant regeneration ability of zygotic embryo-derived callus cultures was studied for 12 A. cepa varieties and accessions, two A. fistulosum varieties, one A. fistulosum x A. cepa interspecific hybrid and two A. porrum varieties. Compact embryogenic callus was induced on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid. The embryogenic calluses of all three Allium species were similar in appearance. For all accessions tested plants could be regenerated at a high frequency from this compact callus through somatic embryogenesis, when using kinetin supplemented MS medium (regeneration medium). Addition of abscisic acid to the regeneration medium stimulated the formation of both somatic embryos and shoots for a number of varieties. Concerning shoot regeneration from callus cultures, significant differences existed between genotypes of all accessions except one.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- VDH
Van Der Have Seed company 相似文献