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1.
M. Szechyńska-Hebda E. Skrzypek G. Dąbrowska J. Biesaga-Kościelniak M. Filek M. Wędzony 《Acta Physiologiae Plantarum》2007,29(4):327-337
As part of work to optimize the regeneration processes of winter wheat callus culture the effects of two auxins (2,4-D, IAA),
two cytokinins (kinetin, zeatin), and the fungal mycotoxin zearalenone, were tested individually in vitro using embryo-, and
inflorescence-derived callus. To determine the role of oxidative stress in cell regeneration, changes in the basic antioxidant
enzymes, superoxide dismutase (SOD), catalase (CAT), and peroxidases (PODs) were investigated. In general, zearalenone (ZEN)
was found to be more effective than cytokinin treatments for inducing shoot production, whereas auxins suppressed the regeneration
process. Regenerating callus showed higher induction of these antioxidant enzymes in comparison with non-regenerating callus.
SOD, CAT and POD activities were higher in callus derived from inflorescence than in callus derived from immature embryo.
Activities of SOD, CAT and POD in culture derived from immature embryos were depending on type of growth regulator in medium.
The highest enzyme activities were observed in non-regenerating tissues after auxins treatment and in regenerating tissues
after cytokinins treatment. The effect of ZEN was similar to that of cytokinins. One MnSOD band and two Cu/ZnSOD bands were
detected in all cultures. Changes in SOD izoform patterns occurred in callus culture on media with auxins and ZEN, but not
on media with cytokinins. Our results suggest that callus regeneration is associated with reactive oxygen species production
induced by specific growth regulators. Reactive oxygen species under the control of cellular antioxidant machinery can mediate
signalling pathways between exogenously applied growth regulators and the induction and/or creation of the direction of morphogenesis. 相似文献
2.
Tuong-Van Nguyen Tran Thanh Thu Martine Claeys Geert Angenon 《Plant Cell, Tissue and Organ Culture》2007,91(2):155-164
An improved regeneration protocol suitable for transformation of sorghum was developed. The improvements focused on limiting
the production of phenolic compounds and the use of suitable culture vessels for each developmental stage in plant regeneration
from immature embryo derived calli. The addition of activated charcoal in the callus induction medium reduced the production
of black pigments, however it also inhibited the callus formation on immature embryo explants. Cold pre-treatment of the immature
seeds from which embryo explants were excised had a positive effect on both explant survival and callus formation. A one-day
4°C treatment of immature seeds significantly improved the callus formation from immature embryos and reduced the need for
frequent subculture. Petri dishes with ventilation were suitable for the callus induction phase, but not for plant regeneration.
Regeneration of plants could be improved by using disposal plastic boxes (250 ml volume) instead of Petri dishes. Agrobacterium-mediated transformation using the improved regeneration protocol and the hygromycin phosphotransferase gene as selectable
marker resulted in the recovery of 15 transgenic plants from 300 initial immature embryos (5% efficiency). The transgenic
nature of the obtained plants was demonstrated by Southern hybridisation and progeny analysis. The transgenes were inherited
in a Mendelian fashion and were integrated at a single locus in the majority of the analysed lines. 相似文献
3.
Rakesh Kumar Harohalli Masthigowda Mamrutha Amandeep Kaur Karnam Venkatesh Anita Grewal Raj Kumar Vinod Tiwari 《Physiology and Molecular Biology of Plants》2017,23(4):945-954
The availability of reproducible regeneration system through tissue culture is a major bottleneck in wheat improvement program. The present study has considered to develop an efficient callus induction and regeneration system using mature and immature embryos as explants in recently released agronomically superior spring wheat varieties. An efficient sterilization process was standardized using 0.1% HgCl2 and 70% ethanol for both seeds and embryos. The maximum possible combinations of plant growth regulators (PGRs) were evaluated for their effect on different wheat regeneration processes through tissue culture starting from callus to root induction. Picloram is found as an effective auxin with 87.63–98.67% callus induction efficiency in both explants. Supplementation of CuSO4 along with 2,4-D, zeatin in regeneration medium significantly enhanced the multiple shoot induction. The shoot development was achieved using full strength Murashige and Skoog’s (MS) medium and root induction using half MS medium without PGRs. The optimized medium and method has resulted up to 100% regeneration irrespective of the genotype used with high reproducibility. Thus, the standardized regeneration system can be used in the regeneration of healthy plants from embryos rescued from interspecies crosses, transgenic production, induced mutation breeding and recently developed genome editing techniques for the procreation of wheat plants having novel traits. 相似文献
4.
A short-term regeneration system from leaf-base-derived callus of wheat (Triticum aestivum L.) was developed. Embryogenic callus formation and shoot regeneration were achieved from the first basal segments of 3–4-day-old seedlings. Callus formation frequency as well as plantlet regeneration frequency was dependent on the composition of basal medium and the concentration of 2,4-dichlorophenoxyacetic acid (2,4-D). MS medium with 2,4-D 4.5–9.0 mol l–1 was optimal for the culture of wheat leaf base. Effects of different combinations of plant growth regulators, which were added in either callus induction medium or shoot regeneration medium, were tested. Adding of BAP in callus induction medium shortened the time of shoot emergence but could not improve the producing of embryogenic calli and green plantlets. Optimal ratio of 2,4-D, BAP and NAA gave similar regeneration frequency to control. Existence of cytokinins in regeneration medium had no effect on increasing the regeneration frequency. The regenerants could grow to normal, fertile plants after they were transferred into soil. 相似文献
5.
Somatic embryogenesis from pea embryos and shoot apices 总被引:3,自引:0,他引:3
Conditions were defined for plant regeneration via somatic embryogenesis in pea, using explants from immature zygotic embryos or from shoot apices. For the induction of somatic embryos, an auxin (picloram or 2,4-dichlorophenoxyacetic acid) was required. Embryogenic callus originated from embryonic axis tissue of immature embryos and from the axillary-bud region and the plumula of shoot apices. A clear effect of embryo size on somatic embryogenesis was shown. There were differences in frequency of somatic embryogenesis among the five genotypes used in the study. Additions of BA to auxin-containing medium reduced embryo production. Histological examinations confirmed the embryogenic nature of the immature embryo cultures and revealed that somatic embryos originated from the meristematic areas near the callus surface.Abbreviations BA
benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- picloram
4-amino-3,5,6-trichloropicolinic acid 相似文献
6.
Tissue cultures of the halophytic saltmarsh grass Sporobolus virginicus were initiated from unemerged immature inflorescence tissue. Typical graminaceous embryogenic and nonembryogenic callus and cell types were noted. Embryogenic callus was compact golden yellow. Histological evidence indicated that proliferation of the ovary tissue of the immature pistil was the source for embryogenic callus. Plants regenerated after first reducing and then eliminating auxin from the culture medium. Regeneration was observed both through the concerted development of bipolar meristems from somatic embryos and by the formation of multiple shoot meristems that were either connected through callus tissue to root meristems or which later adventitiously rooted. The main mode of regeneration appeared to be somatic embryogenesis with additional multiple shoot formation probably due to precocious germination of somatic embryos. Plants recovered from culture were acclimated to soil, grown up in a greenhouse, and planted in field plots with saline irrigation to ensure stability of salt tolerance. 相似文献
7.
Lambé Pascal Mutambel Hity S.N. Deltour Roger Dinant Monique 《Plant Cell, Tissue and Organ Culture》1998,55(1):23-29
Three genotypes of Pearl millet were screened in vitro for induction of embryogenic callus, somatic embryogenesis and regeneration.
Shoot apices excised from in vitro germinated seedlings or immature embryos isolated from green house established plants were
used as primary explants. The frequency of embryogenic callus initiation was significantly higher in shoot apices in comparison
with immature zygotic embryos. Moreover, differences between genotypes were minimal when using shoot apices. Friable embryogenic
calli (type II) developed on the initial nodular calli after 1 to 3 months of culture. The frequency of type II callus is
related to the composition of the maintenance medium and they were more often found in ageing cultures. The transfer of embryogenic
calli onto auxin-free medium was sufficient for inducing somatic embryo development in short-term culture (3 months) while
a progressive loss in regeneration potential was observed with increasing time of subcultures. Maturation of embryogenic calli
on medium supplemented with activated charcoal, followed by germination of somatic embryos on medium supplemented with gibberellic
acid, restored regeneration in long-term cultures.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
8.
Three basal plant tissue culture media, namely, N6, MS, and modified Y3, were compared to optimize micropropagation protocol for E. guineensis. Full strength media were used separately to regenerate plantlets directly using immature zygotic embryos (IZEs), and through somatic embryogenesis of calli obtained from IZEs. The plantlets regenerated by direct regeneration on three media were examined for shoot length and rooting percentage. For the induction of callus, somatic embryogenesis, and rooting modified Y3 medium was the most effective. In conclusion, the results indicate that modified Y3 medium is the most suitable for direct regeneration, callus induction and somatic embryogenesis in E. guineensis. 相似文献
9.
K. Oldach A. Morgenstern S. Rother M. Girgi M. O'Kennedy H. Lörz 《Plant cell reports》2001,20(5):416-421
We report here an in vitro culture system that provides reliable, highly efficient regeneration from immature embryos of pearl millet [Pennisetum glaucum (L.) R. Br.] and sorghum [Sorghum bicolor (L.) Moench]. Immature embryos were isolated 10-20 days after pollination and cultured on various L3 media. The influence of different parameters during the callus induction phase was examined with respect to the regeneration rate: (1) the concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and various cytokinins; (2) the addition of AgNO3; (3) the use of maltose or sucrose as a carbon source. Modifications in the phytohormones alone resulted in the regeneration of fertile sorghum plants at high efficiency. Significant increases in the regeneration rates of pearl millet genotypes were achieved by the combination of sucrose as a carbon source and silver nitrate as a potential ethylene inhibitor. 相似文献
10.
Jinfeng Lü Rong Chen Muhan Zhang Jaime A. Teixeira da Silva Guohua Ma 《Journal of plant physiology》2013
Camellia nitidissima Chi (Theaceae) is a world-famous economic and ornamental plant with golden-yellow flowers. It has been classified as one of the rarest and most endangered plants in China. Our objective was to induce somatic embryogenesis, shoot organogenesis and plant regeneration for C. nitidissima. Three types of callus (whitish, reddish and yellowish) were induced from immature cotyledons on improved woody plant medium (WPM) with different plant growth regulators (PGRs). Among the callus, whitish callus was induced by 4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and reddish and yellowish callus were induced by strongly active cytokinins, thidiazuron (TDZ) or 6-benzylaminopurine (BAP), singly or combined with weakly active auxin, α-naphthaleneacetic acid (NAA). The embryogenic callus could differentiate into somatic embryos, nodular embryogenic structures (large embryo-like structures) or adventitious shoots depending on the PGR used in WPM. BAP was best for adventitious buds and zeatin was best for somatic embryogenesis while kinetin (Kt) was best for the formation of nodular embryogenic structures. The three regeneration pathways often occurred in the same embryogenic callus clumps. Most shoots (80.0%) developed roots in WPM supplemented with 24.6 μM IBA and 0.3 μM NAA while 47.5% of somatic embryos could germinate directly and develop into plantlets on induction medium supplemented with 0.9 μM BAP and 0.1 μM NAA. The nodular embryogenic structures could be sub-cultured and cyclically developed in one of two differentiation pathways: shoot organogenesis or somatic embryogenesis. Plantlets derived from shoot buds rooted and somatic embryos germinated when transplanted into soil in a greenhouse; 66.7% of plantlets from shoot culture and 78.6% of plantlets from somatic embryos survived after 8 weeks’ acclimatization. 相似文献
11.
A procedure was established for the induction of regenerable calli from immature inflorescence segments of high-tannin cultivars of sorghum (Sorghum bicolor (L.) Moench). Murashige & Skoog's medium with several components altered was utilized for inducing, maintaining, and regenerating the cultures. Embryogenic calli formed at a frequency of 8–70% depending on the genotype. During a ten-month period, 3600 plants were regenerated from eight genotypes tested. Among the developmental stages of immature inflorescence tested (from differentiation of secondary branch primordia to floret formation) no critical differences were found in potential for callusing, embryogenesis or regeneration. Genotypic differences were observed in pigment production, embryogenic callus formation, shoot differentiation, and in maintenance of regeneration capacity.Abbreviations 2,4-D
dichlorophenoxyacetic acid
This is Journal Paper Number 11972 from the Purdue University Agricultural Experiment Station 相似文献
12.
Plant regeneration through de novo shoot organogenesis in tissue culture is a critical step in most plant transformation and micropropagation procedures. Establishing an efficient regeneration protocol is an empirical process and requires optimization of multiple factors that influence the regeneration capacity. Here, we review the molecular process of shoot induction in a two-step regeneration protocol and focus on the role of auxins and cytokinins. First, during incubation on an auxin-rich callus induction medium (CIM), organogenic callus is produced that exhibits characteristics of a root meristem. Subsequent incubation on a cytokinin-rich shoot induction medium (SIM) induces root to shoot conversion. Through a detailed analysis of the different aspects of shoot regeneration, we try to reveal hinge points and novel candidate genes that may be targeted to increase shoot regeneration capacity in order to improve transformation protocols. 相似文献
13.
In the present study, interactions between the duration of treatment with auxin and different cytokinins and their effect on shoot regeneration were evaluated with the aim to establish a rapid and efficient in vitro regeneration method applicable to a variety of Populus species. Three different species, Populus angustifolia, P. balsamifera, and P. deltoids, were chosen for that purpose. We were successful in regenerating plantlets from stem and petiole explants from all three chosen species using a four-step simple procedure. The first step was callus induction when the explants were exposed to an auxin-rich medium for 0-20 days. During the second step, they were transferred onto a cytokinin-rich medium for shoot bud induction. In the third step, the shoots regenerated were transferred onto a medium with reduced levels of cytokinins to promote shoot proliferation and elongation; finally, in the fourth step, the shoots were rooted and acclimated. A short period (6-10 days) of time of exposure to auxin was sufficient for shoot regeneration. A culture time longer than ten days in callus induction medium drastically reduced the efficiency of shoot regeneration. Besides, cytokinin type and concentration also affected the frequency of shoot induction. A 0.2 mg/l concentration of 2,4-D for callus induction followed by 0.02 mg/l of Thidiazuron for shoot formation proved to be the best treatment for adventitious shoot bud multiplication, generating a maximum of 10-13 shoots of P. balsamifera and P. angustifolia in ten weeks. In contrast, for P. deltoids, a combination of 1.1mg/l 2,4-D, 1.0mg/l NAA, 0.1mg/l zeatin for callus induction followed by a combination of 1mg/l zeatin plus 1.0mg/l BA for shoot bud induction was found to be the most effective, generating on average 15 shoots over a period of ten weeks. 相似文献
14.
Plant regeneration of Solanum lycopersicoides Dun. from stem explants,callus and suspension cultures
Protocols were established for achieving plant regeneration from stem internode, callus, and cell suspension cultures of Solanum lycopersicoides Dun. Two accessions of S. lycopersicoides exhibited different responses as to callus formation on various media, requirement of gibberellic acid for shoot regeneration, and ability to grow in suspension culture. The optimum medium for initiation and maintenance of cell suspension cultures was Murashige and Skoog [9] medium with 15 mg l– NAA. For shoot regeneration, of three cytokinins tested, zeatin was found most effective relative to number, rapidity of response and overall quality of shoots. Shoot regeneration from stem explants, callus and suspension cultures was optimum on MS + 3.0 mg l–1 zeatin + 0.1 mg l–1 gibberellic acid.Michigan Agricultural Experiment Station Journal Article No. 11589. 相似文献
15.
Genetic variability of somatic embryogenesis in tissue cultures of sugar beet breeding lines 总被引:1,自引:0,他引:1
Golovko A 《T?Sitologii?a i genetika》2001,35(6):10-17
Genetic variability of callus initiation and plant regeneration has been investigated among three sugar beet genotypes. It was found that TDZ has a genotype-independent effect on callus initiation and is responsible for more than a two-fold increase in the friable callus induction rate and more than a three-fold increase in the shoot regeneration rate from this callus. Along with the genotype-independent organogenesis, regeneration from callus occasionally went through the process of somatic embryogenesis in a highly genotype-specific manner. Despite fast and uncontrollable conversion of embryos to normal plants, it was possible to select and maintain repetitive embryogenic culture without loosing regeneration and root formation capabilities. Extensive experimenting with medium composition and culture conditions resulted in an optimal medium for maintenance of repetitive embryos. Comparing with BAP, low concentrations of TDZ provide higher level of adventitious shoot formation and do not induce vitrification of tissues. 相似文献
16.
Sanjay Gupta V. K. Khanna Rameshwar Singh G. K. Garg 《Plant Cell, Tissue and Organ Culture》2006,86(3):379-388
Development of suitable strategy to overcome genotypic limitations of in vitro regeneration in sorghum would help utilize high yielding but poor tissue culture responsive genotypes in genetic manipulation programmes. A factorial experiment was conducted with two explants (immature embryos and inflorescences), eight genotypes (five Sorghum sudanense and three Sorghum bicolor genotypes), three levels of 2,4-D (1 mg l−1, 3 mg l−1, and 5 mg l−1), and two levels of kinetin (0.0 mg l−1 and 0.5 mg l−1). The induced callus was transferred to the regeneration media with factorial combinations of IAA (1.0 mg l−1 and 2.0 mg l−1) and kinetin (0.5 mg l−1 and 1.0 mg l−1). S. sudanense regenerated at significantly higher frequency (38.91%) and produced shoots more intensely (2.2 shoots/callus) than S. bicolor (26.93%, 1.26 shoots/callus). Immature inflorescences regenerated at a much higher frequency (46.48%) and produced significantly more number of shoots (2.71 shoots/callus) than immature embryos (22.35%, 0.99 shoots/callus). Moreover, differences for plant regeneration between genotypes of the same species were minimal when using immature inflorescences. Increase in the 2,4-D concentration in callus induction media exhibited inhibitory effect on callus induction, growth, shoot induction and number of shoots/callus but inclusion of kinetin in callus induction media improved these responses. Use of immature inflorescence explant and inclusion of kinetin in callus induction media could overcome genotypic limitations of plant regeneration to a large extent. The extent of variability, heritability and expected genetic advance was more in plant regeneration traits than in callus induction traits. This indicated that the variability in respect of these attributes in the genotypes may be due to the additive gene action and selection of genotypes for these characters would be rewarding. 相似文献
17.
Efficient callus induction and plant regeneration from mature embryo culture of winter wheat (Triticum aestivum L.) genotypes 总被引:14,自引:0,他引:14
Immature and mature embryos of 12 common winter wheat (Triticum aestivum) genotypes were cultured in vitro to develop an efficient method of callus formation and plant regeneration from mature embryo
culture, and to compare the responses of both embryo cultures. Fifteen days after anthesis, immature embryos were aseptically
dissected from seeds and placed with the scutellum upwards on a solid agar medium containing the inorganic components of Murashige
and Skoog (MS) and 2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). Mature embryos were moved slightly in the imbibed seeds.
The seeds with moved embryos were placed furrow downwards in dishes containing 8 mg/l 2,4-D for callus induction. The developed
calli and regenerated plants were maintained on 2,4-D-free MS medium. Plants regenerated from both embryo cultures were vernalized
and grown to maturity in soil. Regenerated plantlets all maintained the hexaploid chromosome number. A strong genotypic effect
on the culture responses was found for both explant cultures. Callus induction rate, regeneration capacity of callus and number
of plants regenerated were independent of each other. Mature embryos had a high frequency of callus induction and regeneration
capacity, and therefore, being available throughout the year, can be used as an effective explant source in wheat tissue culture.
Received: 4 February 1997 / Revision received: 1 April 1997 / Accepted: 5 May 1997 相似文献
18.
An embryogenic suspension culture of Zea mays, genotype 4C1, was obtained from friable callus that was cultured on solid medium and had been obtained from zygotic embryos. The suspension contained non-dividing elongated cells, clusters of dividing isodiametric cells, and globular, ovoid, and polar stages of somatic embryos. The single somatic embryos were blocked in shoot meristem formation: when transferred to regeneration medium they developed a root and, at the shoot side, a green cap with meristematic cells, but a scutellum and leaf primordia were not formed. In medium containing 2,4-dichlorophenoxy acetic acid, somatic embryos formed embryogenic callus aggregates, consisting of globular stage somatic embryos attached to each other via undifferentiated callus cells. These somatic embryos developed into mature embryos with the zygotic histological characteristics, such as scutellum and leaf primordia, in maturation medium, and then regenerated into plants in regeneration medium. By omitting the maturation phase, regeneration occurred via organogenesis. Polyembryos, i. e. embryos attached to each other without callus tissue in between, behaved as single somatic embryos. It is concluded that the attached callus tissue provides a factor that stimulates scutellum and leaf primordia formation.Abbreviations CMM
callus maintenance medium
- 2,4D
2,4-dichlorophenoxy acetic acid
- PCV
packed cell volume
- MS
Murashige and Skoog medium 相似文献
19.
20.
Cotyledon explants of Brassica tournefortii L. were excised from germinated seedlings and cultured on Murashige & Skoog's [6] basal medium supplemented with various combinations of cytokinins and auxins, Both cytokinin and auxin were required for induction of shoot organogenesis. Of the three cytokinins tested (in combination with a low concentration of IAA), kinetin was found to be the best for shoot regeneration. On this medium, cotyledonary explants invariably underwent callusing followed by multiple shoot formation. NAA in combination with any of the three cytokinins yielded a reduced number of shoots or none, but favoured good callus growth. Callus so produced also regenerated shoots when subcultured on media containing high concentration of KIN or ZEA and low concentration of IAA. Shoots were rooted during prolonged incubation on the same medium or on MS medium free of growth regulators. Mature plants were grown in the greenhouse. 相似文献