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1.
Monika Höfer 《Acta Physiologiae Plantarum》2005,27(4):709-716
Based on optimized protocols for anther and microspore culture in apple (Malus x domestica Borkh.), the regeneration phase and the efficiency of the processes in general were compared by using the same androgenic material of two experimental years. Microspore culture resulted in an increase in embryo induction depending on the genotype (Höfer 2004), however anther culture was superior to microspore culture in the total number of regenerated plants. The regeneration process in anther and microspore culture is similar. Two developmental pathways were observed: 1) secondary embryogenesis followed by adventitious shoot formation and 2) direct adventitious shoot formation from primary embryos. Induction and regeneration processes are delayed in microspore culture as compared with anther culture. The reasons for the reduced regeneration efficiency in microspore culture are discussed. 相似文献
2.
Stress is an essential component during embryogenesis induction in microspore culture. Cold pretreatment has been used in cereal microspore culture but very seldom attempted in Brassica microspore culture. The effect of cold pretreatment of flower buds subjected to a liquid medium on microspore embryogenesis was investigated in spring and winter Brassica napus, as well as in B. rapa and B. oleracea. Cold pretreatment significantly enhanced microspore embryogenesis (by 1–7 fold) compared to commonly used microspore culture protocol in B. napus, while it was less effective in B. rapa or even negative in B. oleracea. The appropriate duration of cold pretreatment was found to be 2–4 days, which stimulated the best microspore embryogenesis. Cold pretreatment was also able to promote embryo development including the improvement of embryo quality and acceleration of embryogenesis. When incorporating with medium refreshing, cold pretreatment could initiate the most microspore embryogenesis than any other treatment used. With further improvement cold pretreatment method may have a positive potential in Brassica breeding programmes. 相似文献
3.
Isolated tobacco (Nicotiana tabacum L.) microspores maturing in vitro can be induced to undergo symmetrical divisions, instead of the normal asymmetrical first pollen mitosis, by addition of anther extracts to the culture medium. The two daughter cells in symmetrically divided pollen resemble vegetative pollen cells in cytological characteristics, nuclear size and chromatin condensation, are separated by a cell wall and remain viable during in vitro maturation. After transfer to a germination medium, only one of the two vegetativelike cells forms a pollen tube in vitro. Therefore, apparently normal gametophytic development can be maintained after symmetrical microspore division. These results are discussed in relation to current models for induction of microspore embryogenesis. 相似文献
4.
Induction of embryogenesis from isolated apple microspores 总被引:2,自引:0,他引:2
We report, for the first time, the induction of embryogenesis and plant formation from isolated apple (Malus domestica Borkh.) microspores in vitro. Different isolation techniques were tested and an optimized protocol was elaborated. Furthermore,
the influence of the induction medium and starvation treatment, using different starvation material, temperatures and time,
were studied. In addition to embryo induction, the number of multicellular structures per divided microspores was found to
be a suitable parameter of assessment and could be used in earlier stages during microspore culture. Although the number of
embryos induced in these first experiments is low, the best frequency of embryo induction was shown to be at least twice as
efficient as that obtained by anther culture.
Received: 9 September 1998 / Revision received: 22 December 1998 / Accepted: 12 January 1999 相似文献
5.
Iwona Żur Ewa Dubas Elżbieta Golemiec Magdalena Szechyńska-Hebda Gabriela Gołębiowska Maria Wędzony 《Plant cell reports》2009,28(8):1279-1287
Isolated microspore cultures of two spring triticale (x Triticosecale Wittm.) cultivars were used to examine the effect of various stress treatments (either high—32°C or low—5°C temperature with
or without nitrogen/carbohydrate starvation) applied to excised anthers on the effectiveness of microspore embryogenesis induction.
To quantify the effects of pretreatment conditions, the activity of antioxidative enzymes (catalase, peroxidase and superoxide
dismutase) together with respiration rate and heat emission were measured. It was observed that heat shock treatment applied
as the only one stress factor increased the activity of antioxidative enzymes which suggests intensive generation of reactive
oxygen species. Such pretreatment effectively triggered microspore reprogramming but drastically decreased microspore viability.
After low temperature treatment, the activity of antioxidative enzymes was similar to the control subjected only with the
stress originated from the transfer to in vitro culture conditions. This pretreatment decreased the number of microspores
entering embryogenesis but sustained cell viability and this effect prevailed in the final estimation of microspore embryogenesis
effectiveness. For both, low- and high-temperature treatments, interaction with starvation stress was beneficial increasing
microspore viability (at 5°C) or efficiency of embryogenesis induction (at 32°C). The latter treatment significantly reduced
cell metabolic activity. Physiological background of these effects seems to be different and some hypothetical explanations
have been discussed. Received data indicate that in triticale, anther preculture conditions could generate oxidative stress
and change the cell metabolic activity which could next be reflected in the cell viability and the efficiency of microspore
embryogenesis. 相似文献
6.
The morphogenic response of anther walls and connective tissue is the greatest obstacle to androgenesis in soybean anther culture. Whereas induction to microspore embryogenesis occurs in the dark in almost all plant species, soybean anthers have been cultured under light. In an attempt to establish culture conditions that simultaneously stimulate microspore embryogenesis and inhibit epidermal and connective cell proliferation, the effect of light and two 2,4-dichlorophenoxyacetic acid (2,4-D) concentrations (2 and 10 mg l–1) on the induction process was investigated. Higher 2,4-D concentration speeded up microspore plasmolysis and did not improve androgenesis. Callogenesis and embryogenesis induction from sporophytic cells were significantly lower in the dark, and some microspores showed major alterations in the sporoderm. Auxin 2,4-D and induction under light contributed to the morphogenic response of the anther walls and connective tissue under the conditions previously recommended to trigger microspore embryogenesis. 相似文献
7.
8.
Efficient production of doubled haploid plants by immediate colchicine treatment of isolated microspores in winter Brassica napus 总被引:1,自引:0,他引:1
The effect of colchicine on embryogenesis induction and chromosomedoubling during microspore culture was evaluated in two F1 hybridsofwinter oilseed rape (Brassica napus L.). Colchicinetreatment (50 and 500 mg/L) of isolated microspores during thefirst 15 h in culture stimulated embryogenesis and produced large amounts ofhealthy-looking embryos. These normal embryos germinated well at 24°C after being transferred to solid regeneration medium and aninitial period of low temperature (2 °C) for 10 days, andcoulddirectly and rapidly regenerate vigorous plants. A high doubling efficiency of84–88% was obtained from 500 mg/L colchicine treatment for15h with low frequency of polyploid and chimeric plants. Acolchicinetreatment duration of 6 h was less effective on embryogenesis anddoubling efficiency. The present experiment also showed that changing of induction medium 15h after microspore isolation produced higher spontaneous doublingefficiency, as compared with medium change 6 h after isolation. 相似文献
9.
María Antonietta GermanàBenedetta Chiancone Diego PadoanIvett Bárány María-Carmen RisuenoPilar S. Testillano 《Environmental and Experimental Botany》2011,71(2):152-157
Prunus armeniaca L. is a worldwide known species, very important particularly in the Mediterranean basin. Microspore embryogenesis through in vitro anther culture is a widely used method to obtain haploid and doubled haploid (DHs) plants which are being routinely used in breeding programmes for new superior cultivar development in many crops. Haploid-diploidization through gametic embryogenesis allows single-step development of complete homozygous lines from heterozygous parents. In the case of fruit crops, with long reproductive cycle, a high degree of heterozygosity, large size, and, often, self-incompatibility, there is no way to obtain haploidization through conventional methods. Induction of microspore embryogenesis in vitro is switched by a stress treatment. In many species, heat or cold stress has been reported to trigger pollen embryogenesis, the response being genotype dependent. In the present work we analyzed whether microspore reprogramming could be induced in apricot cultivars by cold stress through anther culture. We report the development of an in vitro anther culture protocol in P. armeniaca L. and analyse the response of several cultivars to stress treatments and culture media for inducing pollen embryogenesis. Results showed the formation of multicellular pollen and proembryos. The effect of two culture media in the embryogenic response was also analyzed, being the responses genotype-dependent. Monitoring of the cellular changes on the microspores was performed by structural and confocal microscopy analyses. Results indicated that the reprogramming of the microspore and the first steps of the embryogenic pathway have been achieved in different varieties of P. armeniaca, which constitutes a crucial step in the design of protocols for the regeneration of microspore-derived embryos and DH plants, for future potential applications in breeding programmes of this economically important fruit tree. 相似文献
10.
Isolated microspore culture techniques and recent progress for haploid and doubled haploid plant production 总被引:2,自引:0,他引:2
An isolated microspore culture provides an excellent system for the study of microspore induction and embryogenesis, provides
a platform for an ever-increasing array of molecular studies, and can produce doubled haploid (DH) plants, which are used
to accelerate plant-breeding programs. Moreover, isolated microspore cultures have several advantages over anther culture,
wherein presence of the anther walls can lead to the development of diploid, somatic calli and plants. Although protocols
for isolated microspore culture vary from laboratory to laboratory, the basic steps of growing donor plants, harvesting floral
organs, isolating microspores, culturing and inducing microspores, regenerating embryos, and doubling the chromosomes, remain
the same. Over the past few years, a large proportion of the research reports on isolated microspore culture have focused
on cereal and Brassica species. For some of these species, isolated microspore culture protocols are well established and routinely used in laboratories
around the world for developing new varieties, as well as for basic research in areas such as genomics, gene expression, and
genetic mapping. Although these species are considered highly responsive to microspore culture, improvements in efficiency
are still being made. However, with many species, isolated microspore culture is simply not yet efficient enough at producing
DH plants to be cost-effective for breeding programs. There has been a recent resurgence of haploidy research with response
being reported in some species once considered recalcitrant. Future research programs aimed at elucidating pathways involved
in microspore induction and embryogenesis will be of benefit, as will novel approaches to improve the efficiency of microspore
culture for DH production. With many species, anther culture has proven to be more effective than isolated microspore culture,
necessitating more research to clarify the contribution of the anther wall to embryogenesis. The development of molecular
markers for use in determining the gametic origin of regenerated plants, irrespective of their ploidy, would also be beneficial.
In this review, we aim to provide an overview of the basic isolated microspore culture protocol with an emphasis on recent
progress in several crop species. 相似文献
11.
J. Siebel K. P. Pauls 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1989,78(4):473-479
Summary A direct comparison of microspore culture and anther culture was made in Brassica napus using F1 crosses of Regent (canola) by Golden (rapeseed), and their reciprocals, as well as a hybrid between Reston and a highly embryogenic, canola-quality breeding line (G231) as donor plants. The study confirmed that microspore culture can be ten times more efficient than anther culture for embryo production. Embryo yields from cultures initiated from the Reston x G231 were four-fold greater than those initiated from the Regent x Golden crosses, and significant differences were also detected among cultures initiated from the different Regent x Golden crosses. These results illustrate the influence that donor plant genotype has on embryo production. However, superior embryogenic potential among donor material was not always coincident with superior plant production. The average haploid-todiploid ratio in microspore-derived regenerates was 21 for the population obtained from the Regent x Golden crosses but 11 for the Reston x G231 cross. For both types of material, the frequency of diploids increased upon repeated cycles of explanting. A field study showed that there were no differences between the populations of anther-derived and microspore-derived spontaneous diploid and doubled haploid lines, with respect to the days required for them to flower or to mature. The information is valuable for canola breeding programs considering the use of haploidy. 相似文献
12.
Pradeep K. Agarwal Parinita Agarwal Jan B. M. Custers Chun-ming Liu S. S. Bhojwani 《Plant Cell, Tissue and Organ Culture》2006,86(2):201-210
An efficient protocol to improve microspore embryogenesis is established in an important oleiferous crop, Brassica juncea (Indian mustard). Colchicine was used for enhancing microspore embryogenesis and also to obtain doubled haploid embryos. Colchicine at high concentrations (>10 mg l−1), for 24 h, proved convenient for direct recovery of diploid embryos. Higher temperature treatment and an antiauxin PCIB (p-chlorophenoxyisobutyric acid) enhanced microspore embryogenesis significantly as compared to colchicine. An increase in temperature from 32°C to 35°C proved very efficient in increasing embryogenesis by 10-fold. The highest embryogenesis rate was obtained when PCIB was added at 35°C in the culture after 1 day of culture initiation. 20 μM PCIB could enhance microspore embryogenesis by 5-fold. Different abnormal shapes of embryos like lemon, banana, flask and fused cotyledons were observed. Both normal and fused cotyledonous embryos showed normal germination when transferred on the B5 basal medium. 相似文献
13.
M. Cristina Pedroso Salomé Pais 《In vitro cellular & developmental biology. Plant》1993,29(4):155-159
Summary Embryogenesis and callus formation inCamellia japonica L., cv. Elegans and cv. Ville de Nantes (>50 yr old), were higher when anthers at tetrad stage or early uninucleate microspore
stage were used. Direct embryo formation was obtained, both in anthers and anther-petaloids. Embryogenesis only occurred under
light on modified Murashige and Skoog medium supplemented with 6-benzylaminopurine (MS10). Embryo production was higher from
petaloids (2.2 to 3.5 embryos/petaloid) than from anther locules (1.2 to 1.5 embryos/anther). However, petaloid-derived embryos
aborted by Week 10 of culture. The embryogenic efficiency of anthers was 5.3% for cv. Elegans and 4.1% for cv. Ville de Nantes.
Scanning electron microscopy images showed that anther-derived globular embryos were covered by a layer of material of smooth
texture. Shoot regeneration efficiency by organogenesis was 6.8%. Regeneration by embryogenesis was 60% for cv. Ville de Nantes
and 97.5% for cv. Elegans. 相似文献
14.
Shed microspore embryogenesis and fertile plantlet regeneration were observed in a salt susceptible × salt tolerant indica rice F1 hybrid involving IR 24 and CRM 30. The in vitro culture response and regeneration of green plantlets in the hybrid were superior to those of the parents. Direct embryogenesis
and plantlet regeneration with multiple tillers were observed in shed microspore embryos. In intact anther culture, plantlet
development from microspore involved a callus phase. The number of multiple tillers developed through secondary embryogenesis
was almost equal in both the cases. However, the results indicate that regeneration of green plantlets was higher in case
of shed microspore culture in liquid medium containing the synthetic polymer Ficoll 400 than from intact anthers cultured
on a semi-solid system. Shed microspore culture produced a number of double haploids, which may result in far reaching consequences
in genetic improvement of rice.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
15.
The present study involves in vitro androgenesis of Zea mays L. using anther culture. We tested combinations of single factors and their influence on microspore induction. Embryogenic induction of microspores within anthers in in vitro conditions was the best when combination of cold treatment, TIBA (0.1 mg l–1) in media and colchicine (0.02% during first 3 days of culture) was applied, but colchicine alone can be factor, which can stimulate or initiate embryogenesis in anther culture of maize. 相似文献
16.
Scanning electron microscopy was employed to study and compare microspore embryogenesis in vitro with pollen development in planta inBrassica napus andB. oleracea. An exine with its specific pattern had already been formed, when microspores were released from tetrads. During subsequent pollen development, microspores increased in size and continued to strengthen the exine. Upon in vitro culture, all microspores, i.e., embryogenic and nonembryogenic, initially showed the same morphological features. After 24 h in culture, the microspores had increased in size. Thereafter, embryogenesis was indicated in some microspores by two different morphological changes. One featured an expansion in volume of the cell cluster around the germination aperture (type I), the other showed cell cluster volume expansion over the entire microspore surface (type II). Two-thirds of embryogenic microspores in bothB. napus andB. oleracea demonstrated type I development. When followed by fluorescence microscopy, in vitro culture of microspores revealed cultures with a high embryo frequency were those with a high frequency of symmetrical division.Abbreviations
SEM
Scanning electron microscopy
-
TEM
Transmission electron microscopy 相似文献
17.
An isolated microspore culture and green plant regeneration method for rye (Secale cereale L.) was established. Rye isolated microspore androgenesis was genotype-dependent. PG-96M medium supplemented with 6% maltose
gave the highest microspore survival rate after 48 h of culture and the highest embryo/callus yield (930 embryos/calli per
100 anthers from cv. Florida 401). Osmotic pressure in the induction medium played an important role. Pretreatment of the
anthers with mannitol was beneficial for the microspore culture. Embryos/calli of a relatively younger age and smaller size
had a higher regeneration ability, with the best green plant regeneration rate being 6%. Over 150 microspore-derived green
plants have been obtained so far. About 90% of the regenerated plants were spontaneous doubled haploids. This is the first
report of isolated microspore culture in true rye resulting in androgenic embryogenesis and plant regeneration.
Received: 26 April 1999 / Accepted: 23 November 1999 相似文献
18.
Brassica napus L. microspores at the late uninucleate to early binucleate stage of development can be induced in vitro to alter their development from pollen to embryo formation. High temperatures or other stress treatments are required to initiate this redirection process. The critical period for induction of microspore embryogenesis is within the first 8 h of temperature-stress imposition. During this period, which precedes the first embryogenic nuclear division, the process regulating the induction and sustainment of microspore embryogenesis is activated. A number of mRNAs and proteins, some of them possibly heat-shock proteins, appear in microspores during the commitment phase of the induction process.Abbreviations SDS
sodium dodecyl sulfate
- PAGE
polyacrylamide gel electrophoresis 相似文献
19.
Csaba Lantos Lajos Bóna Éva Nagy Ferenc Békés János Pauk 《Plant Cell, Tissue and Organ Culture》2018,133(3):385-393
This is the first report on isolated microspore culture—derived spelt wheat. The efficiency of anther- and isolated microspore was compared using four genotypes (‘Franckenkorn’, ‘GK Fehér’, ‘Mv Martongold’, ‘Oberkulmer Rotkorn’). In anther culture, genotype dependency was observed, and cold pre-treatment enhanced the efficiency of the method. In isolated microspore culture, the ovary co-culture supported the development of embryo-like structures. The presence of growth regulators (0.5 mg/l 2,4-D and 0.5 mg/l kinetin) were not essential for the induction of androgenesis, but these increased the production of embryo-like structures, green and albino plantlets. The low plant regeneration rate and high number of albinos hinder the practical application of isolated microspore culture while anther culture was efficient for in vitro green plantlets production in spelt wheat. The mean of green plantlets production was 41.45/100 anthers (from 20.93 to 83.07 depending on genotype). The phenomenon of albinism was mitigated in anther culture (3.48 albinos/100 anthers). Altogether, 1720 anther culture—derived green plantlets were produced from the four genotypes. 相似文献
20.
Summary. Various stresses such as starvation and cold or heat shocks have been identified as triggers in the induction of the microspore
embryogenesis. This study attempts to quantify the effects of different pretreatment conditions for successful microspore
culture of malting barley (cv. Scarlett). While the sporophytic microspore development could be induced from treated and nontreated
microspores, abiotic stress was essential for embryo formation and plant regeneration. The type of stress treatment applied
affected the numbers and the ratios of albino and green plants regenerated, as well as their fertility. The highest number
of green plants was obtained after the treatment of anthers in 0.3 M mannitol at 32 °C for 24 h before microspore culture.
Correspondence and reprints: Department of Plant Biotechnology and Cytogenetics, Institute of Plant Breeding and Acclimatization,
Radzików, 05-870 Blonie, Poland. 相似文献