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1.
Summary This paper investigates maintenance and proliferation of somatic embryogenesis systems for Ulmus minor and U. glabra. Proliferation occurred with subculture of embryogenic calluses. The calluses were mainly formed by friable nodules composed
of meristematic cells organized into proembryogenic cell masses (PEMs) and thin-walled vacuolated parenchymatic cells. Cotyledonary
embryos, with procambial strands and differentiation of their vascular tissues as well as visible root meristems, were identifiable
after 18d of culture on a proliferation medium with 0.44 μM benzyladenine (BA). The shoot meristem was only occasionally well developed. Somatic embryo multiplication from elm embryogenic
calluses is a clearly asynchronic system, and PEMs as well as embryos at all stages of development are observed simultaneously
at the end of subculture period. Factors affecting the proliferation of elm embryogenic callus, such as culture medium, carbon
source and genotype, were studied. Basal medium (MS) or medium supplemented with 0.44 μM BA produced the highest number of somatic embryos. Somatic embryo production was higher with sucrose or glucose than with
maltose, and significant differences were also found among the four embryogenic lines tested. The use of liquid medium with
filter paper support is an essential step for the survival of isolated somatic embryos during the germination stage. The addition
of 0.22 μM BA′ to liquid MS medium was the best treatment for germination and plantlet conversion of elm somatic embryos. 相似文献
2.
S. Hindson A. R. McElroy C. Portelance 《In vitro cellular & developmental biology. Plant》1998,34(3):181-184
Summary Various preconditioning treatments of alfalfa (Medicago sativa L.) somatic embryos to improve embryo quality and conversion were studied. Four different regenerating genotypes were compared.
Embryogenic cultures were established in liquid culture. Globular embryos were collected and plated on an embryo development
medium until they reached cotyledonary stage. They were then exposed to three treatments: a standard embryo development medium
(control), media supplementation with 1 μM abscisic acid (ABA), 50 mM glutamine and 5% sucrose (T), additional supplementation with 50 μM ABA (TT), and additional supplementation followed by desiccation (TTD). Treatments affected embryo conversion, but not uniformly
for all genotypes. Embryo conversion was increased (P<0.05) by pretreatment (T), while only one exhibited any response to additional ABA (T vs. TT). Desiccation decreased (P<0.05) conversion of pretreated embryos (TT vs. TTD) of all genotypes. The effect of treatments on plantlet weight was less
pronounced and inconsistent across genotypes. 相似文献
3.
A desiccation protocol was developed to evaluate the effect of different levels of desiccation on germination and plantlet
regeneration of black spruce somatic embryos. Large desiccation chambers (80 l) with four liters of saturated salt solutions
provided constant relative humidities (RH) of 63, 79, 88, and 97% (± 2%). Under these conditions, an embryo mass of 10 mg
always dried fast even at 97% RH. In contrast, an embryo mass of 80 mg generated different kinetics of water loss, from fast
drying at 63% RH to slow drying at 97% RH. Drying rates similar to those obtained with 80 mg embryos were also generated by
combining 40 mg embryos with 40 mg water. The effects of drying rate and embryo MC on germination rate, root elongation, and
plantlet regeneration were examined. A fast drying rate to 4–5% embryo MC, obtained under 63% RH, was detrimental to germination
and plantlet development. However slower drying rates, obtained under 79–97% RH and generating 7–19% MC in the embryos, gave
developmental responses similar to the control. Synchronization of root emergence was improved only for embryos desiccated
to approx. 16% MC under 97% RH. The optimal desiccation protocol using large desiccation chamber at 97% RH and a constant
embryo mass of 40 mg embryos plus 40 mg water was applied to five genotypes of black spruce. For all genotypes, desiccated
embryos gave plantlet regeneration rates similar to the control undesiccated embryos.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
4.
Two Phalaenopsis orchids, Phalaenopsis amabilis and Phalaenopsis ‘Nebula’, were used to test the effects of induction period (30, 45 and 60 days), subculture period (30, 45 and 60 days),
and explant length (1, 1.5 and 2 cm) on direct somatic embryogenesis from different regions (leaf tip, adaxial side, abaxial
side and cut end) of leaf explants from in vitro grown seedlings. The results showed that the cut end had a highest competence
to form embryos than the other regions of the leaf explants from both orchids. In addition, the suitable culture conditions
were 60 days for induction period in darkness, 45 days for subculture period in light and 1 cm for explant length. Besides,
the combinations of N6-benzyladenine (BA) and naphthaleneacetic acid were tested on their effects on plantlet conversion and further development
of leaf-derived embryo. It was found that 0.5 mg/l of BA showed the highest response on plantlet conversion rate and the lowest
browning rate of explants. In this communication, the embryo structures and development were proved by scanning electron microscopy. 相似文献
5.
The factors influencing somatic embryo maturation, high frequency somatic embryo germination, and plantlet formation were
studied in Terminalia chebula Retz. Maturation of somatic embryo were influenced by a number of factors such as in vitro culture passage, concentrations
of sucrose, levels of abscisic acid (ABA), basal media and media additive combinations. Maximum frequency of somatic embryo
maturation (57.22 ± 2.02), was obtained on MS medium supplemented with 50 g/l sucrose. Different factors such as strengths
of MS nutrients, plant growth regulators, media additives and their combinations controlling somatic embryo germination and
plantlet formation were studied. High frequency of germination and plantlet formation (58.80 ± 1.47) were achieved by subsequent
subculture of mature somatic embryos on MS medium containing 30 g/l sucrose and 0.5 mg/l benzyladenine (BA). However, although
duration of in vitro passage of the callus tissue was critical, contribution of the combinations of plant growth regulators
and media additives showed nugatory effect on somatic embryo maturation and germination as evident from variable responses. 相似文献
6.
Onay A. Jeffree C.E. Theobald C. Yeoman M.M. 《Plant Cell, Tissue and Organ Culture》2000,60(2):121-129
Embryogenic masses (EMSes) of pistachio (Pistacia vera L.) were proliferated in liquid Murashige and Skoog (MS) medium without growth regulators. To determine the effects of benzylaminopurine
(BAP), racemic (±) abscisic acid (ABA) and sucrose treatments during maturation on the subsequent germination and plantlet
regeneration, clusters of mature somatic embryos were transferred from maturation medium onto the surface of 0.7% agar-solidified
Murashige and Skoog medium. Neither germination nor plantlet development medium contained BAP or ABA. Germination studies
were carried out using 80 somatic embryos at every combination of four sucrose concentrations, three maturation periods and
either five concentrations of BAP or four of ABA, and the numbers germinating were recorded after four durations of culture.
A similar experimental plan was used to study plantlet regeneration. The number of germinated somatic embryos increased markedly
with duration of the culture on germination medium, and was influenced by the concentrations of BAP or ABA in the maturation
medium; the concentration of sucrose in this medium had little influence. Plantlet regeneration also increased with culture
duration and was reduced at the highest levels of BAP or ABA; with ABA, the probability of plantlet regeneration was lower
for longer maturation periods. ABA and BAP have similar effects on somatic embryo germination (except at the highest levels
used), but BAP is superior to ABA for promoting subsequent plantlet regeneration. Linear logistic models were used to investigate
the significance of the treatments, and to estimate the optimum conditions for germination and plantlet regeneration.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
7.
Fernando Pliego-Alfaro Richard E. Litz Pamela A. Moon Dennis J. Gray 《Plant Cell, Tissue and Organ Culture》1996,44(1):53-61
Development of cotyledonary-stage nucellar embryos of mango was arrestedin vitro by exposure to 750–1750 M ABA. The enlargement and germination of nucellar embryos was inhibited for as long as 4 weeks after subculture from ABA-containing medium. Mannitol at concentrations between 7.5 and 12.5% inhibited nucellar embryo development, presumably due to osmotic effects; however, there was no residual effect after subculture of somatic embryos onto medium without mannitol. Temperatures between 22.5 and 37.5°C stimulated embryo development, whereas lower temperatures (7.5 and 15°C) delayed germination. There was no germination 1 month after somatic embryos, pulsed for 8 weeks at 7.5°C, were transferred to 22.5°C; however, after 2 months, 86% of these somatic embryos germinated. These results indicate that it is possible to induce developmental arrest in recalcitrant mango embryos with high concentrations of ABA, mannitol or low temperature (7.5°C).Abbreviations ABA
Abscisic acid
- MM1
Mango maturation medium 相似文献
8.
In vitro somatic embryogenesis from cell suspension cultures of cowpea [Vigna unguiculata (L.) Walp]
We report, an efficient protocol for plantlet regeneration from the cell suspension cultures of cowpea through somatic embryogenesis. Primary leaf-derived, embryogenic calli initiated in MMS [MS salts (Murashige and Skoog 1962) with B5 (Gamborg et al. 1968) vitamins] medium containing 2,4-Dichlorophenoxyacetic acid (2,4-D), casein hydrolysate (CH), and l-Glutamic acid-5-amide (Gln). Fast-growing embryogenic cell suspensions were established in 0.5 mg l–1 2,4-D, which resulted in the highest recovery of early stages of somatic embryos in liquid MMS medium. Embryo development was asynchronous and strongly influenced by the 2,4-D concentration. Mature monocotyledonary-stage somatic embryos were induced in liquid B5 medium containing 0.1 mg l–1 2,4-D, 20 mg l–1 l-Proline (Pro), 5 M Abscisic acid (ABA), and 2% mannitol. B5 medium was found superior for the maturation of somatic embryos compared to MS and MMS media. The importance of duration (5 d) for effective maturation of somatic embryos is demonstrated. A reduction in the 2,4-D level in suspensions increased the somatic embryo induction and maturation with decreased abnormalities. Sucrose was found to be the best carbon source for callus induction while mannitol for embryo maturation and maltose for embryo germination. Extension of hypocotyls and complete development of plantlet was achieved in half-strength B5 medium supplemented with 3% maltose, 2500 mg l–1 potassium nitrate, and 0.05 mg l–1 thidiazuron (TDZ) with 32% regeneration frequency. Field-established plants were morphologically normal and fertile. This regeneration protocol assures a high frequency of embryo induction, maturation, and plantlet conversion. 相似文献
9.
Elena Corredoira Silvia Valladares Ana M. Vieitez Antonio Ballester 《In vitro cellular & developmental biology. Plant》2008,44(4):307-315
For the mass production of chestnut trees with selected, hybrid, or genetically engineered genotypes, one potentially desirable
propagation strategy is based on somatic embryogenesis. Although methods exist for the initiation of embryogenic cultures
of Castanea sativa from immature zygotic embryos or leaf explants, the embryos produced have had low rates of conversion into plantlets. This
study explored the possible benefits for somatic embryos that have already undergone maturation and cold treatments, of (a)
partial slow or fast desiccation, and (b) of the addition of plant growth regulators or glutamine to the germination medium.
Germination response was evaluated in terms of both conversions to plantlets and through embryos developing only shoots (shoot
germination) that could be rooted following the micropropagation protocols developed for chestnut. Two or 3 wk slow desiccation
in sealed empty Petri dishes resulted in a slight reduction in water content that nevertheless increased total potential plant
recovery, shoot length, and the number of leaves per plantlet. However, best results were achieved by 2 h fast drying in a
laminar flow hood, which reduced embryo moisture content to 57–58% and enhanced the potential plant recovery and quality of
regenerated plantlets. Plant yield was also promoted by addition of 0.44 μM benzyladenine and 200–438 mg/l of glutamine to
the germination medium, and plantlet quality (as evidenced by root, shoot, and leaf growth) by the further addition of 0.49 μM
indole-3-butyric acid. 相似文献
10.
Somatic embryogenesis of Cyclamen persicum in liquid medium 总被引:1,自引:0,他引:1
Marc Kreuger Erik Postma Yvon Brouwer Gerrit-Jan van Holst 《Physiologia plantarum》1995,94(4):605-612
A method is described for the production of somatic embryos of Cyclamen persicum Mill. in liquid medium. Five steps are involved; initiation of embryogenic cell lines, proliferation of pro-embryogenic masses (PEMs) on auxin-containing medium, development of somatic embryos on hormone-free medium with high osmolarity, germination and subsequent plantlet formation. Cell lines were initiated by culturing the explant, the seedling tuber, directly in liquid medium. Three parameters were important for obtaining embryogenic cell lines; explant density, hormone concentrations and subculture regime. The rate of uptake of the hormones 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin influenced the formation of PEMs. Highly embryogenic cell lines were obtained only when PEMs had formed within 5–7 weeks. PEMs were proliferated for at least 24 months and could be isolated from each subculture for the production of somatic embryos. A high sucrose content (175 m M ) in the development medium without hormones ensured efficient embryo development from PEMs. A subsequent subculture in low sucrose concentration (58 m M ) induced the formation of a tuber, thus promoting germination. Arabinogalactan-proteins (AGPs) from carrot seeds and AGPs bound by the monoclonal antibody ZUM 18 increased the number of PEMs in a culture, showing that the activity of AGPs is not species specific. 相似文献
11.
Gui Rong Li Wei Ji Gang Wang Jian Xia Zhang Yue Jin Wang 《In vitro cellular & developmental biology. Plant》2014,50(1):110-120
A highly efficient technique of embryo rescue is critical when using stenospermocarpic Vitis vinifera cultivars (female parents) to breed novel, disease-resistant, seedless grape cultivars by hybridizing with wild Chinese Vitis species (male parents) having many disease-resistance alleles. The effects of various factors on the improvement of embryo formation, germination, and plantlet development for seven hybrid combinations were studied. The results indicated that Beichun and Shuangyou were the best male parents. The best sampling time for ovule inoculation differed among the female parents. When hybrid ovules were cultured on a double-phase medium with five different solid medium types, percent embryo formation was highest (11.3–28.3%) on a modified MM3 medium. Percentages of embryo germination (15.4–55.4%) and plantlet development (11.15–44.6%) were all highest when embryos were cultured on Woody Plant Medium?+?5.7 μM indole-3-acetic acid?+?4.4 μM 6-benzylaminopurine?+?1.4 μM gibberellic acid?+?2% sucrose?+?0.05% casein hydrolysate?+?0.3% activated charcoal?+?0.7% agar. In the absence of other amino acids, the addition of proline significantly increased embryo formation (36.1%), embryo germination (64.6%), and plantlet development (90.5%). A highly efficient protocol has been developed for hybrid embryo rescue from seedless V. vinifera grapes?×?wild Chinese Vitis species that results in a significant improvement in breeding efficiency for new disease-resistant seedless grapes. 相似文献
12.
该研究以蔗糖、麦芽糖、山梨醇及PEG(6000)为渗透剂,探讨了不同渗透剂对白刺花体细胞胚发育、胚成熟及萌发的影响。结果表明:白刺花下胚轴形成的胚性愈伤组织接种至MS+2,4-D 0.2 mg·L~(-1)+NAA 1.0 mg·L~(-1)+6-BA 2.0 mg·L~(-1)+TDZ 1.0 mg·L~(-1)+蔗糖40 g·L~(-1)+谷氨酰胺100 mg·L~(-1)+植物凝胶3g·L~(-1)的培养基上,体细胞胚发生率高达66. 21%,总胚数为79个; 7%蔗糖可使体细胞胚成熟率高达64.36%,同时也可提高多子叶畸形胚形成; 2%麦芽糖+2%山梨醇+4%蔗糖组合使体细胞胚成熟率最高达88.89%,畸形胚比例最低; 30 g·L~(-1)PEG培养时,体细胞成熟率最高,为82.35%;鱼雷期的体细胞胚最合适转接,可使体胚萌发率达90.58%,复合糖上培养得到的成熟体细胞胚生根率最高,为87.47%。这为实现白刺花体细胞胚育苗奠定了理论基础,并提供了可行的方案。 相似文献
13.
Effects of media on embryo enlargement,germination and plant development in early-ripening genotypes of Prunus grown in vitro 总被引:1,自引:0,他引:1
Embryo culture of 5–10 mm long embryos of Prunus was investigated. The effects of various media on embryo enlargement, germination and plant formation were compared. Results show that embryos in all genotypes enlarged during stratification on any tested medium. Beneficial embryo enlargement, germination and plant development of peach and nectarine occurred when cultured on WP medium. The embryos of a plum were more responsive to C2d medium for enlargement, germination and plant development. All genotypes germinated well with a large number of embryos growing into plants on WP and C2d media. 相似文献
14.
A protocol for induction of direct somatic embryogenesis, secondary embryogenesis and plant regeneration of Dendrobium cv. Chiengmai Pink was developed. Thidiazuron (TDZ) at 0.3, 1 and 3 mg dm−3 induced 5–25 % of leaf tip segments of in vitro grown plants to directly form embryos after 60 d of culture, and 1 mg dm−3 TDZ was the best treatment. Somatic embryos mostly formed from leaf surfaces near cut ends, and occasionally found on leaf
tips. Higher frequency of embryogenesis was obtained in light than in darkness. During subculture, secondary embryos developed
from outer cell layers of primary embryos. All combinations of NAA (0, 0.1, 1 mg dm−3) and TDZ (0, 0.3, 1, 3 mg dm−3) increased the multiplication rate of embryos. It takes about 8 months from embryo induction, plantlet formation to eventually
acclimatization in greenhouse. 相似文献
15.
In order to study the salinity tolerance of pistachio (Pistacia vera L.), embryos developed from mature seeds were isolated and cultured in vitro and subjected to different NaCl concentrations (0, 42.8, 85.5, 171.1 and 256.6 mM) for 30 days. The results showed that in vitro germination of embryonic axes was not affected by the salt concentration. However, the germinated embryo survival rates decreased from 100% for the control to 62.9% for the highest salt concentration (256.6 mM).In addition, the plantlet growth (length of aerial and root parts, number of leaf produced per embryo, as well as the production of total fresh and dry matter for both aerial parts and roots) showed significant differences according the various salt concentrations. To cite this article: B. Benmahioul et al., C. R. Biologies 332 (2009). 相似文献
16.
17.
M. Danin S. J. Upfold N. Levin B. L. Nadel A. Altman J. van Staden 《Plant Growth Regulation》1993,12(3):245-254
The effect of inhibitors of polyamine biosynthesis on the development of embryogenic cell cultures of celery (Apium graveolus L.) was studied. Several developmental stages of somatic embryos were compared for differences in the content and biosynthesis of free polyamines and for cytokinin content. Cyclohexylamine and particularly methylglyoxal bis(guanylhydrazone), inhibited both cell division and the organization of polar embryos from globular embryos. Difluoromethylornithine slightly promoted embryo development, especially cell division.The free putrescine content of globular embryos was 6-fold that of fully differentiated plantlets, and that of spermidine 2-fold. Only a slight increase in the spermine content was found with embryo development. These differences were confirmed by data from polyamine biosynthesis. Incorporation of 14C-arginine into polyamines was slightly higher than that of 14C-ornithine. Over 96% of this incorporation was detected in the putrescine fraction. Incorporation of 14C into putrescine in globular embryos was 3 to 4-fold that in fully-differentiated plantlets. Incorporation into spermidine and spermine was, however, higher in plantlets than in globular embryos.Cytokinin analysis revealed considerable differences in the biological activity between the developmental stages of embryogenesis. This could be due to endogenous cytokinins and/or BA taken up from the maintenance medium. Cytokinin levels decreased with increased embryo development. Most of the detected cytokinin-like activity co-chromatographed with BA and its metabolites. Some as yet unidentified peaks of activity were recorded in the globular embryos.The results are considered with respect to the possible participation of polyamines and cytokinins in the development of embryogenic cell cultures of celery. It is suggested that the onset of embryogenesis is characterized by a high content of putrescine and cytokinins, while a decrease in putrescine synthesis and cytokinin content, and an increase in spermidine and spermine content, accompany further embryo development and plantlet formation.Abbreviation ADC
arginine decarboxylase
- ODC
ornithine decarboxylase
- 2,4-D
dichlorophenoxyacetic acid
- DFMA
difluoromethylarginine
- DFMO
difluoromethylornithine
- MGBG
methylglyoxal bis(guanylhydrazone)
- CHA
cyclohexylamine
- BA
benzyladenine
- BAR
benzyladenine riboside 相似文献
18.
Summary Somatic embryogenesis has been shown to be an imperfect recapitulation of stages involved to form embryos from vegetative
tissues. Although abscisic acid has been implicated in normalizing development, studies that specifically investigate conversion
(vegetative leaf initiation) in somatic embryos are lacking. This report documents a follow-up of a study that implicated
abscisic acid as a vital factor in allowing embryos ofDaucus carota to progress to the plantlet stage. Abscisic acid was determined to enhance conversion at doses ranging from 1 to 50 µM. Younger embryo stages were more responsive to abscisic acid application with regards to plantlet recovery. Pulses of abscisic
acid were shown to elicit more rapid response with younger embryo stages, indicating more plastic development. Fluridone,
an abscisic acid synthesis inhibitor, was shown to dramatically reduce conversion, even at low doses (<5µM). When abscisic acid was applied concurrently with fluridone, partial restoration of conversion was observed. Histologically,
fluridone was seen to cause pronounced vacuolation in the shoot apical notch which resulted in the loss of meristematic cells,
negating conversion capacity. Quantitation of total cytoplasmic area showed that abscisic acid reduced vacuolar intrusion
into the apical notch, while fluridone caused a significant increase in vacuolation of cells in this region. This report documents
further evidence of a role for abscisic acid in plantlet establishment from somatic embryos ofDaucus carota. 相似文献
19.
Cheng-Hao Li Bao-Guang Liu Tae-Dong Kim Heung-Kyu Moon Yong-Eui Choi 《Plant biotechnology reports》2008,2(4):259-265
Picea koraiensis, called Korean spruce, is an evergreen tree and found mostly in northeast Asia. In this study, plant regeneration via somatic
embryogenesis from open-pollinated immature zygotic embryos of nine genotypes of elite trees was established. Immature zygotic
embryos were cultured onto RJW medium modified from 505 medium with 21.48 μM NAA, 2.22 μM BA, and 2.32 μM KT. The average
frequency for all nine genotypes was 74.2%. Embryogenic calluses of the nine genotypes of elite trees were subcultured on
RJW basal medium containing 8.06 μM NAA, 1.11 μM BA, and 1.16 μM kinetin. The calluses of three lines, 3#, 9#, and 2#, were actively proliferated but others were not. Somatic embryogenesis was induced from the embryogenic callus in genotypes
of 3#, 9#, and 2# on RJW medium with ABA and 60 g l−1 sucrose. Cotyledonary somatic embryos were subjected to a drying process. The drying of embryos by uncapping the culture
bottle for 5 days on a clean bench resulted in a high frequency of germination of somatic embryos (87% in RJW medium). However,
plantlet conversion from germinated embryos was greatly reduced and the optimal medium for plant conversion was 1/2 WPM or
1/2 BMI medium. In conclusion, we have, for the first time, established a plant regeneration system via somatic embryogenesis
in the Korean spruce, which can be applied for rapid micropropagation of elite trees. 相似文献
20.
Embryogenic cultures of lucerne (Medicago sativa L.) cv. Robothave been established and propagated on medium containing yeastextract. These cultures consisted of unorganized callus tissuebearing embryogenic centres which increased in size during subculture,yielding new regenerated somatic embryos at the end of each20-d subculture. A development in the propagation of the embryogenic cultureswas the establishment of single embryo culture in hormone-freemedium where, in selected cases, the process of recurrent somaticembryogenesis (RSE) took place on the hypocotyl of explantedembryos. The process was independent of supporting callus tissueand occurred on simple defined medium. Single embryos underwenteither plantlet development or continued RSE on the hypocotyl.One third of the regenerated plantlets showed RSE after thetwo to three trifoliate leaf stage. In these cases shoot developmentstopped and only somatic embryo production took place. In vitrocloning of regenerated plantlets allowed us to reproduce eachparticular genotype before transplantation into soil. Lucerne (alfalfa), Medicago sativa L., somatic embryogenesis, single embryo culture 相似文献