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We describe a protocol for somatic embryogenesis of Protea cynaroides, with potential for high frequency production of this important horticultural species. Somatic embryos formed directly on
both P. cynaroides mature zygotic embryos and excised cotyledons cultured on MS medium without growth regulators. The addition of growth regulators
such as naphthalene acetic acid (NAA) (5; 13 and 27 μM) and 2,4-dichlorophenoxyacetic acid (2,4-D) (5; 11 and 23 μM), in combination
with thidiazuron (TDZ) (1 μM), benzylaminopurine (BAP) (1 μM) or kinetin (1 μM) suppressed the formation of somatic embryos.
After eight weeks in culture, formation of somatic embryos was observed. Zygotic explants formed the most embryos when cultured
in a 12-h photoperiod in comparison to explants cultured in the dark. Up to 83% of these embryos germinated after transferal
to the germination medium containing 0.3 μM GA3. Significantly fewer embryos germinated in MS medium with no growth regulators, or supplemented with higher concentrations
of GA3, while low germination percentages were also observed in MS media containing casein hydrolysate and coconut water. The germination
of normal somatic embryos (two separate cotyledons and a single radicle) was observed only in media containing either no growth
regulators, 0.3 μM GA3 or 1 μM GA3. All embryos that germinated in high concentrations of GA3 were malformed. 相似文献
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AtLTP1 luciferase expression during carrot somatic embryogenesis 总被引:4,自引:0,他引:4
Marcel A.J. Toonen John A. Verhees Ed D.L. Schmidt Ab van Kammen Sacco C. de Vries 《The Plant journal : for cell and molecular biology》1997,12(5):1213-1221
The carrot (Daucus carota L.) EP2 gene encodes a Lipid Transfer Protein (LTP) which is expressed during protoderm formation in developing embryos. To develop a vital reporter system for gene expression during somatic embryo development a 1.1 kB fragment of the Arabidopsis thaliana LTP1 promoter was fused to the firefly luciferase (LUC) coding sequence. The AtLTP1 luciferase expression pattern in transformed carrot suspension cultures was identical to the expression pattern of the endogenous carrot EP2 gene. Cell tracking experiments revealed that all somatic embryos were derived from AtLTP1 luciferase expressing cell clusters. However, not all cell clusters that expressed the AtLTP1 luciferase reporter gene developed into a somatic embryo, suggesting that initiation of an embryogenic pathway in tissue culture does not always lead to development of a somatic embryo. 相似文献
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Expression of knotted1 ( kn1) and ZmLEC1, a maize homologue of the Arabidopsis LEAFY COTYLEDON1 ( LEC1) was studied using in situ hybridization during in vitro somatic embryogenesis of maize ( Zea mays L.) genotype Hi-II. Expression of kn1 was initially detected in a small group of cells (5-10) in the somatic embryo proper at the globular stage, in a specific region where the shoot meristem is initiating at the scutellar stage, and specifically in the shoot meristem at the coleoptilar stage. Expression of ZmLEC1 was strongly detected in the entire somatic embryo proper at the globular stage, gradually less in the differentiating scutellum at the scutellar and coleoptilar stages. The results of analyses show that the expression pattern of kn1 during in vitro somatic embryogenesis of maize is similar to that of kn1 observed during zygotic embryo development in maize. The expression pattern of ZmLEC1 in maize during in vitro development is similar to that of LEC1 in Arabidopsis during zygotic embryo development. These observations indicate that in vitro somatic embryogenesis likely proceeds through similar developmental pathways as zygotic embryo development, after somatic cells acquire competence to form embryos. In addition, based on the ZmLEC1 expression pattern, we suggest that expression of ZmLEC1 can be used as a reliable molecular marker for detecting early-stage in vitro somatic embryogenesis in maize. 相似文献
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Zanin Fabiana Couto Freitas Natália Chagas Pinto Renan Terassi Máximo Wesley Pires Flausino Diniz Leandro Eugenio Cardamone Paiva Luciano Vilela 《Molecular biology reports》2022,49(3):1973-1984
Molecular Biology Reports - Small auxin-up RNA (SAUR) genes form a wide family supposedly involved in different physiological and developmental processes in plants such as leaf senescence, auxin... 相似文献
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Hernández-Castellano Sara Andrade-Marcial Mauricio Aguilar-Méndez Edder D. Loyola-Vargas Víctor M. de Folter Stefan De-la-Peña Clelia 《Plant Cell, Tissue and Organ Culture》2022,148(1):177-188
Plant Cell, Tissue and Organ Culture (PCTOC) - The diploid cotton species G. arboreum offers a better opportunity to elucidate gene structure and function as opposed to the allotetraploid cotton... 相似文献
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Isozyme patterns of carrot (Daucus carota L.) zygotic embryos between the torpedo stage up to 5-day-old seedlings have been compared with those of the similar stages from the embryogenic cell suspension culture to the late somatic plantlet. Somatic embryos blocked at the torpedo stage by -cyclodextrine have also been analyzed. All these stages have been analyzed with respect to seven different enzyme systems: arylesterase, glucosephosphate isomerase, phosphogluconate dehydrogenase, alcohol dehydrogenase, isocitrate dehydrogenase, aspartate aminotransferase and phosphoglucomutase (EC 2.7.5.1, PGM). The relationships between the different stages of both types of embryogenesis have been visualized using an unrooted tree. Generally, profiles of somatic embryos were different from those of zygotic embryos. Interestingly however, a typical zygotic embryo pattern was found in the cyclodextrine-blocked somatic embryos. Only aspartate aminotransferase patterns revealed a similarity between zygotic and somatic torpedo embryos. Both plantlet types showed close patterns with common isozymes. Moreover, similarities were evident between somatic plantlets and cell suspensions. A few isozymes appeared to be stage specific markers: esterase 10–11 were specific to achenes and early germination, phosphogluconate dehydrogenase 8 was specific to 4–5 day-old seedlings and phosphoglucomutase 1 and 7 and alcohol dehydrogenase 4 were markers for zygotic embryos. No somatic embryogenesis specific isozyme could be found. We show that patterns can be associated with particular tissue formation: mainly, aspartate aminotransferase 2 and 1, phosphoglucomutase 8 and 9 and phosphogluconate dehydrogenase 7 coincided with apical meristem initiation and phosphoglucomutase 4 and 5, zones b and d of esterase and zone b of phosphogluconate dehydrogenase coincided with vascular bundle formation.Abbreviations ADH
alcohol dehydrogenase
- CD
-cyclodextrine
- CS
cell suspension culture
- 2,4-D
2,4-dichlorophenoxyacetic acid
- EDTA
ethylenediaminetetraaeetie acid
- LiBo
lithium hydroxide/boric acid
- PEG
polyethylene glycol
- PVP
polyvinylpyrrolidone
- SEg
somatic embryo at the globular stage
- SEh
heart stage
- SEte
early torpedo stage
- SEtl
late torpedo stage
- SEce
early cotyledonary stage
- SEcl
late cotyledonary stage
- SECD
somatic embryo blocked at the torpedo stage with -cyclodextrine
- EST
esterase
- GOT
aspartate aminotransferase
- IDH
isocitrate dehydrogenase
- MTT
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide)
- PMS
phenazonium methosulfate
- PGD
phosphogluconate dehydrogenase
- PGI
glucosephosphate isomerase
- PGM
phosphoglucomutase
- SO
dry seed
- S1–3
seed after 1–3 days of germination
- SP1–2
young and old somatic plantlets
- ZE
zygotic embryo
- ZP4–5
4–5 day-old seedlings 相似文献
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Xiuming Wu Fuguang Li Chaojun Zhang Chuanliang Liu Xueyan Zhang 《Journal of plant physiology》2009,166(12):1275-1283
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L. Alemanno M. Berthouly N. Michaux-Ferriere 《In vitro cellular & developmental biology. Plant》1997,33(3):163-172
Summary In order to improve the late phases of Theobroma cacao L. embryogenesis from tissues of maternal origin, zygotic embryogenesis and somatic embryogenesis were compared, with respect
to morphological, histological, and physiological parameters. Zygotic embryogenesis could be divided into three steps: (a)
embryogenesis sensu stricto, (b) a growth period in which cotyledonary embryos reached their final dimensions, and (c) a maturation period in which embryos
accumulated protein and starch reserves, dehydrated to a water content equal to 30%, and underwent a modification in soluble
sugar composition. Monosaccharides and sucrose contents decreased to the benefit of the oligosaccharides raffinose and stachyose.
The formation of somatic embryos by use of basic protocols was studied to define the limiting factors that could lie behind
their poor development. Morphological abnormalities of somatic embryos, which represented 80% of the total population, were
described. A histological study showed that somatic embryos lacked starch and protein reserves; moreover, their water content
was much higher than that of their zygotic counterparts. Introducing a growth period into the culture protocol made for better
embryo development. Adding sucrose and abscisic acid to the maturation medium was effective in increasing reserve synthesis
and resulted in higher germination, conversion, and acclimatization rates. 相似文献
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Rina Fujihara Naoyuki Uchida Toshiaki Tameshige Nozomi Kawamoto Yugo Hotokezaka Takumi Higaki Rüdiger Simon Keiko U Torii Masao Tasaka Mitsuhiro Aida 《Plant Biotechnology》2021,38(3):317
The shoot organ boundaries have important roles in plant growth and morphogenesis. It has been reported that a gene encoding a cysteine-rich secreted peptide of the EPIDERMAL PATTERNING FACTOR-LIKE (EPFL) family, EPFL2, is expressed in the boundary domain between the two cotyledon primordia of Arabidopsis thaliana embryo. However, its developmental functions remain unknown. This study aimed to analyze the role of EPFL2 during embryogenesis. We found that cotyledon growth was reduced in its loss-of-function mutants, and this phenotype was associated with the reduction of auxin response peaks at the tips of the primordia. The reduced cotyledon size of the mutant embryo recovered in germinating seedlings, indicating the presence of a factor that acted redundantly with EPFL2 to promote cotyledon growth in late embryogenesis. Our analysis suggests that the boundary domain between the cotyledon primordia acts as a signaling center that organizes auxin response peaks and promotes cotyledon growth. 相似文献