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1.
Immature and mature zygotic embryos of Paspalum scrobiculatum L. cv. PSC 1 cultured on MS or N6 nutrient medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), formed embryogenic callus. Induction of embryogenic callus and subsequent somatic embryogenesis was possible at a lower concentration of 2,4-D on N6 than MS medium. Immature embryos were highly totipotent, forming somatic embryos at a higher frequency than mature embryos. Addition of amino acids (L-proline or L-tryptophan) to 2,4-D medium resulted in significant enhancement of embryogenesis on culture of mature embryos. Silver nitrate also supported an increased frequency of embryogenesis. Thus it is possible to have high frequency of somatic embryogenesis on culture of mature embryos, which are available in abundance and with ease than immature embryos. The somatic embryos readily germinated and formed plantlets on hormone-free regeneration medium. The regenerated plantlets were successful on transfer to soil and set seed. 相似文献
2.
Induction of somatic embryogenesis using side chain and ring modified forms of phenoxy Acid growth regulators 总被引:2,自引:0,他引:2
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The induction of somatic embryo development in cell cultures of alfalfa (Medicago sativa), celery (Apium graveolens), and lettuce (Lactuca sativa) was compared for 2,4-dichlorophenoxy-acetic acid (2,4-D) and various phenoxy acid growth regulators. Tests using a series of straight chain extensions to the phenoxy acid side chain indicate that phenoxybutanoic acid is active, whereas the phenoxypropanoic and phenoxypentanoic analogs are inactive for the induction of alfalfa embryogenesis. Side branching on the carbon adjacent to the phenoxy group results in optically active compounds. Racemic mixtures and the (+) enantiomers of the compounds are active for alfalfa embryo induction, whereas the (−) enantiomers are inactive and apparently do not inhibit embryogenesis in any way. Development of alfalfa embryos, as measured by plantlet formation from individual embryos, is improved by 4-(2,4-dichlorophenoxy)butanoic acid and with side branching at the carbon adjacent to the phenoxy group compared with induction with 2,4-D. Similarly, substituted phenoxy acids also enhance somatic embryo development in celery and lettuce when compared with 2,4-D. These results are discussed with reference to earlier studies on the structure activity of various synthetic auxins during cell elongation and with reference to the possible importance of auxin metabolism on subsequent somatic embryo development. 相似文献
3.
An efficient in vitro plant regeneration system characterized by rapid and continuous production of somatic embryos using leaf and petiole expiants has been developed in sweetpotato [Ipomoea batatas L. (Lam.)]. The optimal somatic embryogenic response was obtained in the genotype PI 318846-3 with a two-step protocol: (1) stage I-incubation of expiants in the dark for 2 weeks on Murashige Skoog (MS) medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) (2.5 mg/l) and 6-benzylaminopurine (0.25 mg/l) and, (2) stage II-culture in the light on MS medium with abscisic acid (ABA) (2.5 mg/l). The addition of ABA was critical for enhanced production of somatic embryos. Secondary somatic embryos were produced from the primary embryos cultured on MS medium with 2,4-D at 0.2 mg/l. The somatic embryos were converted into normal plantlets when cultured on basal MS medium. Upon transfer to soil, plants grew well and appeared normal with no mortality. The system of somatic embryogenesis described here will facilitate tissue culture, germplasm conservation and gene transfer research of sweetpotato due to its rapidity (6 to 10 weeks), prolific plant production by direct embryogenesis, ease of secondary somatic embryo production and reproducibility.Abbreviations ABA
abscisic acid
- BAP
6-benzylaminopurine, 2,4-D-2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- KIN
kinetin
- MS
medium of Murashige and Skoog (1962)
- NAA
1-naph-thaleneacetic acid
- PIC
picolinic acid
- TDZ
thidiazuron 相似文献
4.
Levels of wheat germ agglutinin have been determined by radioimmunoassay in tissues of immature wheat embryos cultured under different conditions in order to determine the suitability of the lectin as a marker for somatic embryogenesis. Embryos cultured on media favouring continued embryo development accumulated lectin in a similar manner to zygotic embryos in planta unless precocious germination occurred. Embryos cultured on media containing 2,4-D produced callus, and some of this developed somatic embryos. Both embryogenic and non-embryogenic callus contained WGA, that in non-embryogenic callus possibly arising from developmentally arrested root primordia.Abbreviations ABA
abscisic acid
- dpa
days post anthesis
- PBS
phosphate buffered saline, (10 mM KH2PO4 K2HPO4, 145 mM NaCl, pH 7.4)
- RIA
radioimmunoassay
- WGA
wheat germ agglutinin
- 2,4-D
2,4-dichlorophenoxyacetic acid 相似文献
5.
Efficient plant regeneration via somatic embryogenesis has been developed in chickpea cultivar C235. Leaf explants, on MS medium supplemented with 1.25 mg/l 2,4-D and 0.25 mg/l kinetin, yielded somatic embryos with high efficiency during dark incubation. MS medium supplemented with B5 vitamins, 0.125 mg/l IBA and 2 mg/l BAP was found suitable for embryo maturation. The well formed embryos germinated into plantlets on basal B5 medium supplemented with 0.25 mg/l BAP. Further development into healthy plantlets was obtained on basal B5 medium. Hardened plantlets produced normal, fertile plants upon transfer to soil.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
6-Benzyl-aminopurine
- IAA
IndoIe-3-acetic acid
- IBA
Indole-3-butyric acid
- NAA
1-Naphthalene acetic acid
- Kinetin
6-furfuryl aminopurine
- Zeatin
6-(4-hydroxy-3-methylbut-2-enylamino)-purine 相似文献
6.
Jing Li Yang Yu Da Niu Chuan Ping Yang Gui Feng Liu Cheng Hao Li 《Plant Cell, Tissue and Organ Culture》2011,106(3):391-399
Somatic embryogenesis (SE) was induced in female flower buds from mature Schisandra chinensis cultivar ‘Hongzhenzhu’. Somatic embryo structures were induced at a low frequency from unopened female flower buds and excised
unopened on Murashige and Skoog (MS) agar medium containing 4.0 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Friable embryogenic calli were induced from somatic embryo structures after three
to four subcultures on initiation medium. The frequencies of mature somatic embryo germination and plantlet conversion were
low, but increased in the presence of gibberellic acid (GA3). Some germinated somatic embryos could form friable embryogenic calli on medium without plant growth regulators (PGRs).
The germination and conversion frequencies of somatic embryos from embryogenic calli induced using PGR-free medium were higher
than for somatic embryos from embryogenic calli induced on medium containing 2,4-D. Most somatic embryos from 2,4-D-induced
embryogenic calli had trumpet-shaped embryos, and most somatic embryos from PGR-free medium–induced embryogenic calli had
two or three cotyledons. Histological observation indicated that two- and three-cotyledon embryos had defined shoot primordia,
but most of the trumpet-shaped embryos yielded plantlets that lacked or had poorly developed meristem tissue. Cytological
and random amplification of polymorphic DNA (RAPD) analyses indicated no evidence of genetic variation in the plantlets of
somatic embryo origin. 相似文献
7.
Embryogenic callus was obtained from bulb segments of Iris
pseudacorus on Murashige and Skoog (MS) medium with 2,4-dichlorophenoxyacetic acid (2,4-D) alone or in combination with kinetin. When
early globular somatic embryos were subcultured onto MS medium with 4.52 μM 2,4-D, high frequency of somatic embryogenesis
was obtained. Deprivation of 2,4-D was required for maturation. Mature somatic embryos had an elongated scutellum with a notch
on the base of scutellum. Separation of embryos from embryo clusters was necessary to enhance the frequency of germination.
Germination was stimulated by separation of embryos from embryo clusters and transfer onto fresh half-strength MS medium with
3% sucrose. After acclimation in artificial soil in greenhouse for 2 months, 96.4% of plantlets survived. 相似文献
8.
Somatic embryogenesis was obtained from mature cassava cotyledons explants. A two-step medium sequence was developed for efficient embryogenesis. Application of 2,4-D (4 mg l-1) yielded proembryogenic masses which developed into somatic embryos after transfer to a medium containing NAA (0.01 mg l-1), BA (0.1 mg l-1) and GA3 (0.1 mg l-1). The 2,4-D concentrations used for embryo initiation strongly influenced embryo development. Among the cultivars tested, TMS 30395 was most responsive. Full strength MS basal medium alone or with 4 x MS micro salts was efficient for the formation of somatic embryos. Casein hydrolysate, adenine sulfate, nicotinic acid, glycine, tryptophan, and serine were ineffective for embryo development. High sucrose concentration (6%, w/v) inhibited the induction of somatic embryos, while 6% sucrose was optimal concentration for the development of somatic embryos after an induction treatment using 2% sucrose. Addition of 0.52 mg l-1 ABA to the induction media resulted in an increase in somatic embryos production. The ploidy levels of the regenerated plantlets were determined by flow cytometry analysis. Fifty regenerants tested were all tetraploids as the source plants and were morphologically normal. The implications of these results are discussed in relation to genetic transformation using the cotyledons as the explant source.Abbreviations ABA
abscisic acid
- BA
6-benzylaminopurine
- DAPI
4,6-diamidino-2-phenylindole
- SR
101 sulforhodamine
- GA3
gibberellic acid
- MCPA
methyl- chlorophenoxyacetic acid
- NAA
naphthalen-acetic acid
- PCPA
P-chlorophenoxyacetic acid
- 2,4-d
2,4-dichlorophenoxyacetic acid
- 2,4,5 T
2,4,5-trichlorophenoxyacetic acid 相似文献
9.
A rapid protocol for somatic embryogenesis from immature leaflets of groundnut (Arachis hypogaea L.)
P. Venkatachalam P. B. Kavi Kishor N. Geetha M. Thangavelu N. Jayabalan 《In vitro cellular & developmental biology. Plant》1999,35(5):409-412
Summary Plant regeneration via somatic embryogenesis was developed in two groundnut varieties. Somatic embryogenesis was induced from
immature leaflets on MS medium with different concentrations of the auxins 2,4-dichlorophenoxyacetic acid (2,4-D) or naphthaleneacetic
acid (NAA) in combination with 0.5 mg/l of the cytokinin BA. The highest frequency of somatic embryo formation occurred on
MS medium fortified with 20 mg 2,4-D per l. Of the two auxins tested individually 2,4-D was more effective for induction of
embryogenesis as well as production of embryos. Embryo development and maturation was achieved on MS medium supplemented with
N6-benzyladenine (BA) (0.5–2.0 mg/l) and 2,4-D (0.5 mg/l). Plant conversion frequency from somatic embryos was highest in presence
of 2.0 mg BA per l and 0.5 mg NAA per l. The frequency of embryogenesis and plant regeneration was higher in the VRI-2 cultivar
than in the other cultivar tested. Regenerated plants were transferred to soil, grown to maturity, and produced viable seeds. 相似文献
10.
Jietang Zhao Jin Cui Juanxu Liu Feixiong Liao Richard J. Henny Jianjun Chen 《Plant Cell, Tissue and Organ Culture》2012,110(2):239-249
This study established a method of regenerating Spathiphyllum ??Supreme?? through direct somatic embryogenesis. Somatic embryos occurred in leaf and petiole explants cultured in the dark on a Murashige and Skoog basal medium supplemented with 2.27, 4.54, or 9.08???M N-phenyl-N??-1,2,3-thiadiazol-5-ylurea (TDZ) in combination with 1.08???M ??-naphthalene acetic acid or 2.26???M 2,4-dichlorophenoxyacetic acid (2,4-D). Explants with somatic embryos were transferred to fresh medium containing the same concentrations of growth regulators under lighted conditions for embryo conversion. The highest frequencies of leaf explants with somatic embryos and embryo conversion were both 84.4?%, which were induced by 9.08???M TDZ with 2.26???M 2,4-D. The frequencies for somatic embryo induction and embryo conversion were both 100?% when petiole explants were induced by 4.54???M TDZ with 2.26???M 2,4-D. The number of plantlets produced per leaf explant and petiole explant were as high as 67.4 and 74.4, respectively. Plantlets after transplanting to a soilless substrate grew vigorously in a shaded greenhouse. Liners were stable without phenotypic variation. Flow cytometry analysis of randomly selected plants showed that they all had a single identical peak. The mean nuclear DNA index for ??Supreme?? was 1.568, and the nuclear DNA content was 14.222?pg 2C?1. The estimated genome size for ??Supreme?? was 6,954.5?Mbp 1C?1 with a CV at 4.008?%. The results suggest that the regenerated plants have a stable ploidy level and this established regeneration method can be used for highly effective propagation of uniform Spathiphyllum ??Supreme??. 相似文献
11.
Summary Pepper (cv. New Mexico — 6 and Rajur Hirapur) plants were regenerated from immature zygotic embryos via direct somatic embryogenesis. Somatic embryos were formed directly, without any intervening callus, on the zygotic embryo apex, embryo axis and cotyledons on Murashige and Skoog's (MS) medium containing 2,4-D (418 M), thidiazuron (10 M) and a high concentration of sucrose (6–10%). The best response was observed on MS medium containing 2,4-D (9 M), coconut water (10%) and high sucrose (8%). The entire process of induction and maturation of the embryos was completed on the same medium. Histological examination indicated that secondary embryogenesis also occurred directly from the primary somatic embryos. Differentiation of embryos was nonsynchronous, and some embryos were swollen and distorted with fasciation. More than 70% of the mature normal somatic embryos germinated readily on MS medium containing GA3 or TDZ, alone and in combination, and following transfer to pots developed into normal plants.Abbreviations CM
Coconut milk
- 2,4-D
2,4-dichlorophonoxyacetic acid
- GA3
gibberellic acid
- MS
Murashige and Skoog (1962) medium
- NAA
napthaleneacetic acid
- TDZ
thidiazuron 相似文献
12.
《Plant science》1987,52(3):229-235
A proliferating embryonal-suspensor mass (ESM) was initiated from immature embryos of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco), 4–5 weeks after fertilization, on modified MS medium with 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin, and N6-benzylaminopurine (BAP). ESMs were maintained for over 6 months as cell suspension cultures on modified DCR media with low 2,4-D and with kinetin and BAP. The development of individual somatic embryos was initiated in suspension culture by the gradual reduction of plant growth regulators and by addition of abscisic acid. The early stages of zygotic embryogenesis in Douglas-fir are unique among conifers and cleavage polyembroyogenesis is unknown. In somatic embryogenesis, characteristic stages of zygotic embryonic development were recapitulated and complete embryos were recovered by inhibiting cleavage polyembryony with abscisic acid and culturing individual embryos without growth regulators. Histological examination confirmed bipolar organization of somatic embryos. While conversion is low, plantlets with multiple cotyledons have been transferred to soil and continue to grow with production of a new shoot. 相似文献
13.
Callus cultures of Encephalartos cycadifolius were established from zygotic embryo explants on a modified B5 medium containing 1 mg l–1 2,4-D and 1 mg l–1 kinetin. Callus was transferred to media containing various combinations of 2,4-D and kinetin for improvement of somatic embryogenesis. Somatic embryos were produced on media with several growth regulator combinations. The somatic embryos developed from proembryos, which developed long suspensors. A dicotyledonary embryo formed at the distal end of the suspensor. The embryos turned green in light. When transferred to a medium containing 1 mg l–1 ABA the somatic embryos matured. The suspensors desiccated and these embryos rooted when transferred to a medium without phytohormones.Abbreviations ABA
abscisic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid 相似文献
14.
Direct regeneration of somatic embryos was obtained from immature zygotic embryos of Dalbergia latifolia. Immature embryos dissected from green pods 90 d after flowering gave the highest frequency of somatic embryo formation. Preculture on high 2,4-D medium for 4 weeks induced direct somatic embryogenesis, which was expressed during the second culture phase in the presence of low 2,4-D along with a high sucrose concentration. Embryos were separated and transferred to the maturation medium containing MS + 0.5–1.0 mg/L BAP, where embryos developed into plantlets. Somatic embryos failed to convert into complete plants without BAP treatment. This method of direct regeneration of somatic embryos without a callus phase has direct application for genetic manipulation studies.Abbreviations MS
Murashige and Skoog (1962) medium
- 2,4-D
2,4-Dichlorophenoxyacetic acid
- BAP
6-benzylaminopurine
- ABA
Abscisic acid
- KIN
Kinetin 相似文献
15.
T. H. Lan P. I. Hong C. C. Huang W. C. Chang C. S. Lin 《In vitro cellular & developmental biology. Plant》2009,45(1):44-47
Whole plants were regenerated from excised leaves of Drimiopsis kirkii Baker (Lily of the Valley) through direct somatic embryogenesis. An initial exposure to a low level of 2,4-dichlorophenoxyacetic
acid (2,4-D, 0.45 μM) in the medium was essential in inducing the direct formation of somatic embryos. A high concentration
of 2,4-D (4.52 μM) in the proliferation medium reduced embryogenesis and enhanced callus formation. The presence of kinetin
in the medium enhanced the somatic-embryogenesis-inducing effect of 2,4-D (0.45 μM). The maximum embryogenesis rate (4,026
somatic embryos per gram of leaf) was obtained in explants cultured for 30 d in medium supplemented with 2.33 μM kinetin and
0.45 μM 2,4-D (embryo induction medium). Kinetin (4.65 μM) also enhanced embryo germination (97.6%), but the presence of α-naphthalene
acetic acid in the medium drastically reduced embryo germination. Following conversion, the regenerated plantlets were transferred
to soil and showed normal morphological characteristics. 相似文献
16.
Somatic embryogenesis of <Emphasis Type="Italic">Myrciaria aureana</Emphasis> (Brazilian grape tree)
Sergio Yoshimitsu Motoike Edson Santana Saraiva Marilia Contin Ventrella Crislene Viana Silva Luiz Carlos Chamhum Salomão 《Plant Cell, Tissue and Organ Culture》2007,89(1):75-81
The aim of this research was to establish a long-term somatic embryogenic cultures that could be used for cryopreservation.
For the induction of somatic embryogenesis, different levels of 2,4-D as well as the combination of 2,4-D and indole-3-acetyl-l-aspartic acid (IASP) were tested on cotyledons of zygotic embryos. The somatic embryogenic cultures were established and
maintained up to 2 years through frequent subculturing on a medium containing 2,4D + IASP. Light, activated charcoal, and
polyethylene glycol (PEG) were tested for the regeneration and maturation of somatic embryos, and the mature embryos were
germinated in JADS medium. The combination of light and PEG provided the highest number of mature embryos. The somatic embryos
obtained were smaller than zygotic embryos and lacked starch. There was an interaction between 2,4-D and IASP on the induction
and regeneration of somatic embryo in Myrciaria aureana. The combination of light and PEG increased the number of mature embryos; however, charcoal was detrimental to the process. 相似文献
17.
Somatic embryos were induced on peanut (Arachis hypogaea) leaflets from aseptically germinated embryo axes. Leaflet size influenced percent somatic embryogenesis; 5–8 mm long cut leaflets were superior to 2–3 mm long uncut leaflets. Maximum embryogenesis of 14.6% was obtained after a 15 d incubation on induction medium (modified MS with B5 vitamins, 30 g/l sucrose, 4 g/l Gel-Gro, 40 mg/l 2,4-D +0.2 mg/l kinetin) followed by transfer to a secondary medium with 5 mg/l 2,4-D+0.2 mg/l kinetin. Primary somatic embryos were fused along the axes with no distinct cotyledons, but secondary embryos had single axes with two cotyledons. Other treatments had lower percent embryogenesis, no secondary embryogenesis, and embryos with single axes with two cotyledons. Some somatic embryos converted into normal plants capable of greenhouse survival.Abbreviations MS
Murashige and Skoog (1962) medium
- B5
Gamborg et al. (1968) B5 medium
- 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
6benzylaminopurine
- NAA
1-naphthaleneacetic acid 相似文献
18.
Somatic embryogenesis was achieved from cell suspension cultures of niger (Guizotia abyssinica Cass.). Initially, friable embryogenic calluses were induced from cotyledonary leaves of niger on Murashige and Skoog (MS) agar medium containing 5 μM 2,4-Dichlorophenoxyacetic acid (2,4-D) and 0.5 μM kinetin (KIN). Cell suspension cultures were established by using embryogenic calluses in MS liquid medium containing 5 μM 2,4-D and 0.5 μM KIN. Initiation of somatic embryogenesis and development up to globular stage from embryogenic cell clumps occurred in the liquid medium itself. Thereafter embryogenic cell aggregates were transferred to MS agar medium supplemented with 3 μM KIN for embryo differentiation, whereas maturation of somatic embryos occurred in MS agar medium containing 10 μM abscisic acid. 相似文献
19.
Summary In leek (Allium ampeloprasum L.) a cyclic system of somatic embryogenesis was developed. Somatic embryos used for cyclic embryogenesis were able to develop the same type of embryogenic callus as zygotic embryos in the primary cycle. For the first time a comparison of the efficiencies of both expiants was made. Ten families were investigated for somatic embryogenesis. There was a genetic relationship with respect to somatic embryo production between the reciprocal crosses. From each family one genotype was selected for investigating cyclic somatic embryogenesis. Different levels of somatic embryo production were found between the expiants of zygotic and somatic embryos. The two best genotypes, 92.001-03 and 92.002-33 produced twice as many somatic embryos as the overall average. On average, 56% of the somatic embryos finally developed into greenhouse plantlets.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- BA
6-benzyladenine
- MS medium
Murashige and skoog (1962) basal medium 相似文献
20.
Kanniah Rajasekaran John W. Pellow 《In vitro cellular & developmental biology. Plant》1997,33(2):88-91
Summary Regeneration of several varieties of soybean [Glycine max (L.) Merrill] by somatic embryogenesis from cultured epicotyls and primary leaves has been demonstrated. Somatic embryogenesis
was induced from epicotyls and primary leaves when cotyledon halves with the intact zygotic embryo axes were cultured on Murashige
and Skoog (MS) medium supplemented with 10 mg 1−1 (45.2 μM) 2,4-D. Stable, continuously proliferating globular embryo cultures (GEC) were established from small groups of somatic embryos
on MS medium supplemented with 20 mg 1−1 (90.5 μM) 2,4-dichlorophenoxyacetic acid (2,4-D). Rapid multiplication of shoot tips from germinating somatic embryos was achieved
on Cheng’s basal medium (CBO) containing 2.5 mg 1−1 (11.3 μM) 6-benzyladenine. Fertile plants were obtained from individual somatic embryos and in vitro propagated adventitious shoot bud cultures. 相似文献