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1.
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
2.
《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
3.
Haeng S. Lee Suk W. Kim Kwang -W. Lee Tage Eriksson Jang R. Liu 《Plant cell reports》1995,14(9):545-549
Cotyledonary expiants of ginseng zygotic embryos were cocultured with Agrobacterium tumefadens strain LBA4404 harboring the binary vector pBI121 for 48 h and transferred onto MS medium supplemented with 1 mgl–1 2,4-dichlorophenoxyacetic acid (2,4-D), 0.1 mgl–1 kinetin, and 100 mgl–1 kanamycin. After 8 weeks of culture, kanamycin-resistant calli formed on the cut surfaces of cotyledonary expiants and subsequently they gave rise to numerous somatic embryos. Eight weeks after transfer onto medium containing 1 mgl–1 each of 6-benzyladenine (BA) and gibberellic acid, most of them developed into plantlets. Southern analysis confirmed that the -glucuronidase (GUS) gene was incorporated into the genomic DNA of regenerants. Protoplasts were enzymatically isolated from transformed somatic embryo segments and cultured in liquid medium containing 60 gl–1 myo-inositol, 1 mgl–1 2,4-D, 0.5 mgl–1 BA, and 0.5 mgl–1 kinetin. Plants were regenerated from protoplasts via somatic embryogenesis. The polymerase chain reaction method revealed that 92% of the regenerants retained the GUS gene. When treated with X-glucuronide, 78% of the regenerants showed a GUS-positive response. The overall results indicate that the transgene is stably transmitted during somatic ontogeny and stably expressed in most the regenerants, whereas it may be deleted or impaired in some portion of them.Abbreviations BA
6-benzyladenine; 2,4-D,2,4-dichlorophenoxyacetic acid
- DIG
digoxigenine
- GA3
gibberellic acid
- X-gluc
X-glucuronide
- GUS
-glucuronidase
- MS
Murashige and Skoog (1962) 相似文献
4.
Callus cultures were established from immature embryos of Calotropis gigantea (Linn.) R. Br. on a modified basal medium of Murashige & Skoog supplemented with 1 mgl-1 2,4-D. In addition to 0.1 mgl-1 of NAA the optimal BAP concentration for promoting shoot bud formation and growth was 2 mgl-1. Rooting was induced when shoots were transferred to auxin-supplemented Bonner's solution or half-strength MS basal salt solutions.Abbreviations NAA
-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-butyric acid
- BAP
6-benzylaminopurine
- Kin
kinetin 相似文献
5.
D. Q. Binh L. E. Heszky G. Gyulai E. Kiss A. Csillag 《Plant Cell, Tissue and Organ Culture》1989,18(2):195-200
Callus was induced from seeds of Puccinellia distans (L.) Parl. on MS medium supplemented with 2 mgl-1 2,4-dichlorophenoxyacetic acid and 0.5 mgl-1 kinetin. Morphogenesis initiation was achieved during subculture on medium containing 0.1 mgl-1 2,4-D. From the point of morphogenetic capacity, 3 types of callus were selected. High frequency of plant regeneration was obtained by selection of embryogenic type of callus, and culture on N6 medium and N6 medium supplemented with kinetin (5–10 mgl-1), or kinetin (2 mgl-1) and IAA (0.5 mgl-1). A high ratio of albinos among regenerants was observed. 相似文献
6.
Somatic embryogenesis and subsequent plant regeneration of Euonymus europaeus L (European Spindle Tree) were obtained from square pieces of mature zygotic embryos with an intervening callus phase. Callus and somatic embryos were induced using a Murashige and Skoog's semi-solid basal medium supplemented with several combinations of auxins and cytokinins. The greatest number of somatic embryos was obtained with a continuous exposure to 22.8 M indoleacetic acid and 0.046 M kinetin. The frequency of somatic embryogenesis from zygotic embryos depends on the cold conservation time of seeds. The embryos frequently germinated on the same medium. Further development of somatic embryos into plantlets was achieved on a medium devoid of growth regulators.Abbreviations MS
Murashige and Skoog's medium
- IAA
indole-3-acetic acid
- NAA
-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IBA
indole-3-butyric acid
- BAP
6-benzylaminopurine
- RH
relative humidity 相似文献
7.
Somatic embryos isolated from mature seed-derived cotyledon cultures of cassava (Mannihot esculenta Crantz) underwent direct secondary somatic embryogenesis or plant development under appropriate incubation conditions. Isolated somatic embryos were subjected to a two-stage culture procedure similar to that which induced their development on cotyledon explants. This involved incubation for 24–30 days on Murashige and Skoog basal medium supplemented with 2–8 mgl-1 2,4-dichlorophenoxyacetic acid (2,4-D) (Stage I medium) before transfer to medium supplemented with 0.01 mgl-1 2,4-D and 0.1 mgl-1 6-benzylamino purine (BAP) (Stage II medium). Under these conditions, secondary somatic embryos developed directly from the cotyledons and shoot-tip region of primary somatic embryos by a developmental process morphologically very similar to that occurring on zygotic cotyledon explants. Apical shoot extension and adventitious root formation occurred when somatic embryos were isolated from parental cultures and incubated on Stage II medium. Somatic embryo-derived plants growing in greenhouse conditions appeared morphologically normal when compared with non-regenerated plants. 相似文献
8.
Lucia Martinelli Paola Bragagna Valentino Poletti Attilio Scienza 《Plant cell reports》1993,12(4):207-210
Somatic embryogenesis from leaf- and petiole-derived calli of Vitis rupestris was obtained with an efficiency of 3.2% and 4.2% of plated explants, respectively on two combinations of 6-benzyladenine and 2,4-dichlorophenoxyacetic acid (1/0.1 and 1/1 mgl–1) added to MS medium. Embryogenic callus, embryo subcultures and somatic embryogenesis from somatic embryos were obtained either in the presence of 1 mgl–1 indole-3-acetic acid or 0.1 mgl–1 indole-3-butyric acid added to MS or NN media. Within a 4-month culture, embryo germination occurred at a frequency of 13% of explanted embryos when chilling at 4°C was provided for two weeks and a combination of 6-benzyladenine (1 mgl–1) with indole-3-butyric acid (0.1 mgl–1) was added to NN medium supplemented with casein hydrolysate (250 mgl–1). A higher frequency (51%) was obtained in a longer culture time (9 months) when only indole-3-butyric acid was present in the medium and in absence of chilling.Abbreviations BA
6-benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- MS
Murashige and Skoog (1962)
- NN
Nitsch and Nitsch (1969)
- NOA
2-naphthoxyacetic acid 相似文献
9.
An efficient protocol was developed for in vitro plant regeneration via somatic embryogenesis from cell suspension cultures
of metal tolerant grass Echinochloa colona (L.) Link. Callus was obtained by culturing leaf base on MS medium supplemented
with 0.5 mg dm-3 of 6-benzylaminopurine (BAP) and 2.0 mg dm-3 of 1-naphthaleneacetic acid (NAA). Cell suspensions were initiated and established in MS liquid medium containing 0.5 mg
dm-3 BAP, 1.0 mg dm-3 NAA and 2.0 mg dm-3 2,4-dichlorophenoxyacetic acid (2,4-D). A reduction in the concentration of 2,4-D to 0.5 mg dm-3 induced formation of somatic embryos. The embryos developed and grew into normal plants in the presence of half strength
MS medium without growth regulators. The regenerated plants were hardened in the greenhouse and subsequently grown in the
open. This system may be also used for isolation and culture of protoplasts as a first step in somatic hybridization.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
10.
Somatic embryogenesis and plantlet formation were obtained from 60–75 day old cell cultures of carnation. Callus was generated on MS basal medium supplemented with 2,4-dichchlorophenoxy acetic acid (2,4-D). Removal of 2,4-D during subsequent subculturing of cell suspensions resulted in formation of embroids. These somatic embryos originated from single cells and their early development proceeded normally with clearly defined apical and root meristems. Some embryos developed into plants and were acclimatized to ex vitro conditions.Abbreviations BAP
6-benzylaminopurine
- Kinetin
6-furfurylamino purine
- 2,4-D
2,4-dichlorophenoxy acetic acid
- MS
Murashige and Skoog 相似文献
11.
A. Nordine C. R. Tlemcani A. El Meskaoui 《In vitro cellular & developmental biology. Plant》2014,50(1):19-25
A protocol for somatic embryogenesis was developed for Thymus hyemalis, a wild species in the Mediterranean region. First, the effects of explant type, plant growth regulators [kinetin (KIN) and 2,4-dichlorophenoxyacetic acid (2,4-D)], and genotype on callus induction were tested. For callus induction, the node was the best explant; Murashige and Skoog (MS) medium supplemented with 1.8 μM 2,4-D and 0.5 μM KIN was the best medium, and the genotype had a highly significant effect. To induce production of somatic embryos, the effects of KIN, 6-benzylaminopurine (BAP), and naphthalene acetic acid (NAA) were evaluated. After 5 wk of culture in the dark, MS medium supplemented with 4.44 μM BAP, 0.54 μM NAA, and 4.65 μM KIN gave the highest percentage (85%) of embryogenic callus and the highest number of somatic embryos (27.00) per 45 mg of callus. For germination and plant recovery, somatic embryos were transferred to MS medium without plant growth regulators and plantlet conversion from developed somatic embryos was 90%. In vitro plants with adequate growth and sufficient root systems were subsequently transplanted into a mixture of peat and vermiculite (2:1?v/v) under greenhouse conditions. The survival rate of the plantlets under ex vitro conditions was 80%. 相似文献
12.
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
13.
Induction of somatic embryogenesis and adventitious shoots from immature leaves of cassava 总被引:4,自引:0,他引:4
Direct somatic embryogenesis was successfully achieved from immature leaves of cassava (Manihot esculenta Crantz) cultured on induction medium containing 2,4-dichlorophenoxyacetic acid or naphthaleneacetic acid. Changing the duration of induction or changing plant growth regulators resulted in differences in regeneration of somatic embryos or adventitious shoots. The results showed that auxin was a key factor for inducing embryogenic cells. The embryogenic cells were mainly induced within 4–12 days. Only if the embryogenic cells were induced, the auxin enhanced formation of somatic embryo whereas 6-benzylaminopurine stimulated development of adventitious shoots. Histological examinations supported the conclusion. 相似文献
14.
A protocol has been established for rapid, high frequency plant regeneration from protoplasts of the wild tomato species Lycopersicon chilense Dun. Cell suspension cultures were obtained from calli initiated from seedling stem explants. Protoplasts were isolated from cell suspensions by an overnight one-step enzyme digestion, purified by washing in salts solution and cultured in liquid medium. Dilution of liquid medium every 3 days, with medium containing low levels of growth regulators and sucrose, was critical for sustained colony formation. Up to 70% of protoplast-derived calli regenerated shoots when cultured on agar-solidified medium with Murashige & Skoog (1962) salts and vitamins, 2.0 mg l-1 zeatin and 0.1 mg l-1 indole acetic acid for 21 days, followed by transfer to the same medium lacking indole acetic acid.Abbreviations BAP
6-benzylaminopurine
- IAA
indole acetic acid
- IBA
indole butyric acid
- MES-2
(N-morpholino)-ethane sulfonic acid
- NAA
-naphthaleneacetic acid
- 2,4-d
2,4-dichlorophenoxyacetic acid 相似文献
15.
Sarvesh Adda T. P. Reddy P. B. Kavi Kishor 《In vitro cellular & developmental biology. Plant》1994,30(2):104-107
Summary Induction of somatic embryogenesis, shoot organogenesis, and subsequent plant regeneration in niger seem to be dependent on
genotype, choice of explant, and composition of media growth regulators. Two distinct regeneration protocols have been developed
for somatic embryogenesis and shoot organogenesis. Somatic embryogenesis was induced from epicotyls and cotyledonary explants
(9 to 35%) (but not from hypocotyls and roots) in presence of 2,4-dichlorophenoxyacetic acid, 2,4,5-trichlorophenoxyacetic
acid, and 2,4,5-trichlorophenoxypropionic acid. These embryos matured in MS medium containing Kinetin plus naphthalene acetic
acid (NAA), Kinetin plus Zeatin, and Kinetin plus abscisic acid (ABA). Matured embryos could be germinated on LS and MS basal
media without hormones. Non-embryogenic callus initiated on Linsmaier and Skoog’s (LS) medium from cotyledons of six
different genotypes produced shoots (9 to 32%) on Murashige and Skoog’s (MS) medium fortified with 6-benzylaminopurine
(BAP, 0.5 mg · liter−1), and BAP (1 mg · liter−1) plus NAA (0.1 mg · liter−1). These shoots were rooted with 100% frequency by using indole-3-acetic acid or NAA and transferred successfully to the soil. 相似文献
16.
Somatic embryogenesis and plant regeneration in chrysanthemum (Dendranthema grandiflorum (Ramat.) Kitamura) 总被引:2,自引:0,他引:2
Somatic embryogenesis was observed in ray-floret explants of Dendranthema grandiflorum (Ramat.) Kitamura cv. Aboukyu on Murashige and Skoog medium containing high concentrations of 3-indoleacetic acid (IAA) and
kinetin. 1-Naphthaleneacetic acid also induced somatic embryogenesis but indole-3-butyric acid or 2,4-dichlorophenoxy acetic
acid did not. Other cytokinins, such as 6-benzylaminopurine (BAP) and thidiazuron, were also not effective. No embryos were
seen at lower IAA concentrations with kinetin and various concentrations of BAP, although higher BAP concentrations yielded
many adventitious shoots. In contrast, no somatic embryogenesis was observed from leaves using any combination of plant growth
regulators. Histologically, primordia showed a typical embryo shape with a well-developed vascular bundle between the shoot
and the root primordia. Embryos had both stomata cells and a root system with polarity. Plants were efficiently regenerated
from ray floret-derived embryos subcultured in the appropriate medium.
Received: 30 April 1999 / Revision received: 7 October 1999 / Accepted: 27 January 2000 相似文献
17.
G. Jogeswar D. Ranadheer V. Anjaiah P. B. Kavi Kishor 《In vitro cellular & developmental biology. Plant》2007,43(2):159-166
This study describes a protocol for the induction of high frequency somatic embryogenesis directly from immature inflorescence
explants in three sorghum genotypes (SPV-462, SPV-839, and M35-1). The effect of various growth regulators on somatic embryogenesis
was investigated. High frequency somatic embrogenesis was obtained on Murashige and Skoog (MS) medium supplemented with 2 mg
l−1 2,4-dichlorophenoxyacetic acid (2,4-D), and addition of 0.5 mg l−1 kinetin (KN) in the medium further improved the formation of somatic embryos per explant in all genotypes. The presence of
1.5 mg l−1 6-benzylaminopurine plus 1.0 mg l−1 KN in MS medium was most efficient for maturation and germination of somatic embryos. The genotype SPV-462 performed better
than SPV-839 and M35-1 in terms of induction and germination of somatic embryos. Organogenesis also occurred in callus of
all genotypes at the frequency of 20–25%. Regenerated plants from somatic embryos were successfully acclimatized in soil in
the greenhouse where plants were grown to maturity, flowered, and set seeds. Regenerated plants appeared normal like that
of the seed-raised plants. 相似文献
18.
Summary A method for the production of somatic embryos and subsequent plant regeneration for Anthurium andraeanum Linden ex André (Monocotyledonae) hybrids is described. Whole leaf blade explants, derived from plantlets grown in vitro, formed translucent embryogénic calli at their basal ends within one month of culture in the dark. Secondary somatic embryos formed frequently and without an intervening callus on surfaces of primary embryos. Embryogenesis was induced with three genotypes using a modified half-strength Murashige and Skoog (MS) medium supplemented with 1.0 to 4.0 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.33 to 1.0 mg l–1 kinetin. A combination of 2% sucrose with 1% glucose in the medium favored embryogenesis over 3% sucrose alone. Whole leaf blades on medium solidified with 0.18% Gelrite produced more somatic embryos than leaves on medium with 0.7% Bacto-agar. Within two to three months after culture initiation, embryos were transferred to modified MS medium containing 0.2 mg l–1 6-benzyladenine (BA) and 2% sucrose and placed in the light for conversion into plantlets. Rooted plantlets were recovered and transferred into pots with tree fern fiber medium and grown in the greenhouse.Abbreviations MS
Murashige and Skoog (1962)
- 2,4-D
2,4 dichlorophenoxyacetic acid
- BA
6-benzyladenine 相似文献
19.
Summary A procedure leading to the regeneration of whole plants from protoplasts of melon is described. Protoplasts were isolated from cotyledons and leaves of plants grown in vitro. After 14 days of culture, average viability and division rates were respectively 60% and 30% for the two organs, considering total initial protoplasts plated. The manipulation of the exogenous auxin / cytokinin balance in regeneration media enabled to direct morphogenesis towards somatic embryogenesis (1 mg·l–1 2,4-dichlorophenoxyacetic acid and 0.1 mg·l–1 6-benzylaminopurine) or caulogenesis (0.5 mg·l–1 6-benzylaminopurine and 0.5 mg·l–1 kinetin). Contrary to division ability, regeneration capacity was genotype-dependent under our conditions, but the two organs expressed similar division and regeneration capacities. Maltose was superior to sucrose for the development of caulogenic nodules into buds. Some plants were transplanted to soil, where they appeared to be fertile and produced seeds.Abbreviations BAP
6-benzylaminopurine
- CPW
Cell and Protoplast Washing medium
- KIN
kinetin
- MES
2-(N-morpholino) ethanesulfonic acid
- MS
Murashige and Skoog (1962)
- NAA
1 — naphthaleneacetic acid
- PAS
H (staining), Periodic Acid-Schiff / Hematoxylin
- 2,4-D
2,4-dichlorophenoxyacetic acid 相似文献
20.
Jiahua Xie Mingwei Gao Qihua Cai Xiongying Cheng Yuwei Shen Zhuqing Liang 《Plant Cell, Tissue and Organ Culture》1995,42(3):245-250
The influence of maltose and growth regulators on microspore culture response was investigated in japonica rice. High frequency of callus induction of isolated microspores was obtained with liquid medium containing MS salts, 100 mg l–1 myo-inositol, 1 mg l–1 thiamine-HCl, 500 mg l–1 glutamine, 60 g l–1 maltose, and several growth regulators. The effect of maltose on promoting callus formation was associated with keeping a high proportion of swollen microspores after 5 day preculture and increasing the microspore division rate on the 3rd day after culture initiation. No significant effect of maltose in place of sucrose on plantlet regeneration was seen in regeneration medium. Among the growth regulators tested, the combination of auxin 2,4-dichlorophenoxyacetic acid (1 mg l–1), naphthaleneacetic acid (1 mg l–1), and cytokinin (6-benzyl-aminopurine 1 mg l–1) in the medium proved to be much better for callus formation than in the other media, and the percentage of callusing microspores of that medium reached 0.86%. Indole-3-acetic acid (0.5 mg l–1) and kinetin (2 mg l–1) in regeneration medium were beneficial for green plantlet differentiation. The results also showed that the frequencies of microspores initial division, callus formation and green plant regeneration varied among genotypes no matter what kind of growth regulator and sugar were used. Xiushui 117 was the best variety for callusing followed by 02428 & Taipei 309. Taipei 309 showed a good ability for green plantlet regeneration.Abbreviations 2,4-d
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- 6-BA
6-benzylaminopurine
- KT
kinetin
- IAA
indole-3 acetic acid 相似文献