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1.
G. B. Cabral V. T. C. Carneiro A. L. Lacerda C. B. do Valle A. P. Martinelli D. M. A. Dusi 《Plant Cell, Tissue and Organ Culture》2011,107(2):271-282
Brachiaria brizantha (syn. Urochloa brizantha) is an important tropical forage grass widely cultivated in Brazil. In order to optimize tissue culture conditions for B. brizantha, in vitro culture of mature seeds, basal segments and leaf segments from in vitro plants of an apomictic and a sexual genotype
of B. brizantha was performed. When cultured on different media, leaf segments yielded non-embryogenic calluses which formed several roots.
Friable calluses from mature seeds and basal segments explants incubated on Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic
acid and 6-benzyladenine yielded 80% compact and nodular embryogenic structures. Calluses with such compact embryogenic structures
were highly regenerable upon transfer to medium supplemented with kinetin and naphthalene acetic acid. They produced isolated
somatic embryos, multiple fused scutelli or isolated scutellum with polyembryos that germinated into isolated or multiple
shoots. Green and morphologically normal plants were obtained for the two genotypes. Changing the media from pH 5.8 to pH
4.0 increased the number of explants that formed calluses as well as the number of shoots per explant. When embryogenic calluses
from mature seeds were successively sub-cultured for 4 months, aiming at repetitive somatic embryogenesis, all the regenerated
plants were albinos. The embryogenic nature of the compact structure was confirmed by scanning electron microscopy. 相似文献
2.
Plant Regeneration from Immature Zygotic Embryo-Derived Embryogenic Calluses and Cell Suspension Cultures of Catharanthus roseus 总被引:2,自引:0,他引:2
Kim Suk Weon In Dong Su Choi Pil Son Liu Jang R. 《Plant Cell, Tissue and Organ Culture》2004,76(2):131-135
Culture conditions for plant regeneration in immature zygotic embryo-derived embryogenic cell suspension cultures of Catharanthus roseus (Madagascar periwinkle) Little Bright Eye are described. Immature zygotic embryos formed off-white, friable calluses at a frequency of 20% on Murashige and Skoog (MS) medium supplemented with 4.52 µM 2,4-dichlorophenoxyacetic acid (2,4-D) after 8 weeks of culture. After a second subculture using MS basal medium at 4-week intervals, off-white friable calluses formed a small quantity of yellowish, compact embryogenic calluses. Upon transfer to MS basal medium, embryogenic calluses gave rise to numerous somatic embryos. Cell suspension cultures were established with embryogenic calluses using liquid MS medium supplemented with 4.52 µM 2,4-D. Embryogenic cell clumps from cell suspension cultures developed into plantlets at a frequency of 56.7% when plated onto MS basal medium. Plantlets were transplanted to potting soil and grown to maturity in a growth chamber. 相似文献
3.
This work presents the preliminary results of in vitro studies with Araujia sericifera, which is cultivated for ornamental purposes. Immature seeds from wild plants were used to start the cultures. Somatic embryos and friable embryogenic calluses were obtained from white cotyledons in media containing naphthaleneacetic acid and benzyladenine or 2,4-dichlorophenoxyacetic acid. Plants were regenerated from these somatic embryos.Cell suspensions obtained from friable calluses cultured in M1 modified medium showed a considerable growth capacity. The packed cell volume was doubled in about 15 days of culture at the exponential phase. the results obtained may be used to design further experiments with the aim of improving somatic embryogenesis.Abbreviations NAA
-naphthaleneacetic acid
- BA
benzyladenine
- IBA
indolebutyric acid
- 2,4-d
2,4-dichlorophenoxyacetic acid 相似文献
4.
Summary Establishment of fast-growing, highly regenerable callus cultures was examined in Muscari armeniacum Leichtl. ex Bak. in order to develop an efficient genetic transformation system. High-frequency callus formation was obtained
from leaf explants of cv. Blue Pearl on media containing 2,4-dichlorophenoxyacetic acid (2,4-D), α-naphthaleneacetic acid
(NAA) or 4-amino-3,5,6-trichloropicolinic acid (picloram, PIC). Fast-growing, yellowish nodular callus lines and white friable
callus lines containing a few somatic embryos were established on initiation medium supplemented with 4.5 μM 2,4-D and with 54 μM NAA, respectively. The yellowish nodular calluses vigorously produced shoot buds after transfer to media containing 0.44–44
μM 6-benzyladenine (BA), whereas the white friable calluses produced numerous somatic embryos upon transfer to plant growth
regulator-free (PGR-F) medium. Histological observation of shoot buds and somatic embryos indicated that the former consisted
of an apparent shoot meristem and several leaf primordia, and the latter had two distinct meristematic regions, corresponding
to shoot and root meristems. Both shoot buds and somatic embryos developed into complete plantlets on PGR-F medium. Regenerated
plants showed no observable morphological alterations. High proliferation and regeneration ability of these calluses, were
maintained for over 2 yr. 相似文献
5.
Decai Cui J. R. Myers G. B. Collins P. A. Lazzeri 《Plant Cell, Tissue and Organ Culture》1988,15(1):33-45
The origin and development of zygotic and somatic embryos of Trifolium rubens L. was studied with the aid of paraffin sections and light microscopy. Zygotic embryos were collected, fixed and prepared daily from one to ten days after cross-pollination. Somatic embryos were obtained by plating petiole sections on modified L2 medium with 0.015 mgl-1 picloram and 0.1 mgl-1 6-BAP. Cultured petioles were collected and fixed daily from one to 25 days after plating. Two regions in the vascular bundle sheath of cultured petioles gave rise to callus. The first region was adjacent to the phloem fibers and produced friable callus. The second region gave rise to compact callus that was connected to the fascicular cambium. Somatic embryos originated from single cells in the cortex directly without intervening callus formation and from single cells in the friable callus. In addition, embryos arose from meristematic regions in compact callus. Many early stages of embryogenesis (one, two and four-celled stages) were observed in the cortex and friable callus. Zygotic embryogenesis in Trifolium differs from other legumes in that the suspensor is short and has a broad attachment. This arrangement was observed in zygotic embryos of T. rubens and in many somatic embryos. However, a continuum of somatic embryogenesis was observed where some young embryos had a Trifolium suspensor-like arrangement while others were attached to a long narrow suspensor-like structure more characteristic of Medicago. 相似文献
6.
Neo-formation of flower buds and other morphogenetic responses in tissue cultures of Melia azedarach 总被引:1,自引:0,他引:1
Melia azedarach has great interest because of its insecticidal properties. Recently, the occurrence of precocious flowering in tissue cultures of this species was reported. This paper describes some in vitro morphogenetic responses using hypocotyl segments as explants and MS basal medium. Amongst the results we report are: (a) in basal medium, 5% of the explants neo-formed floral buds and flowers, and 80% formed vegetative shoots; (b) flower neo-formation could not be controlled or increased by addition of benzyladenine, or lowering the nitrogen level; (c) benzyladenine increased the regeneration of vegetative shoots; (d) compact green calluses were eventually formed in basal medium, and vigorous friable calluses can be easily induced with 0.5 mM 2,4-D; (e) green calluses could be subcultured and regenerated into plants, and, from friable calluses, cell suspensions were started; (f) histological studies showed that neo-formations originate in the wound tissue or from the inner tissue of the hypocotyl. 相似文献
7.
Glória Pinto Helena Valentim Armando Costa Sílvia Castro Conceição Santos 《In vitro cellular & developmental biology. Plant》2002,38(6):569-572
Summary Somatic embryos were obtained from a 60-yr-old Quercus suber L. tree. Leaf explants were cultivated on Murashige and Skoog medium with 30 gl−1 sucrose, 3 gl−1 gelrite, pH adjusted to 5.8, and different growth regulator combinations. Callus induction took place at 24±1°C in the dark
during the first 3 wk. After 3 mo, calluses that showed embryogenic structures were transferred to the same medium without
growth regulators. Somatic embryogenesis was only observed in calluses induced on E3 medium (supplemented with 4.5 μM 2,4-dichlorophenoxyacetic acid and 9.0 μM zeatin). On average, 7.5% of the initial explants formed embryogenic calluses in this medium. Somatic embryo proliferation
was high due to secondary embryogenesis. On average, 10% of the somatic embryos germinated and 40% of these germinated embryos
converted into plants. Plants were elongated on the same medium without growth regulators and acclimated to greenhouse conditions. 相似文献
8.
Root explants excised from carnation plants maintained in vitro formed off-white, friable calluses after three weeks of culture
on Murashige and Skoog (MS) medium supplemented with 1 mg l−1 thidiazuron (TDZ) and 1 mg l−1 α-naphthalaneacetic acid (NAA). These calluses were subsequently transferred to MS basal medium where, after an additional
four weeks of culture, approximately 50% of the calluses formed somatic embryos. However, calluses formed on root explants
that had been cultured on MS medium supplemented with 2,4-dichlorophenoxyacetic acid did not produce somatic embryos upon
transfer to MS basal medium. Somatic embryos developed into plantlets and subsequently were grown to maturity. These results
indicate that root explants have a high competence for somatic embryogenesis in carnation.
J. Seo and S.W. Kim contributed equally to this work. 相似文献
9.
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
10.
F. X. Côte R. Domergue S. Monmarson J. Schwendiman C. Teisson J. V. Escalant 《Physiologia plantarum》1996,97(2):285-290
There are very few reports on the establishment of long-term embryogenic cell cultures of banana, especially of triploid cultivars of commercial interest. Embryogenic cell suspensions were prepared using the cultivar Grand nain, the most widely grown dessert banana in the world. After culture for 5 or 6 months of immature male flowerbuds adjacent to the floral apex, yellow, compact calluses and white, friable embryogenic tissues were induced. Suspension cultures were initiated from embryogenic tissues placed in liquid medium. The packed cell volume (PCV) of the suspensions increased 2- to 5- fold with each monthly culture cycle. Plating of the embryogenic suspensions resulted in approximately 370×103 embryos per ml of PCV. Depending on the size of embryos, 3 to 20% germination was observed. A histological survey of cell suspensions and embryo development was carried out. Cellular aggregates with cells displaying typical embryogenic features were formed. Most of the somatic embryos were probably of unicellular origin. 相似文献
11.
An efficient somatic embryogenesis system has been established in Catharanthus roseus (L.) G. Don in which primary and secondary embryogenic calluses were developed from hypocotyls and primary cotyledonary somatic
embryos (PCSEs), respectively. Two types of calluses were different in morphology and growth behaviour. Hypocotyl-derived
embryogenic callus (HEC) was friable and fast-growing, while secondary callus derived from PCSE was compact and slow-growing.
HEC differentiated into somatic embryos which proliferated quickly on medium supplemented with NAA (1.0 mg l−1) and BA (1.5 mg l−1). Although differentiation and proliferation of somatic embryos were faster in primary HEC, maturation and germination efficiency
were better in somatic embryos developed from primary cotyledonary somatic embryo-derived secondary embryogenic callus (PCSEC).
At the biochemical level, two somatic embryogenesis systems were different. Both primary and secondary/adventive somatic embryogenesis
and the role of plant growth regulators in two modes of somatic embryo formation have been discussed. 相似文献
12.
A procedure for plant regeneration from immature seed-derived calli of rugosa rose (Rosa rugosa Thunb.) via somatic embryogenesis is described. Embryogenic calli were initiated from immature seeds 2–3 weeks after anthesis on Murashige and Skoog (MS) medium without growth regulators. Induced calli had a white, friable and nodular appearance with several proembryos. These calli were subcultured at 20-day intervals on MS medium containing 0.1–0.2 M galactose on which they grew rapidly; but somatic embryogenesis was inhibited. Somatic embryos were again induced from the subcultured calli after transferring to MS medium containing 0.1 M M fructose or sucrose but lacking growth regulators. After transferring these embryos (1–2 mm) to MS medium containing 0.1 M sorbitol, 3% of them germinated and grew into plantlets which showed sustained growth on the MS medium containing only 0.1 M sorbitol as the sole carbon source. 相似文献
13.
F. A. Redway V. Vasil D. Lu I. K. Vasil 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1990,79(5):609-617
Summary Immature embryos, inflorescences, and anthers of eight commercial cultivars of Triticum aestivum (wheat) formed embryogenic callus on a variety of media. Immature embryos (1.0–1.5 mm long) were found to be most suitable for embryogenic callus formation while anthers responded poorly; inflorescences gave intermediate values. Immature embryos of various cultivars showed significant differences in callus formation in response to 11 of the 12 media tested. No significant differences were observed when the embryos were cultred under similar conditions on MS medium with twice the concentration of inorganic salts, supplemented with 2,4-D, casein hydrolysate and glutamine. Furthermore, with inflorescences also no significant differences were observed. Explants on callus formation media formed two types of embryogenic calli: an off-white, compact, and nodular callus and a white compact callus. Upon successive subcultures (approximately 5 months), the nodular embryogenic callus became more prominent and was identified as aged callus. The aged callus upon further subculture, formed an off-white, soft, and friable embryogenic callus. Both the aged and friable calli maintained their embryogenic capacity over many subculture passages (to date up to 19 months). All embryogenic calli (1 month old) from the different callus-forming media, irrespective of expiant source, formed only green shoots on regeneration media that developed to maturity in the greenhouse. There were no significant differences in the response of calli derived from embryos and inflorescences cultured on the different initiation media. Also, the shoot-forming capacity of the cultivars was not significantly different. Anther-derived calli formed the least shoots. Aged and friable calli on regeneration media also formed green shoots but at lower frequencies. Plants from long-term culture have also been grown to maturity in soil.Florida Agricultural Experiment Station Journal Series No. R-00494 相似文献
14.
Unopened leaves, petioles and fully opened leaves from micropropagation cultures of five Vitis rotundifolia Michx. varieties were cultured on induction medium to study their embryogenic response. Among the various explants tested,
the maximum number of varieties produced embryogenic cultures from unopened leaves followed by fully opened leaves and petioles.
Based on morphological differences, two types of embryogenic cultures were identified. Friable cultures typically arose as
proembryonic masses (PEM) on induction medium, whereas somatic embryo production without an intervening PEM stage was observed
in compact cultures. Of the five varieties tested, the highest frequency of embryogenic response was observed from fully opened
leaves of ‘Supreme’ and unopened leaves and petioles of ‘Delicious’. Attempts to initiate suspension cultures from varieties
resulted in proliferation and maintenance of ‘Alachua’ and ‘Carlos’ cultures in liquid medium for 16 weeks. Embryogenic potential
of varieties was studied on cultures growing on embryo development medium. The maximum number of cotyledonary stage somatic
embryos from 0.2 g proembryonic masses were observed in ‘Carlos’ (379.3) followed by ‘Alachua’ (350.0) and ‘Delicious’ (305.0).
Cotyledonary stage somatic embryos germinated when cultured on Murashige and Skoog medium containing 1 μM Benzyladenine (BA).
Although high embryo germination rates (80–100%) were observed in the varieties tested, plant recovery from germinated somatic
embryos ranged from 6–47%. Embryogenic cultures could be maintained on X6 medium and used in genetic engineering studies. 相似文献
15.
K. P. Martin A. Shahanaz Beegum C.-L. Zhang A. Slater P. V. Madhusoodanan 《Biologia Plantarum》2007,51(4):769-772
In vitro propagation of an anticancerous drug synthesizing plant, Ophiorrhiza prostrata D. Don, was established through indirect somatic embryogenesis. Friable embryogenic calluses were initiated from O. prostrata leaf and internode explants on Murashige and Skoog (MS) media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) either
alone or in combination with N6-benzyladenine (BA) or kinetin (KIN). Somatic embryos were developed after subculture of the friable calluses onto half strength
MS media containing 0.45 or 2.26 μM 2,4-D alone or in combination with BA or KIN. Medium supplemented with 2.26 μM 2,4-D and
2.22 μM BA was optimal, supporting the production of a mean of 5.8 globular embryos. Subculture of globular embryo-bearing
calluses on half strength MS medium without growth regulators produced the highest embryo frequency, and the majority of them
developing to early torpedo stage. Somatic embryos underwent maturation and converted to plantlets at high frequency (90 %)
on half strength MS medium supplemented with 0.44 μM BA. Somatic embryo-derived plantlets with well-developed roots were established
in field conditions with a 90 % survival rate. 相似文献
16.
Coffea arabica leaf explants cultured on medium with 5 µM6-benzyladenine (BA) as the sole plant growth regulator producedwhite friable calluses that formed somatic embryos. These calluseshave been subcultured on the same medium for more than 2 yearsand maintain the ability to produce somatic embryos. (Received October 3, 1984; Accepted January 18, 1985) 相似文献
17.
Callus cultures were initiated from mature excised caryopses of bahiagrass (Paspalum notatum Flugge) on Murashige & Skoog medium supplemented with 20 gl–1 sucrose and 2 mg l–1 2,4-D. Excised mature caryopses readily germinated and callus developed at the base of coleoptiles. There was considerable variation in the amount of non-embryogenic callus among the cultures. Most of the explants produced non-embryogenic translucent callus consisting of thin-walled cells and unorganized tissue. Some of these calli gave rise only to roots. Other explants formed embryogenic calli which were distinguished morphologically as white, globular and friable. Somatic embryos developed and germinated precociously when embryogenic calli were transferred to a 2,4-D-free medium. Somatic embryogenesis was confirmed by histological sections and scanning electron microscopy. Of the 300 cultures, 35 were embryogenic but only 10 produced plants that were successfully grown to maturity. 相似文献
18.
A reproducible system for somatic embryogenesis and plantlet formation of
sandalwood has been developed. A high frequency (100%) of somatic embryos
were induced directly from various explants in MS (Murashige and Skoog,
1962) medium with thidiazuron (1 or 2 M) or
indirectly in medium containing 2,4-D plus thidiazuron. Within 8 weeks,
white globular somatic embryos or friable embryogenic tissue developed on
cultured explants. In S. album the globular somatic
embryos were transferred to MS medium supplemented with IAA (6 M) and kinetin (1 and M) where they
developed further, multiplied and maintained friable embryogenic tissue.
After 15-30 d, mature somatic embryos (1-2 mm) with well-developed
cotyledons were separated and subcultured on to medium containing GA3 (6
M) for germination. Once germinated, elongated somatic embryos
(10-20 mm long) grew further in MS supplemented with lower GA3 (3
M). In S. spicatum, the addition of casein
hydrolysate and coconut milk was necessary for plantlet development from
somatic embryos. From histological studies, it appeared that primary
somatic embryos arose from single cells or had a multicellular origin from
the epidermis or cortical parenchyma. Secondary somatic embryos and friable
embryogenic tissue differentiated from groups of proembryogenic cells from
a superficial layer of the primary somatic embryos.Keywords:
Santalum album, Santalum spicatum, somatic
embryogenesis, histological studies.
相似文献
19.
H. -S. Lin C. van der Toorn K. J. J. M. Raemakers R. G. F. Visser M.J. De Jeu E. Jacobsen 《Plant cell reports》2000,19(5):529-534
Stem segments of seedlings from two Alstroemeria breeding lines, cultured on media supplemented with 4 mg/l 2,4-dichlorophenoxyacetic acid and 0.5–1.0 mg/l 6-benzylaminopurine
(BA), initiated soft callus, which became compact after subculture on a medium with only 0.5 mg/l BA. Friable embryogenic
calli were initiated from compact callus on a medium supplemented with 10 mg/l picloram. Proembryos developed from friable
embryogenic calli via embryos into plants after subculture on medium supplemented with 0.1 mg/l BA. The proembryos formed
friable embryogenic calli again after culture on medium supplemented with 10 mg/l picloram. The total time needed to regenerate
a complete plantlet from friable callus was approximately 6 months. This system for the production of embryogenic material
is considered to have valuable applications for genetic transformation in Alstroemeria.
Received: 22 April 1999 / Revision received: 16 July 1999 · Accepted: 20 July 1999 相似文献
20.
Summary
In vitro regeneration of plants via somatic embryogenesis through cell suspension culture was achieved in horsegram. Embryogenic calluses
were induced on leaf segments on solid Murashige and Skoog (MS) medium with 9.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Differentiation of somatic embryos occurred when the embryogenic calluses were transferred
to liquid MS medium containing 2,4-D. Maximum frequency (33.2%) of somatic embryos was observed on MS medium supplemented
with 7.9 μM 2,4-D. Cotyledonary-torpedo-shaped embryos were transferred to liquid MS medium without growth regulators for maturation
and germination. About 5% of the embryos germinated into plants, which grew further on solid MS medium. The plants were hardened
and established in soil. Effects of various auxins, cytokinins, carbohydrates, amino acids, and other additives on induction
and germination of somatic embryos were also studied. A medium supplemented with 7.9 μM 2,4-D, 3.0% sucrose, 40 mg l−1
L-glutamine, and 1.0 μM abscisic acid was effective to achieve a high frequency of somatic embryo induction, maturation, and further development. 相似文献