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1.
During the phase of exponential growth in chlorophyllous calluscultures derived from Haplopappus gracilis, Hypochaeris radicata,and Acer pseudoplatamus, cells double their number on the average,and also their volume, in about 4.3, 6.6, and 92 daysrespectively. The two rates decline subsequently but cell expansioncontinues for a short time after division has ceased. With culturesof Oxalis dispar, however, which have an average cell generationtime of about 10 days, there is first a short exponential phasedominated by division, and this is followed by a series of phasesdominated alternately by either division or expansion. Chlorophyll accumulation does not occur in Haplopappus duringthe exponential phase (chlorophyll a decreases) but there isa slow accumulation of caro-tenoids. The bulk of the pigmentsaccumulate during the declining phase of growth mainly afterdivision has ceased. With Hypochaeris and Acer, on the otherhand, accumulation is most intense during the exponential phase,and few pigments are added later. With Oxalis, most of the accumulationoccurs after the exponential phase; carotenoids accumulate untilthe cessation of growth whereas chlorophylls start to declinebefore this. With all species, pigments decline after the cessationof growth. The loss is small in Haplopappus and the tissuesare still bright green when the medium dries out: Hypochaerisand Oxalis, in contrast, eventually become colourless. The data are discussed in relation to the changes in pigmentcontent that accompany the growth and development of a singlecell in each species. 相似文献
2.
Somatic Embryogenesis from Clonal Leaf Tissues of Cassava 总被引:3,自引:0,他引:3
Leaf lobes were isolated from palmate leaves of clonal cassava(Manihot esculenta Crantz) material growing in vitro or in glasshouseconditions and subjected to a two-stage culture procedure involvingincubation on Murashige and Skoog (MS2) basal medium supplementedwith 212 mg l1 2,4-D for 20 d (Stage I) beforetransfer to MS2 basal medium supplemented with 0.01 mg l12,4-D and 0.1 mg l1 6-benzylamino purine (BAP) (StageII medium). Embryogenetic tissues, foliose structures and somatic embryosdeveloped from leaf lobes at all Stage I 2,4-D concentrations,except on those explants isolated from shoot-tip cultures incubatedon MS2 basal medium supplemented with 0.1 mg l1 NAA and1.0 mg l1 BAP. Leaf lobes isolated directly from glasshouse plants showed optimalembryogenetic competence when subjected to a Stage I cultureperiod of 17 d, although foliose structure initiation was optimalwith shorter Stage I durations. Leaf lobes of 24 mm lengthand those isolated from phyllotaxic leaf numbers 4 and 5 showedthe greatest embryogenetic competence. Manihot esculenta, cassava, somatic embryogenesis, tissue culture, morphogenetic competence 相似文献
3.
Female gametophytes (at the archegonial stage) excised fromyoung ovules of Ephedra foliata Boiss, were cultured on a basalmedium (Murnshige and Skoog's combinations of major and minorsalts, Iron source, vitamins, myo-inositol along with 2 percent sucrose and 10 per cent coconut milk) under aseptic conditions.Growth and morphogenetic responses of the explants to auxinswere compared at different concentrations and a study of theirinteractions with cytokinins has also been made. At 2 mg 11,2, 4-D induced profuse callusing which subsequently producedroots. NAA at 4 mg 11 was optimal for callus growth androoting. Combinations of 2,4-D and kinetin were more effectivein inducing roots and shoot buds than those of 2,4-D and benzylamino-purine (BAP). Addition of BAP (0.05 mg 11) to themedium containing optimal concentrations of NAA resulted information of a large number of roots. Kinetin induced only rootingin the presence of 4 mg 11 NAA. A high concentrationof BAP (8 mg 11), stimulated shoot bud formation. Forthe further development of shoot buds, neither auxin nor cytokininwas needed. Cytological observations revealed the presence ofhaploid number of chromosomes, i.e. seven. Ephedra foliata, tissue culture, callus, regeneration, 2,4-dichlorophenoxyacetic acid, naphthalene acetic acid, kinetin, benzyl amino-purine 相似文献
4.
Protoplasts were successfully isolated from internodal callustissues of both Oxalis glaucifolia and O. rhombeo-ovata whenthey were digested in a solution containing 0.1% (w/v) MacerozymeR-10, 0.5% (w/v) cellulase Onozuka R-10 and 0.3 mmol m3sucrose. Protoplasts proliferated to give cell colonies on Gamborget al.'s B5 medium supplemented with 0.3 mmol m3 mannitol,0.5 mg dm32, 4-D, and 2.0 mg dm3 kinetin. Calluswas produced upon transfer of cell colonies to Murashige andSkoog medium containing 2.0 mg dm3 l-naphthaleneaceticacid (NAA) and 0.1 mg dm3 kinetin for O. glaucifolia,or with 5.0 mg dm3 NAA and 0.5 mg dm3 6-benzylaminopurine,for O. rhombeo-ovata. Plants were regenerated from O. glaucifoliaprotoplasts on a medium containing 0.1 mg dm3 NAA, 1.0mg dm3 kinetin and 1.0 mg dm3 gibberellic acid,but only vascular nodules were differentiated by O. rhombeo-ovataprotoplast-derived calli. Key words: Tissue culture, protoplasts, plant regeneration, Oxalis spp 相似文献
5.
Explants derived from Zamia pumila embryos were cultured ona Murashige and Skoog basal medium supplemented with naphthaleneaceticacid (NAA), N4-benzylaminopurine (BAP), or combinations of thetwo at 27 °C in darkness. NAA was invariably required forcallus initiation, and its minimal effective concentration was0.1 mg l1. BAP was not always required, and dependingon the explant type and NAA concentration, BAP either enhanced,suppressed, or had little effect on the frequency of callusinitiation. High frequency of callus initiation occurred with1.0 mg l1 NAA combined with 0.01 or 1.0 mg l1BAP. When the concentration of NAA was high relative to thatof BAP, friable callus was produced. As the relative BAP concentrationwas increased, a more compact callus formed. Compact-nodularcallus developed at equal concentrations of NAA and BAP overa wide range of absolute concentrations. Friable callus formedroots only. Compact-nodular callus formed roots, shoots andembryo-like structures. Root and shoot formation predominatedand were of nearly equal frequency. Formation of embryo-likestructures was infrequent. Zamia pumila, callus differentiation, callus formation, embryo culture, naphthaleneacetic acid, N4-benzylaminopurine 相似文献
6.
Callus production along with caulogenesis and rhizogenesis were obtained from internodal stem explants of kenaf (Hibiscus cannabinus L.) after 4 weeks in culture. Murashige and Skoog medium was used for two 4×4 matrix experiments designed to determine suitable growth regulator combinations (NAA/BAP or 2,4-D/kinetin) and concentrations (0.1, 0.3, 1.0, 3.0 mg/L). The most abundant callus production was observed at 0.3/3.0 and 1.0/3.0 mg/L 2,4-D/kinetin and at 1.0/1.0 and 3.0/1.0 mg/L NAA/BAP. Rhizogenesis was most extensive with NAA/BAP at concentrations of 0.1/3.0 and 0.3/ 3.0 mg/L. Adventitious shoots developed on both auxin/cytokinin matrixes when each concentration was at 0.3 mg/L or less. These protocols will facilitate the development of in vitro approaches to kenaf improvement and the study of certain host-pathogen interactions.Abbreviations MS
Murashige and Skoog (1962) medium
- 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
6-benzylaminopurine
- NAA
1-naphthyleneacetic acid
- SDS
sodium dodecyl sulfate 相似文献
7.
Excised leaf discs of three Brassica species, B. oleracea, B.napus, and B. campestris were induced to produce adventitiousbuds and subsequently entire plants by culture on media withspecific combinations of 6-benzylaminopurine (BAP) and -naphthylaceticacid (NAA). Each species required a particular hormone concentrationfor optimum growth and differentiation: B. oleracea, BAP 10mg1 and NAA 1 mg 11; B. napus, BAP 10 mg 11and NAA 10 mg 11; B. campestris, BAP 1 mg 11 andNAA 10 mg 11. In a more detailed study on one of these species, namely B.oleracea, the relative influence of other media components suchas amino acids and other organic additives was examined. Itwas also found that the source and size of the explant greatlyaffected the growth response, as did the size of the culturevessel. The regenerated plants dislayed a range of ploidy as well asphenotypic abnormalities. Findings are discussed in relation to results from other tissueculture systems. 相似文献
8.
We have developed in vitro culture systems for both “grain” and “vegetable” species of the genus Amaranthus. Leaf discs and hypocotyl segments from 2- to 3-wk-old seedlings of A. hypochondriacus, A. cruentus, and A. tricolor were cultured in B5 and MS media supplemented with 2,4-dichlorophenoxyacetic acid, α-naphthaleneacetic acid, benzyladenine and zeatin in various combinations. Rapidly growing callus and abnormal roots formed on leaf discs of A. hypochondriacus and A. cruentus in the presence of 0.1-1.0 mg/l 2,4-D. At higher levels (1.0-10.0 mg/l) of 2,4-D, embryo-like structures arose from the surface and veins of the leaf discs. Shoots formed on hypocotyl-derived callus of both grain species in B5 medium + 0.1 mg/l NAA and 0.1-1.0 mg/l zeatin. Lower ratios of zeatin/NAA stimulated root formation from hypocotyl segments. Viable mesophyll protoplasts were isolated from primary leaves of all three species, with an enzyme solution of 1% Cellulysin and 0.05% Pectolyase Y-23, producing a yield of more than 106 protoplasts/g fresh weight. 相似文献
9.
Callus cultures of 12 temperate grasses were established, somefor the first time, by incubating detached roots or whole seedlingson a Linsmaier and Skoog basal medium which contained 2, 4-dichlorophenoxyaceticacid (2, 4-D) at 1.0 mg 11 as the only growth hormone.The callus, which developed in the pericycle of the roots andon the embryo of the seeds, was subcultured on the same medium;further growth of the callus varied from good in the case ofDactylis glomeraia, Agrostis tennis, Cynosurus cristatus, andPoa trivialis, to poor in several Lolium species and varieties.Most cultures developed root primordia which sometimes grewinto visible roots, but shoot primordia, none of which grewinto shoots, were found only in the callus of Lolium multiflorumvar. westerwoldicum. Cell suspension cultures were also readily established and maintainedusing the same culture medium. Most cultures contained a highproportion of round or oval cells, which ranged from 18 to 77µm in diameter or length, while many also had a significantproportion of larger, more elongated cells which varied in lengthfrom 46 to 182 µm. The cells of Dactylis glomerata werecharacteristically larger and more convoluted than the cellsof other grass species that were examined. The addition of kinetinat 0.1 mg 11 to the 2, 4-D-containing culture mediumincreased the proportion of irregular-shaped cells and reducedthe dispersion of the cells, perhaps by improving cellular contactand adhesion; in some species, such as Agrostis tenuis and Phleumpratense, the presence of kinetin promoted the deposition ofstarch granules in cells. 相似文献
10.
Explants of leaves, basal plates, petals, anthers and ovariesof young growing corms of Crocus chrysanthus var. E. P. Bowleswere cultured on MS basal media with 20 different combinationsof either kinetin and NAA or BAP and 2, 4-D in the dark. Nomajor change was observed except on ovary explants. The ovaryexplants produced callus at 5.0 mg 11 and 10 mg1BAP and subsequently stigma-like structures formed on the surfaceof the callus. Transfer to light resulted in the stigma-likestructures developing a yellow pigmentation whereupon they cameto resemble the naturally-grown stigmas. Corm formation andshoot regeneration was obtained from the callus when the ovaryexplants were cultured on media containing 5.0 and 10 mg I1BAP with 0.5 mg 11 2, 4-D. Increasing the level of 2,4-D markedly reduced the number of shoots produced per explant. Key words: Crocus chrysanthus, callus, ovary explants 相似文献
11.
Growth Hormones and Propagation of Cymbidium in vitro 总被引:2,自引:0,他引:2
MOGENS FONNESBECH 《Physiologia plantarum》1972,27(3):310-316
Protocorms of Cymbidium (Orchidaceae) were grown on solid or liquid medium with macro-nutrients according to Wimber (van Raalte 1967) and iron, micro-nutrients and vitamins according to Nitsch (1968) the medium also contained 2% sucrose. The effects of 1) the auxins; indol-3yl-acetic acid (IAA), α-naphthaleneacetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D); 2) the cytokinins; 6-furfurylaminopurine (kinetin) and benzyladenine (BA) and 3) the gibberellin; gibberellic acid (GA) were examined alone or in combinations. IAA had no effect alone. NAA resulted in optimal fresh weight at 10 μM and the protocorms were vigorous, but lighter green than usual. 2,4-D caused a high weight increase at 1 μM, but the protocorms were abnormal. Higher concentrations of NAA and 2,4-D inhibited chlorophyll synthesis. On solid medium kinetin (100 μM) induced a growth of many small shoots, but had no effect on the fresh weight. In liquid medium, kinetin promoted a callus formation and fresh weight increase. BA had effects similar to kinetin, but at lower concentrations. GA alone promoted shoot and leaf growth. Combinations of kinetin and NAA resulted in a maximal fresh weight increase at kinetin concentrations one tenth of the NAA concentrations. The optimal growth and the best development occurred at 10 μM NAA and 1 μM kinetin. NAA and kinetin together could limit the shoot and leaf growth induced by GA. 相似文献
12.
Leaf sections of Browallia viscosa and B. speciosa were placedon Murashige and Skoog (1962) salts and vitamins medium (MS)containing auxins and cytokinins, singly or in combination,to elicit morphogenetic responses. B. viscosa developed extensiveroots in 4 weeks on media supplemented with indolebutyric acid(IBA), indol-3-yl acetic acid (IAA) or naphthalene acetic acid(NAA) (0·01, 0·1, 1·0, 5·0 and 10·0mg1), but with 2, 4-D (0·1 mg1) only lightyellow friable callus was obtained. Shoot initiation and elongationoccurred consistently in 46 weeks on leaf sections inthe presence of 6---dimethylallyl amino purine (2iP). Similarly,shoot regeneration from leaf-derived callus, initiated and sub-culturedon MS + benzyladenine (BA) + NAA only induced callus on leafexplants of both species. B. speciosa did not respond exceptfor moderate and prolific callus formation on MS + BA + NAAand Uchimiya and Murashige (1974) media respectively. Browallia viscosa, Browallia speciosa, tissue culture, regeneration, morphogenetic potential 相似文献
13.
Embryogenic cell suspension cultures were established from calliderived from young leaves of sugarcane (Saccharum officinarumL.) by placing them in liquid medium containing 5 per cent coconutwater (CW), 23 mg 11 2, 4-D and 500 mg 11casein hydrolysate (CH). The cultures were maintained by transferring2.55.0 ml of the suspension to 35 ml of fresh mediumevery 45 days. Organized structures resembling the earlystages of embryogeny were formed when 2, 4-D in the medium waslowered (0.11.0 mg 11) but these did not developbeyond the globular or early scutellar stages. High levels ofsucrose (610 per cent) promoted the formation of proembryoids.Plating of the suspension on MS agar medium supplemented with0.252.0 mg 11 2, 4-D, 5 per cent CW, 500 mg 11CH, with or without activated charcoal, resulted in the formationof embryogenic calli. A large number of embryoids were formedin media containing lower 2, 4-D concentrations. Transfer ofembryoids to half-strength MS medium with 6 per cent sucroseestablished plantlets which were successfully transferred tosoil. Saccharum officinarumL, sugarcane, suspension culture, embryogenesis, regeneration 相似文献
14.
Dragan V. Vinterhalter 《Plant Cell, Tissue and Organ Culture》1989,17(1):13-19
Callus tissue was induced in young stem segments cultured on MS based media supplemented with 0.25–0.5 mg l-1 2, 4-D. Shoots were differentiated on media containing 0.5–1.0 mg l-1 BA and 0.5–2.0 mg l-1 IBA or 0.1–0.2 mg l-1 NAA. The same media were suitable for shoot multiplication. Shoot elongation and rooting were strongly inhibited by BA and stimulated by auxins IBA and NAA. Medium containing 0.5 mg l-1 IBA was optimal for rooting. Root elongation was stimulated by light and inhibited in darkness. Transfer of rooted plantlets to outdoor conditions was feasible and special hardening procedures were not required. Among more than 5000 plants produced by this procedure only 9 off-type plants with variegated leaves were found. 相似文献
15.
Flow cytometric determination of DNA levels in embryos of fullymatured seeds of various plant species revealed large amountsof 2C DNA signals, indicating that most cells had arrested thecell cycle at the presynthetic G1 phase of nuclear division.The accumulation of cells at G1 was found both in orthodox andin recalcitrant (i.e. Castanea sativa) seed species. As recalcitrantseeds are characterized by the absence of maturation drying,the arrest of the cell cycle in the presynthetic phase may notbe linked to the seed water status. Apart from the 2C signal, 4C values were found in the embryoof some seed species (e.g. Raphanus sativus) indicating thatcells were arrested in G2 Cells arrested in G2 were primarilylocated in the root-tip region of the embryo. In addition, combinationsof higher C values (i.e. 8C, 12C, 16C and 64C) were observedin the endosperm of Solanum melongena and Lycopersicon esculentum,and in the root-tip cells of Phaseolus vulgaris and Spinaciaoleracea. These mixtures of polyploid nuclei (also called 'polysomaty')may arise from a developmentally controlled cellular endoreduplicationand indicates that in each cell type of the seed the amountof DNA is regulated both spatially and temporally.Copyright1993, 1999 Academic Press Endive, Cichorium endiva, lettuce, Lactuca sativa, egg-plant, Solanum melongena, pepper, Capsicum annuum, tomato, Lycopersicon esculentum, radish, Raphanus sativus, bean Phaseolus vulgaris, spinach, Spinacia oleracea, chestnut, Castanea sativa, beech, Fagus sylvatica, pine, Pinus nigra, DNA content, flow cytometry, seed, nuclear replication stage, C levels, storage 相似文献
16.
Yasunari Ogihara 《Journal of plant research》1979,92(3):163-171
Effects of three different auxins and kinetin in various combinations on greening and redifferentiation of the callus ofHaworthia setata were investigated. All auxins at the concentration of 50 mg/l inhibited callus greening. NAA in combination with kinetin promoted both callus greening and production of redifferentiated
shoots. Low concentrations of IAA without kinetin promoted redifferentiation of shoots, but not callus greening. Addition
of 2,4-D completely inhibited both greening and redifferentiation regardless of the level of kinetin except for the effects
on shoot formation in the medium with 0.1 mg/l 2,4-D added. The calluses with the highest chlorophyll content were observed in the medium containing 2.0 mg/l kinetin without any auxins or with 0.1 mg/l NAA added. Most frequent shoot redifferentiation was observed in the medium containing 0.1 mg/l IAA without kinetin (redifferentiation rate; 67%), followed by the medium containing 10 mg/l NAA with 2.0 mg/l kinetin (44%), and 0.1 mg/l 2,4-D with 0.2 mg/l kinetin (33%). Generally, higher degrees of greening were associated with better growth. However, the auxins (IAA, NAA and
2,4-D) given at concentrations optimal for growth did not exhibit the highest degree of callus greening. Differences of the
three auxins in their actions and interaction with kinetin were disclosed.
Contribution from the Laboratory of Genetics, Faculty of Agriculture, Kyoto University, Japan, No. 423 相似文献
17.
Callus of Orthosiphon stamineus could be induced successfully from petiole, leaf and stem tissues but not roots when cultured on MS medium containing different concentration of NAA (0–4.0 mg l–1) and 2,4-D (0–2.0 mg l–1). Highest fresh weight callus production was obtained from leaf explants and those with best friability were obtained on MS medium plus 1.0 mg l–1 2,4-D plus 1.0 mg l–1 NAA. Cell suspension cultures were established from these cultures. The appropriate cell inoculum size for the best cell growth was 0.75 g of cells in 20 ml culture medium. Cell suspension culture using MS medium supplemented with 1.0 mg l–1 2,4-D promoted the best cell growth with maximum biomass of 8.609 g fresh weight and 0.309 g dry weight 24 days after inoculation. Cells that grew in MS medium supplemented with 1.0 mg l–1 2,4-D reached the stationary growth phase in 15 days as compared to the cells that grew in MS medium supplemented with 1.0 mg l–1 2,4-D + 1.0 mg l–1 NAA reached the stationary phase in 24 days. MS medium supplemented with 1.0 mg l–1 2,4-D was considered as the maintenance medium for maintaining the optimum cell growth of O. stamineus in the cell suspension cultures with 2-week interval subculture. 相似文献
18.
The longitudinal and lateral transport of 2, 4-D-[1-14C] hasbeen studied in 6-mm horizontally disposed segments of the hypocotylof Helianthun annuus. The technique involved the asymmetricapplication of the auxin in agar donor blocks to either theupper or lower halves of the cut surface of the segments andthe measurement of 14C accumulating with time in split receiverblocks on the upper and lower halves of the cut surfaces atthe opposite ends. A highly significant effect of gravity inducinga polar migration of 2, 4-D to the lower side has been establishedand represents, under optimal conditions, about 10 per centof the total 2, 4-D in transit. This lateral polar movementshows a tendency to saturate at higher donor concentrations(5 mg/1) and is affected by temperature in precisely the sameway as the basipetal longitudinal transport. The optimal flux(intensity of transport) occurs at about 35 °C or slightlyabove, and above 40 °C both transport systems are seriouslyimpaired. There is some evidence that gravity increases thevelocity of 2, 4-D transport in the lowermost tissues of thehorizontal hypocotyl but does not affect the transport intensity. 相似文献
19.
Further studies have been made on the interactions of plant-growthregulators during uptake by Avena sativa coleoptile and Phaseolusradiatus hypocotyl segments. 2, 4-Dichlorophenoxyacetic acid(2, 4-D) had no effect on the uptake of either indol-3yl-aceticacid (IAA) or -naphthylacetic acid (NAA) by Avena. On the otherhand, a-(i-naphthylmethylthio)-propionic acid (NMSP) stronglyinhibited IAA uptake non-competitively but was much less effectiveon NAA uptake by Avena. The metabolic uptake ofIAA by hypocotyl segments of Phaseolus radiatus was very stronglyinhibited by 2, 3, 5-tri-iodobenzoic acid (TIBA). 相似文献
20.
为了建立火龙果愈伤组织诱导与植株再生体系,以火龙果茎段、幼苗和子叶为外植体进行离体培养试验。结果表明:茎段诱导愈伤组织的最优培养基为1/2MS+2,4-D2.0mg·L^-1+6-BAO.5mg·L^-1,诱导子叶愈伤组织的最适培养基是1/2MS+2,4-D2.0mg·L^-1+6-BA1.0mg·L^-1,诱导愈伤组织分化的最优培养基为1/2MS+6-BA4.0mg·L^-1+NAA0.5mg·L^-1,最佳生根培养基为1/2MS+6.BA1mg·L^-1+NAA0-3mg·L^-1。 相似文献