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1.
Embryogenic callus cultures were initiated from mature embryosof Lasiurus scindicus on Murashige and Skoog's medium supplementedwith 6 mg l1 2,4-Dichlorophenoxyacetic acid (2,4-D).These cultures were maintained on 2 mg l1 2,4-D. Plantletswere regenerated via somatic embryogenesis when the calli weretransferred onto hormone-free MS basal medium. Young plantswere successfully transplanted to pots and grown to maturityin a greenhouse. Grass, Lasiurus scindicus, Thar Desert, drought tolerant, somatic embryogenesis, plant regeneration 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.
By manipulation of various growth regulators and physical conditions,plants have been regenerated from excised roots, stem segments,cotyledons, leaves, and callus cultures of red cabbage (Brassicaoleracea var. capitata) grown under in vitro conditions. Shootbuds were induced on isolated root segments (1 cm long) culturedon Murashige and Skoog's medium and the frequency of bud formationwas greatly enhanced by the addition of kinetin (0.5 part 106).Callus obtained from the seeds, cotyledons, and hypocotyl segmentscultured on a medium fortified with 2,4-D (1 part 106),kinetin (0.1 part 106), and coconut milk (10%, v/v) hasbeen repeatedly subcultured. The callus is slow growing, andon transference to a kinetin (2 parts 106) and IAA (2parts 106) medium underwent morphogenesis to give riseto plants. The significance of the propagation of red cabbageby in vitro culture is pointed out. 相似文献
4.
Somatic Embryogenesis in Cassava: The Anatomy and Morphology of the Regeneration Process 总被引:2,自引:0,他引:2
Anatomical and morphological studies demonstrated that somaticembryos developed similarly on mature seed and clonal leaf explantsof cassava (Manihot esculenta Crantz) cultured for 2024d on Murashige and Skoog (MS2) basal medium supplemented with4.0 mg l1 2,4-D (Stage 1) before transfer to MS2 basalmedium supplemented with 001 mg l1 2,4-D and 01mg l1 6-benzylaminopurine (Stage II medium). Within 7d of inoculation onto Stage I medium, cell divisions occurredin the adaxial tissues of cotyledon-piece and leaf-lobe explants,and associated with this was the development of embryogeneticprotusions and ridges on the adaxial surface. Foliose structuresand somatic embryo initials developed from these tissues oncotyledon, embryonic axis and leaf-lobe explants and, when cultureswere transferred to Stage II medium, further somatic embryodevelopment occurred. Somatic embryos apparently originatedfrom groups of cells and were identified by the presence ofa closed root axis, a shoot axis and cotyledons of similar shapeand venation to those of zygotic embryos. Somatic embryos hadno vascular connection with parental cultures. Manihot esculenta, cassava, somatic embryogenesis, tissue culture, anatomy, morphology, morphogenesis 相似文献
5.
Shoot bud formation was induced in the stem callus of Sisymbriumirio L., a Cruciferous plant. The callus was established onMurashige and Skoog medium with IAA (1?0 mg l1) and kinetin(0?5 mg l1). The effect of three purines (kinetin, 6-benzylaminopurine,and 6-methylaminopurine) incorporated singly along with IAAin MS medium was investigated. It was found that kinetin orMAP (35 mg l1) along with IAA (0?5 mg l1)were the most effective in inducing shoot bud formation. Adeninesulphate (10 mg l1) with kinetin (1?0 mg l1) alsoinduced bud differentiation. The morphogenetic potential of the callus to differentiate shootbuds was seemingly lost in 2 year old callus cultures. However,on successively subculturing on a regeneration medium shootbuds differentiated and the number of buds formed improved onfurther subculture. Two types of meristematic outgrowths were recognized: (i) arisingfrom superficial cells and (ii) arising from deep-seated cellsin the vicinity of tracheidal elements. However, both typesformed meristematic nodules on the surface of which shoot budsdifferentiated. Some embryoids were also recognized arisingsuperficially. 相似文献
6.
Embryogenic callus was induced from immature inflorescence segmentsof Java citronella (Cymbopogon winterianus) and maintained for2 years on Murashige and Skoog's medium supplemented with 2,4-D(l mg l1). The callus cells retained the original chromosomenumber of 2n = 20. The somatic embryos germinated into plantletson MS basal medium or medium with IAA, NAA, BAP or KN individually(l mg l1). The regenerated plantlets developed a goodroot system on full strength solid MS inorganics medium withIAA (1 mg l1). The regenerated plants were similar tothe donor plant in morphology and had the same chromosome number,but showed some variation in the essential oil content. Java citronella, Cymbopogon winterianus, somatic embryogenesis, regeneration, inflorescence culture 相似文献
7.
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
8.
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
9.
Ethylene and Ethane Production in 2,4-D Treated and Salt Treated Tobacco Tissue Cultures 总被引:1,自引:0,他引:1
Gas chromatography was used to measure ethylene (ethene) andethane production by tobacco (Nicotiana tabacum cv. Wisconsinno. 38) callus tissues grown on media containing inorganic saltsaccording to Murashige and Skoog (1962), sucrose, myo-inositol,thiamine-HCl kinetic according to Linsmaier and Skoog (1965),and either 2,4-dichiorophenoxyacetic acid (2,4-D) in the range0100 mgl1 or 2 mgl1 indoi-3-ylacetic acidplus NaCl in the range 0200 Meq l1. Ethylene productionrates were high (> 500 nl h1 g1 fresh weight)initially in all treatments. Subsequently, ethylene productiondeclined in rapidly growing cultures but remained high in moderatelyand severely 2,4-D (> 0·5 mgl1) stressed andin severely NaCl (150 Meql1) stressed cultures. Highinitial rates of ethane production (> 200 nl h1 g1fresh weight) were obtained under conditions of severe stresscaused by 2,4-D or NaCl but not in control or moderately inhibitedcultures. With further incubation ethane production declinedin the severely stressed cultures. It is concluded that ethyleneproduction can be used as an index of moderate 2,4-D stressand severe NaCl stress by virtue of the high persisting ratesof ethylene production in stressed cultures. Ethane productioncan be used as an early index of severe stress caused by either2,4-D or NaCl in vitro. Nicotiana tabacum L., tobacco, ethylene, ethenen, ethane, 2,4-dichlorophenoxyacetic acid, auxin, stress, callus tissue 相似文献
10.
The Growth, Anatomy and Morphogenetic Potential of Callus and Cell Suspension Cultures of Hevea brasiliensis 总被引:1,自引:0,他引:1
Callus cultures have been initiated from stem explants of youngplants of Hevea brasiliensis and maintained over long periodsat 30 ?C by serial subculture in Murashige and Skoog mediumcontaining 2 mg 11 2,4-D and 0.5 mg 11 kinetin.Newly-initiated cultures spontaneously initiated roots but,on serial subculture, this property was lost and the culturesbecame heterogeneous (consisting of proliferating light segmentsand darker compact non-growing segments). Serially propagatedcultures continued to differentiate a few scattered latex vesselscontaining particulate material similar to that in the rootlaticifers. This callus (O callus) did not yield a growing cellsuspension when transferred to agitated liquid medium. However,the large cell aggregates which could be recovered after twopassages in liquid medium, when again grown on solid mediumyielded a highly friable light-coloured fast-growing homogeneouscallus (R callus) which retained its distinctive character onsubculture. This callus when transferred back to agitated liquidmedium yielded a fine rapidly growing cell suspension culturewhich could be serially propagated at 30 ?C in the same mediumas that used for callus culture. Both the O and R cultures were2,4-D dependent, but differed in their responses to 2,4-D. Bothretained their diploid character when serially propagated. Serially-propagatedsuspensions came to contain a proportion of polyploid cells.When the suspensions were maintained for several months withoutsubculture the larger cell aggregates which developed gave riseto embryo-like structures. Attempts to promote the further developmentof these embryo-like structures into plantlets were unsuccessful. 相似文献
11.
Agrobacterium rhizogenes Mediated Transformation of the Forage Legumes Medicago sativa and Onobrychis viciifolia 总被引:3,自引:0,他引:3
GOLDS T. J.; LEE J. Y.; HUSNAIN T.; GHOSE T. K.; DAVEY M. R. 《Journal of experimental botany》1991,42(9):1147-1157
Three cultivars of M. sativa and one cultivar of O. viciifoliawere evaluated for their response to inoculation with A. rhizogenesstrain A4T (containing pRiA4b). A cultivar-dependent responsewas observed in M. sativa with 94%, 25%, and 4% of infectedstem explants producing transformed roots in the cultivars Vertus,Regen-S, and Rangelander, respectively. In O. viciifolia cv.Hampshire Giant, an explant-dependent response was observedwith 78% and 50% of seedling cotyledon and hypocotyl explantsresponding, respectively. Leaf explants failed to produce transformedroots. Transformed roots showed plagiotropic and negativelygeotropic growth on hormone-free agar MS medium. Productionof transgenic shoots from O. viciifolia root cultures occurredspontaneously. Recovery of transgenic plants from M. salivacv. Rangelander was achieved by transfer of callus (inducedon UM medium containing 2·0mg dm3 2,4-D and 0·25mg dm3 kinetin) to MS medium containing 0·5 ingdm3 BAP and 0·05 mg dm3 NAA. Cultured rootsof both species synthesized opines (agropine and mannopine).Extensive morphological variation was observed in plants ofM. sativa (clone Al) and O. viciifolia (clone A4Tl) establishedin the glasshouse. DNA sequences homologous to TL-DNA and TR-DNAwere present in root clones and regenerated plants. Key words: Agrobacterium rhizogenes, Medicago sativa, Onobrychis viciifolia, transformed roots, transgenic plants 相似文献
12.
Callus cultures of Nardostachys jatamansi DC. maintained onMurashige and Skoog's medium containing 3.0 mg 11 of-naphthaleneacetic acid and 0.25 mg 11 of kinetin whenshifted to medium containing 0.251.0 mg 11 ofindole-3-acetic acid or indole-3-butync acid showed profuserhizogenesis. The callus-regenerated roots when transferredto medium containing 2.06.0 mg 11 of kinetin producedshoot buds. The de novo shoot bud regeneration took place eitherdirectly from cortical cells or from the inner stelar region.In addition, direct, concomitant root-shoot development wasalso observed. Nardostachys jatamansi, organogenesis, root-buds 相似文献
13.
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
14.
Explants obtained from the basal portion of leaves of Hordeumvulgare (cv. Karan 92) gave rise to callus when cultured onMurashige and Skoog (MS) basal medium supplemented with 2, 4-dichlorophenoxyaceticacid (2, 4-D). Initially, the callus was friable, shiny-whiteand watery but subsequently some compact, nodular callus appeared.The latter were cultured on MS medium containing 0.05 mg l12, 4-D and 0.1 mg l1 N6-furfurylaminopurine (kinetin),when plantlets were generated. Histological studies showed thatplantlet regeneration occurred by the formation of somatic embryos.The regenerated plants had the normal diploid chromosome number(2n = 14). Hordeum vulgare, barley, somatic embryogenesis, tissue culture, plant regeneration 相似文献
15.
An 18-h treatment of synchronously-grown Chlorella pyrenoidosawith 2,4-D did not significantly alter the size, dry weight,degree of synchrony, or pigment content of the cells, nor weredetectable quantities of ethylene produced. When Chlorella pyrenoidosawas treated with 5?104 M 2,4-D, there was a statisticallysignificant stimulation of both net oxygen uptake and productionwhile 5?10 M 2,4-D inhibited both processes. When Chlorellapyrenoidosa was treated with 5?104 M and 5?103M 2,4-D, significantly greater amounts of glycollate were presentin the culture medium, even though an assay for glycollate dehydrogenaseshowed that the activity of this enzyme from 2,4-D-treated Chlorellapyrenoidosa was three times greater than in control cells. Looselybound 2,4-D was partitioned from a nonaqueously isolated chloroplastfraction, while other cell fractions failed to show detectablequantities of 2,4-D. It is postulated that in Chlorella pyrenoidosathe chloroplast is a target for 2,4-D action and that interferencein photorespiratory processes may underlie the observed responses. 相似文献
16.
Callus cultures of Ipomoea pes-caprae and I. batatas were establishedon MS medium containing 105 M 2,4-D and 108 Mbenzyladenine. Ipomoea pes-caprae calli exhibited green pigmentationand grew better in the light than in darkness. Callus tissuesof I. batatas showed a pale-yellow colouration and they grewat the same rate in light as in dark conditions. I. pes-capraeand I. batatas callus cultures were sensitive to the presenceof 60 mM NaCl in the culture medium, the growth of the formerbeing more sensitive in light than in darkness. The significanceof the responses of I. pes-caprae callus cultures in relationto the mechanism of salt tolerance is discussed. Ipomoea batatas, Ipomoea pes-caprae, sweet potato, railroad vine, callus cultures, salinity, light 相似文献
17.
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
18.
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
19.
Somatic Embryogenesis and Plant Regeneration from Cultured Leaf Explants of Zea mays 总被引:4,自引:0,他引:4
Young leaf segments of Zea mays L. seedlings were cultured onMurashige and Skoog's basal nutrient medium supplemented with2 mg l1 2, 4-D and sub-cultured on medium containing8 mg l1 2,4-D. Two types of callus tissues appearedembryogenicand non-embryogenic. The embryogenic callus tissue producednumerous somatic embryos which on transfer to media containinglow amounts of 2,4-D or ABA produced plantlets. Callus tissuesexhibited embryogenic potential for more than 1 year. Zea mays L. cv. Ageti-76, Zea mays L. cv. N-L-D-Comp., maize, leaf, callus, somatic embryogenesis, regeneration 相似文献
20.
Wheat Callus Culture: the Initiation, Growth and Organogenesis of Callus Derived from Various Explant Sources 总被引:2,自引:0,他引:2
Callus was obtained from mature excised embryos of wheat, fromnodal and internodal stem segments and from rachis segmentsusing the medium of Murashige and Skoog(1962)(M medium), containing1-0mg l1 2,4-D, and from immature embryos using the mediumof Green and Phillips (1975) containing 2 mg l1 2,4-D.Callus yield from mature embryos depended upon the cultivarused. No callus could be obtained from leaf segments. Callusderived from mature embryos and nodal stem segments was successfullymaintained by serial sub-culture on the M medium containing2,4-D for up to 3 years although its growth rate declined toa lower level as culture proceeded. Such cultures consistently produced roots when transferred toa medium containing a low level of 2,4-D or no 2,4-D. The presenceof the auxin was essential for continued proliferation of thecallus tissue. Shoot initiation was infrequent, did not occurafter the first few sub-cultures and could not be enhanced byvarious auxin and cytokinin additions to the medium. Callusderived from immature embryos did not have an enhanced potentialfor shoot initiation. Triticum aestivum, wheat, callus culture, organogenesis 相似文献