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1.
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
2.
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
3.
Callus of sainfoin (Onobrychis viciifolia Scop.) was initiatedfrom stem and root explants which were obtained from seedlingsgrowing in vitro, on Linsmaier Skoog (LS) medium supplementedwith 1 mg l1 2, 4-D and 1 mg l1 BA or only 1 mgl1 BA, and the Vacin and Went medium without hormones.Somatic embryos were formed on LS medium containing 1 m l1BA. Embryos developed into complete plants on filter paper saturatedwith hormone-free LS medium. Onobrychis viciifolia, somatic embryogenesis, callus culture, plant regeneration 相似文献
4.
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
5.
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
6.
Qian Zhang Jianjun Chen Richard J. Henny 《Plant Cell, Tissue and Organ Culture》2006,84(2):100161-100168
A simple and effective method of regenerating Syngonium podophyllum ‘Variegatum’ via direct somatic embryogenesis has been established. Leaf and petiole explants were cultured on Murashige
and Skoog (MS) medium supplemented with N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU) or N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ) with either α-naphthalene acetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D). Somatic embryos directly formed at one or two sides of petiole explants on MS medium supplemented 2.5 mg l−1 TDZ with 0.5 mg l−1 NAA or 2.0 mg l−1 TDZ with 0.2 mg l−1 NAA or with 0.2 and 0.5 mg l−1 2,4-D, respectively. The frequency of petiole explants with somatic embryos produced was as high as 86% when cultured on medium
containing 2.5 mg l−1 TDZ with 0.5 mg l−1 NAA. Up to 85% of somatic embryos were able to germinate after transferring onto medium containing 2.0 mg l−1 6-benzylaminopurine (BA) and 0.2 mg l−1 NAA. Approximately 50–150 plantlets were regenerated from a single petiole explant. However, there was no somatic embryo
formation from leaf explants regardless of growth regulator combinations used. Regenerated plantlets from petiole explants
were stable and grew vigorously after transplanting to a soilless container substrate in a shaded greenhouse. 相似文献
7.
A new method was established for somatic embryogenesis and plant regeneration from callus cultures of Dioscorea zingiberensis C.H. Wright. Primary callus was induced by culturing stems, leaves and petioles on Murashige and Skoog (MS) medium supplemented with 0.5–2.0 mg l–1 N6-benzyladenine (BA) and 0–2.0 mg l–1 -naphthaleneacetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D) for 1 month. The highest frequency (87%) of callus formation was achieved from stem explants treated with 0.5 mg l–1 BA and 2.0 mg l–1 2,4-D. Somatic embryos were obtained by subculturing embryogenic calli derived from stem explants on MS medium supplemented with 2.0–4.0 mg l–1 BA and 0–0.4 mg l–1 NAA or 2,4-D for 3 weeks. The optimum combination of 4.0 mg l–1 BA and 0.2 mg l–1 NAA promoted embryo formation on one-third of the calli. After a further month of subculture on the same medium, mature embryos were transferred to MS medium supplemented with 0–4.0 mg l–1 BA, NAA or indole-3-butyric acid (IBA) for further development of plantlets and tuber formation. Plant growth regulators had a negative effect on the development of mature embryos. 相似文献
8.
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
9.
Somatic embryogenesis can be induced in tissue cultures of Freesiarefracta either directly from the epidermal cells of explants,or indirectly via intervening callus. These two pathways ofsomatic embryogenesis can be controlled and regulated by varyingthe combinations and levels of exogenous hormones. When younginflorescence segments were cultured in vitro on modified N4(MN4) medium supplemented with 2 mg l1 indoleacetic acid(IAA) and 3 mg l1 6-benzylaminopurine (BAP), some ofthe epidermal cells began to exhibit the features of embryogeniccells. These cells produced embryoids and developed into newplants through direct somatic embryogenesis. If the same explantswere placed on Murashige and Skoog's (MS) medium containing2 mg l1 IAA, 05 mg l1 BAP and 05 mg l1naphthaleneacetic acid (NAA), pale-yellow translucent nodularcalluses appeared on the surface of the explants. When thiskind of callus was transferred to MN6 medium with 2 mg l1IAA and 3 mg l1 BAP, embryoids formed which further developedinto plantlets. The regenerated plants were morphologicallynormal and possessed the normal diploid chromosome number of2n = 22. A similar result has also been obtained with youngleaf explants of this plant. The early segmentations of embryogeniccells and the development of embryoids were studied using histologicaland scanning electron microscopic techniques, and the resultshave been discussed in association with the ontogeny and originof the embryoids. Freesia refracta Klatt, somatic embryogenesis, plant regeneration, exogenous hormones 相似文献
10.
Somatic embryos and embryogenic callus were initiated from immature zygotic embryos of ginseng (Panax ginseng C.A. Meyer). These somatic embryos were multiplied by adventitious (secondary and tertiary) embryogenesis and their growth and development were dependent on growth hormones in the medium. Auxins, 2,4-d, NAA, and IAA at 1.0 mg l-1 were effective in inducing secondary and tertiary somatic embryos, which proliferated directly from the apical or cotyledonary portions of the primary somatic embryos. Single somatic embryos or clusters or embryos developed from the explanted primary embryos. Cytokinin (Kn, BA) inhibited adventitious embryogenesis. Secondary somatic embryos developed to maturation and later regenerated into plantlets in two stage process; firstly elongation of the shoot axes on MS +1.0 mg l-1 Kn, secondly formation of root on 1.0 mg l-1 Kn+1.0 mg-1 GA3 medium.Abbreviations 2,4-d
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- IAA
in-doleacetic acid
- Kn
kinetin
- BA
benzylaminopurine
- PSE
primary somatic embryo
- SSE
secondary somatic embryo
- TSE
tertiary somatic embryo 相似文献
11.
Somatic embryogenesis in peanut (Arachis hypogaea L.) using immature cotyledonary explants was induced on a wide range of 2,4-dichlorophenoxyacetic acid (2,4-D) (5 to 60mg l–1) and naphthaleneacetic acid (NAA) (20 to 50 mg l–1) levels. Percent embryogenesis ranged from 31 to 94%. As auxin level increased in induction medium, percent embryogenesis decreased and was associated with browning of explants. However, with higher 2,4-D induction levels (40 mg l–1 and over), embryogenic explants had dense masses of embryogenic areas and repetitive embryogenesis was enhanced. Higher auxin concentrations during induction decreased precocious germination of embryos, but had no marked effect on somatic embryo morphology. The use of 2,4-D compared to NAA in the induction medium resulted in greater per cent embryogenesis and mean number of embryos. Embryos induced on NAA were harder, less pliant, and less succulent; cultures exhibited more extensive root development and nonembryogenic callus proliferation.Abbreviations B5
Gamborg et al. (1968)
- BA
benzyladenine
- 2,4-D
dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- MS
Murashige & Skoog (1962)
- NAA
naphthaleneacetic acid
- picloram
4-amino-3,5,6-trichloropicolinic acid 相似文献
12.
Jing Li Yang Bo Zhao Eun Soo Seong Myong Jo Kim Won Hee Kang Na Young Kim Chang Yeon Yu Cheng Hao Li 《Plant biotechnology reports》2010,4(4):261-267
We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver
nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when
petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg l−1 α-naphthaleneacetic acid (NAA) and 0.1 mg l−1 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously
from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing
1.0 mg l−1 BA and 1.0 mg l−1 NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg l−1 GA3 had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully
acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the
primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction
of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained
by 1% sucrose. Most secondary embryos (87.2–94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary
secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose. 相似文献
13.
A procedure is described to regenerate plants from embryogenic suspension-derived protoplasts of ginger (Zingiber officinale Rosc.). Somatic embryogenic calli were induced from ginger shoot tips on solid MS medium with half the concentration of NH4NO3 and supplemented with 1.0 mg l−1 2,4-Dichloroacetic acid (2, 4-D) and 0.2 mg l−1 Kin. Rapid-growing and well-dispersed suspension cultures were established by subculturing the embryogenic calli in the same
liquid medium. Protoplasts were isolated from embryogenic suspensions with an enzyme solution composed of 4.0 mg l−1 cellulase, 1.0 mg l−1 macerozyme, 0.1 mg l−1 pectolyase, 11% mannitol, 0.5% CaCl2 and 0.1% 2-(N-morpholino) ethane sulphonic acid (MES) for 12–14 h at 27°C with a yield of 6.27 × 106 protoplasts g−1 fresh weight. The protoplasts were cultured initially in liquid MS medium with 1.0 mg l−1 2, 4-D and 0.2 mg l−1 Kin. Then the protoplast-derived calli (1.5 cm2) were transferred to a basal MS medium containing 0.2 mg l−1 2, 4-D, 5.0 mg l−1 benzyladenine (BA), 3% sucrose and 0.7% agar. The white somatic embryos were transferred to MS medium lacking growth regulators
for shoot development. Shoots developed into complete plantlets on a solid MS medium supplemented with 2.0 mg l−1 BA and 0.6 mg l−1 α-Naphthaleneacetic acid (NAA). In addition, the effects of AgNO3, activated charcoal (AC) and ascorbic acid (AA) on browning of protoplast-derived calli are discussed. 相似文献
14.
Friable embryogenic callus and somatic embryo formation from cotyledon explants of African marigold (Tagetes erecta L.) 总被引:3,自引:0,他引:3
Embryogenic callus and somatic embryos were induced from cotyledonary explants of African marigold (Tagetes erecta L.). Cotyledons were first cultured on MS medium supplemented with 2.0 mg l–1 2,4-D and 0.2 mg l–1 kinetin. After 5 weeks, calli were transferred to MS medium supplemented with 0.02 mg l–1 thidiazuron where compact embryogenic callus developed. Friable embryogenic callus developed when the compact embryogenic
callus was transferred to medium containing 2,4-D and subcultured every 2 weeks. Friable embryogenic callus has been maintained
for more than 2 years without losing the capacity to generate embryos. Embryo development was obtained when friable embryogenic
callus was transferred to MS medium supplemented with 3 mg l–1 ABA and 60 g l–1 sucrose. The addition of 10–30 mM
l-glutamine improved embryo development.
Received: 13 May 1997 / Revision received: 24 February 1998 / Accepted: 28 March 1998 相似文献
15.
Hypocotyl explants of S. melongena showed three types of regenerationthrough callus formation depending on the concentration of NAAin the medium. At 0.8 mg l1, only callus was produced.Lower concentrations resulted in callus, adventitious roots(optimum, 0.016 mg 11 NAA), and adventitious shoots (noNAA). Roots and shoots developed during the early stages ofculture. Higher concentrations of NAA depressed callus growthand stimulated embryoid formation (optimum 8.0 mg 11NAA), Embryoids were identifiable after about 6 weeks as greenspots on the surface of callus: Addition of 6-BA enhanced shootproduction but inhibited both root and embryoid production.Whole plants were obtained from embryogenic callus after transferto NAA free medium. Genotypic differences in response were observed. In general,the potential for embryogenesis was independent of or inverselyrelated to the potential for organogenesis. 相似文献
16.
Yongxue Yang Guofeng Liu Manzhu Bao 《In vitro cellular & developmental biology. Plant》2006,42(6):520-524
Summary A protocol of somatic embryogenesis and plant regeneration from petiole segments of Parthenocissus tricuspidata Planch. has been developed. Embryogenic tissue was induced on B5 (Gamborg) basal medium supplemented with 2.25–9.0 μM 2,4-dichlorophenoxyacetic acid, 500 mg l−1 casein hydrolysate (CH), and 0.1 gl−1 activated charcoal. Somatic embryos were induced on B5 medium containing various concentrations of benzyladenine (BA) (4.44,
6.66, and 8.88 μM) and α-naphthaleneacetic acid (NAA) (0, 0.54, and 1.61 μM) plus 500 mg l−1 CH. Ninety percent of normal somatic embryos were converted into plantlets directly on Murashige and Skoog (MS) medium free
of plant growth regulators. Shoots could be induced from abnormal somatic embryos on MS medium containing 4.44 μM BA, 0.05 μM NAA, and 500 mg l−1 CH. Genotypic differences were found in the process of somatic embryogenesis and plant regeneration. Histological analysis
confirmed the process of somatic embryogenesis. Regenerated plantlets with well-developed roots were successfully acclimatized
in greenhouse and all plants showed normal morphological characteristics. 相似文献
17.
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
18.
Ai Hua Chen Jing Li Yang Yu Da Niu Chuan Ping Yang Gui Feng Liu Chang Yeon Yu Cheng Hao Li 《Plant Cell, Tissue and Organ Culture》2010,102(3):357-364
We developed a new protocol for highly efficient somatic embryogenesis and plantlet conversion of Schisandra chinensis. Friable embryogenic callus was induced from cotyledonary leaves and hypocotyls of germinated zygotic embryos on Murashige
and Skoog (MS) agar medium containing 2,4-dichlorophenoxyacetic acid (2,4-D). Preculture of zygotic embryos on 2,4-D-containing
medium increased embryogenic callus induction efficiency. The highest embryogenic callus induction frequency of 56.7% was
obtained from shoot apical meristem-containing hypocotyl explants from 1-week-old germinated embryos on MS medium containing
4.0 mg l−1 2,4-D. Embryogenic callus proliferation, somatic embryo (SE) formation, and subsequent plantlet conversion occurred under
optimal culture conditions. The effects of MS medium strength, sucrose, gibberellic acid (GA3), and 6-benzyladenine (BA) on SE formation and plantlet conversion were evaluated. Low MS medium strength (1/4 to 1/2) was
necessary for SE formation, and the optimal sucrose concentration was 2.0%. Supplementing medium with GA3 negatively impacted SE formation and subsequent development. BA significantly increased the number of SEs and the plantlet
conversion capacity. One-third-strength MS medium with 1.0% sucrose and 0.5 mg l−1 BA produced the highest number of SEs (309 embryos from 9 mg embryogenic callus) and the highest frequency of plantlet conversion
from germinated SEs (52.6%). When transplanted to soil, 90% of the regenerated plants developed into normal plants. 相似文献
19.
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
20.
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