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1.
Archana Giri Paramvir Singh Ahuja P. V. Ajay Kumar 《Plant Cell, Tissue and Organ Culture》1993,32(2):213-218
Plants were obtained via somatic embryogenesis in callus derived from in vitro raised leaf and petiole explants of Aconitum heterophyllum Wall. Callus was induced on a Murashige-Skoog medium supplemented with either 2,4-dichlorophenoxy acetic acid (2,4-d 1 mg l-1) and kinetin (KN 0.5 mg l-1) with coconut water (CW 10% v/v) or naphthalene acetic acid (NAA 5 mg l-1) and benzylaminopurine (BAP 1 mg l-1). Somatic embryos appeared after 2–3 months or 2 subculture passages when 2,4-d or NAA induced source of the callus was transferred to a MS medium containing BAP (1 mg l-1) and NAA (0.1 mg l-1). For successful plantlet formation, the somatic embryos were transferred to a medium containing 1/4 strength MS nutrient with indole-3-butyric acid (IBA 1 mg l-1). Alternatively, the somatic embryos were dipped in a concentrated solution of IBA for 5 min and placed on a hormone free medium. Complete plantlets were formed after 4 weeks and were transferred successfully to soil.CIMAP Publication No. 1020. 相似文献
2.
A protocol has been established for rapid, high frequency plant regeneration from protoplasts of the wild tomato species Lycopersicon chilense Dun. Cell suspension cultures were obtained from calli initiated from seedling stem explants. Protoplasts were isolated from cell suspensions by an overnight one-step enzyme digestion, purified by washing in salts solution and cultured in liquid medium. Dilution of liquid medium every 3 days, with medium containing low levels of growth regulators and sucrose, was critical for sustained colony formation. Up to 70% of protoplast-derived calli regenerated shoots when cultured on agar-solidified medium with Murashige & Skoog (1962) salts and vitamins, 2.0 mg l-1 zeatin and 0.1 mg l-1 indole acetic acid for 21 days, followed by transfer to the same medium lacking indole acetic acid.Abbreviations BAP
6-benzylaminopurine
- IAA
indole acetic acid
- IBA
indole butyric acid
- MES-2
(N-morpholino)-ethane sulfonic acid
- NAA
-naphthaleneacetic acid
- 2,4-d
2,4-dichlorophenoxyacetic acid 相似文献
3.
Plant rgeneration occurred on leaf-and stem-derived callus of Cuphea ericoides Cham. & Schlechtd obtained in Murashige and Skoog medium supplemented with auxins [indole-3-acetic acid (IAA), -naphthaleneacetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-d)] plus cytokinins [6-benzyladenine (BA) or kinetin]. These calluses were subcultured and showed vigorous growth. When subcultured on medium containing 2.22 or 4.44 M BA, the calluses showed profuse regeneration of shoots whereas those subcultured on medium supplemented with 2.69 M NAA or 0.226 M 2,4-d produced numerous roots. Isolated shoots rooted on Murashige and Skoog medium lacking growth regulators or containing 0.54 M NAA or 0.49 M indole-3-butyric acid (IBA). Plantlets were acclimatized to greenhouse conditions.Abbreviations BA
6-benzyladenine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- MS
Murashige & Skoog medium
- NAA
1--naphthaleneacetic acid 相似文献
4.
Joseph C. V. Vu Randall P. Niedz George Yelenosky 《Plant Cell, Tissue and Organ Culture》1993,35(1):75-80
Anthers of niger (Guizotia abyssinica. Cass) were inoculated onto five different media differing mainly in their inorganic and organic constituents and plant growth regulators to study their influence on callus induction (embryogenic/non-embryogenic) and plant regeneration. LS medium supplemented with 2 mg 1-1 2,4-d, and 0.3 mg 1-1 KN favoured the production of EC, whereas 2 mg 1-1 BAP and 0.5 mg 1-1 KN promoted the NEC from anthers. Different types of embryos were initiated upon transfer of EC to Chaleff's R-2 medium containing 2 mg 1-1 NAA and 0.3 mg 1-1 KN and/or 5 mg 1-1 ABA. NEC when transferred onto the medium supplemented with 1 mg 1-1 BAP and 0.1 mg 1-1 NAA produced on an average 8–12 shoots/callus mass. Embryoids developed from the EC and shoots differentiated from NEC when cultured onto the Chaleff's R-2 and MS media respectively lacking growth regulators, they transformed into whole plantlets. The plantlets thus obtained were successfully hardened and grown to maturity for analysis of various plant characters.Abbreviations EC
embryogenic callus
- NEC
non-embryogenic callus
- 2,4-d
2,4-dichlorophenoxyacetic acid
- NAA
-naphthaleneacetic acid
- ABA
abscisic acid
- BAP
6-benzylaminopurine
- KN
kinetin
- MS
Murashige and Skoog's medium
- LS
Linsmaier and Skoog's medium 相似文献
5.
Leaf, stem and root explants of Mandevilla velutina were cultured in vitro and produced vigorous callus in LS basal medium containing one auxin (2,4-D or NAA) plus BAP. Calli can be subcultured indefinitely with vigorous growth. Subculture of calli to NAA (1.0 mg/l) plus BAP (5.0 mg/l) caused profuse regeneration of shoots. Isolated shoots were rooted in basal medium plus NAA (5.0 mg/l) or IBA (8.0 mg/l). Rapidly growing cell suspensions can be easily obtained from friable callus cultured in liquid medium.Abbreviations LS
Linsmaier & Skoog
- 2,4-D
2,4 dichlorophenoxi-acetic acid
- NAA
-naphthalene-acetic acid
- GA3
gibberellic acid
- IAA
indole-3-acetic acid
- BAP
6-benzylaminopurine
- IBA
indole-3-butyric acid 相似文献
6.
Statistical analyses of the data revealed very significant differences in androgenesis induction ofA. carnea Hayne anther culture depending on the bud length, nutrient medium composition and age of the parental tree. Significant mutual influence of all these factors was also observed. The highest number of androgenic anthers was obtained when 4 mm long buds were used. Older trees (60 and 100 yrs) gave a higher number of androgenic anthers than the younger ones (20 and 40 yrs). MS medium supplemented with 2,4-d and Kin (1 mg l–1, each) was the most favourable for androgenesis induction. Pollen embryos (haploids and aneuploids) were formed by the division of uninuclear microspores.The highest percentage of germinated embryos and further synchronous development of the shoot and root was achieved in MS medium supplemented with IAA, GA3 (1 mg l–1) and activated charcoal (1%). When other germination media were used, malformations of androgenic embryos were observed.Abbreviations AC
activated charcoal
- H
casein hydrolysate
- 2,4-d
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- BAP
6-benzylaminopurine
- GA3
gibberelic acid
- Kin
6-furfurylaminopurine
- MS
Murashige and Skoog
- T
thidiazurone
- N
phenyl-N'-1,2,3-thiadiazol-5-ylurea
- Z
zeatin-6-(4-hydroxy-3-methyl-trans-2-butenylamino)purine 相似文献
7.
Root, hypocotyl and cotyledonary explants of niger (Guizotia abyssinica Cass) CV. Sahyadri were aseptically cultured on Murashige and Skoog's basal medium (MS) containing BAP and kinetin. Multiple shoot regeneration was induced from hypocotyl and cotyledonary explants while root explants produced only callus on MS medium supplemented with BAP. BAP (1 mg l-1) was optimum for shoot regeneration. Regenerated shoots were transferred to MS medium without auxins, with auxins and with increasing concentrations of sucrose for rooting. Complete plantlets were obtained in all cases; however, 0.5 mg l-1 NAA was the best for induction of roots. Ninety-seven per cent of the plantlets survived and completed their life cycle when transferred to natural conditions.Abbreviations BAP
6-benzylamino purine
- NAA
-naphthaleneacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid 相似文献
8.
C. L. Webb M. R. Davey J. A. Lucas J. B. Power 《Plant Cell, Tissue and Organ Culture》1994,38(1):77-79
Cultured protoplasts of young, unexpanded leaves of the wild lettuce, Lactuca perennis, divided to produce cell colonies in an agarose-solidified, modified MS medium with reduced levels of inorganic salts, together with 2,4-d, NAA and zeatin at 0.2, 0.1 and 0.5 mg 1-1 respectively. Organogenesis followed the initial transfer of protoplastderived colonies to modified MS medium with 2,4-d, NAA and zeatin (0.1, 1.0 and 0.2 mg 1-1 respectively) and then to full-strength MS medium with 6-BA and NAA (0.4 and 0.05 mg 1-1). Shoots were rooted on agar-solidified MS medium lacking growth regulators. Regenerated shoots were established ex vitro, 21 weeks after protoplast isolation.Abbreviations 6-BA
6-benzyladenine
- BSA
bovine serum albumin
- d
days
- 2.4-d
2,4-dichlorophenoxyacetic acid
- f. wt.
fresh weight
- IAA
indoleacetic acid
- MES
2 [N-morpholino]ethane sulphonic acid
- MS
Murashige & Skoog (1962) 相似文献
9.
Callus cultures were established from immature embryos of Calotropis gigantea (Linn.) R. Br. on a modified basal medium of Murashige & Skoog supplemented with 1 mgl-1 2,4-D. In addition to 0.1 mgl-1 of NAA the optimal BAP concentration for promoting shoot bud formation and growth was 2 mgl-1. Rooting was induced when shoots were transferred to auxin-supplemented Bonner's solution or half-strength MS basal salt solutions.Abbreviations NAA
-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-butyric acid
- BAP
6-benzylaminopurine
- Kin
kinetin 相似文献
10.
Callus cultures of Solanum paludosum were established from roots, hypocotyles, cotyledons and leaf limbs of plantlets cultivated in sterile conditions on a Murashige and Skoog's modified medium. Non organogenous calluses were obtained with addition of BA or kinetin (10-5M to 10-6M) as the cytokinin and 2,4-d or NAA (10-5M to 10-6M) as the auxin. These calluses permitted the establishment of a cell suspension culture with BA (10-6M) and 2,4-d (10-6M). Zeatin (10-6M) with IAA (10-6M) gave rise to organogenous calluses. These organogenous callus cultures developed multiple shoots which either proliferated if they were cultivated on a medium containing zeatin with IAA or IBA or were able to regenerate into whole plants when zeatin was used as the only hormone. The different plant material produced solamargine, the main steroidal glycoalkaloid present in the unripe fruits. The best production was obtained with the fruits of regenerated plants from organogenous callus cultures after reintroduction of these plants in their brasilian biotope. The solamargine content of the two types of plant materials was about 0.06% and 2.5% (dry weight) respectively for the callus cultures and the fruits from in vitro plants. The fruits were harvested a year after the beginning of the plantlet regeneration step.Abbreviations HPTLC
high performance thin layer chromatography
- HPLC
high performance liquid chromatography
- 2,4-d
2,4-dichlorophenoxyacetic acid
- BA
benzylaminopurine
- IAA
3-indolebutyric acid
- NAA
-naphthaleneacetic acid
- IBA
3-indolebutyric acid
- IPA
isopentenyladenine 相似文献
11.
Induction of haploid callus and embryogenesis in in vitro cultured anthers of mulberry (Morus indica) 总被引:1,自引:0,他引:1
Anthers of Morus indica L., with microspores at the uninucleate stage were cultured; and the influence of temperature and kinetin pretreatment on induction of androgenic calluses was examined. The effects of various pretreatments revealed that 24 h cold pretreatment increased the percentage of cultures inducing callus. First microspore division was observed after 16 to 20 days of culture. Th anthers split and developed embryogenic calluses on MB medium supplemented with NAA (0.5 mg l–1 and BA (1.0 mg l–1)) using 8% sucrose. Rhizogenesis was induced on medium supplemented with NAA and BA (each 0.5 mg l–1) with reduced myo-inositol (75 mg l–1). Cytological study of induced roots confirmed the haploid nature of calluses. Different type of embryos were initiated upon transfer of calluses to medium supplemented with NAA, BA (each 0.5 mg l–1), 2,4-d (1.0 mg l–1) and PVP (600 mg l–1). These embryoids further developed roots on removal of 2,4-d from the medium and developed precociously without developing cotyledons and formed elongated shoots.Abbreviations BA
6 benzylaminopurine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- FAA
formalin: Acetic acid: Alcohol
- GA3
gibberellic acid
- IBA
indole-3-butyric acid
- MB
modifed Bourgin (Qian et al., 1982)
- NAA
1-naphthalene acetic acid
- PVP
polyvinylpyrrolidone
- RFS-135
rainfed selection 135
- SE
standard error 相似文献
12.
The number of dividing microspores of Coffea arabica L. cv. Catuai and Catimor could be drastically increased in microspore media containing 16% (w/v) coconut milk, allowing cell divisions to continue in the microspore and multicellular microspores to survive until day 60. After a cold treatment, the microspores were mechanically isolated prior to cultivation in Murashige and Skoog medium supplemented with sucrose and maltose and (mg l-1) 2,4-d: 2, BAP: 1 or a combination of kinetin: .5, 2,4-d: .5 and NAA: .5 as stationary suspension at a density of 1,200/ml. The crucial stage during microsporogenesis suitable for in vitro androgenesis proved to be mid uninucleate till early binucleate in flowerbuds with the size of 13–15 mm two to three days before anthesis. The initial steps of androgenesis were determined.Abbreviations BAP
6-benzylaminopurine
- 2,4-d
2,4-dichlorophenoxy-acetic acid
- NAA
1-naphthaleneacetic acid 相似文献
13.
Summary Protoplasts were isolated from immature cotyledons of Vigna sinensis and cultured in a modified MS Liquid medium containing 0. 2 mg/l 2, 4-dichlorophenoxyacetic acid (2, 4-D), 1 mg/l naphthaleneacetic acid (NAA) and 0. 5 mg/l 6-benzylaminopurine (BAP) in the dark at a density of 1 × 105/ml. The protoplasts began to divide in 3–5 days. Sustained cell division resulted in formation of cell clusters and small calli, with the cell division frequency and plating efficiency of cell colonies reaching 27. 7% and 1. 7% respectively. When calli of 2 mm in size were transferred onto MSB medium (MS salts and B5 vitamins) containing 500mg/l NaCl, 500 mg/ 1 casein hydrolysate (CH), 2 mg/l 2,4-D and 0. 5 mg/l BAP for further growth, approximately 5% of the calli developed embryogenically. The embryogenic calli were selected and subcultured on the same composition of MSB medium and were able to maintain somatic embryogenesis capacity in subculture for a long time. When the calli were moved to MSB medium with 0. 1 mg/l indole-3-acetic acid (IAA), 0. 5mg/l kinetin(KT), 3–5% mannitol and 2% sucrose in the light, many somatic embryos formed from the calli. Only part of the embryoids developed further to the cotyledonary stage, and the others died at the globular, heart-shaped or torpedo stages. Finally, some cotyledonary embryoids germinated and developed into plants or shoots. The shoots were readily rooted on 1/2 strength MS medium with 0. 1–0.3 mg/l indole-3-butyric acid (IBA). The plants grew well in soil and were fertile.Abbreviations 2, 4-D
dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- BAP
6-benzylaminopurine
- IAA
indole-3-acetic acid
- KT
kinetin
- IBA
indole-3-butyric acid
- CH
casein hydrolysate
- CM
coconut milk
- ZT
zeatin 相似文献
14.
A procedure for rapid in vitro multiplication of Tylophora indica (Burm. f.) Merrill., an important indigenous medicinal plant, has been developed. Addition of ascorbic acid was essential to induce sprouting of axillary buds. Optimum multiplication was observed on MS medium containing 6-benzylamino purine (5.0 mg l–1), -naphathalene-acetic acid (0.5 mg l–1) and ascorbic acid (100 mg l–1). Rooting of in vitro produced shoots was readily achieved with indole-3-acetic acid alone (1.0 mg l–1) in MS. The plantlets thus obtained were successfully transferred to pots in large numbers which grew normally.Abbreviations BAP
6-benzylamino purine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- 2ip
2-isopentenyladenine
- Kn
kinetin
- MS
Murashige & Skoog media
- NAA
-naphthalene acetic acid 相似文献
15.
The effects of different combinations of plant growth regulators and light intensity on the formation of multiple shoots of Catharanthus roseus (L.) were studied. By composing three dimension surfaces and their topo views from experimental data, it was clear that Murashige-Shoog (MS) medium supplemented with 7.0 mg l-1 BA and 1.0 mg l-1 NAA strongly stimulated the formation of shoots, whereas medium supplemented with 2,4-d suppressed the formation of shoots or caused shoot dedifferentiated. Light intensities of 550–700 Lux were found to be beneficial to the formation of shoots when MS medium was supplemented with 2 mg l-1 6-BA and 0–1.0mg l-1 NAA.Abbreviations BA-6
benzyladenine
- NAA
-naphthalenacetic acid
- 2,4-d
2,4-dichlorophenoxyacetic acid 相似文献
16.
A yield of 2–4×106 protoplasts/g F.W. could be obtained when fresh cauliflower inflorescence segments were digested with 2% cellulase Onozuka R-10, 1% cellulase RS and 0.4% Macerozyme R-10 in CPW18S for 7 to 10 h. Purified protoplasts were cultured in K8p liquid and agarose medium. Although protoplasts in liquid medium divided earlier than in agarose, protoplast-derived cells in liquid culture could not avoid browning. With agarose culture, sustained division and callus formation could be achieved. After 20 days, calli were transferred onto B5 agar medium with ZT 1.5 mg l-1, BA 0.5 mg l-1 and IAA 0.1 mg l-1 for shoot formation. The frequency of bud formation varied from 56.7% for calli of 1mm in size to 5.6% for 5mm calli. The shoots formed were rooted in B5 medium containing 0.5 mg l-1 IBA, and the regenerated plants were transplanted to pots and grew normally. It took about two months from protoplasts to the regenerated plants.Abbreviations Ade
adenine
- BA
6-benzyl aminopurine
- CH
casein hydrolysate
- CM
coconut milk
- 2,4-D
2,4,-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- Gln
glutamine
- NAA
-naphthylacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- ZT
zeatin 相似文献
17.
Shoot regeneration,re-flowering and post flowering survival in bamboo inflorescence culture 总被引:1,自引:0,他引:1
In vitro grown inflorescences of Bambusa edulis were used to investigate the process of vegetative shoot growth in detail. The findings revealed that auxins and ACC could be significant growth regulators in this process. Overall, auxins [NAA, indolebutyric acid (IBA), and 2,4-dichlorophenoxyacetic acid (2,4-D)] induced inflorescences to grow vegetative shoots. However, the efficiency of shoot regeneration varied. A greater percentage (27.3–34.5) of inflorescences in the 5 mg l−1 NAA, 10 mg l−1 NAA, and 1 mg l−1 2,4-D treatments formed more vegetative shoots than those exposed to other treatments. IBA promoted shoot regeneration less effectively than NAA and 2,4-D. Fifty percent of regenerated vegetative shoots flowered after 2 months when the medium was supplemented with 5 mg l−1 NAA. All shoots that received 1 mg l−1 1-amino-cyclopropane-1-carboxylic acid (ACC) flowered in 5 mg l−1 NAA medium. Rooted plantlets were used to examine their survival following in vitro flowering. All plantlets with vegetative shoots, even those with inflorescences, survived and grew. 相似文献
18.
Callus cultures of Prosopis tamarugo Phil (Leguminosae, Sub family-Mimosoideae) were established from hypocotyls and cotyledons on MS medium supplemented with NAA (2.0 mg l-1) and BAP (0.2 mg l-1). Regeneration through various juvenile explants was obtained on hormone-free and high cytokinin containing Murashige and Skoog's medium. Multiple shoot buds formation was observed from the embryonic axis on MS medium incorporated with BAP (5.0 mg l-1)). Elongation of shoot buds was observed on subsequent transfer to MS medium with BAP (1.0–2.5 mg l-1) or without BAP. Explants containing apical meristem showed higher number of shoot formation at an early period. De novo shoot buds formation through callus morphogenesis was observed at the base of differentiated shoots on high cytokinin containing medium. All the manipulations of salt strength of MS, nitrogen, carbon, ascorbic acid and polyamines failed to induce organogenesis in isolated callus. In vitro produced shoots were rooted on MS medium supplemented with IBA or NAA singly or in combination.Abbreviations HC
high cytokinin (BAP 5.0 mg l-1)
- BAP
6-benzyl amino purine
- IBA
indole-3-butyric acid
- HF
hormone free
- NAA
I-naphthalene acetic acid
- MS
Murashige & Skoog 相似文献
19.
Summary Cotyledon and hypocotyl protoplasts of Helianthus annuus inbred line 47 302 bcd were embedded in alginate and plated on L4 medium (Lenée and Chupeau 1986). After one month, the calli were transferred on MSSH regeneration medium (Murashige and Skoog 1962; Schenk and Hildebrandt 1972) where they regenerated shoots (overall efficiency 10–2%). The shoots were elongated on B5 (Gamborg et al. 1968) medium first without hormones, then supplemented with GA3 and BAP (both 0.05 mg/l). In order to overcome the difficulty to induce rooting by classical methods, the elongated shoots were grafted on a sunflower rootstock. The grafted shoots produced flowers and seeds. Different factors have been shown to have an important influence on the capacity to regenerate shoots: the genotype, the physical culture conditions at the callus regeneration step (e.g. protoplasts embedded in alginate), and the media composition.Abbreviations BAP
6-benzylaminopurine
- GA3
gibberellic acid
- IBA
indole-3-butanoic acid
- IAA
indole acetic acid
- MES
2-N-morpholinoethane sulfonic acid
- NAA
1-naphthalene acetic acid
- 2,4D
2,4 dichlorophenoxyacetic acid 相似文献
20.
Alejandro Vazquez-Tello Makoto Hidaka Takeshi Uozumi 《Plant Cell, Tissue and Organ Culture》1995,40(2):169-177
Embryogenic cell suspensions of Lavatera thuringiaca L. were established from leaf petiole and shoot regeneration was achieved when cells were plated on medium without growth regulators. We tested three methods for protoplast culture, isolated from a one-year old embryogenic cell suspension, to determine the best conditions for L. thuringiaca protoplast culture and shoot regeneration. The highest protoplast plating efficiency was obtained with the agaroseembedded method, reaching 30%, while the nursing culture method gave 5% when the protoplasts were plated over Whatman paper No. 2. However, the same nursing culture failed to produce protoplast-derived microcalluses when the protoplasts were plated on a nitrocellulose filter. The liquid thin layer method gave the lowest plating efficiency with only 0.5%. Shoot regeneration from protoplast-derived microcalluses was achieved in two steps; first, globular embryo development was favored in medium low in auxin (2,4-d and BA at 0.01 and 0.05 mg 1-1, respectively), second, the globular embryos further differentiate into shoots in medium without growth regulators or in medium containing GA3 (0.5 to 1.0 mg 1-1).Abbreviations 2,4-d
2,4-dichlorophenoxyacetic acid
- BA
benzyladenine
- GA3
gibberellic acid
- NAA
-naphthaleneacetic acid
- IBA
indole-3-butyric acid 相似文献