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1.
Embryogenic callus in Catharanthus roseus was initiated from hypocotyl on Murashige and Skoog’s (MS) medium supplemented with 1.0–2.0 mg dm−3 of 2,4-dichlorophenoxyacetic acid (2,4-D) or chlorophenoxyacetic acid (CPA). Calli from other sources were non-embryogenic.
Numerous somatic embryos were induced from primary callus on MS medium suplemented with naphthalene acetic acid (NAA) within
two weeks of culture. Embryo proliferation was much faster on medium supplemented with 6-benzylaminopurine (BAP). After transfer
to medium with gibberellic acid (GA3, 1.0 mg dm− 3) mature green embryos were developed and germinated well into plantlets on MS liquid medium supplemented with 0.5 mg dm−3 BAP. Later, embryos with cotyledonary leaves were subjected to different auxins treatments for the development of roots.
Before transfer ex vitro, plantlets were cultivated on half strength MS medium containing 3 % sucrose and 0.5 mg dm−3 BAP for additional 2 weeks. Additionally, the effect of liquid medium has been evaluated at different morphogenetic stages. 相似文献
2.
A. Muthusamy K. Vasanth D. Sivasankari B. R. Chandrasekar N. Jayabalan 《Biologia Plantarum》2007,51(3):430-435
The embryogenic calli (EC) were obtained from hypocotyl explants of groundnut (Arachis hypogaea L.) cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid
(2,4-D) in combination with 0.5 mg dm−3 6-benzylaminopurine (BAP). The EC were exposed to γ-radiation (10–50 Gy) or treated with 1–5 mM of ethyl methane sulphonate
(EMS) or sodium azide (SA). The mutated EC were subcultured on embryo induction medium containing 20 mg dm−3 2,4-D. Somatic embryos (SE) developed from these calli were transferred to MS medium supplemented with BAP (2.0 mg dm−3) and 0.5 mg dm−3 2,4-D for maturation. The well-developed embryos were cultured on germination medium consisting of MS salts with 2.0 mg dm−3 BAP and 0.25 mg dm−3 naphthaleneacetic acid (NAA). Well-developed plantlets were transferred for hardening and hardened plants produced normal
flowers and set viable seeds. The fresh mass of the EC, mean number of SE per explant and regeneration percentage were higher
at lower concentrations of mutagens (up to 30 Gy/3 mM). Some abnormalities in regenerated plants were observed, especially
variations in leaf shape. 相似文献
3.
M. Ganesan R. Chandrasekar B. D. Ranjitha Kumari N. Jayabalan 《Biologia Plantarum》2007,51(3):414-420
A simple and reliable protocol for regeneration of okra through somatic embryogenesis from suspension cultures has been developed.
Embryogenic callus was obtained from hypocotyl explants cultured on media with Murashige and Skoog (MS) salts, Gamborg (B5)
vitamins, 2.0 mg dm−3 2,4-dichlorophenoxyacetic acid (2,4-D), 1.0 mg dm−3 naphthaleneacetic acid (NAA), 25 mg dm−3 polyvinylpyrrolidone and 30 g dm−3 sucrose. More number and high frequency of healthy embryoids appeared individually in suspension culture containing MS salts,
B5 vitamins, 2.0 mg dm−3 2,4-D and 1.0 mg dm−3 kinetin. Formation of cell clusters from the single cells was clearly noticed during ontogeny. Matured embryos at the cotyledonary
stage were transferred to agar solidified medium for germination. The best conversion of embrya into plantlets (67.3 %) was
recorded on media with half strength MS salts, B5 vitamins, 0.2 mg dm−3 benzylaminopurine (BAP) and 0.2 mg dm−3 gibberellic acid (GA3). The plantlets were transferred to soil and hardened in the plastic pots. After proper acclimatization, the plantlets regenerated
through somatic embryogenesis were compared to seed grown plants to observe any variation. 相似文献
4.
In the present study, in vitro regeneration system for a recalcitrant woody tree legume, Leucaena leucocephala (cvs. K-8, K-29, K-68 and K-850) from mature tree derived nodal explants as well as seedling derived cotyledonary node explants
was developed. Best shoot initiation and elongation was found on full-strength Murashige and Skoog (MS) medium supplemented
with 3 % (m/v) sucrose, 100 mg dm−3 myoinositol, 100 mg dm−3 glutamine, 20.9 μM N
6-benzylamino-purine (BAP) and 5.37 μM 1-naphthalene acetic acid (NAA). Rooting was induced in half-strength MS medium containing
2 % (m/v) sucrose, 100 mg dm−3 myoinositol, 14.76 μM indole-3-butyric acid (IBA) and 0.23 μM kinetin. The cultivar K-29 gave the best response under in vitro conditions. Rooted plantlets were subjected to hardening and successfully transferred to greenhouse. Further, somatic embryogenesis
from nodal explants of cv. K-29 via an intermittent callus phase was also established. Pronounced callusing was observed on full-strength MS medium containing
3 % (m/v) sucrose, 100 mg dm−3 myoinositol, 40.28 μM NAA and 12.24 μM BAP. These calli were transferred to induction medium and maximum number of globular
shaped somatic embryos was achieved in full-strength MS medium fortified with 3 % (m/v) sucrose, 100 mg dm−3 myoinositol, 15.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 5.0 μM BAP and 1.0 mM proline. Moreover, an increase in endogenous
proline content up to 28th day of culture in induction medium was observed. These globular shaped somatic embryos matured in full-strength MS medium
with 3 % (m/v) sucrose, 100 mg dm−3 myoinositol, 10.0 μM BAP, 2.5 to 5.0 μM IBA and 0.5 mM spermidine. 相似文献
5.
Mingxi Liu Jing Yang Shaoyun Lu Zhenfei Guo Xiping Lin Hong Wu 《In vitro cellular & developmental biology. Plant》2008,44(2):100-104
Centipedegrass (Eremochloa ophiuroides [Munro] Hack.) is an important warm-season turfgrass and pasture grass. To explore the potential use of biotechnical tools
in breeding of centipedegrass, we established an efficient plant regeneration system for this species. Four basal media and
24 combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BAP) were examined for their effects on callus
induction from mature seed explants. Twenty combinations of naphthaleneacetic acid (NAA) and BAP were tested for their effect
on plant regeneration. Results indicated that Murashige and Skoog basal medium supplemented with 4.5 mg l−1 2,4-D and 1 mg l−1 BAP was the best medium for callus induction, while the combination of 2 mg l−1 BAP and 1 mg l−1 NAA induced the highest rate of regeneration and development of shoots and roots. This work provides a basis for the breeding
of centipedegrass through somaclonal variation and genetic transformation. 相似文献
6.
Seeds of Citrus reticulata were germinated efficiently when they were sown directly after their extraction from fruits harvested in January, and incubated
at constant temperature (25 °C). Seed drying decreased both the percentage of seed germination and the number of seedling
per seed. Germination of seeds was better on Murashige and Skoog (MS) medium supplemented with 0.5 mg dm−3 benzylaminopurine (BAP) than in a soil. Shoot cuttings obtained from germinated seeds were subcultured on B5 medium supplemented
with 1 mg dm−3 BAP, 0.5 mg dm−3 kinetin (KIN) and 0.5 mg dm−3 naphthalene acetic acid (NAA), where shoots grew and multiplied. They were rooted on half strength MS medium supplemented
with 0.25 mg dm−3 BAP, 0.5 mg dm−3 NAA and 1 mg dm−3 isobutyric acid (IBA). Rooting under light was better than under dark. Seedlings and shoot cuttings with roots were transferred
successfully to the soil after three weeks of acclimatization.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
7.
Experiments were performed to evaluate the capacity of quince (Cydonia oblonga Mill.) leaves to regenerate somatic embryos and shoots and/or roots simultaneously. Leaves, treated for 2 d in liquid medium
containing 2.5 mg dm−3 2,4-dichlorophenoxyacetic acid were cultured for 0, 3, 6, 9, 12, 15, 18, 21 d on a gelled medium supplemented with 1 mg dm−3 kinetin (Kin) and 0.1 mg dm−3 naphthalenacetic acid (NAA) and were transferred to a medium either without growth regulator (GR-) or containing 0.6 mg dm−3 6-benzylaminopurine (BA) + 0.2 mg dm−3 gibberellic acid (GA3) + 0.06 mg dm−3 indole-3-butyric acid (IBA) (GR+). Leaves producing somatic embryos (SEs) only, or adventitious roots (Rs) only, or SEs+Rs
simultaneously, were detected on GR- culture medium; on GR+ medium, leaves producing adventitious shoots (Ss) only, SEs+Ss
or SEs+Rs+Ss simultaneously, also appeared. Leaves producing both Ss+Rs were never detected. Proportions among the various
types of regenerating leaves varied according to both the length of Kin+NAA treatment and the presence or absence of GR in
the transfer medium. The greatest variations, both on GR− and on GR+, took place within the first 9 d of culturing on Kin+NAA.
After this period, no further substantial differences in the trend of each type of regenerating leaf were observed. The length
of the treatment with Kin+NAA also modified the proportions between the different types of morphogenic structures.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
8.
Plants of two cytotypes (2n=2x=20, and 2n=3x=30) of pinto peanut (Arachis pintoi Krapov. & W.C. Gregory) were regenerated through somatic embryogenesis. Embryogenic calli were induced from shoot tips or
immature leaves dissected from in vitro growing plants. In the case of the diploid peanut the best somatic embryogenesis was achieved when shoot tips were cultured
on Murashige and Skoog (MS) medium supplemented with 10 mg dm−3
Picloram (PIC) and 0.1 mg dm−3 6-benzylaminopurine (BAP) or when explants from immature leaves were cultured on MS + 10 mg dm−3 PIC + 0.01 mg dm−3 BAP. In the case of triploid peanut the highest number of somatic embryos was obtained when shoot tips were cultured on MS
+ 10 mg dm−3 PIC + 0.01 mg dm−3 BAP or when immature leaves were cultured on MS + 20 mg dm−3 PIC + 0.01 mg dm−3 BAP. Somatic embryos were converted into plants by culture on MS + 0.01 mg dm−3 naphthaleneacetic acid + 0.01 mg dm−3 BAP. Plants were successfully transferred to pots in greenhouse. 相似文献
9.
The endosperms of Carthamus tinctorius cv. HUS-305, excised at globular to heart-shaped stages of zygotic embryo development, were cultured on Murashige and Skoog’s
medium (MS) supplemented with different concentrations of 6-benzylaminopurine (BAP), kinetin, thidiazuron (TDZ), 2,4-dichlorophenoxyacetic
acid (2,4-D) or α-naphthalene-acetic acid (NAA). The highest incidence of callusing was on 2,4-D supplemented media. However,
embryos differentiated only from the calli developed on media supplemented with BAP, kinetin or TDZ with the last eliciting
maximum embryogenic response. The addition of a reduced nitrogen source, casein hydrolysate to MS medium supplemented with
BAP and/or NAA, did not stimulate the response. However, adenine sulphate (100 mg dm−3) promoted the induction of somatic embryos. Upon transfer to MS basal medium or the same supplemented with 0.61 μM gibberellic
acid (GA3), plumular poles of few embryos elongated resulting in the development of shoots. 相似文献
10.
Anin vitro procedure for large scale multiplication ofBoswellia serrata Roxb. has been developed using cotyledonary node segments. In average 4 shoots per node were obtained on Murashige and Skoog's
(MS) medium containing 0.5 mg dm−3 6-benzylaminopurine (BAP) and 0.05 mg dm−3 napthaleneacetic acid (NAA) within 22 d. By repeated subculture technique 90–100 shoots per node could be obtained after
88 d of initial culture. Shoots could be rooted on MS medium containing 1/4 salts, 1% saccharose, and a combination of 0.5
mg dm−3 indole-3-butyric acid (IBA) and 0.25 mg dm−3 indole-3-acetic acid (IAA). Addition of antioxidants like polyvinylpyrrolidone (PVP-50 mg dm−3) and ascorbic acid (100 mg dm−3) in both multiplication and rooting media prevented browning of cultures. Approximately 80% of shoots rooted within 8–10
d. Rooted plantlets were kept for 15 d in culture bottles containing SoilriteTM irrigated with a nutrient solution containing 1/4 MS salts and finally transferred to pots containing soil: SoilriteTM (1∶1), mixture with 70% transplantation success. 相似文献
11.
The effect of auxin, GA and BAP on potato shoot growth and tuberization was investigated under in vitro condition. The shoot length of potato explants increased with the increasing of concentrations (0.5 – 10 mg dm−3) of IAA treatment especially with the addition of GA3 (0.5 mg dm−3), but was inhibited by BAP (5 mg dm−3). The root number and root fresh weight of potato explants increased with the increasing of IAA levels either in the presence
of GA3 (treatment IAA+GA) or not (IAA alone). However, no root was observed in the treatment IAA+BAP, instead there were brown swollen
calli formed around the basal cut surface of the explants. The addition of GA3 remarkably increased the fresh weight and diameter of calli. Microtubers were formed in the treatments of IAA+BAP and IAA
+ GA + BAP but not observed in the treatments of IAA alone or IAA + GA. IAA of higher concentrations (2.5 – 10 mg dm−3) was helpful to form sessile tubers. With the increasing of IAA levels, the fresh weight and diameter of microtubers increased
progressively. At 10 mg/L IAA, the fresh weight and diameter of microtubers in the treatment of IAA + GA + BAP were 409.6
% and 184.4 % of that in the treatment of IAA + BAP respectively, indicating the interaction effect of GA and IAA in potato
microtuberization. 相似文献
12.
The effect of various hormonal combinations on regeneration of shoots and roots from meristem-derived callus of Crocus sativus L. and activities of antioxidant enzymes have been studied. The most efficient regeneration occurred with 1.0 mg dm−3 1-naphthaleneacetic acid (NAA) + 1.0 mg dm−3 thidiazuron and 1.0 mg dm−3 NAA + 2.0 mg dm−3 kinetin. For sprouting, regenerated shoot were subcultured on Murashige and Skoog medium containing 1.0 mg dm−3 NAA + 1.0 mg dm−3 benzylaminopurine (BAP). Protein content and superoxide dismutase activity decreased in regenerated shoots and roots and
increased in sprouting shoots, while catalase (CAT), peroxidase (POX) and polyphenol oxidase (PPO) activities increased during
organogenesis and decreased in sprouting shoots. High CAT and PPO activities were detected in regenerated roots, whereas high
POX activity was observed in regenerated shoot. 相似文献
13.
Immature zygotic embryos were cultured on Murashige and Skoog's medium (MS) supplemented with various combinations of 2,4-dichlorophenoxyacetic
acid (2,4-D), naphthaleneacetic acid (NAA), benzyladenine (BA) and zeatin or with various concentrations of 2,4-D alone. The
maximum number (8 per embryo) of adventitious buds formed from cotyledons of heart stage embryos cultured on MS medium with
1 mg dm−3 BA and 0.01 mg dm−3 NAA. The adventitious buds originated from procambial strands of immature embryo cotyledons and then developed into adventitious
bud primordia within 20 d. Adventitious buds transferred to hormone free MS medium grew into shoots, but did not produce plantlets
because the shoots failed to root.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
14.
Influence of boron on somatic embryogenesis in papaya (Carica papaya L.) cv. Honey Dew was investigated. Immature zygotic embryos were grown in the induction medium containing Murashige and
Skoog basal salts, with B5 vitamins, picloram (1 mg dm−3) or 2,4-dichlorophenoxy acetic acid (2 mg dm−3) and different concentrations of boric acid (30 to 500 mg dm−3). Maximum somatic embryo initiation was observed at 62 mg dm−3 boric acid irrespective of the growth regulator used. The cotyledonary stage somatic embryos were germinated on MS basal
medium devoid of growth regulators. The regenerated plantlets were hardened under greenhouse conditions and transferred to
field.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
15.
The anthers of three genotypes ofLycopersicon esculentum, viz. cv. HS-101, cv. HS-102 and an F1 hybrid (Montfavet 63-4xHS-101) in different stages of development were cultured in various defined nutritive media. Only
anthers containing microspores in the early uninucleate stage were found to respond with the culture medium in the formation
of androgenic callus. The DGII medium with 2 mg l−1 NAA and 1 mg 1−1 kinetin was found to be best for callus induction but MS medium supplemented with 2 mg l−1 2,4-D and 0.1 mg 1−1 BAP favoured proliferation and growth of the callus. The androgenic microspores followed the ‘B’ type pathway of androgenesis
in the formation of callus.
Induction of tracheids in the callus could be achieved by supplementing the basal medium with NAA and kinetin or 2,4-D and
BAP. Initiation of vessel elements and cambium were favoured by addition of NAA and kinetin and that of the phloem in the
presence of 2,4-D and BAP in the basal medium, suggesting that the hormonal requirements for production of different elements
of the vascular system in androgenic callus are different. Although roots could be induced from the callus, shoot differentiation
could not be achieved under cultural conditions. 相似文献
16.
A protocol was developed for plant regeneration of Melia azedarach L. by in vitro culture of apical meristem (0.5 mm in length). The influence of six clones was investigated. The culture procedure comprised
two sequential steps: 1) Induction of shoots by in vitro culture of axillary buds from adult trees (10–15 years old) by culture on Murashige and Skoog (1962) medium (MS) supplemented
with 0.5 mg·dm−3 BAP (6-benzylaminopurine), 0.1 mg·dm−3 IBA (indolebutyric acid), and 0.1 mg·dm−3 GA3 (gibberellic acid). The Multiplication of the regenerated shoots was achieved in MS + 0.5 mg·dm−3 BAP + 0.1 mg·dm−3 GA3. 2) In vitro culture of the apical meristems from the regenerated shoots in MS medium (0.7 %) supplemented with various combinations of
BAP and IBA.
Maximum shoot proliferation was obtained on MS medium supplemented with 0.5 mg·dm−3 BAP and 0.1 mg·dm−3 IBA.
Regenerated shoots were rooted on MS + 3.5 mg·dm−3 IBA (4 days) followed by subculture on MS lacking growth regulators (30 days). Complete plants were transferred to soil. 相似文献
17.
A method for plant regeneration in Robinia pseudoacacia L. from cell suspension culture was established. Non regenerative friable callus from hypocotyls and cotyledon explants from
in vitro raised seedling induced on solid Murashige and Skoog (MS) medium supplemented with 0.05 mg dm−3 2,4-dichlorophenoxyacetic acid (2,4-D) was used for initiation of cell suspension cultures on same MS medium but without
agar. Single cells were isolated after 3 d and the optimum cell density was 1–3 × 104 cells per cm3 of the liquid MS medium. Plating efficiency was 29.6 % and callus formed within 4 weeks was subcultured and transferred to
solid MS medium supplemented with 0.6 mg dm−3 benzyladenine (BA) along with 0.05 mg dm−3 α-naphthalene-1-acetic acid (NAA) for the induction of adventitious bud primordia. The shoots developed were isolated and
re-cultured on MS medium containing 0.6 mg dm−3 BA. These microshoots after dipping in 1–2 cm3 of 10 mg dm−3 indole-3-butyric acid (IBA) for 24 h in dark were cultured on half strength solid MS medium supplemented with 0.05 % charcoal
and showed 80–82 % rooting within 4 weeks. 相似文献
18.
Oncidium cultivars gave different embryogenic responses of leaf explants when affected by auxins (2,4-D, IAA, IBA and NAA), cytokinins
(2iP, BA, kinetin, TDZ and zeatin), sucrose, NaH2PO4, casein hydrolysate, peptone, and glutamine. The best embryogenic responses of cv. Sweet Sugar were at 20 g dm−3 sucrose, 85 mg dm−3 NaH2PO4 and 3 mg dm−3 kinetin, respectively. The development of somatic embryos on leaf explants of cv. Sweet Sugar was delayed for about 10 –
20 d in comparison with cv. Gower Ramsey. On growth regulator-free medium, about 40 % of leaf derived embryos of cv. Gower
Ramsey were fused together in their basal parts and so called multiple-state embryos. However, under the same condition, the
embryos of cv. Sweet Sugar were all in multiple-state form. 相似文献
19.
An efficient regeneration protocol via somatic embryogenesis was optimized for mung bean [Vigna radiata (L.) Wilczek; cv. Vamban 1]. Primary leaf explants were used for embryogenic callus induction in MMS medium (Murashige and
Skoog salts with B5 vitamins) containing 2.0 mg dm−3 2,4-dichlorophenoxyacetic acid (2,4-D), 150 mg dm−3 glutamine and 3 % sucrose. Fast growing, highly embryogenic cell suspensions were established from 21-d-old calli in MMS
medium supplemented with 0.5 mg dm−3 2,4-D and 50 mg dm−3 proline (Pro), and maximum recovery of globular (39.0 %), heart-shaped (26.3 %) and torpedo-stage (21.0 %) somatic embryos
were observed in this medium. Mature cotyledonary-stage somatic embryos were cultured for 5 d in half strength B5 liquid medium
containing 0.05 mg dm−3 2,4-D, 20 mg dm−3 Pro, 5 μM abscisic acid, 1000 mg dm−3 KNO3, 50 mg dm−3 polyethylene glycol (PEG 6000) and 30 g dm−3 D-mannitol. Mature somatic embryos were germinated after dessication for 3 d and complete development of plantlets accomplished
in MMS medium containing 30 g dm−3 maltose, 0.5 mg dm−3 benzyladenine and 500 mg dm−3 KNO3. Profuse lateral roots, and regeneration frequency (up to 60 %) were observed in half-strength MMS medium containing 0.5
mg dm−3 indolebutyric acid (IBA). The regenerated plants were grown to fruiting and were morphologically normal and fertile. 相似文献
20.
S. Kiran Ghanti K. G. Sujata M. Srinath Rao P. B. Kavi Kishor 《Biologia Plantarum》2010,54(1):121-125
A protocol for plant regeneration via somatic embryogenesis was developed in two chickpea (Cicer arietinum L.) cultivars ICCV-10 and Annigeri. Somatic embryos were induced from immature cotyledons on Murashige and Skoog’s (MS) medium
supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), 2,4,5-trichlorophenoxyacetic acid (2,4,5-T),
α-naphthaleneacetic acid (NAA) and picloram alone or in combination with 0.5 — 2.0 mg dm−3 N6-benzylaminopurine (BA) or kinetin (KIN). NAA was better for somatic embryo induction compared to other auxins. The well formed,
cotyledonary shaped embryos germinated into plantlets with 36.6 % frequency on MS medium supplemented with 2.0 mg dm−3 BA + 0.5 mg dm−3 abscisic acid (ABA). The frequency of embryogenesis and plantlet regeneration was higher in cv. ICCV-10 as compared to cv.
Annigeri. Regenerated plants were transferred to soil (40 % survival) and grown to maturity. Histological studies of explants
at various developmental stages of somatic embryogenesis reveled that somatic embryos developed directly from the cotyledon
cells and they were single cell origin. 相似文献