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1.
Summary
Haworthia comptoniana specimens were cultured to determine how benzyladenine (BA) level and in vitro selection for shoot and callus production
affected regeneration capacity and plant phenotype. Leaf explants were cultured on Murashige and Skoog medium containing 0
to 10 mg·liter−1 of BA. The highest number of shoots was obtained with 0.5 mg·liter−1 of BA.H. comptoniana stock cultures (hc) maintained with 0.5 mg·liter−1 of BA produced clumps of small shoots interspersed with friable, white, tan, and green callus. A clump of very large shoots
was isolated and designated cell line Rhc; it differed from the original hc culture in shoot size, the lack of callus growth,
and higher water content. A line of green callus (designated Gc), a line of white callus (Wc), and a line of soft tan callus
(Tc) were also isolated from hc. Optimal BA levels for shoot regeneration from lines Gc and Wc were 2 and 5 mg·liter−1, respectively. No normal shoots could be regenerated from Tc. The phenotypes of these cell lines remained stable for 24 subculture
generations. The hc line that initially required BA for growth became hormone autotrophic whereas the other lines did not.
Culturing using Gelrite and sealing vessels with parafilm promoted vitrification of the hc line. Culturing using GIBCO agar
and unsealed vessels reduced vitrification. The ex-vitro greenhouse survival rates for hc and Rhc plantlets were 10 and 80%,
respectively. The large size of the Rhc shoots apparently resulted in significantly higher survival rates under greenhouse
conditions, but did not result in any phenotypic whole plant changes. 相似文献
2.
Teodora Janković Branka Vinterhalter Dijana Krstić-Milošević Radomirka Nikolić Dragan Vinterhalter Slobodan Milosavljević 《Acta Physiologiae Plantarum》2011,33(4):1515-1520
Shoot cultures of Gentianella bulgarica established from seedling epicotyls were maintained on MS medium supplemented with BA 0.2 mg l−1 + NAA 0.1 mg l−1. Cultures were prone to precocious flowering requiring the use of small shoot buds for multiplication purposes. The contents
of three xanthone compounds identified as DGL, BGL, and DMB, in different plant material were determined by HPLC. The analysis
revealed that the production of xanthones was affected by different concentrations of BA in medium. Shoot cultures grown at
higher BA concentrations contained more DGL than material grown in nature. The concentrations of other two xanthones were
lower in shoot cultures than in plants from nature. The radical scavenging activity of plant extracts and xanthones was investigated
by DPPH test. Samples from plants grown in nature showed the highest activity (IC50 = 0.26 mg ml−1), while the extracts of shoot cultures grown in media with higher concentrations of BA showed moderate activities (IC50 from 1.6 to 4.4 mg ml−1). 相似文献
3.
Ma Dolores Lledó Manuel B. Crespo Juan Bta Amo-Marco 《In vitro cellular & developmental biology. Plant》1995,31(4):199-201
Summary
Vella lucentina M. B. Crespo is a threatened Spanish species that is endemic to a small area in eastern Alicante Province (SE Spain). Micropropagation
techniques were applied forex situ conservation of this plant. Aseptic epicotyls bearing the apical bud were grown in Murashige and Skoog medium supplemented
with 6-furfurylaminopurine (Kin), N6-benzyladenine (BA) or 6-(γ,γ,-dimethilalylamino) purine (2iP). High multiplication rates were obtained with 0.5, 1, or 2
mg·liter−1 BA, or 1 or 2 mg·liter−1 2iP. Indole-3-acetic acid and indole-3-butyric acid were utilized for rooting in half-strength Murashige and Skoog medium.
Regenerated plants were transferred to a potting mix and gradually acclimated to field conditions. No morphological differences
were observed amongin vitro andin vivo plants. 相似文献
4.
An efficient regeneration protocol for rapid multiplication of Melia azedarach, an economically as well as medicinally important timber-yielding tree, was developed. Nearly 90% of the culture exhibited
axillary bud sprouting and multiple shoot formation from nodal segments derived from 20-year-old candidate plus tree on Murashige
and Skoog (MS) medium supplemented with 5 μM 6-benzyladenine (BA). The highest shoot regeneration frequency (92%), maximum
number of multiple shoots (19.7 ± 0.31) as well as shoot length (4.9 ± 0.08 cm) was induced from nodal explants on MS medium
amended with 5.0 μM BA, 0.5 μM indole-3-acetic acid (IAA) and 30 μM adenine sulfate (AdS). Addition of 250 mg l−1 ammonium sulphate, (NH4)2SO4, and 100 mg l−1 K2SO4, prevented defoliation and tip burning without affecting the number of shoots. The explant harvest period also influenced
the bud break and shoot sprouting from nodal segments. Repeated subculturing of nodal explants on fresh MS medium containing
lower concentration of BA (2.5 μM) along with IAA (0.5 μM), AdS (30 μM) and additives was found most suitable growth regulator
regime for achieving 1.2-fold increase in shoot multiplication rate. The percentage of shoot multiplication as well as the
number of shoots per node remained the same during first three subculture passages, afterwards a decline was recorded. About
90% of the in vitro regenerated shoots were successfully rooted ex vitro by giving a pulse treatment of 250 μM indole-3-butyric
acid for 15 min, followed by their transfer to thermocol cups containing soilrite. The raised plantlets were successfully
acclimatized first under culture room conditions, then to green house with 85% survival rate. 相似文献
5.
Hippolyte Kodja Isabelle Robene-Soustrade Jacques Figier 《Acta Physiologiae Plantarum》1997,19(3):359-366
Callus cultures of Tabernaemontana persicariaefolia, (Apocynaceae), an endangered species endemic to the Mascarene Islands, were established from leaf explants on MS medium containing either
5 mg·l−1 2,4-D and 0.5 mg·l−1 BA or 5 mg·l−1 2,4-D, 0.5 mg·l−1 BA and 200 mg·l−1 DFMO. Histological studies showed regenerating nodules resembling globular embryos in calli after 4 weeks on the DFMO medium.
Green shoot formation was achieved by sequential subculture of the induced calli on media with gradually decreasing 2,4-D
concentrations (5→1→0 mg·l−1). Regeneration was greatly stimulated in the presence of DFMO. The first emergence of shoots occured 3 weeks earlier than
in untreated callus cultures. 相似文献
6.
Micropropagation of Sesbania rostrata from the Cotyledonary Node 总被引:5,自引:1,他引:4
Multiple shoots were induced from the cotyledonary nodes derived from seedling of Sesbania rostrata on Nitsch (1969; N) medium supplemented with various concentrations of benzyladenine (BA). 1 mg dm−3 BA proved to be the best, eliciting 5.8 ± 1.0 shoots per explant in 100 % cultures. The elongation of shoots was best at
2.0 mg dm−3 BA. The shoot proliferation capacity increased to 7.5 shoots per explant following transfer of explants to the fresh shoot
multiplication medium (MS + 1.0 mg dm−3 BA), after an initial incubation of 30 d. To further enhance number of shoots per explant an alternative strategy of cultivation
of mother explant on fresh shoot multiplication medium after excision of shoots was adopted. Following the repeated harvesting
of shoots an average of 33 shoots per explant could be obtained. The in vitro regenerated shoots produced roots when transferred to half-strength MS medium supplemented with 3 % sucrose and 1 mg dm−3 IBA. The developed plantlets were planted in the soil and transferred to the field after an acclimatization period of 3 –
4 months. These plants produced flowers and fruits in the field and exhibited the development of prominent and more organized
stem nodules as compared to the in vivo raised plants of the same age.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
7.
Summary Endogenous and exogenous volatile substances were analyzed during 30 days' incubation of four cultivars of thePrunus avium species grown in vitro on a proliferation medium. Cultivars Bigarreau Moreau and Bigarreau Burlat show photosynthetic capability
at 35 μmol·m−2·s−1; oxygen concentration slightly increased (22 to 24%), carbon dioxide was lowered to less than 300 μ·liter−1, and low ethylene (0.8 to 1.2·liter−1) accumulation was recorded. Quite different headspace evolution was observed during growth of cultivars Victoria and Casavecchia:
a large oxygen concentration decrease was accompanied by a sharp carbon dioxide increase (19%) and ethylene boost (4 to 5
μl·liter−1). The evolution of these gaseous metabolites has been correlated to photosynthetic incapability and respiratory stress responsible
for leaf yellowing and tissue softening observed when acetaldehyde and ethanol started to form in cultivars Victoria and Casavecchia.
Dry and fresh weight were measured, and no substantial difference was recorded among cultures with low and high photosynthetic
capability. Evidence is reported that different genotypes within the same species may follow different metabolic pathways. 相似文献
8.
Anwaar Ahmad Heng Zhong Wengling Wang Mariam B. Sticklen 《In vitro cellular & developmental biology. Plant》2002,38(2):163-167
Summary We report a less genotype-dependent in vitro regeneration system capable of producing multiple shoot clumps and whole plants in four different wheat genotypes. Shool
apical meristems from 7-d-old-seedlings produced axillary and adventitious shoots and somatic embryos on media containing
N6-benzyladenine (BA) and 2,4-dichlorophenoxyacetic acid (2,4-D). All four genotypes responded positively to shoot multiplication
depending upon media composition. Scanning electron microscopies of cultures showed a proliferating budding state that gave
rise to adventitious shoots and somatic embryos on further multiplication. The percentage of relative shoot apical meristem
multiplication was 80–90%, and the average number of shoot meristems per multiplied shoot was 40–50 in all genotypes. Among
different concentrations of phytohormones, 2 and 4 mgl−1 BA (8.8 and 17.7 μM) in combination with 0.5 mg l−1 2,4-D (2.26 μM) gave the best results. Actively multiplying shoot clumps were recovered with high frequency among 3-mo.-old cultures. These
shoot clumps regenerated normally and produced fertile plants containing viable seeds. This in vitro system might prove useful for the production of transgenic plants of wheat in a relatively genotype-independent manner. 相似文献
9.
Direct somatic embryogenesis from axes of mature peanut embryos 总被引:2,自引:0,他引:2
A. H. McKently 《In vitro cellular & developmental biology. Plant》1991,27(4):197-200
Summary Plant regeneration via somatic embryogenesis was obtained in peanut (Arachis hypogaea L.) from axes of mature zygotic embryos. The area of greatest embryogenic activity was a 2-mm region adjacent to and encircling
the epicotyl. Somatic embryogenesis was evaluated on Murashige and Skoog media supplemented with a variety of auxin treatments.
Maximum production occurred on medium supplemented with 3 mg · liter−1 4-amino-3,5,6-trichloropicolinic acid. Explant cultures were transferred to half-strength medium supplemented with 1 mg ·
liter−1 gibberellic acid for somatic embryo germination and early plantlet growth. Plantlets, transferred to soil, were placed in
a greenhouse and grown to maturity. 相似文献
10.
A. M. Vieitez E. Corredoira A. Ballester F. Muñoz J. Durán M. Ibarra 《Plant Cell, Tissue and Organ Culture》2009,98(2):135-145
North American oak species, with their characteristic strong episodic seasonal shoot growth, are highly problematic for clonal
micropropagation, resulting in the inability to achieve a stabilized shoot multiplication stage. The potential for initiating
and proliferating shoot cultures derived from Quercus alba, Q. bicolor and Q. rubra explants was investigated, and a micropropagation method for these species was developed. Branch segments from 6 to 7-year-old
trees were forced-flushed and the forced shoots were used as source of explants for culture initiation. A consistent shoot
multiplication stage was achieved, in 13 of the 15 genotypes established in vitro, although marked differences occurred in
explants from different genotypes/species. The control of efficient shoot multiplication involved the culture of decapitated
shoots in a stressful horizontal position on cytokinin-containing medium with a sequence of transfers within a 6-week subculture
cycle, which was beneficial to overcoming the episodic character of shoot growth. During each subculture cycle, the horizontally
placed explants were cultured on media containing 0.2 mg l−1 benzyladenine (BA) for 2 weeks with two successive transfers (2 weeks each) to fresh medium with 0.1 mg l−1 BA, giving a 6-week subculture cycle. The general appearance and vigor of Q. alba and Q. bicolor shoot cultures were improved by the inclusion of both 0.1 mg l−1 BA and 0.5 mg l−1 zeatin in the medium used for the second transfer within the 6-week subculture cycle. Addition of AgNO3 (3 mg l−1) to the shoot proliferation medium of Q. rubra had a significant positive effect on shoot development pattern by reducing deleterious symptoms, including shoot tip necrosis
and early senescence of leaves. The three species showed acceptable in vitro rooting rates by culturing microcuttings in medium
containing 25 mg l−1 indolebutyric acid for 48 h with subsequent transfer to auxin-free medium supplemented with 0.4% activated charcoal. Although
an initial 5-day dark period generally improved the rooting response, it was detrimental to the quality of regenerated plantlets.
However, activated charcoal stimulated not only the rooting frequencies, but it also enhanced plant quality, as evidenced
by root, shoot and leaf growth. 相似文献
11.
This study investigated the distribution of bacteria in groundwater from 16 different levels in five boreholes in granite
bedrock down to a maximum of 860 m. Enrichment cultures were used to assay the groups of bacteria present. Autoradiographic
studies with14C- or3H-labeled formate, methanol, acetate, lactate, glucose, sodium bicarbonate, leucine, glutamine, thymidine, orN-acetyl-glucosamine were used to obtain information about bacteria active in substrate uptake. The biofilm formation potential
was studied in one borehole. The chemical environment in the groundwater was anaerobic with an Eh between −112 and −383 mV, a pH usually around 8, and a temperature range of 10.2 to 20.5°C, depending on the depth. The organic
content ranged between <0.5 and 9.5 mg total organic carbon liter−1. Carbon dioxide, hydrogen, hydrogen sulfide, and methane were present in the water. The nitrate, nitrite, and phosphate concentrations
were close to, or below, the detection limits, while there were detectable amounts of NH
4
+
in the range of 4 to 330 μg liter−1. The average total number of bacteria was 2.6×105 bacteria ml−1, as determined with an acridine organge direct-count (AODC) technique. The average number of bacteria that grew on a medium
with 1.5 g liter−1 of organic substrate was 7.7×103 colony-forming units (CFU) ml−1. The majority of these were facultatively anaerobic, gram-negative, nonfermenting heterotrophs. Enrichment cultures indicated
the presence of anaerobic bacteria capable of growth on C-1 compounds and hydrogen, presumably methanogenic bacteria. Most
probable number assays with sulfate and lactate revealed up to 5.6×104 viable sulfate-reducing bacteria per ml. A biofilm development experiment indicated an active attached microbial population.
Active substrate uptake could not be registered with the bulk water populations, except for an uptake of leucine not associated
with growth. The bulk water microbial cells in deep groundwater may be inactive cells detached from active biofilms on the
rock surface. 相似文献
12.
Vishwanath M. Patil 《In vitro cellular & developmental biology. Plant》1998,34(3):240-243
Summary The purpose of this study was to developin vitro techniques for conserving wild and endemic species ofCeropegia by mass multiplication for subsequent reintroduction in their natural habitat. Micropropagation involving a combination of
axillary bud culture, shoot multiplication, somatic embryogenesis andin vitro tuber formation forCeropegia jainii, a rare plant of the Indian sub continent,C. bulbosa var.bulbosa andC. bulbosa var.lushii, common species, was developed. Nodal explants from all species were cultured on 0.5 MS medium with 8.8 μM (2 mg·l−1) N6-benzyl aminopurine (BA) to regenerate the axillary buds. These produced multiple shoots when transferred to multiplication
medium consisting of 0.5 MS medium with 2.2 μM (0.5 mg·l−1) BA, or microtubers when transferred to 0.5 MS medium with 22.2 μM (5 mg·l−1) BA and 23.2 μM (5 mg·l−1) kinetin.In vitro flowering occurred inC. jainii and not in the other two varieties when the plants were cultured on multiplication media with spermine at 0.25 μM (50 μg·l−1) as an additive. Shoot pieces produced callus on MS medium with 9.05 μM (2 mg·l−1) 2,4-dichlorophenoxy acetic acid. Regeneration of the calli by somatic embryogenesis was achieved when they were transferred
to 0.5 MS medium with 2.2 μM (0.5 mg·l−1) BA. Rooting of the shoots was possible both byin vitro andex vitro means. 相似文献
13.
U. A. Jo H. N. Murthy E. J. Hahn K. Y. Paek 《In vitro cellular & developmental biology. Plant》2008,44(1):26-32
An efficient, simple micropropagation method was developed for Alocasia amazonica using corms in semisolid and liquid cultures. Explants were cultured onto Murashige and Skoog (MS) medium (Murashige and
Skoog, Physiol. Plant. 15:473–497, 1962) supplemented with different cytokinins (Benzyladenine [BA, 2.22–13.32 μM], kinetin [2.32–13.95 μM], Thidiazuron [TDZ, 0.45–4.54 μM])
and cytokinin in combination with auxins [naphthalene acetic acid (NAA, 0.54–5.37 μM)/indole acetic acid (IAA, 0.57–5.71 μM)/indole
butyric acid (IBA, 0.49–4.9 μM)]. All supplementary-induced shoot proliferation and the optimal results was on the medium
supplemented with 2.27 μM TDZ, which induced 5.1 shoots per explant. Among the different concentrations of sucrose (0–120 g
l−1) tested for shoot proliferation, 30 g l−1 was found suitable for corm cultures of Alocasia amazonica. The optimal shoot proliferation and biomass values were with the plantlets grown at 30 μmol m−2 s−1 photosynthetic photon flux (PPF) and 25°C. Liquid cultures found suitable for shoot proliferation and biomass accumulation
was compared to semisolid cultures. Comparative studies of bioreactor systems [continuous immersion (with or without net)
and temporary immersion in liquid media using ebb and flood] revealed that shoot multiplication and growth were greatest with
the raft bioreactor system. Plantlets (cormlets) from the bioreactor were hydroponically cultured for 30 days, and 100% of
plants were acclimatized successfully. The simple efficient method of production of plantlets (cormlets) is useful for large-scale
multiplication of this important ornamental plant.
An erratum to this article can be found at 相似文献
14.
Paul I. Boon 《Microbial ecology》1991,22(1):27-52
Billabongs, lentic waterbodies common to the floodplain of Australian rivers, differ considerably from the lotic riverine
environment in terms of hydrology, physiochemical characteristics, and biological assemblages present. As little is known
regarding the bacterial ecology of billabong habitats, a comparison was made of the bacterial assemblages in the water column
of seven paired river/billabong sites in the Murray-Darling Basin of southeastern Australia. Billabongs supported larger populations
of bacteria (1–157×109 cells liter−1; 11–10,270 μg C liter−1) than did rivers (1–10×109 cells liter−1; 6–143 μg C liter−1). Phospholipid analyses confirmed that billabongs (14–111 μg phospholipid fatty acid liter−1) had larger bacterial populations than rivers (<12 μg liter−1). Bacterial production, measured with3H-leucine, was also greater in billabongs (0.28–3.05 μg C liter−1 hour−1) than rivers (0.05–0.62 μg C liter−1 hour−1). Production calculated from the frequency of dividing cells confirmed this conclusion, and suggested bacterial production
in some billabongs could exceed 100 μg C liter−1 hour−1. An INT-formazan method indicated that usually <25% of bacterial cells were active in either habitat, but this was probably
an underestimate of the bona fide value. Turnover times of glucose were usually shorter in billabongs, and the cell-specific
activity greater for billabong than river assemblages. The factors most likely to be responsible for the differences between
the bacterial assemblages in rivers and billabongs relate to hydrological regime and the availability of organic carbon substrates. 相似文献
15.
Martha E. Pedraza-Santos Ma. Cristina López-Peralta Víctor A. González- Hernández E. Mark Engleman-Clark Prometeo Sánchez-García 《Plant Cell, Tissue and Organ Culture》2006,84(2):100169-100178
The common techniques for the in vitro production of Alstroemeria plants are based on rhizomes as explants, which have low multiplication rates and a high risk of carrying viral diseases.
To overcome these problems, we developed a protocol for the in vitro regeneration of Alstroemeria cv.‘Yellow King’, by testing for shoot induction several explant sources (leaf, stem apices, rhizomes and immature inflorescence
apices), temperature and light/dark regimes, hormone and salt concentrations. For shoot multiplication and rooting, several
hormone concentrations were tested. We found that only the young floral apices produced adventitious shoots by direct organogenesis.
The highest shoot induction rate (10.4 shoots per explant) was obtained by incubation in the dark for 15 days at 8 °C followed
by 15 days at 25 °C and a 16-h/8-h light/dark regime, on a Murashige and Skoog (1962) liquid medium at 50% of the salt concentration,
supplemented with 2.5 mg l−1 KIN, 1.5 mg l−1 BA and 1.0 mg l−1 NAA, using a piece filter paper to support the explant. The highest shoot multiplication rate (9 shoots per explant) was
obtained on a liquid MS medium at full strength supplemented only with BA at 1.0 mg l−1. In vitro rooting of shoots was induced also on a liquid MS medium, either with or without plant hormones. 相似文献
16.
Lakshmanan Venkatachalam R. V. Sreedhar Neelwarne Bhagyalakshmi 《Plant Growth Regulation》2007,51(3):193-205
Use of high levels of growth regulators during micropropagation results in undesirable clonal variability in important commercial
crops such as banana. The present study investigated the effects of high levels of cytokinins on micropropagation in banana
(genotype AAB), and the genetic stability of plantlets was assessed using RAPD and ISSR markers. Cytokinins, such as BA and
kinetin were added to the routine shoot multiplication medium at concentrations up to 10 mg l−1. After 12 weeks of culture involving three subcultures, the maximum number of shoot buds were produced in cultures receiving
either 5 mg l−1 BA (80 shoot buds) or 4 mg l−1 kinetin (62 shoot buds). Certain morphological abnormalities observed during proliferation of shoot buds in vitro were not
observed during acclimatization ex vitro. To check the genetic stability, RAPD and ISSR profiles of micropropagated plantlets
obtained from different cytokinin-treatments were compared with control microplants maintained on MS medium as well as the
field-grown mother plant. A total of 50 RAPD and 12 ISSR primers resulted in 625 distinct and reproducible bands. Thus a total
of 17,400 bands were generated showing homogeneous RAPD and ISSR patterns. Band intensity histogram of each gel confirmed
their monomorphic nature with no genetic variation in all the plantlets analysed. Based on these results a protocol for high
rate shoot multiplication was worked out leading to uniform shoot production. 相似文献
17.
An efficient regeneration system for Phaseolus vulgaris was developed from mature seeds germinated on Murashige and Skoog (MS) medium supplemented with thidiazuron or N6-benzylaminopurine (BA) for 6 d. Using cotyledonary nodes, multiple buds were induced on the MS medium supplemented with 5.0
mg dm−3 BA with the induction frequency 71.9 % after 4-week culture. The buds were then transferred onto shoot formation medium containing
1.0 mg dm−3 BA, 0.1 mg dm−3 gibberellic acid and 2.0 mg dm−3 silver nitrate. The addition of AgNO3 enhanced the frequency of the shoot formation from 61.3 to 87.6 %. Root induction medium was half-strength MS medium with
0.75 mg dm−3 indolebutyric acid and 0.02 mg dm−3 BA. The average root frequency was 84.3 %. The regenerated plantlets with healthy roots grew successfully when transferred
to soil. Using this system we obtained over 10 regenerated plantlets from one explant. 相似文献
18.
H. Cao J. Yang Z. S. Peng C. Y. Kang D. C. Chen Z. C. Gong X. Tan 《In vitro cellular & developmental biology. Plant》2007,43(2):149-153
This study reports a protocol for successful micropropagation of Penthorum chinense using nodal explants on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA) or kinetin (Kn). The presence
of BA promoted a higher rate of shoot multiplication than Kn. Maximum multiple shoot formation was observed in 59.2% of nodal
explants cultured on MS medium supplemented with 2.0 mg l−1 BA after 6 wk. After subculture for 4 wk, the maximum number of shoots (6.4) was obtained on a medium with 2.0 mg l−1 BA, but shoots were too short and not suitable for micropropagation. The taller shoots that regenerated in the presence of
lower BA concentration (1.0 mg l−1) were selected for root induction study. Most shoots (98.8%) rooted in the presence of 0.5 mg l−1 indole-3-acetic acid after 3 wk, with each shoot forming an average of 10.0 roots. Plantlets were transferred to soil and
successfully acclimatized. 相似文献
19.
A simple protocol for direct shoot organogenesis and plant regeneration in Lessertia frutescens using hypocotyl and cotyledon segments is reported. l-canavanine content in the derived shoots is also quantified. Media containing different concentrations and combinations of
the cytokinins kinetin (K) and benzyladenine (BA) were tested for shoot induction potential. The best shoot regeneration rate
(83%) was obtained from hypocotyl segments cultured in Murashige and Skoog (MS) medium supplemented with 1 mg l−1 K; these hypocotyls also produced the largest number of shoots per explant (3.5) and the longest shoots per explant (13.3
mm). The best shoot regeneration rate (46%) using cotyledons as explant material was obtained in MS medium supplemented with
1 mg l−1 K and 1 mg l−1 BA or with 5 mg l−1 K and 0.5 mg l−1 BA. The highest number of cotyledon-derived shoots (1.5) was obtained in MS medium containing 2 mg l−1 K and 0.5 mg l−1 BA, and the longest cotyledon-derived shoots (6.1 mm) were obtained in MS medium containing 1 mg l−1 K and 0.5 mg l−1 BA. Shoots derived from hypocotyls cultured on media containing 1 mg l−1 K contained the highest quantity of l-canavanine (1.42 mg g−1) relative to the control (0.52 mg g−1). Shoots derived from cotyledons cultured on media containing 2 mg l−1 K contained the highest quantity of l-canavanine (2.07 mg g−1) compared to the control. Scanning electron microscopy revealed that shoots regenerated directly from the wounded epidermal
tissue, although callus formation was observed in most cultures. Young shoot clusters proliferated into healthy adventitious
shoots that were subsequently transferred directly onto rooting medium (MS medium containing 4 mg l−1 indole-3-butyric acid), eliminating the need for an additional multiplication or elongation phase. The in vitro plants were
successfully acclimatized in a growth chamber, achieving an 85% survival rate. 相似文献
20.
Maria de Fátima P. S. Machado José Prioli 《In vitro cellular & developmental biology. Plant》1996,32(3):199-203
Summary Currently,Cereus peruvianus plants can be rapidly clonedin vitro via adventitious organogenesis using callus cultures; however, somaclonal variation is a problem. A method is described herein
using lateral bud explants to produce multiple shoots for clonal propagation. Apical and lateral explants were cultured on
MS (Murashige and Skoog, 1962) media with factorial combinations of the auxins indole-3-acetic acid (IAA), 1-naphthaleneacetic
acid (NAA), and cytokinins 6-ben-zyladenine (BA) and N-(2-furanyl-methyl)-1-purine-6 amine (kinetin) at the concentrations
0.0, 0.01, 0.1, 1.0 mg“l−1. Positive results were obtained from the lateral explants in all conditions tested, but apical explants did not respond toin vitro multiplication ofC. peruvianus cactus at all growth regulator combinations tested. Formation of axillary shoots inC. peruvianus seems most frequent in medium containing BA at 1.0 mg·l−1 (4.44 μM) and IAA or NAA at 1.0 mg·l−1 (5.71 μM or 5.37 μM respectively), but the frequency of shoot formation in the BA or kinetin and NAA or IAA combinations indicated that any of
the combinations tested can be used for multiplication ofC. peruvianus plants regenerated from callus tissue culture. Root formation occurred in all (100%) of the cactus shoots after 9 wk in the
same culture medium. All the cacti that developed at the different auxin and cytokin combinations continued growth after transfer
to a potting mix of red earth (Paleudult) and ground river sand (1∶1). 相似文献