scholarly journals Clonal fidelity of chrysanthemum regenerated from long term cultures

Genetika ◽  
2006 ◽  
Vol 38 (3) ◽  
pp. 243-249 ◽  
Author(s):  
Sladjana Jevremovic ◽  
Milana Trifunovic ◽  
Marija Nikolic ◽  
Angelina Subotic ◽  
Ljiljana Radojevic

Morphological characteristics of flowers of long term regenerated chrysanthemum, cv. "White Spider", after ten years of micropropagation are investigated. Shoot cultures are established and maintained more than ten years by stem segment culture on MS medium supplemented with BAP and NAA (1.0, 0.1 mgL-1, respectively). Rooting of shoots (100 %) has done on MS medium without hormones and it was very successful after ten years, as well as, after two or eight years of micropropagation. Acclimation of rooted chrysanthemum plantlets at greenhouse conditions was excellent and after appropriate photoperiod "in vitro" plants flowered 90.3 % and have the same flower color, shape and size as mother plants. Flower color changes of "in vitro" plants are observed during another flowering cycle one year after acclimatization. Observed variations of chrysanthemum flowers could be attributed to epigenetic factors.

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 873G-874
Author(s):  
D. Sankhla ◽  
T.D. Davis ◽  
N. Sankhla ◽  
A. Upadhyaya

This report describes an efficient in vitro regeneration protocol for H. patens (firebush), a heat-tolerant ornamental shrub native to tropical and subtropical America. Shoot cultures were initially established using shoot tips placed on MS-revised medium containing 2.3 μM 2,4-D, 2.3 μM kinetin, and 0.25% polyvinylpyrrolidone. Other types of explants (nodal and internodal segments, leaf pieces, floral buds) did not regenerate shoots when placed on this medium. Two-month-old plantlets derived from the shoot tips were subcultured on MS medium supplemented with 0.5 μM thidiazuron (TDZ), and within 3 to 4 weeks, some callus was produced at the root–shoot junction. When this callus, with a small portion of the root and shoots, was placed on MS medium with 0.05 μM TDZ and 0.01 μM ABA, prolific shoot formation occurred within 3 to 4 weeks followed by root formation. By regular subculturing every 5 to 6 weeks, hundreds of plantlets have been obtained over the past 3 years with no apparent decline in regeneration potential. Addition of activated charcoal (0.5%) to the culture medium has greatly improved growth of the plantlets.


2021 ◽  
Author(s):  
Jose Javier Regalado González ◽  
Manuel López Granero ◽  
Carlos Lopez Encina

Abstract We present the data on proliferation corresponding to 10 years of continuous incubation in vitro of papaya shoots, and propose a reliable method for long-term micropropagation for papaya, using two types of explants: Microshoots from somatic embryos, and from axillary buds of papaya. Three different media were assayed. The proliferation medium (PPRM) allowed to maintain papaya shoots under continuous proliferation during 20 years, maintaining a consistent behaviour. Most of the shoots developed in PPRM rooted during the incubation, and after acclimated easily, maintaining the morphological characteristics of the parental plants, flowering and setting fruits normally. The PPRM medium consist in MS medium supplemented with NAA (0.1 mg l-1), BA (0.5 mg l-1), GA3 (0.5 mg l-1) and Adenine sulphate (40 mg l-1). The average multiplication rate was higher than 20 shoots per explant along the long-term assay. The elongation medium (PELM), was designed to recover shoots with a poor growth, and allowed the development of high quality shoots ready for rooting, and consist in a MS basal medium supplemented with NAA (0.1 mg l-1), Kin (0.5 mg l-1) and GA3 (1 mg l-1). The rooting medium (PROM) was designed to induce high quality roots from non-rooted shoots and consist in a half strength MS medium plus IBA (1mg l-1). On PROM, agar can be exchanged for expanded vermiculite. Acclimation took place inside an acclimatization tunnel under progressive hydric stress. After 4 weeks, the plant recovery rate was 90% for plants maintained under continuous proliferation during ten years.


1970 ◽  
Vol 18 (2) ◽  
pp. 157-164 ◽  
Author(s):  
Rituparna Kundu Chaudhuri ◽  
Timir Baran Jha

In vitro germplasms of ipecac (Cephaelis ipecacuanha Rich.), an important medicinal plant, maintained through reduced growth conditions for more than 12-years, were used as source material for micropropagation. MS with different combinations of Kn (2 mg/l), BAP (2 mg/l), 2iP (2-3 mg/l), NAA (0.2 mg/l) and adenine (40-100 mg/l) as additive was used to induce fresh multiplication of shoots from the nodal meristems and direct shoot bud initiation on the internodal segments. Complete plant regeneration has been achieved from such long term cultures. Regenerated plants maintained their phenotypic and chromosomal stability. Eighty per cent hardened plants, survived in the field condition, are growing well and 25% of them produced flowers within one year. Long term preservation through reduced growth conditions and successful regeneration of morphologically stable plants with stable chromosome numbers (2n = 22) from such long term cultures of ipecae plants.  Key words: Long term culture, Ipecac, micropropagation, flowering D.O.I. 10.3329/ptcb.v18i2.3646 Plant Tissue Cult. & Biotech. 18(2): 157-164, 2008 (December)


HortScience ◽  
1995 ◽  
Vol 30 (7) ◽  
pp. 1443-1444 ◽  
Author(s):  
Arthur M. Richwine ◽  
Jimmy L. Tipton ◽  
Gary A. Thompson

Shoot cultures of Aloe, Gasteria, and Haworthia species were initiated directly from immature inflorescences. Explants placed on a modified MS medium containing 5.4 mm zeatin riboside initiated shoots within 8 to 12 weeks. Long-term shoot cultures were established and maintained on media containing either 5.4 μm zeatin riboside or 4 μm BA. Shoots easily rooted in vitro, and rooted plantlets were esablished in soil. Chemical names used: N-(phenylmethyl)-1H-purin-6-amine (BA); 6-[4-hydroxy-3-methyl-but-2-enylamino]purine riboside (zeatin riboside).


2018 ◽  
Vol 17 (5) ◽  
pp. 405-411
Author(s):  
Jiraporn PALEE

To evaluate an efficient protocol for the micropropagation of Tupistra albiflora K. Larsen, the effects of N6-benzylaminopurine (BA) and naphthalene acetic acid (NAA) concentrations on multiple shoot and root induction were examined. In vitro shoots were used as the explant materials which were cultured on Murashige and Skoog (MS) agar medium supplemented with 0, 1, 2, 3 and 4 mg/L BA for 4 weeks to induce multiple shoots. It was found that the MS medium containing 3 mg/L BA induced 100 % shoot formation with the highest number of 3.2 shoots per explant (2.4-fold significantly higher than the control). For root induction, in vitro shoots were cultured on MS agar medium supplemented with 0, 1, 2, 3 and 4 mg/L NAA for 8 weeks. The results showed that the MS medium containing 1 mg/L NAA induced 100 % root formation with the highest number of 6.6 roots per explant (1.8-fold significantly higher than the control).


2021 ◽  
Author(s):  
Paulo Hercilio Viegas Rodrigues ◽  
Emerson Oliveira ◽  
Christian Demetrio ◽  
Guilherme Ambrosano ◽  
Sônia Maria Stefano Piedade

Abstract Maintaining updated in vitro plant subcultures is essential for commercial micropropagation and tissue culture research. In unusual situations, the subcultures can be delay and the slow-growth in vitro storage technic could be applied to reduce the loss of plant material. The present study aimed to evaluate the slow-growth in vitro storage of banana plantlets (‘Prata Catarina’; group AAB) under different light spectra. Shoot cultures in MS medium without plant growth regulators were maintained under blue (B), red (R), red plus blue (R2B), and white (CW) light spectra (25°C ± 2°C; 50 µmol m -2 s -1 ) for up to 140 days. The plantlets maintained under the R, CW, and R2B spectra did not survive after 140 days of in vitro slow-growth storage. The plantlets maintained under the B spectrum survived after 140 days of in vitro slow-growth storage and showed little browning.


2005 ◽  
Vol 3 (1) ◽  
pp. 15-25 ◽  
Author(s):  
Renata Filkorn-Kaiser ◽  
Konrad Botzenhart ◽  
Albrecht Wiedenmann

A recently described quantitative rapid cycle real time PCR (LightCycler™) assay detects Cryptosporidium parvum after in vitro excystation, which is a surrogate marker for the viability of the organisms. In the original assay the quantification standard is a dilution series of C. parvum oocysts with a microscopically determined excystation rate. The need to keep suspensions of viable oocysts in stock and to continuously monitor their excystation rate, however, renders the assay impracticable for routine application. A synthetic standard was developed to replace the in vivo standard and was calibrated using oocysts with known excystation rates. The standard consists of a 486 bp DNA segment ranging from 229 bp upstream to 79 bp downstream of the actual PCR target site. Aliquots of the standard were frozen and stored at −20 °C and at −70 °C or lyophilised and stored at room temperature in the dark. For a period of one year samples preserved with each of the three methods were restored every four or five weeks. They were amplified in the LightCycler™ and the crossing points (CP) were monitored. No significant trend in the raw CP values could be observed for any of the three storage methods. However, when the methods were compared to each other by calculating the CP ratios (−20 °C/−70 °C; −20 °C/lyophilised; −70 °C/lyophilised) at the 10 monitoring dates, the CP ratios −20 °C/−70 °C and −20 °C/lyophilised showed a highly significant positive trend (p<0.0001) while the CP ratio −70 °C/lyophilised did not differ from the null hypothesis (p=0.53). It can be concluded that the latter two preservation methods are both appropriate, while storage at −20 °C is less advisable. Calculations based on the molecular weight of the standard and on the assumption of an average yield of three sporozoites per oocyst led to the conclusion that the target sequence is probably located on a double copy gene


1993 ◽  
Vol 67 (2) ◽  
pp. 145-150 ◽  
Author(s):  
Nobuaki Akao ◽  
Yoshihisa Goto ◽  
Kaoru Kondo ◽  
Yoshisuke Tsuda

AbstractSecond-stage larvae of Toxocara canis were maintained in vitro for one year. Susceptibility of the larvae to drugs was evaluated by means of minimal larvicidal concentration (MLC) and larval bursting percentage. MLCs of citral and decanoic acid were almost constant throughout all stages of incubation. However, bursting percentage markedly varied within the first 20 weeks of incubation. Therefore, while larvae are available for use in the MLC assay at any stage of incubation, those beyond the first 20 weeks after incubation should be used for the bursting assay to obtain reproducible results.


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