Competence, determination, and meristemoid plasticity in tobacco organogenesis in vitro

2003 ◽  
Vol 81 (6) ◽  
pp. 611-621 ◽  
Author(s):  
Harbinder S Dhaliwal ◽  
Nicole S Ramesar-Fortner ◽  
Edward C Yeung ◽  
Trevor A Thorpe
Keyword(s):  
Root Formation ◽  
Leaf Explants ◽  
Basal Medium ◽  
Shoot Formation ◽  
Culture Conditions ◽  
Vascular Bundles ◽  
Mesophyll Cells ◽  
Tobacco Leaf ◽  
Organogenesis In Vitro

Tobacco leaf explants can produce both shoots and roots depending on the phytohormones in the medium. These arise directly via meristemoids (meristematic centers), which form distinct primordia and then organs. In this study it was found that shoot primordia arose from the palisade mesophyll cells at the adaxial surface, while root primordia arose from the rib parenchyma cells, near the existing vascular bundles. In studies on competency and determination, it was found that the tobacco leaf explants required 4–6 days in culture on a shoot-inducing medium (SIM) to become determined for shoot formation, while the explants were competent for rooting at excision and needed only 1 day on the root-inducing medium (RIM) to become determined for root formation. Transfer of explants from SIM or RIM to basal medium (BM without phytohormones) and vice versa supported the above findings. Transfer of explants from SIM to RIM and vice versa, delayed the timing of root and shoot formation, but not the position in the explant from which the organs arose. On transfer from SIM to RIM or vice versa, meristemoids that were already determined for shoot or root formation continued to develop, while those not yet determined were inhibited and (or) reverted to parenchymatous tissue. Thus under our culture conditions meristemoids in tobacco leaf explants are not plastic.Key words: competence, determination, meristemoid plasticity, organogenesis, tobacco.

scholarly journals Regeneration of Chlorophytum amaniense ‘Fire Flash’ Through Indirect Shoot Organogenesis

HortScience ◽  
2011 ◽  
Vol 46 (3) ◽  
pp. 466-469
Author(s):  
Jin Cui ◽  
Juanxu Liu ◽  
Jianjun Chen ◽  
Richard J. Henny
Keyword(s):  
Callus Formation ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Culture Method ◽  
Shoot Formation ◽  
Photon Flux ◽  
Naphthalene Acetic Acid ◽  
Foliage Plant

Chlorophytum amaniense Engl. ‘Fire Flash’ is a popular exotic ornamental foliage plant as a result of its unique coral-colored midribs and petioles and tolerance to interior low light levels. Currently, demand for propagative materials exceeds the availability of seeds. This study was intended to develop an in vitro culture method for rapid propagation of this cultivar. Leaf and sprouted seed explants were cultured on a Murashige and Skoog basal medium supplemented with different cytokinins with 1.1 μM α-naphthalene acetic acid (NAA) or 2.3 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Leaf explants showed poor responses in callus production and no adventitious shoots were obtained. Callus formation frequencies from sprouted seeds were 71% and 85% when induced by 9.8 μM N6-(2-isopentyl) adenine (2iP) with 1.1 μM NAA and 9.1 μM N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ) with 1.1 μM NAA, respectively. Adventitious shoots occurred after the induced calluses were subcultured on the same concentrations of TDZ or 2iP with NAA. Shoot formation frequencies from calluses cultured on TDZ with NAA and 2iP with NAA were 92% and 85%, and the corresponding mean shoot numbers were 37 and 31 per piece of callus (1 cm3), respectively. Adventitious shoots rooted at 100% after transferring to the basal medium containing 4.4 μM 6-benzylaminopurine (BA) with 2.7 μM NAA. Plantlets, after transplanting to a soilless substrate were easily acclimatized in a shaded greenhouse under a photosynthetic photon flux (PPF) density of 200 μmol·m−2·s−1. Regenerated plants grew vigorously without undesirable basal branching or distorted leaves. This newly established regeneration method can provide the foliage plant industry with a means for rapidly propagating ‘Fire Flash’ liners in a year-round fashion.

scholarly journals Morphogenetic Response of Lilium michiganense to Four Auxins In Vitro

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 813B-813
Author(s):  
Jim Ault* ◽  
Sandy Siqueira
Keyword(s):  
North American ◽  
Callus Induction ◽  
Root Formation ◽  
Callus Formation ◽  
Basal Medium ◽  
Shoot Formation ◽  
Response To Treatment ◽  
The North ◽  
Cut Surface

Shoot, root, and callus induction were examined in the North American lily, Lilium michiganense, in response to treatment with four auxins. Seed from controlled crosses were aseptically excised from slightly immature capsules and cultured in vitro on Murashige and Skoog basal medium and vitamins with 30 g/l sucrose, 7.0 g/l agar, and a pH = 5.7. Seed were maintained at 20 °C with a 14-h photoperiod. After 5.0-5.5 months, leaves and roots were removed from seedlings, the bulbs transversely sectioned, then the bulb sections cultured cut-surface down on the identical medium supplemented with 0.0, 1.0, 2.0, 4.0, or 8.0 μm dicamba, picloram, K-NAA, or 2,4-D. PGRs were added to medium prior to autoclaving except dicamba which was dissolved in 50% ethanol and added after medium autoclaving. 16 explants were utilized for each treatment. The experiment was conducted three times. Morphogenetic response (# of shoots produced, % of explants forming roots, and % of explants forming callus) was tabulated 4 months after treatment. Shoot formation was promoted by treatment with dicamba, picloram, and K-NAA in comparison to the control (2.5 shoots/explant). Shoot formation varied significantly in response to individual dicamba, picloram, and 2,4-D concentrations. A maximum of 7.9 shoots per explant was promoted by 4.0 μm K-NAA and 1.0 μm dicamba, respectively. Root and callus formation also varied significantly between auxin treatments. Root formation was inhibited by dicamba, picloram, and 2,4-D treatments in comparison with the control (100% rooting); callus formation was promoted by dicamba, picloram, and K-NAA treatments in comparison with the control (15% callusing).

scholarly journals In vitro Regeneration, Rooting, and Cloning of Artemisia tridentata

2020 ◽  
Author(s):  
Rachael Barron
Keyword(s):  
Adventitious Roots ◽  
Root Formation ◽  
Callus Formation ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Adventitious Root Formation ◽  
Big Sagebrush ◽  
Nodal Cuttings ◽  
Wyoming Big Sagebrush

Artemisia tridentata (big sagebrush) is an ecologically important shrub found in western North America. In vitro techniques can be applied to big sagebrush for the purpose of studying gene function, genotypic and phenotypic plasticity studies, cloning, genotypic preservation, and restoration. I performed experiments to develop an indirect organogenesis protocol to regenerate whole Wyoming big sagebrush plants from leaf explants. Callus formation frequency was 88% (±4.0%) in leaf explants cultured on medium containing 0.5 mg/l BAP and 1.0 mg/l NAA. Shoot formation frequency was variable between replicates and was the highest when callus tissue was cultured on medium containing 1.5 mg/l BAP and 0.1 mg/l NAA, 37% to 80%. I tested several auxin treatments to induce root formation and concluded the best to be 0.5mg/l IBA, which yielded 42% to 60% rooting. Taking into account all these variables, I estimate the total regeneration efficiency to range between 14% to 43% on this set of treatments. This protocol was also applied to basin big sagebrush. Callus formation was 100% in leaf explants. Shoot formation was 34% (±14.6%), but shoots exhibited a hyperhydric phenotype and were not transferred to root induction medium. The in vitro regeneration protocol developed is a crucial element that would be required to transform big sagebrush using molecular approaches. Experiments were also conducted to determine the feasibility of shoot tip and nodal cuttings to develop adventitious roots in vitro. This method can provide genetically identical material much faster than in vitro regeneration. Adventitious root formation in Wyoming big sagebrush cuttings cultured on two media types was inconsistent, ranging from 10% in some experiments to 80% in others. Limited success was achieved in nodal cuttings cultured on modified MS medium containing auxin and cytokinin 12.5% (±5.6%). No root formation was achieved in mature plant tissue collected in the field. Results indicated that genotypic influences were likely more responsible for variations in rooting than the medium or vessel conditions tested. Cloning experiments in basin big sagebrush further supported this notion. All material for these experiments came from half-sibling individuals that was maintained separately throughout the course of the experiments. Some half-siblings formed no adventitious roots on any treatments tested whereas others had high rates of formation on all treatments. Further studies, utilizing exogenous PGRs, such as auxins, may provide more successful adventitious root formation in shoot tips from both big sagebrush subspecies.

scholarly journals Somatic Embryogenesis and Plant Regeneration from in-vitro-grown Leaf Explants of Rose

HortScience ◽  
2004 ◽  
Vol 39 (6) ◽  
pp. 1378-1380 ◽  
Author(s):  
C.K. Kim ◽  
J.Y. Oh ◽  
J.D. Chung ◽  
A.M. Burrell ◽  
D.H. Byrne
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Basal Medium ◽  
Shoot Formation ◽  
Ms Medium ◽  
Regeneration Frequency

Somatic embryogenesis was initiated from in vitro-grown leaf explants of rose using an induction period of 4 weeks on MS basal medium supplemented with auxin followed by several subcultures on MS basal medium with cytokinin. `4th of July' showed the highest regeneration frequency (24.4%) on 5.3 μm NAA followed by culture on medium containing 18.2 μm zeatin. `Tournament of Roses' produced somatic embryos when cultured for 4 weeks on medium containing dicamba, 2.3 μm followed by three subcultures on medium containing 18.2 μm zeatin. Embryogenic callus matured on MS media containing 0.5 μm NAA, 6.8 μm zeatin, and 2.9 μm GA3. Long-term cultures were established for both cultivars. Somatic embryos germinated on MS medium containing IBA and BA. Silver nitrate (58.8 μm) enhanced shoot formation and germination of somatic embryos. Plants derived from somatic embryos were acclimatized and successfully established in the greenhouse.

scholarly journals IN VITRO REGENERATION AND MASS PROPAGATION OF AZIMA TETRACANTHA [LAM.] FROM THE LEAF EXPLANTS THROUGH CALLUS CULTURE

2017 ◽  
Vol 4 (2) ◽  
pp. 52-56
Author(s):  
Mallika Devi T
Keyword(s):  
Callus Induction ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Ms Medium ◽  
Apical Leaf ◽  
Half Strength ◽  
Regenerated Plantlets ◽  
Azima Tetracantha

In the present study the protocol for callus induction and regeneration in Azima tetracantha has been developed in culture medium. The young apical leaf explants were used for callus induction on MS medium containing BAP and NAA at 1.0 and 0.4mgl-1 respectively showed maximum callus induction (73%). The amount of callus responded for shoot formation (74%) was obtained in the MS medium containing BAP (1.5 mgl-1) and NAA (0.3mgl-1).The elongated shoots were rooted on half strength medium supplemented with IBA (1.5 mgl-1) and Kn (0.4 mgl-1) for shoots rooted. Regenerated plantlets were successfully acclimatized and hardened off inside the culture and then transferred to green house with better survival rate.

scholarly journals In vitro Regeneration of the Medicinal Plant, Plumbago zeylanica L. with Reference to a Unique Population in Maruthamalai, the Western Ghats, India

1970 ◽  
Vol 18 (2) ◽  
pp. 173-179 ◽  
Author(s):  
T. Mallikadevi ◽  
P. Senthilkumar ◽  
S. Paulsamy
Keyword(s):  
Medicinal Plant ◽  
In Vitro Regeneration ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Shoot Formation ◽  
Garden Soil ◽  
Plumbago Zeylanica ◽  
Adventitious Shoot Formation ◽  
The Western Ghats

The in vitro regeneration of Plubago zeylanica exhibited that the callus was initiated in the basal medium containing BAP, NAA, 2, 4-D, and IBA.  The high amount (90%) of organic calli was induced in the basal medium supplemented with 2, 4-D, alone at 2.0 mg/l. In the subculture the adventitious shoot formation was prominently higher (83%) in the basal medium containing BAP, and NAA at 3.5 and 0.3 mg/l, respectively. IAA (1.0 mg/l)effectively produced higher percen-tage (90) of roots and root growth. After sequential hardening, survivability rate was observed to be significantly higher (80%) in the hardening medium containing garden soil, sand and vermicompost in the ratio of 1 : 1 : 1 by volume under greenhouse condition.  Key words: Plumbago zeylanica, In vitro regeneration, Medicinal plant D.O.I. 10.3329/ptcb.v18i2.3648 Plant Tissue Cult. & Biotech. 18(2): 173-179, 2008 (December)

scholarly journals In Vitro Effects of Different Combinations of Phytohormones on Callus Induction from Different Explants of Cabbage Brassica Oleracea Var. Capitata L. Seedlings

2021 ◽  
pp. 3476-3486
Author(s):  
Alaa. M. Hasan ◽  
Ekhlas. A.J. ElKaaby ◽  
Rakad. M.Kh. AL-Jumaily
Keyword(s):  
Brassica Oleracea ◽  
Callus Induction ◽  
Basal Medium ◽  
Culture Conditions ◽  
Control Treatment ◽  
Fresh Weight ◽  
Superior Result ◽  
Significant Difference ◽  
In Vitro Effects

    The leading purpose of this work is the development of efficient culture conditions to induce calli from cabbage (Brassica oleracea var. capitata L.) under in vitro conditions. The mature seeds were surface sterilized with combinations of different concentrations of ethanol and NaOCl in different time durations and  were germinated on MS basal medium. The results revealed that the best sterilization method of cabbage seeds was by using 70% ethanol for one minute, followed by 15 min in 2% (NaOCl). Seedlings were used as donor sources for hypocotyls, cotyledon leaves, true leaves, and shoot tip explants. These explants were cultured on different combinations of cytokinins (TDZ, BAP, Ad) and auxins (IAA, NAA, 2, 4-D) then implanted in Murashige and Skoog (MS) media. 4 weeks after culturing, a significant difference was found among the explants in response to plant hormones. The maximum percentage of callus induction (100%) was using the combinations of 1 BAP + 1 2, 4-D, 1 BAP + 1 NAA, and 1 BAP + 2 2,4-D mg. l-1. In addition, explants responses varied and the hypocotyls showed a superior result (85.71 %) as compared to other explants. For callus fresh weight, the combination of 0.22 TDZ + 79.9 Ad mg. l-1    had a significant effect, causing the highest fresh weight (0.2745g), while control treatment gave the lowest mean of 0.0066 g. Data showed that cotyledon explants were significantly superior in giving highest callus fresh weight with the mean of 0.1723 g. On the other hand, hypocotyl explants gave the lowest mean, reaching 0.1542 g.

scholarly journals Adventitious Shoot Regeneration from In Vitro Leaf Explants of the Peach Rootstock Hansen 536

Plants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 755
Author(s):  
Angela Ricci ◽  
Luca Capriotti ◽  
Bruno Mezzetti ◽  
Oriano Navacchi ◽  
Silvia Sabbadini
Keyword(s):  
In Vitro Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Shoot Cultures ◽  
Efficient System ◽  
Regeneration Rate ◽  
Factors Affecting

In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 μM N6-benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 μM STS or carbenicillin (238 μM) combined with cefotaxime (210 μM) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in Prunus spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars.

scholarly journals Total Polyphenol Content, in vitro Antifungal and Antioxidant Activities of Callus Cultures from Inula crithmoides

2013 ◽  
Vol 8 (11) ◽  
pp. 1934578X1300801 ◽  
Author(s):  
Anahi Bucchini ◽  
Laura Giamperi ◽  
Donata Ricci
Keyword(s):  
Methanol Extract ◽  
Antioxidant Activities ◽  
Leaf Explants ◽  
Basal Medium ◽  
Alternaria Solani ◽  
Antimicrobial Activities ◽  
Callus Cultures ◽  
Naphthaleneacetic Acid ◽  
Stable Free Radical

This is the first report on the antioxidant and antifungal activities of callus cultures from Inula crithmoides L. (Asteraceae). Callus cultures were initiated from leaf sections, on initial culture MS basal medium supplemented with various concentrations of 2,4-D (2,4-dichlorophenoxyacetic acid), NAA (1-naphthaleneacetic acid) and IBA (indole-3-butyric acid) and a 72% survival was achieved. Significant differences between the various auxins used as phytohormones on callus growth were found. Maximum callusing was noticed on the leaf explants grown on MS basal medium supplemented with 1 mgL–1 2,4-D. Subsequently the antioxidant and antimicrobial activities of the methanol extract from calli were investigated. Antioxidant studies suggested that the methanol extracts of dark-grown and light-grown callus were able to reduce the stable free radical 2,2-diphenyl-1-picrilhydrazyl (DPPH). In the inhibition against lipid peroxidation, extracts of dark-grown callus showed the strongest effect with IC50 values better than those of the standards. The methanol extract of callus cultures had significant antifungal activity only against two of the fungi tested: Alternaria solani and Phytophthora cryptogea. Against all the other tested fungi, the I. crithmoides calli extracts showed fungistatic activity.

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