scholarly journals Acacia Mangium × A. Auriculiformis Micropropagation in a Non-Sterile Environment

2021 ◽  
Author(s):  
Yanping Lu ◽  
Liejian Huang ◽  
Hong Wang ◽  
Muhammad Qasim Shahid
Keyword(s):  
Tissue Culture ◽  
Simple Procedure ◽  
Acacia Mangium ◽  
Proliferation Rate ◽  
Process Time ◽  
Induction Medium ◽  
Contamination Rate ◽  
Indolebutyric Acid ◽  
Bud Induction ◽  
Chemical Sterilization

Abstract Background: Autoclaving is used to eliminate contamination during tissue culturing, however, it is a complicated process, time-consuming and costly. Chemical sterilization of tissue culture can effectively eliminate contamination, is a simple procedure, and cost effective. However, studies on the chemical sterilization mostly focus on bud induction, while the effects of chemical sterilization overall process of tissue culture, including bud induction, proliferation, and rooting, remain to be determined. Here, we investigate the effect of chemical sterilization on bud induction, proliferation, and rooting of Acacia mangium × A. auriculiformis.Results: The results showed that chlorothalonil (0.2 g/L) was a suitable chemical sterilant, and bud induction medium was 1/8 Murashige and Skoog medium + agar 7 g/L + chlorothalonil 0.2 g/L + 6-benzylaminopurine 0.5 mg/L The highest induction rate (99.54%) was observed in the third to fifth buds’ stem segments collected in October treated with 0.8 g/L carbendazim for 3 min, with a contamination rate of 0. The rooting medium was agar 7 g/L + chlorothalonil 0.2 g/L+ indolebutyric acid 1.5 mg/L + naphthylacetic acid 0.5 mg/L, and the rooting rate was 97.62%. The proliferation rate and subculture duration showed a positive correlation, while the proliferation rate was 3.58 times higher at the fourth subculture rooting. Conclusions: Our results suggest that chlorothalonil can effectively replace autoclaving during bud induction, proliferation, and rooting of A. mangium × A. auriculiformis. The findings of this study provide technical support for rapid seedlings propagation, accelerates the breeding process of Acacia, and can be applied in other tree species.

scholarly journals The Effect of Plant Growth Regulators on Callus Induction and Regeneration of Amygdalus communis

2011 ◽  
Vol 3 (3) ◽  
pp. 97-100
Author(s):  
Naimeh SHARIFMOGHADAM ◽  
Abbas SAFARNEJAD ◽  
Sayed Mohammad TABATABAEI
Keyword(s):  
Tissue Culture ◽  
Plant Growth ◽  
Plant Growth Regulators ◽  
Growth Regulators ◽  
Base Material ◽  
Culture Technique ◽  
Induction Medium ◽  
Root Induction ◽  
Amygdalus Communis

The Almond (Amygdalus communis) is one of the most important and oldest commercial nut crops, belonging to the Rosaceae family. Almond has been used as base material in pharmaceutical, cosmetic, hygienically and food industry. Propagation by tissue culture technique is the most important one in woody plants. In the current research, in vitro optimization of tissue culture and mass production of almond was investigated. In this idea, explants of actively growing shoots were collected and sterilized, then transferred to MS medium with different concentrations and combinations of plant growth regulators. The experiment was done in completely randomized blocks design, with 7 treatment and 30 replications. After 4 weeks, calli induction, proliferation, shoot length and number of shoot per explants were measured. Results showed that the best medium for shoot initiation and proliferation was MS + 0.5 mg/l IAA (Indol-3-Acetic Acid) + 1 mg/l BA (Benzyl Adenine). Autumn was the best season for collecting explants. The shoots were transferred to root induction medium with different concentrations of plant growth regulators. The best root induction medium was MS + 0.5 mg/l IBA (Indol Butyric Acid).

scholarly journals The Effect of Daminozide, Dark/Light Schedule and Copper Sulphate in Tissue Culture of Triticum timopheevii

Plants ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2620
Author(s):  
Dmitry Miroshnichenko ◽  
Anna Klementyeva ◽  
Sergey Dolgov
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Callus Induction ◽  
Copper Ions ◽  
Induction Medium ◽  
Tissue Cultures ◽  
Triticum Timopheevii ◽  
Dark Light ◽  
Green Plants ◽  
Callus Induction Medium

Triticum timopheevii Zhuk. is a tetraploid wheat that is utilized worldwide as a valuable breeding source for wheat improvement. Gene-based biotechnologies can contribute to this field; however, T. timopheevii exhibits recalcitrance and albinism in tissue cultures, making this species of little use for manipulation through genetic engineering and genome editing. This study tested various approaches to increasing in vitro somatic embryogenesis and plant regeneration, while reducing the portion of albinos in cultures derived from immature embryos (IEs) of T. timopheevii. They included (i) adjusting the balance between 2,4-D and daminozide in callus induction medium; (ii) cultivation using various darkness/illumination schedules; and (iii) inclusion of additional concentrations of copper ions in the tissue culture medium. We achieved a 2.5-fold increase in somatic embryogenesis (up to 80%) when 50 mg L−1 daminozide was included in the callus induction medium together with 3 mg L−1 2,4-D. It was found that the dark cultivation for 20–30 days was superior in terms of achieving maximum culture efficiency; moreover, switching to light in under 2 weeks from culture initiation significantly increased the number of albino plants, suppressed somatic embryogenesis, and decreased the regeneration of green plants. Media containing higher levels of copper ions did not have a positive effect on the regeneration of green plants; contrarily, the elevated concentrations caused albinism in plantlets. The results and relevant conclusions of the present study might be valuable for establishing an improved protocol for the regeneration of green plants in tissue cultures of T. timopheevii.

scholarly journals A Simple Procedure for Mesophyll Protoplast Culture and Plant Regeneration in Brassica oleracea L. and Brassica napus L.

2011 ◽  
Vol 42 (No. 3) ◽  
pp. 103-110 ◽  
Author(s):  
N.D. Kaur ◽  
M. Vyvadilová ◽  
M. Klíma ◽  
M. Bechyně
Keyword(s):  
Brassica Napus ◽  
Plant Regeneration ◽  
Brassica Oleracea ◽  
Growth Regulators ◽  
Protoplast Culture ◽  
Treatment Time ◽  
Simple Procedure ◽  
Enzyme Treatment ◽  
Induction Medium ◽  
Brassica Napus L

An improved protocol for Brassica protoplast culture and plant regeneration was developed. Isolated protoplasts from four-weeks-old in vitro shoot tip culture of Brassica oleracea var. botrytis cv. Siria F1 and Brassica napus doubled haploid of breeding line OP-1 were cultured at a density of 9.8&ndash;11.2 &times; 10<sup>4 </sup>protoplasts/ml in darkness at 25&deg;C in a modified medium containing 2% glucose, 0.25 mg/l 2,4-D, 1 mg/l BAP and 1 mg/l NAA. The first divisions of protoplasts were observed on the third day of culture in B. oleracea and on the fourth day in B. napus. The protoplast cultures were diluted with low osmotic medium on 7<sup>th</sup> and 11<sup>th</sup> day. The frequency of dividing cells was about 80% in B. oleracea and 50% in B. napus. After one month, the microcalli of approximately 0.5&ndash;1 mm in size were transferred into an induction medium with various combinations of growth regulators. Minimum duration of enzyme treatment time and extended dark period in the initial phase of culture increased the survival rate of protoplasts. Organogenesis started when the calli enlarged in size on an induction medium (1 mg/l NAA, 0.02 mg/l GA<sub>3</sub>, 1 mg/l 2iP) with 2% sucrose and 0.8% agar. Regeneration frequency of calli was found to be 69&ndash;75% in B. oleracea and 2&ndash;3% in B. napus. Well-developed shoots were transferred for rooting to a half-strength MS medium without growth regulators. More than 100 B. oleracea regenerants were transferred into soil, and they produced normal heads and set seeds. This very simple procedure is efficient and suitable mainly for B. oleracea var. botrytis and represents a background for fusion experiments. &nbsp;

scholarly journals Nutrient optimization for improved in vitro plant regeneration in Eclipta alba (L.) Hassk. and assessment of genetic fidelity using RAPD analysis

2015 ◽  
Vol 24 (2) ◽  
pp. 223-234
Author(s):  
Shruti Bardar ◽  
Varsha Khurana Kaul ◽  
Sumita Kachhwaha ◽  
SL Kothari
Keyword(s):  
Basal Medium ◽  
Genetic Fidelity ◽  
Ex Situ ◽  
Nutrient Level ◽  
Induction Medium ◽  
Eclipta Alba ◽  
Shoot Bud ◽  
Bud Induction ◽  
Regeneration Response

This study highlights the effect of different inorganic micronutrients like copper, cobalt, molybdenum, zinc, boron, iodine, iron and manganese in accelerating and amplifying in vitro shoot bud induction and proliferation of a medicinally important plant, Eclipta alba (L.) Hassk. Direct shoot bud induction was observed on MS fortified with Kn (2 mg/l). However, maximum number of shoots was achieved when GA3, 0.5 mg/l was added to induction medium along with 1?M copper sulphate (ten times the normal MS level). Optimization of nutrient level in the basal medium promoted maximum regeneration response from both shoot tips and nodal explants. Elongated shoots were rooted in MS supplemented with IBA, 1.0 mg/l. Healthy, green plantlets with well developed roots, flowered normally in the field. Genetic stability of micropropagated plantlets was evaluated using RAPD markers. The amplification products were monomorphic in micropropagated plantlets and similar to those of mother plant revealing the genetic uniformity of plantlets. The regeneration protocol is highly efficient and reproducible so would be useful for mass multiplication, ex situ conservation and genetic transformation of E. alba (L.) Hassk.Plant Tissue Cult. & Biotech. 24(2): 223-234, 2014 (December)

scholarly journals HISTOLOGICAL EXAMINATION OF IN VITRO ADVENTITIOUS BUD DEVELOPMENT ON HYPOCOTYLS OF FRASER FIR

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1102a-1102
Author(s):  
Carole H. Saravitz ◽  
Frank A. Blazich ◽  
Henry V. Amerson
Keyword(s):  
Growth Regulators ◽  
Induction Medium ◽  
Adventitious Bud ◽  
Naphthaleneacetic Acid ◽  
Abies Fraseri ◽  
Fraser Fir ◽  
Cell Clusters ◽  
Cell Divisions ◽  
Bud Induction

Hypocotyls of Fraser fir (Abies fraseri (Pursh) Poir.) were excised from seeds germination 9 days and placed on bud induction medium containing 10 mg/liter benzyladenine (BA) and 0.01 mg/liter naphthaleneacetic acid (NAA) or medium without growth regulators. After 3 days on medium containing growth regulators, cell divisions were localized in epidermal and subepidermal layers of the hypocotyl while similar cell divisions were not observed in control-treated hypocotyls. Cell clusters consisting of two to five cells were present after 7 days in hypocotyls placed on bud induction medium. In control-treated hypocotyls, stomata continued to develop and cells within the cortex became vacuolated during the first 2 weeks in culture. All hypocotyls were transferred to secondary medium after 3 weeks. Cell clusters continued to enlarge into meristemoids in hypocotyls initially placed on bud induction medium. Gradually, meristemoids developed into buds and cataphylls were observed covering bud meristems.

scholarly journals Establishment of Tissue Culture Regeneration System for Medicago ruthenica L. cv. ‘Zhilixing’

2021 ◽  
Author(s):  
Bo Xu ◽  
Rina Wu ◽  
Cuiping Gao ◽  
Fengling Shi
Keyword(s):  
Tissue Culture ◽  
Stress Resistance ◽  
Callus Induction ◽  
Combination Method ◽  
Induction Medium ◽  
Naphthaleneacetic Acid ◽  
Regeneration System ◽  
Medicago Ruthenica ◽  
Rooting Medium ◽  
Drought And Salt Tolerance

Background: Medicago ruthenica L. ‘Zhilixing’ is a new variety with superior forage and seed yield compared to the wild type. The cold, drought and salt tolerance of Zhlixing are better than those of alfalfa, suggesting that this variety can serve as a high-quality genetic resource for improving the stress resistance of alfalfa. However, because of the lack of tissue culture regeneration system, it is difficult to perform genetic transformation studies on stress resistance genes. This study aimed to establish an efficient tissue culture regeneration system for Zhilixing variety. Methods: Three types of explants were selected and tested on four types of basal media supplemented with different combinations of auxin and cytokinin for callus induction and differentiation, based on orthogonal tests to select the combinations of auxin and cytokinin suitable for callus induction and differentiation. Two-factor combination method was used to formulate a suitable rooting medium. Result: The hypocotyledonary axis was found to be an excellent explant for callus induction on MS medium. The optimum callus induction medium contained thidiazuron (TDZ, 0.5 mg/L), 2,4-dichlorophenoxyacetic acid (2,4-D, 1.0 mg/L) and naphthaleneacetic acid (NAA, 0.5 mg/L) where the callus induction rate was 93.33%. The differentiation medium was supplemented with TDZ (0.75 mg/L), 2,4-D (0.25 mg/L) and 6-benzyladenine (6-BA, 1.5 mg/L) where the differentiation rate was 63.33%. Thidiazuron played the key role in both processes of callus induction and differentiation. Half-strength MS containing 0.1 mg/L of NAA was the most efficient rooting medium.

scholarly journals PLANT REGENERATION OF AVOCADO (PERSEA AMERICANA MILL) IN VITRO

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 696d-696
Author(s):  
Yaseen Mohamed-Yaseen ◽  
Raymond J. Schnell ◽  
Robert J. Knight ◽  
T.L. Davenport
Keyword(s):  
Tissue Culture ◽  
Multiple Shoots ◽  
Persea Americana ◽  
Naphthalene Acetic Acid ◽  
Root Induction ◽  
Ms Medium ◽  
Indolebutyric Acid ◽  
Embryonic Axes ◽  
Phenolic Oxidation

A procedure was developed to regenerate plants via tissue culture from embryonic axes of mature avocado seeds. Explants were cultured in Murashige and Skoog (MS) medium supplemented with benzyladenine (BA) and naphthalene-acetic acid (NAA) or thidiazuron (TDZ) and NAA. Culture were kept in the dark for 7-10 days to reduce browning resulting from phenolic oxidation. Multiple shoots (5-8) were formed after transfer to light. Further multiplication were achieved using different combination of BA and NAA or TDZ and NAA. Shoots were cultured in MS supplemented with 2mg/l indolebutyric acid (IBA) for 2 weeks then transferred to MS supplemented with lg/l activated charcoal for root induction. Complete plants were obtained in vitro.

scholarly journals Effects of Constant Magnetic Field to the Proliferation Rate of Human Fibroblasts Grown onto Different Substrates: Tissue Culture Polystyrene, Polyacrylamide Hydrogel and Ferrogels γ-Fe2O3 Magnetic Nanoparticles

Nanomaterials ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1697 ◽  
Author(s):  
Felix A. Blyakhman ◽  
Grigory Yu. Melnikov ◽  
Emilia B. Makarova ◽  
Fedor A. Fadeyev ◽  
Daiana V. Sedneva-Lugovets ◽  
...  
Keyword(s):  
Magnetic Field ◽  
Cell Proliferation ◽  
Tissue Culture ◽  
Magnetic Nanoparticles ◽  
Regenerative Medicine ◽  
Constant Magnetic Field ◽  
Dermal Fibroblasts ◽  
Proliferation Rate ◽  
Tissue Culture Polystyrene ◽  
The Magnetic Field

The static magnetic field was shown to affect the proliferation, adhesion and differentiation of various types of cells, making it a helpful tool for regenerative medicine, though the mechanism of its impact on cells is not completely understood. In this work, we have designed and tested a magnetic system consisting of an equidistant set of the similar commercial permanent magnets (6 × 4 assay) in order to get insight on the potential of its experimental usage in the biological studies with cells culturing in a magnetic field. Human dermal fibroblasts, which are widely applied in regenerative medicine, were used for the comparative study of their proliferation rate on tissue culture polystyrene (TCPS) and on the polyacrylamide ferrogels with 0.00, 0.63 and 1.19 wt % concentrations of γ-Fe2O3 magnetic nanoparticles obtained by the well-established technique of laser target evaporation. We used either the same batch as in previously performed but different biological experiments or the same fabrication conditions for fabrication of the nanoparticles. This adds special value to the understanding of the mechanisms of nanoparticles contributions to the processes occurring in the living systems in their presence. The magnetic field increased human dermal fibroblast cell proliferation rate on TCPS, but, at the same time, it suppressed the growth of fibroblasts on blank gel and on polyacrylamide ferrogels. However, the proliferation rate of cells on ferrogels positively correlated with the concentration of nanoparticles. Such a dependence was observed both for cell proliferation without the application of the magnetic field and under the exposure to the constant magnetic field.

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