In vitro rooting of hybrid hazelnuts (Corylus avellana × Corylus americana) in a temporary immersion system

Commercial micropropagation of hybrid hazelnuts (Corylus avellana L. × C. americana Marshall) has been limited, owing to their poor rooting ability in vitro as well as ex vitro, leading to high mortality of plantlets transplanted in the greenhouse. The objective of this study was to develop an efficient and cost-effective protocol for rooting and plantlet acclimation of in vitro grown hazelnut shoots. Efficient in vitro rooting was accomplished in a rocker-based temporary immersion bioreactor system. The use of a temporary immersion system (TIS) in combination with the inert substrate Oasis® In Vitro Express (IVE) significantly improved the in vitro rooting efficiency (100%) compared with semi-solid medium (27%) after four weeks of culture. A higher density (36 explants/vessel) of shoot explants in the TIS was found to support a significantly greater shoot height, chlorophyll content, and longest root length, compared with the lowest density treatment (12 explants/vessel). Efficiency of rooting and the number of roots formed were similar for both the high and low density of explants in the culture vessels, and the resulting plantlets exhibited > 80% survival in the greenhouse. These results demonstrate the usefulness of rocker-based TIS for commercial micropropagation of hazelnuts and, potentially, other tree species.

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A comparative study between temporary immersion system and semi-solid cultures on shoot multiplication and plantlets production of two Moroccan date palm (Phoenix dactylifera L.) varieties in vitro

Notulae Scientia Biologicae ◽

10.15835/nsb12210610 ◽

2020 ◽

Vol 12 (2) ◽

pp. 277-288

Author(s):

Larbi ABAHMANE

Keyword(s):

Date Palm ◽

New Technologies ◽

Shoot Multiplication ◽

Handling Time ◽

Cost Effective ◽

Phoenix Dactylifera ◽

Temporary Immersion System ◽

Temporary Immersion ◽

Semi Solid

Date palm micropropagation is commonly performed on gelled media. However, it’s typically a labour-intensive system and consequently plantlets production cost is very high. Therefore, it is necessary to develop cost effective alternatives without compromising the quality of produced plant material. New technologies based on liquid media in bioreactors have been developed to reduce the handling time, while increasing the multiplication rates and plant quality. The present research focuses on the comparison between Temporary Immersion System (TIS) and gelled media (GM) culture systems of two Moroccan date palm varieties ‘Mejhool’ and ‘Boufeggous’. Obtained results indicated that shoot and root lengths as well as shoot fresh and dry weights were significantly (P < 0.05) higher in TIS compared to GM. Moreover, the vigour of obtained shoots was better in TIS compared to GM. Therefore, TIS-derived plantlets have shown an acclimatization rate of 95% while this rate for GM-derived plantlets was 82%. Hence, bioreactors, as a growing system based on TIS, can be a valid alternative to conventional systems for in vitro culture, resulting in a reduction of cost, shelving area requirements, labour and time for the mass propagation of date palm cultivars.

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In Vitro Propagation of Rheophytic Orchid, Epipactis flava Seidenf.—A Comparison of Semi-Solid, Continuous Immersion and Temporary Immersion Systems

Biology ◽

10.3390/biology8040072 ◽

2019 ◽

Vol 8 (4) ◽

pp. 72

Author(s):

Boworn Kunakhonnuruk ◽

Phithak Inthima ◽

Anupan Kongbangkerd

Keyword(s):

Large Scale ◽

Growth Parameters ◽

Natural Habitat ◽

Ex Situ ◽

Temporary Immersion System ◽

Shoot Height ◽

Temporary Immersion ◽

Shoot Buds ◽

Semi Solid

Epipactis flava Seidenf. is an endangered Thai rheophytic orchid that has recently shown a rapid decrease in its natural habitat, prompting an urgent need for conservation using ex situ reintroduction methods. Temporary immersion system (TIS) has been successfully applied for large-scale propagation in various plants species. Propagation efficiency of E. flava using TIS was investigated and compared with conventional semi-solid system (SSS) and liquid continuous immersion system (CIS). The highest percentage of new shoot and shoot bud formation was obtained from TIS, followed by CIS and SSS, respectively. Growth parameters as indicated by number of new shoots, shoot buds, shoot height and leaves per explant were significantly higher using TIS than with SSS and CIS. Moreover, the maximum number of new shoots and shoot buds per replication were reliably obtained from TIS higher than SSS and CIS. After acclimatization, the highest survival percentage of plantlets was observed in TIS (76.7%), with 60% surviving after eight weeks of transplantation in artificial stream. TIS was determined as the most suitable culture system for in vitro mass propagation of E. flava compared to CIS and SSS.

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In Vitro Propagation of Easter Island Curcuma longa from Rhizome Explants Using Temporary Immersion System

Agronomy ◽

10.3390/agronomy11112121 ◽

2021 ◽

Vol 11 (11) ◽

pp. 2121

Author(s):

María José Marchant ◽

Paula Molina ◽

Miriam Montecinos ◽

Leda Guzmán ◽

Cristobal Balada ◽

...

Keyword(s):

In Vitro Propagation ◽

Large Scale ◽

Curcuma Longa ◽

Cost Effective ◽

Rapa Nui ◽

Temporary Immersion System ◽

Term Survival ◽

Temporary Immersion ◽

Large Scale Cultivation

Curcuma longa (C. longa) is widely known for its medicinal properties. However, the potential overexploitation of this plant raises doubts about its long-term survival on Rapa Nui. Micropropagation using a temporary immersion system (TIS) could be the basis for developing a cost-effective and highly productive method of large-scale cultivation of this plant. Our objective was to develop and refine the in vitro multiplication system for mass propagation of C. longa, and thus help restore the fragile ecosystem of Rapa Nui. Three parameters were evaluated: number of explants per flask, flask capacity, and LEDs spectrum. For each parameter evaluated, four aspects were analyzed: fresh weight per plant, number of shoots, percentage of non-sprouting explants, and the proliferation rate. The use of 30 explants per two-liter flask results in more plants with high fresh biomass than other configurations. In addition, LEDs with a red:blue ratio of 2:1 provided the best lighting conditions for in vitro propagation and positively affected C. longa proliferation and rooting. Therefore, our results show that 30 explants per two-liter flask and an LED source with a red:blue ratio of 2:1 allow a higher number of C. longa plants to be obtained using TIS.

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In Vitro Culture of Rosmarinus officinalis L. in a Temporary Immersion System: Influence of Two Phytohormones on Plant Growth and Carnosol Production

Pharmaceuticals ◽

10.3390/ph14080747 ◽

2021 ◽

Vol 14 (8) ◽

pp. 747

Author(s):

Eder Villegas Sánchez ◽

Mariana Macías-Alonso ◽

Soraya Osegueda Robles ◽

Lisset Herrera-Isidrón ◽

Hector Nuñez-Palenius ◽

...

Keyword(s):

High Performance ◽

Emerging Infectious Diseases ◽

Rosmarinus Officinalis ◽

Wild Plant ◽

Array Detector ◽

Phase System ◽

Naphthaleneacetic Acid ◽

Temporary Immersion System ◽

Temporary Immersion

Emerging infectious diseases have become a major global problem with public health and economic consequences. It is an urgent need to develop new anti-infective therapies. The natural diterpene carnosol exhibit a wide variety of interesting antibacterial and antiviral properties, and it is considered a theoretical inhibitor of COVID-19 Mpro. However, this compound is present in the family Lamiaceae in low quantities. To obtain carnosol in concentrations high enough to develop pharmacological studies, we evaluated the efficiency of a micropropagation protocol of Rosmarinus officinalis using a solid medium and a temporary immersion system (TIS), as well as the effect of 6-benzylaminopurine (6-BAP) and α-naphthaleneacetic acid (NAA) on the growth of shoots. Moreover, we developed and validated an analytical method to quantify carnosol using the H-point standard additions method in the high-performance liquid chromatography diode array detector (HPLC-DAD). After 30 days of culture, TIS produced the maximum number of shoots per explant (24.33 ± 1.15) on a liquid medium supplemented with 6-BAP at 5.0 mg L−1. Next, we also evaluated the effect of immersion time and frequency for TIS. After 72 days of culture, the best results were obtained with an immersion cycle of 1 min every 12 h, yielding 170.33 ± 29.40 shoots. The quantification of carnosol on the samples was performed at a flow rate of 1.2 mL min−1 using binary isocratic mobile phase system 60:40 (v/v) 10 mM formic acid (pH 3.0) (A) and acetonitrile (B) on a reverse-phase column. The content of carnosol in the in vitro cultures was around 8-fold higher than in the wild plant. The present study represents an efficient alternative method to obtain carnosol for its pre-clinical and clinical development.

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