In vitro leaf anatomy, ex vitro photosynthetic behaviors and growth of Calathea orbifolia (Linden) Kennedy plants obtained from semi-solid medium and temporary immersion systems

bstractTo achieve Indonesian sugar self-sufficiency in2014, the national production needs to be escalatedthrough land extensification that requires a largenumbers of cane planting materials. This can be achievedby mass propagation of sugarcane through in vitroculture. Solid medium is commonly used for callusproliferation in sugarcane tissue culture. However, solidmedium is considered inefficient in terms of plantletproduction level, labour and space. The use of liquidmedium may solve the problem by allowing automationto increase plantlet production scale and uniformity.Temporary immersion system (TIS) is based on a shortperiodic immersion of explants in a liquid medium for aspecific frequency and duration. Research on in vitromass propagation of sugarcane using TIS was conductedat the Indonesian Biotechnology Research Institute forEstate Crops. Callus initiated from immature unfoldedleaves of PSJT 941 and PS 881 was cultured on liquidMS medium in TIS with different frequencies (12 and24 h) and durations (1 and 3 min) of immersion. Eachtreatment was replicated three times. The callus biomassof two elite cane varieties (PSJT 941 and PS 881)cultured in TIS for six weeks was higher (2 – 4 times fold)than that of on solid medium. The PSJT 941 varietyreached the highest calli biomass with immersion forthree min every 24 h. However, PS 881 variety reachedits highest biomass with immersion for one minute every24 h. The propagation of sugarcane using TIS culturewas proven to produce higher calli biomass up to fourfolds and to form more numbers and uniform shootscompared to the solid medium culture. The callus wassuccesfully regenerated to shoots and plantlets.AbstrakUntuk mencapai swasembada gula, perlu dilakukanpeningkatan produksi gula nasional melalui perluasanareal pertanaman tebu sehingga diperlukan bibit dalamjumlah besar. Hal tersebut dapat diatasi antara laindengan perbanyakan tebu melalui kultur in vitro. Peng-gunaan medium padat pada perbanyakan kalus tebumelalui kultur in vitro merupakan teknik yang umumdigunakan saat ini. Akan tetapi penggunaan mediumpadat dianggap kurang efisien dalam hal jumlah planletyang diproduksi, tenaga kerja dan ruang digunakan.Penggunaan medium cair dapat mengatasi kelemahantersebut dengan dimungkinkannya otomatisasi sehinggadapat meningkatkan skala produksi secara massal dankeseragaman planlet. Sistem perendaman sesaat (SPS)merupakan teknik kultur in vitro dalam medium cairmenggunakan bejana bersekat dimana kontak antaraeksplan dan medium terjadi hanya secara sesaat danperiodik. Penelitian perbanyakan massal bibit tebumelalui SPS dilakukan di Balai Penelitian BioteknologiPerkebunan Indonesia. Kalus diinisiasi dari daun meng-gulung varietas PSJT 941 dan PS 881 yang ditumbuhkanpada media MS cair dalam kultur SPS dengan frekuensiyang berbeda (12 dan 24 jam) dan lama perendaman (1dan 3 menit). Setiap perlakuan diulang tiga kali. Bobotbasah (biomassa) kalus dari dua varietas tebu (PSJT 941dan PS 881) yang ditumbuhkan dengan metode SPSsetelah enam minggu menunjukkan pening-katan yanglebih tinggi yaitu antara 2 - 4 kali lipat dibandingkandengan kontrol (media padat). Peningkatan biomassatertinggi pada varietas PSJT 941 diperoleh pada per-lakuan SPS dengan interval perendaman 24 jam dan lamaperendaman tiga menit. Sedangkan pada PS 881,peningkatan tertinggi biomassa diperoleh pada intervalperendaman 24 jam dan lama perendaman satu menit.Perbanyakan dengan metode SPS terbukti dapat mening-katkan biomassa kalus lebih dari empat kali lipat danpembentukan tunas yang lebih seragam dibandingkandengan pada media padat. Kalus yang dihasilkan dapatdiregenerasikan menjadi tunas dan planlet.

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In vitro callogenesis induction of Guarianthe skinneri (Bateman) Dressler & W.E. Higgins (Orchidaceae)

Acta Agronómica ◽

10.15446/acag.v66n2.57982 ◽

2017 ◽

Vol 66 (2) ◽

Cited By ~ 1

Author(s):

Ana Gabriela Coutiño-Cortés ◽

Vincenzo Bertolini ◽

Leobardo Iracheta Donjuan ◽

Lorena Ruíz-Montoya ◽

Javier Francisco Valle-Mora

Keyword(s):

Statistical Analysis ◽

Solid Medium ◽

Callus Formation ◽

Natural Habitat ◽

Leaf Explants ◽

Lower Percentage ◽

Production Technique ◽

Flowering Season ◽

Semi Solid

Guarianthe skinneri (Bateman) Dressler & W.E. Higgins., is a native orchid from Mexico, considered as threatened species by NOM-ECOL-059-SEMARNAT-2010, mainly due to the disappearance of its natural habitat and the illegal collection during its flowering season. The aim of this research was to induce in vitro callogenesis from different type of explants, using phytoregulators, in order to look for a massive production technique to contribute to its conservation. We evaluated the leaf and pseudobulb marrow explants growing in semi-solid medium MS adding BAP, 2, 4-D, Kin, the interaction of BAP/2, 4-D/Kin/Sad and a control without any type of plant growth regulators. Statistical analysis showed the pseudobulb marrow explants are more suitable for in vitro introduction in comparison to leaf explants, since they perform a lower percentage of contamination (18.8% in marrow and 73.2% in leaves). Likewise, the pseudobulb marrow explants increased callus formation (10.8%) in comparison to leaf explants (7.6%). Regarding the phytoregulators employed, BAP have allowed to increased callus formation (17%) compared to other phytoregulators (7-10%). This is the first report, which proposes the use of pseudobulb marrow as explant for callus induction in G. skinneri.

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Análisis bioinformático de las rutas pks y fas en plantas y uso de inductores exógenos sobre la producción de acetogeninas en guanábana cultivada in vitro

Revista de Ingeniería Biomédica y Biotecnología ◽

10.35429/jbeb.2019.9.3.19.28 ◽

2019 ◽

pp. 19-28

Author(s):

Sanjuana Sánchez-Ramos ◽

Felipe Guadalupe Carrillo-Landell ◽

Gustavo Cruz-Jimenez ◽

Alan Alexander González-Ibarra

Keyword(s):

Amino Acids ◽

Metabolic Pathways ◽

Fatty Acid Synthetase ◽

Bioinformatic Analysis ◽

Annona Muricata ◽

Ex Vitro ◽

Temporary Immersion ◽

The Third ◽

Analysis Methodology

Goals. To analyze the effect on the production of acetogenins of Annona muricata cultivated in vitro by means of Temporary Immersion Bioreactors (BITs) promoted by the use of exogenous inducers proposed from a bioinformatic analysis. Methodology. A bioinformatic analysis was carried out at the KEGG site of the metabolic pathways of polyketide synthetase (PKS) and fatty acid synthetase (FAS) plants from which acetogenins are derived. Five treatments were performed in the BITs with soursop explants; 1) BAP, 2) BAP + AAs 3) BAP + AAs + Putrescine, 4) BAP + IAA and 5) BAP + IAA + Citrate. BAP and IAA are plant hormones, AAs is a mixture of three ketogenic amino acids, Putrescine and Citrate are inducers of PKS and FAS. Contribution. The cultivation of soursop explants was monitored using BITs of the 5 treatments for 8 weeks without apparent contamination. Sprout observation started from week 6 in treatment 1 and 2 but not in 3 over 8 weeks, with putrescine being more than an inducer a stress factor for the explant as they became necrotic. Treatments 4 and 5 were only monitored until the third week and no information on sprouting was collected. Differences in the profile of acetogenins by HPLC-PDA were observed between soursop leaves to ex vitro and in vitro and between treatments 1, 2 and 3.

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Emetine and cephaeline production and regulation by in vitro propagation of Psychotria ipecacuanha (Brot.) Stokes in semi-solid media and temporary immersion bioreactor

Revista Fitos ◽

10.32712/2446-4775.2020.799 ◽

2020 ◽

Vol 14 (01) ◽

pp. 45-55

Author(s):

Simone Da Silva ◽

Danielle Cardoso Alencar ◽

Paulo José Coelho Benevides ◽

Spartaco Astolfi-Filho

Keyword(s):

Natural Populations ◽

Root Induction ◽

Relative Area ◽

Temporary Immersion Bioreactor ◽

Temporary Immersion ◽

Mean Values ◽

Solid Media ◽

Medicinal Properties ◽

Semi Solid

Psychotria ipecacuanha, is a plant species with known medicinal properties that is critically endangered due to overexploitation of natural populations. Although the difficulties in conventional propagation by seed and by vegetative propagation are generally understood, the present study enhances our knowledge by describing efficient plant regeneration and root induction protocols for P. ipecacuanha while comparing alkaloid content (emetine and cephaeline) in in vitro-derived tissues. Stem node explants were cultured on MS medium MS supplemented with indolbutiric acid (IBA) in semi-solid media and the RITA® temporary immersion bioreactor. The highest root formation (81%) was in MS + 1.5 mg L−1 IBA in the bioreactor. After 24 months of acclimatization, the plants cultivated in MS + 0.50 and 1.0 mg L-1 of IBA had the highest number of roots (3), with mean values of 10.47 and 9.40 cm, respectively. The cultures coming from 1.0 mg L−1 and 0.5 mg L−1 IBA in the bioreactor contained higher cephaeline content, with a relative area of 14.2 and 14.9%, respectively. For emetine, the 1.0 mg L−1 IBA cultures in the bioreactor, 0.5 mg L−1 IBA and MS0 cultures contained higher content than the other treatments, with a relative area of 10.2, 10.2 and 10.1%, respectively.

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Influence of Polyethylene Glycol on Leaf Anatomy, Stomatal Behavior, Water Loss, and Some Physiological Traits of Date Palm Plantlets Grown In Vitro and Ex Vitro

Plants ◽

10.3390/plants9111440 ◽

2020 ◽

Vol 9 (11) ◽

pp. 1440

Author(s):

Amal F. M. Zein El Din ◽

Mohamed F. M. Ibrahim ◽

Reham Farag ◽

Hany G. Abd El-Gawad ◽

Ahmed El-Banhawy ◽

...

Keyword(s):

Polyethylene Glycol ◽

Photosynthetic Pigments ◽

Leaf Anatomy ◽

Water Loss ◽

Date Palm ◽

High Survival ◽

Ex Vitro ◽

Stomatal Behavior ◽

Ex Vitro Transfer

Few reports explain the mechanism of PEG action on stomatal behavior and anatomical structure and analyze the photosynthetic pigments of in vitro date palm plantlets for better tolerance to ex vitro exposure. The main challenge for in vitro micropropagation of date palm techniques remains restricted to high survival rates and vigorous growth after ex vitro transplantation. In vitro hardening is induced by Polyethylene glycol PEG (0.0, 10, 20, 30 g L−1) for 4 weeks. Leaf anatomy, stomatal behavior, water loss %, photosynthetic pigments, and reducing sugars were examined in date palm plantlets (Phoenix dactylifera L.) cv. (Sewi) after 4 weeks from in vitro PEG treatment and after 4 weeks from ex vitro transplanting to the greenhouse. Leaf anatomy and the surface ultrastructure of in vitro untreated leaves showed a thin cuticle layer, wide opened malfunctioning stomata, and abnormal leaf anatomy. Furthermore, addition of PEG resulted in increasing cuticle thickness, epicuticular wax depositions, and plastids density, improving the stomatal ability to close and decreasing the stomatal aperture length while reducing the substomatal chambers and intercellular spaces in the mesophyll. As a result, a significant reduction in water loss % was observed in both in vitro and ex vitro PEG treated leaves as compared to untreated ones, which exhibited rapid wilting when exposed to low humidity for 4 h. PEG application significantly increased Chlorophylls a, b and carotenoids concentrations, especially 10, 20 g L−1 treatments, which were sequentially reflected in increasing the reducing sugar concentration. However, leaves of plantlets treated with PEG at 30 g L−1 became yellow and had necrosis ends with death. In vitro hardening by 20 g L−1 PEG increased the survival rate of plantlets to 90% after ex vitro transfer compared to 63% recorded for the untreated plantlets. Therefore, this application provides normal date palm plantlets developed faster and enhances survival after ex vitro transfer.

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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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Comparison of Different Semi-Automated Bioreactors for In Vitro Propagation of Taro (Colocasia esculenta L. Schott)

Plants ◽

10.3390/plants10051010 ◽

2021 ◽

Vol 10 (5) ◽

pp. 1010

Author(s):

Eucario Mancilla-Álvarez ◽

Juan Antonio Pérez-Sato ◽

Rosalía Núñez-Pastrana ◽

José L. Spinoso-Castillo ◽

Jericó J. Bello-Bello

Keyword(s):

Culture Medium ◽

Solid Medium ◽

Survival Rates ◽

Shoot Multiplication ◽

Control Treatment ◽

Multiplication Rate ◽

Solid Culture ◽

Solid Culture Medium ◽

Semi Solid

Taro is important for its nutritional content, medicinal use, and bioethanol production. The aim of the present study was to compare different semi-automated bioreactors (SABs) during in vitro multiplication of C. esculenta. The SABs used were temporary immersion bioreactors (TIBs), SETIS™ bioreactors and ebb-and-flow bioreactors; semi-solid culture medium was used as a control treatment. At 30 d of culture, different developmental variables, determination of chlorophyll, stomatal content, and survival percentage during acclimatization were evaluated. SABs increased the shoot multiplication rate relative to the semi-solid medium; however, the SETIS™ bioreactor showed the highest shoot production, with 36 shoots per explant, and the highest chlorophyll content. The stomatal index was higher in the semi-solid medium compared to the SABs, while the percentage of closed stomata was higher in the SABs than in the semi-solid culture medium. The survival rate during acclimatization showed no differences among the culture systems assessed, obtaining survival rates higher than 99%. In conclusion, the SETIS™ bioreactor showed the highest multiplication rate; however, other bioreactor alternatives are available for semi-automation and cost reduction for micropropagation of C. esculenta.

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