Development of a medium and culture system for in vitro propagation of the seagrass Halophila engelmannii

1994 ◽  
Vol 72 (10) ◽  
pp. 1503-1510 ◽  
Author(s):  
K. T. Bird ◽  
J. Jewett-Smith

An in vitro culture system was developed for axenic culture of the seagrass Halophila engelmannii Aschers. The culture system had a rhizome layer solidified with agar (0.8% w/v) and a liquid overlay layer. The rhizome layer contained artificial seawater (20%0, inorganic nutrients, plant growth regulators, sucrose (1% w/v), and activated charcoal (1% w/v). The overlay layer contained artificial seawater and inorganic nutrients. The medium f/4 provided the best source of inorganic nutrients of the five media compared. Axenic cultures of H. engelmannii died if NO3 was the only N source. They grew best when supplied with glutamic acid at a concentration of 3.4 mM as the N source. A combination of α-naphthaleneacetic acid at a concentration of 0.25 mg ∙ L−1 and 6-benzylaminopurine at a concentration of 10 mg ∙ L−1 provided the most rapid rates of propagation as measured by the numbers of pseudowhorls and branches produced. Key words: tissue culture, marine plants, media, plant growth regulators.

Agronomy ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 320
Author(s):  
Nisar Ahmad Zahid ◽  
Hawa Z.E. Jaafar ◽  
Mansor Hakiman

Ginger (Zingiber officinale Roscoe) var. Bentong is a monocotyledon plant that belongs to the Zingiberaceae family. Bentong ginger is the most popular cultivar of ginger in Malaysia, which is conventionally propagated by its rhizome. As its rhizomes are the economic part of the plant, the allocation of a large amount of rhizomes as planting materials increases agricultural input cost. Simultaneously, the rhizomes’ availability as planting materials is restricted due to the high demand for fresh rhizomes in the market. Moreover, ginger propagation using its rhizome is accompanied by several types of soil-borne diseases. Plant tissue culture techniques have been applied to produce disease-free planting materials of ginger to overcome these problems. Hence, the in vitro-induced microrhizomes are considered as alternative disease-free planting materials for ginger cultivation. On the other hand, Bentong ginger has not been studied for its microrhizome induction. Therefore, this study was conducted to optimize sucrose and plant growth regulators (PGRs) for its microrhizome induction. Microrhizomes were successfully induced in Murashige and Skoog (MS) medium supplemented with a high sucrose concentration (>45 g L−1). In addition, zeatin at 5–10 µM was found more effective for microrhizome induction than 6-benzylaminopurine (BAP) at a similar concentration. The addition of 7.5 µM 1-naphthaleneacetic acid (NAA) further enhanced microrhizome formation and reduced sucrose’s required dose that needs to be supplied for efficient microrhizome formation. MS medium supplemented with 60 g L−1 sucrose, 10 µM zeatin and 7.5 µM NAA was the optimum combination for the microrhizome induction of Bentong ginger. The in vitro-induced microrhizomes sprouted indoors in moist sand and all the sprouted microrhizomes were successfully established in field conditions. In conclusion, in vitro microrhizomes can be used as disease-free planting materials for the commercial cultivation of Bentong ginger.


2021 ◽  
Author(s):  
Yuan-yuan Meng ◽  
Shi-jie Song ◽  
Sven Landrein

Abstract Passiflora xishuangbannaensis (Passifloraceae) is endemic to a few sites of Mengyang nature reserve in Yunnan, Xishuangbanna and less than 40 individuals have been recorded. Nine Passiflora species are endemic to Yunnan with most species occurring in South America, making P. xishuangbannaensis highly significant and emblematic to the conservation work in the region. This study is designed to provide the first protocol for in vitro organogenesis and plant regeneration for ex situ conservation and reintroduction for an Asian Passiflora species. Using internodes, petioles and tendrils we optimize calli formation and root elongation using several plant growth regulators, individually or in combination. We also assess the genetic stability of regenerated cells. The maximum callus induction and shoot bud differentiation were both achieved on half Murashige and Skoog basal medium supplemented with 4.44 µM 6-Benzylaminopurine and 1.08 µM 1-Naphthaleneacetic acid. The best rooting was achieved from 30 days old, regenerated shoots on half Murashige and Skoog basal medium supplemented with 1.08 µM 1-Naphthaleneacetic acid. Micropropagated plants were subjected to inter simple sequence repeat markers analyses. Collectively, 86 bands were generated from 6 primers of which 12 bands were polymorphic, showing genetic variation between the regenerated plantlets and the original plant. Response to plant growth regulators was more specific than most other studies using South American species, which could be explained by the morphological and physiological differences between South American and Asian Passiflora species


Author(s):  
Ileana MICLEA ◽  
Rita BERNAT

The aim of the current research was to find the best plant growth regulators for the multiplication of Sarracenia purpurea. Murashige and Skoog medium (MS) was prepared with macronutrients and micronutrients at 1/3 strength, full strength vitamins, supplemented with 30 g/l sucrose and 5 g/l phytagel and autoclaved. After cooling 0.5 mg\l α-naphthaleneacetic acid (NAA), 5 mg\l 6-benzyladenine (BA) or 0.5 mg\l NAA + 3 mg\l BA were added. Young S. purpurea plants were selected and transferred to media with or without plant growth regulators and cultured for 12 weeks. At the end of this time frame number of roots, root length (cm) and number of shoots were evaluated and differences were analysed by the analysis of variance and interpreted using the Tuckey test. The largest number of roots grew in medium supplemented with 0.5 mg\l NAA but the the absence of plant growth regulators increased their length. The best conditions for shoot multiplication were provided by supplementing 1/3MS with 5 mg\l BA.


2012 ◽  
Vol 12 ◽  
pp. 1-6 ◽  
Author(s):  
Shambhu P. Dhital ◽  
Hak T. Lim ◽  
Hira K. Manandhar

Response of widely grown potato cv. Superior and newly developed cvs. Gui valley and Bora valley to plant growth regulators (PGRs) for direct plant regeneration from internode, leaf blade and petiole explants were investigated. The explants were cultured on a MS solid medium supplemented with different concentrations and combinations of 6-benzylaminopurine (BAP), 1-naphthaleneacetic acid (NAA), zeatin, indole-3-acetic acid (IAA) and gibberellic acid (GA3). Potato cv. Superior, regenerated direct shoot without callus and root formation on MS solid medium supplemented with BAP or zeatin, proliferous roots were produced on NAA or IAA supplemented medium and only some calli were produced on GA3 supplemented medium. The regeneration response varied with different concentrations of PGRs, singly and also in combinations. In the case of combined application of PGRs, the highest shoot regeneration (75.3%) and number of shoot per explant (11.5) and number of roots per explant (7.0) were obtained from the MS solid medium supplemented with zeatin (2 mg l-1), NAA (0.1 mg l-1) and GA3 0.1 mg l-1). Among the three types of explants evaluated, internodes produced the highest number of shoots and roots for both potato cvs. Gui valley and Bora valley, and petiole produced the least number of shoots and roots. The regenerated shoots were rooted in PGRs-free MS solid medium and successfully established under glasshouse condition. Leaf, flower, and tuber morphology were identical to in vitro control and mother plants in the same conditions. This optimized regeneration system can be used for rapid shoot proliferation and also for gene transformation.DOI: http://dx.doi.org/10.3126/njst.v12i0.6471 Nepal Journal of Science and Technology 12 (2011) 1-6 


Agronomy ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1181
Author(s):  
María Eugenia Martinez ◽  
Lorena Jorquera ◽  
Paola Poirrier ◽  
Katy Díaz ◽  
Rolando Chamy

Taraxacum officinale (L.) Weber ex F.H. Wigg, commonly known as dandelion, is a cosmopolitan and perennial weed, which has medicinal properties. In vitro propagation methods are widely used on plants that have difficulties in cultivation and, consequently, low extraction yields of active metabolites. Thus, callus culture has been considered to be useful for the accumulation of several metabolites. In this study, we aimed to establish an efficient protocol for callus induction and maintenance of T. officinale, for which explant type, carbon source, light conditions, and nine different combinations of plant growth regulators (PGRs), such as 1-naphthaleneacetic acid (NAA) (from 0.05 to 0.5 mg/L) and 6-benzylaminopurine acid (BAP) (from 0.5 to 3.0 mg/L), were evaluated. The results showed that hypocotyls and roots from sterile seedlings are the best sources for callus induction, with 100% of callogenesis at every condition tested, and more than 95% of viability and friability. Complete darkness and a medium supplemented with sucrose at 2.3% (w/v) and 0.5 mg/L of NAA and 0.5 mg/L of BAP were the best conditions for callus induction, showing callus with low organogenesis and high friability. This study provides a basis for future studies on improving large-scale callus propagation and further establishment of suspension culture systems for commercial purposes.


Horticulturae ◽  
2021 ◽  
Vol 7 (11) ◽  
pp. 486
Author(s):  
María Eugenia Martínez ◽  
Lorena Jorquera ◽  
Paola Poirrier ◽  
Katy Díaz ◽  
Rolando Chamy

There are several studies on the medicinal properties of dandelions (Taraxacum officinale), but few studies are aimed at understanding the in vitro germination process of this plant to improve its propagation. This research was focused on studying in vitro seed germination and development of seedlings under different carbon sources, glucose (GLU) or sucrose (SUC) and its concentrations (1.0–5.5%). Additionally, the effect of supplementation with plant growth regulators (PGRs) was studied, measuring the germination capacity, uncertainty and synchrony. Germination was promoted under low carbon source concentrations (≤2.3%), whereas higher concentrations (≥3.2%) had a detrimental effect on this process. GLU allowed the final germination percentages to be slightly better than SUC. Uniformity and synchrony values improved with the presence of PGRs. Results suggested that the best condition to assess T. officinale seed germination is in a medium containing GLU 2.3%, 0.5 mg/L 1-Naphthaleneacetic acid (NAA) and 0.5 mg/L 6-Benzylaminopurine acid (BAP). After germination, the best condition for optimal growth of T. officinale seedlings was 1.0% SUC supplemented with 0.225 mg/L of NAA and 3.0 mg/L of BAP for initial shoot development. The survival rate was 97% after greenhouse acclimatization. This new method of germination was implemented for the massive propagation of T. officinale for further medicinal studies.


1995 ◽  
Vol 22 (2) ◽  
pp. 135-141
Author(s):  
Q. L. Feng ◽  
H. E. Pattee ◽  
H. T. Stalker

Abstract Research on in vitro embryo culture in Arachis has the objective of rescuing interspecific hybrid embryos which abort before they reach maturity. This study explored effects of the three exogenous plant growth regulators 1-naphthaleneacetic acid (NAA), gibberellic acid (GA3), and 6-benzylaminopurine (6-BAP); sucrose; and medium pH on in vitro fruit and embryo development of A. hypogaea L. by culturing 10-d-old peg tips. Results indicated that medium containing 0.5 to 1.0 mg L-1 NAA was optimal for in vitro pod formation and embryo development. GA3 did not have a significant influence and 6-BAP had negative effects on both in vitro fruit and embryo development. High concentrations of 6-BAP and NAA induced callus which inhibited ovary enlargement and embryo development. Sixty g L-1 sucrose was the best concentration for ovary enlargement and embryo development. Acidic medium was needed for in vitro reproductive development with pH 4.5–6.5 the most favorable. A pod formation frequency of 81%, a seed production rate of 90% (from pods recovered in vitro), and plant recovery of 33% were obtained for a medium containing 1.0 mg L-1 NAA and 0.5 mg L-1 GA plus 60 g L-1 sucrose at pH 5.8. In vitro-recovered cotyledonary embryos between 4 and 10 mm long germinated precociously into seedlings at relatively higher frequencies than morphologically mature embryos which produced more vigorous plants.


HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1024D-1024
Author(s):  
Guochen Yang ◽  
Zhongge (Cindy) Lu ◽  
Carl E. Niedziela

This research was initiated to study different culture media and plant growth regulators for their influences on callus initiation and production, with a research goal of developing an efficient in vitro callus regeneration protocol for guava (Psidium guajava L.). Guava is an important tropical fruit species that is rich in vitamins and vitamin precursors, minerals, organic acids, and pectins. Seventy-nine phytochemicals provide guava with many unique properties and actions, including anti-microbial, astringent, bactericidal, cicatrizant, emmenagogue, hypoglycemic, laxative, nutritive, and spasmolytic. Different concentrations of various plant growth regulators (PGR), such as 6-benzyladenine (BA), kinetin, or 2,4-dichlorophenoxyacetic acid (2,4-D), and naphthaleneacetic acid (NAA) were added to basic Murashige and Skoog (MS) and woody plant medium (WPM) and tested for their influences. Differences in callus initiation and morphology were noticed between MS and WPM, and among PGR concentration treatments.


HortScience ◽  
2015 ◽  
Vol 50 (3) ◽  
pp. 374-382 ◽  
Author(s):  
Piyada Alisha Tantasawat ◽  
Atitaya Sorntip ◽  
Paniti Pornbungkerd

The effects of exogenous application of plant growth regulators (PGRs) were evaluated on both field performance and in vitro gynogenesis of ‘Chai Lai’ and ‘Big C’ cucumber (Cucumis sativus L.). Plants were sprayed with two concentrations of N6-furfuryladenine [kinetin (KIN); 2 and 20 ppm], 2,3,5-triiodobenzoic acid (TIBA; 1 and 10 ppm), naphthaleneacetic acid (NAA; 10 and 100 ppm), abscisic acid (ABA; 2 and 20 ppm), thidiazuron (TDZ; 1 and 10 ppm), and maleic hydrazide (MH; 10 and 100 ppm) to assess their effects on vegetative growth and floral and yield related traits in the Winter of 2013 and in the Summer of 2014 compared with distilled water control. Meanwhile, the effects of two PGRs (KIN and TIBA) on cucumber gynogenesis were also investigated in vitro. Growth parameters and floral and yield-related traits were significantly affected by the various PGRs in both cultivars during both seasons. In both cultivars, the highest yield was obtained with the application of 10 ppm NAA during the Winter of 2013 (1.5- to 1.8-fold over control) and with 1 ppm TIBA during the Summer of 2014 (2.1- to 2.2-fold over control). With regard to the ovary culture response, exogenous application of KIN and TIBA on floral buds tended to enhance callus formation in ‘Chai Lai’ cultured on I7 medium, whereas no effect was observed in ‘Big C’. The embryo-like structure (ELS) formation efficiencies also tended to increase with 2 and 20 ppm KIN and 1 ppm TIBA application in ‘Chai Lai’ and with 20 ppm KIN and 1 ppm TIBA application in ‘Big C’ when cultured on I7 medium. Nevertheless, TIBA at high concentrations (10 ppm) decreased the percentages of ELS formation and the number of ELSs/piece in both cultivars. These results suggest that the polar auxin transport may play a major role on growth, floral and yield-related traits, yield as well as in vitro gynogenesis in cucumber. However, the success of exogenous applications of these PGRs depended on several factors including plant genotypes, growing seasons, types and concentrations of PGRs, and for ovary culture, the responses also varied according to the induction media used. Chemical names: abscisic acid (ABA); maleic hydrazide (MH); naphthaleneacetic acid (NAA); N6-furfuryladenine (kinetin; KIN); thidiazuron (TDZ); 2,3,5-triiodobenzoic acid (TIBA).


2020 ◽  
Vol 33 (3) ◽  
pp. 660-670
Author(s):  
VANESSA FERNANDES FONSECA WELZ ◽  
JÉSSICA REZENDE TRETTEL ◽  
ANDRESSA BEZERRA NASCIMENTO ◽  
HÉLIDA MARA MAGALHÃES

ABSTRACT Sweet basil is a perennial herb. Studies on in vitro cultivation of these plant species are scarce and inconclusive. This study was carried out to investigate the effect of culture medium concentration in combination with antioxidants and plant growth regulators on the in vitro growth and biochemical activity of sweet basil seedlings. Seeds of the ‘Genovese’ cultivar were inoculated into Murashige and Skoog culture medium supplemented with activated carbon and plant growth regulators 6 -benzylaminopurine and a-naphthaleneacetic acid. The seedlings were grown under controlled conditions for 80 days and their biometric and biochemical characteristics evaluated. More abnormal seedlings were observed in the 100% medium with 30 g L-1 sucrose, 0.4 g L-1 6-benzylaminopurine, and 0.2 g L-1 a-naphthaleneacetic acid (T4) and the medium without regulators (T1). However, the T4 culture medium resulted in a higher leaf number and shoot dry mass. Antioxidant activity was higher in the seedlings grown in the culture medium composed of 100% medium + 3.0 g L-1 activated carbon + 0.4 mg L-1 6-benzylaminopurine + 0.2 mg L-1 a-naphthaleneacetic acid (T5) and that composed of 70% medium + 3.0 g L-1 activated carbon + 0.1 mg L-1 6-benzylaminopurine (T3). The enzyme superoxide dismutase showed higher activity in all culture media than catalase or ascorbate peroxidase. Sweet basil seedlings growing in T4 and T1 medium showed the highest growth rate of shoots and the lowest antioxidant activity, whereas seedlings grown in T3 medium had the highest catalase and ascorbate peroxidase activity.


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