Acadian Marine Plant Extract Powder Research Articles

Effects of biostimulants, AMPEP and Kelpak on the growth and asexual reproduction of Pyropia yezoensis (Bangiales, Rhodophyta) at different temperatures

Acadian marine plant extract powder (AMPEP) and Kelpak are commercial biostimulants derived from brown algae Ascophyllum nodosum. This study was to determine if AMPEP and Kelpak can induce thermal resistance in Pyropia yezoensis. P. yezoensis blades were exposed to different concentrations (control: 0, low: 0.001, high: 1 ppm) of AMPEP and Kelpak at 10°C for 6 and 7 days, respectively. Those blades were then cultivated in von Stosch enriched seawater medium at different temperatures (10, 15, 20, and 25°C) with 12 : 12 L : D photoperiod and 100 μmol m-2 s-1 of photosynthetically active radiation for additional 15 days. Results showed that P. yezoensis reproduced archeospores at 20 and 25°C at all biostimulant conditions within 15 days. At lower temperatures (10 and 15°C), only AMPEP-treated P. yezoensis reproduced archeospores. P. yezoensis exposed to 1 ppm Kelpak exhibited higher phycoerythrin and phycocyanin contents than control and 0.001 ppm conditions at 15°C. AMPEP-treated conditions showed higher phycoerythrin and phycocyanin contents than control at 10°C. These results suggest that AMPEP and Kelpak may not enhance the thermal resistance of P. yezoensis. However, AMPEP stimulated archeospores release at lower temperatures. The treatment of AMPEP and Kelpak also increased the pigment contents in P. yezoensis. These results suggest that the use of seaweed-derived biostimulants can provide some economic benefits in P. yezoensis aquaculture. The enhancement of archeospores formation by AMPEP at lower temperature may also increase the productivity since Pyropia farming relies on the accumulation of secondary seedings via asexual reproduction.

Effect of Acadian Marine Plant Extract Powder (AMPEP) on the growth of the Seaweed Kappaphycus alvarezii “Kulisusu“ seedlings cultivated in encircling tanks

Acadian Marine Plant Extract Powder (AMPEP) is a commercial organic fertilizer extract from brown seaweed, Ascophyllum nodosum. This study aimed to examine the effect of different concentrations of AMPEP extract on K. alvarezii “Kulisusu” seedlings. We investigated the daily growth rate (DGR), ratio of wet weight and dry weight (FW: DW ratio), and the survival rate of seedlings in encircling tanks. Concentrations of AMPEP used in this study were 0.2, 0.4, 0.6, 0.8, 1.0 g L-1 at 30 min immersion time and without AMPEP as control for a 1-month cultivation period. The results showed that the 0.2 g L-1 of AMPEP showed higher DGR in 6-18 days of culture period (3.43-4.25%/day), the 0.6 g L-1 was higher in the 24 days culture period (3.50%/day), and the 0.8 g L-1 concentration was higher in the 30 days of culture period (3.76%/day). For FW: DW ratio, 0.2 g L-1 of AMPEP concentration showed the highest ratio (8.73:1) and was significantly different from the other treatments (p<0.05). In addition, all treatments showed a high survival rate (>93.33%) with minor variation amongst the treatments (p>0.05). The results concluded that using 0.2 g L-1 of AMPEP in encircling tanks was suitable for seedling development.

Efeito do extrato de algas no desempenho germinativo e crescimento radicular em sementes de feijão BRS Estilo em resposta a diferentes métodos de aplicação

RESUMO The biostimulating potential of algae extracts represents an opportunity to increase efficiency in food production. Cultivable microalgae are viable candidates due to the high production capacity. In this work, the chemical composition and effect of the microalgae extract Scenedesmus acuminatus (Lagerh.) Chodat and commercial product AMPEP (Acadian Marine Plant Extract Powder) derived from the macroalgae Ascophyllum nodosum (L.) Le Jol. on the germination of BRS Estilo bean seeds were evaluated. The treatments analyzed the methodology of exposure of seeds to extracts: soaking for 5 hours (T1) and applications every 48 hours (T2). The parameters evaluated were: germination percentage (PG), speed index (IVG) and mean germination time (TMG). The values of PG and IVG were higher in T2, representing about 50% higher yield than T1. In relation to chemical composition, Scenedesmus acuminatus extract obtained higher levels of proteins and carbohydrates. Thus, S. acuminatus is a potential producer of biostimulant extracts, while beans obtained lower germinative performance when submitted to the soaking process.

A review of multiple biostimulant and bioeffector benefits of AMPEP, an extract of the brown alga Ascophyllum nodosum, as applied to the enhanced cultivation and micropropagation of the commercially important red algal carrageenophyte Kappaphycus alvarezii and its selected cultivars

Commercially manufactured extracts of a variety of seaweeds have been used extensively for several decades for the relief of abiotic and biotic stresses for (terrestrial) agricultural crops and horticulture, to improve yield and quality. However, the use of seaweed extracts for their beneficial properties, as applied to marine macroalgae, only began in the mid to late 2000s. Kappaphycus alvarezii is an important red seaweed on the coasts of tropical to sub-tropical waters, mainly because of the various applications of kappa carrageenan, which is the major industrial colloid extracted from this extensively cultivated biomass. Cultivation of this seaweed has brought economic benefits to tens of thousands of seaweed farmers in Southeast Asia and other minor producing countries. Recently, Ascophyllum (aka. Acadian) Marine Plant Extract Powder (AMPEP), a commercial seaweed extract from the brown intertidal, macroalga A. nodosum was used in steps taken during the micropropagation and field cultivation of K. alvarezii. The reasons for utilizing this treatment included addressing the current problems facing the industry, such as decreased productivity, loss of vigor and diminished crop quality. This was brought about by shortages in the availability of good quality propagules (seedlings) and also disease and endo-epiphyte infestations, which affected the ability to grow and harvest saleable biomass, thus decreasing the income, and therefore the interest and participation of the potential seaweed farmers (based on the factor of repetitive, “drudge” labor and their derived income per unit effort). This paper reviews studies on Kappaphycus including the use of AMPEP specifically and also some alternative extracts to mitigate both biotic and abiotic stressors, using examples from micropropagation, field cultivation, endophyte mitigation and impacts on the resulting carrageenan qualities. Taken together, this body of evidence provides proof of concept and very promising results which may lead to further studies more specifically to identify the modes of action and the metabolic pathways by which the complex AMPEP extract might improve stress tolerances in K. alvarezii in order to obtain higher productivity and enhanced quality characteristics (i.e., exposure to increasing surface seawater temperature, salinity fluctuations and photo-inhibitory irradiance as well as attacks by pathogenic and opportunistic organisms).

The comparative efficiency of a brown algal-derived biostimulant extract (AMPEP), with and without supplemented PGRs: the induction of direct, axis shoots as applied to the propagation of vegetative seedlings for the successful mass cultivation of three commercial strains of Kappaphycus in Sabah, Malaysia

Three strains of Kappaphycus spp. (viz. K. alvarezii tambalang brown and green and K. striatus sacol green) were used in the present study to optimize the use of Ascophyllum (Acadian) marine plant extract powder (AMPEP) as a culture medium ingredient acting as a biostimulant, applied with, and without, the addition of terrestrial plant growth regulators (PGRs). This was undertaken in order to develop management tools and best practice recommendations for the mass production of new plantlets (seedlings) for industrial, nursery, and out-planting purposes in eastern Malaysia, Sabah, and Peninsular Malaysia (i.e., Langkawi, Kedah and Batu Maung, Penang). After 45 days of laboratory incubation, the three strains tested demonstrated their best performances at 3 mg L−1 of AMPEP, supplemented with PGR. This evaluation was based on the longest direct axes formed, the shortest time to their appearance, and also their highest percentage emergence. Kappaphycus alvarezii (tambalang green) had the longest direct axes (7.0 ± 0.23 mm), followed by K. alvarezii (tambalang brown) at (6.4 ± 0.48 mm) and finally K. striatus (sacol green). In terms of the highest percentage of direct axes formed, K. alvarezii (tambalang brown), K. alvarezii (tambalang green), and K. striatus (sacol green) were recorded as follows: 100 ± 0.00, 99 ± 1.34, and 98 ± 2.66, respectively. The shortest duration taken for the emergence of direct axes was observed in K. alvarezii (tambalang green) followed by tambalang brown and K. striatus (sacol green) on days 9, 10, and 15, respectively. The use of a brown seaweed-derived extract acting as a biostimulant and as the main ingredient of the culture medium for the micropropagation of three strains of Kappaphycus was highly encouraging and one which may be promoted as a protocol for the economic and commercial mass production of new plantlets (asexual seedlings) which are an urgent requirement for Malaysian seaweed farming to meet its full potential.

Direct formation of axes in new plantlets of Kappaphycus alvarezii (Doty) Doty, as influenced by the use of AMPEP K+, spindle inhibitors, and plant growth hormones

In order to induce the direct formation of erect dark brown vegetative shoots of Kappahycus alvarezii, under micro-propagation conditions, the agricultural bio-stimulant “Acadian Marine Plant Extract Powder” (or AMPEP) K+ (a Canadian commercial extract of the brown seaweed Ascophyllum nodosum with enhanced levels of potassium) was used singly or in combination with colchicine or oryzalin at different concentrations plus (synthetic) plant growth regulators (i.e., PGR, IAA, and kinetin) over 45 days of incubation. Amongst the treatments tested, 5 mg L−1 AMPEP K+ with PGR produced the longest direct axis shoots (9.6 ± 0.33 mm), followed by 0.1 mg L−1 AMPEP K+ + 1.0 mg L−1 oryzalin with PGR (8.7 ± 0.00 mm), 10 mg L−1 AMPEP K+ + 0.5 mg L−1 colchicine with PGR (8.6 ± 1.20 mm), and 0.5 mg AMPEP K+ only (5.3 ± 1.8 mm). The shortest, direct axis shoots measured were the following: 8.3 ± 0.33 mm with 0.1, 1 and 10 mg L−1 AMPEP K+ with PGR, 6.0 ± 0.58 mm with 0.5 mg L−1 AMPEP K+ + 0.1 mg L−1 oryzalin with PGR, 5.3 ± 0.89 mm with 0.5 mg L−1 AMPEP K+ + 0.1 mg L−1 colchicine with PGR, and finally, 2.7 ± 0.33 mm with 0.1 mg L−1 AMPEP K+ only. The percentage of direct shoots formed ranged from 93 to 100 % and 87 to 100 % in AMPEP K+ with PGR and AMPEP K+ only, respectively, while 62–100 % and 87–100 % were observed in AMPEP K+ + colchicine with PGR and AMPEP K+ + oryzalin with PGR, respectively. The use of AMPEP K+ with the PGR resulted in the longest and highest percentage formation of direct axis shoots. These observations, taken together, suggested the efficacy of AMPEP K+ used as a protocol to induce direct, erect shoots for the micro-propagation of K. alvarezii plantlets under laboratory conditions.

The effect of Ascophyllum nodosum (Ochrophyta) extract powder on the epibiosis of Kappaphycus alvarezii (Rhodophyta) commercially cultivated on floating rafts

Acadian marine plant extract powder (AMPEP), a product derived from Ascophyllum nodosum, has been widely used as a plant growth stimulant and recently has been tested on seaweeds with successful results on increasing growth and preventing epibiosis in Kappaphycus alvarezii cultivation. Although, the AMPEP ability to deal with epibiosis on seedlings cultivated in tubular nets and its commercial implications are still unknown. Between July 2012 and September 2013, three sampling periods were randomly selected at a Brazilian commercial farm in order to evaluate the effect of AMPEP on K. alvarezii daily growth rate, carrageenan yield and quality, and epibiont settling. Ten tubular nets containing three 100 g seedlings each (five treated with AMPEP and five not) were tied on floating rafts. Control samples displayed higher biomass of epibionts [0.76 ± 0.52 g (dry mass) tubular net−1] when compared to the AMPEP-treated samples [0.26 ± 0.27 g (dry mass) tubular net−1]. Carrageenan yield (44.65 ± 7.09 %) showed no significant difference between treatments (t = 0.46; p = 0.64), with larger percentages by the end of the third sampling period (F = 130.96, p < 0.001). When comparing AMPEP treatment with the control, carrageenan gel strength (252.14 ± 82.4 g cm−3) and viscosity (106.72 ± 45.11 cP) showed opposite results. The gel strength of AMPEP-treated samples was higher than their controls (t = 2.88; p = 0.007), while the gel viscosity of samples treated with AMPEP was lower than samples from their control (t = −2.4; p = 0.02). K. alvarezii daily growth rates between treatments were not different (t = 0.92; p = 0.36), exhibiting low commercial values probably due to herbivory. It was concluded that AMPEP was efficient against epibiont settlement on K. alvarezii cultivated on tubular nets tied on floating rafts maintaining favorable commercial results for carrageenan yield.

Performance of Red Seaweed (Kappaphycus sp.) Cultivated Using Tank Culture System

Little is known on the performance of seaweed cultivated in land-based facilities. The present study was conducted to determine the performance of Kappaphycus sp. cultivated in tank culture system with the effect of seaweed density, usage of Acadian Marine Plant Extract Powder (AMPEP), AMPEP fertilizer and the observation on the disease occurrence. Two red seaweed species (K. alvarezii variety Brown Tambalang, BT and K. striatum variety Green Flower, GF) were selected and mutually cultivated using hanging culture method. Three 40 days trials were conducted with different seaweed density (Trial 1:2.40, Trial 2:1.92 and Trial 3:0.96 g L−1). Besides, four treatments were performed in each trial: T1 (filtered seawater only), T2 (filtered seawater enriched with 13 mg L−1 AMPEP fertilizer), T3 (filtered seawater enriched with sands and corals and T4 (filtered seawater enriched with 13 mg L−1 AMPEP fertilizer, sands and corals). Based on the result, the maximum daily growth rate (2.00±0.03% day−1 for K. striatum and 1.46±0.06% day−1 for K. alvarezii) was recorded during Trial 3 with the lowest seaweed density using T4 treatment under natural culture parameters of light intensity, DO level, pH, temperature and salinity. In the present study, the performance of seaweed culture in tank was challenged by diseases such as “ice-ice� and seawater epiphyte infection that might cause bleached stems and weight loss to the seaweed. These findings are significant to provide a baseline data and facilitate the land-based seaweed farming in the future. © 2015 Academic Journals Inc.

Effect of the commercial extract of the brown alga Ascophyllum nodosum Mont. on Kappaphycus alvarezii (Doty) Doty ex P.C. Silva in situ submitted to lethal temperatures

Kappaphycus alvarezii is being introduced in several countries and in some of them there is a need to adapt this cultivation to periods with lethal temperatures, such as the 16–18 °C that occurs in the winter in southern Brazil. Moreover, there is the need to maintain the seedlings during this lethal temperature period. Considering the promising results obtained with the commercial powder extract of Ascophyllum nodosum (Acadian marine plant extract powder—AMPEP) treatment in the cultivation of K. alvarezii in vitro and in the sea allowing more resistance to epiphytes and increasing the growth rate and carrageenan yield, it was hypothesized that seedlings previously subjected to an AMPEP treatment could be more resilient to lethal temperatures. The daily growth rate and carrageenan yield and gel quality (gel strength and gel viscosity) of K. alvarezii in vitro previously treated with AMPEP were analyzed under temperature stress. The daily growth rates and the gel strengths of the AMPEP-treated samples were increased. In spite of the lower carrageenan yield and lower gel viscosity, the values were within the ones accepted by the carrageenan industry. Thus, the treatment of the seedlings of the K. alvarezii with AMPEP solution can be used as an alternative to lower temperature effects on crops as a preventive action for the cultivation of the seedlings in tanks and in the sea in periods of low temperatures at sea.

The ameliorating effect of Acadian marine plant extract against ionic liquids-induced oxidative stress and DNA damage in marine macroalga Ulva lactuca

Ionic liquids (ILs) are generally considered as the green replacement for conventional volatile organic solvents. Nonetheless, their high solubility in water with proven toxic effects on aquatic biota has questioned their green credentials. In the present study, the detoxification potential of Acadian marine plant extract powder (AMPEP) prepared from the brown alga Ascophyllum nodosum was investigated against the 1-alkyl-3-methylimidazolium bromide [C12mim]Br ionic liquid-induced toxicity and oxidative stress in marine macroalga Ulva lactuca. The IL ([C12mim]Br) at LC50 (70 μM) exposure triggered the generation of reactive oxygen species (ROS) such as O2·−, H2O2 and OH· causing membrane and DNA damage together with inhibition of antioxidant systems in the alga. The supplementation of AMPEP (150 μg mL−1) to the culture medium significantly reduced the accumulation of ROS and lipid peroxidation together with the inhibition of lipoxygenase (LOX) activity specially LOX-2 and LOX-3 isoforms. This is for the first time wherein comet assay was performed to ascertain the protective role of AMPEP against DNA damage in algal tissue grown in medium supplemented with IL and AMPEP. The AMPEP showed protective role against DNA damage (5–45 % tail DNA) when compared to those of grown in IL alone (45–70 % tail DNA). Further, specific isomorphs of different antioxidant enzymes such as superoxide dismutase (Mn-SOD-1, ~150 kDa), ascorbate peroxidase (APX-4, ~55 kDa), glutathione peroxidase (GSH-Px-2, ~55 kDa) and glutathione reductase (GR-1, ~180 kDa) responded specifically to AMPEP supplementation. It is evident from these findings that AMPEP could possibly be used for circumventing the negative effects arising from ILs-induced toxicity in marine ecosystem.

Investigation of the application of Acadian Marine Plant Extract Powder (AMPEP) to enhance the growth, phenolic content, free radical scavenging, and iron chelating activities of Kappaphycus Doty (Solieriaceae, Gigartinales, Rhodophyta)

An optimization study on concentration (viz. 0.01, 0.1, and 1.0 g L−1) and dipping time (i.e., 30 and 60 min) was conducted on three different color morphotypes (i.e., reddish brown, yellowish brown and purple) of the commercial carrageenophyte Kappaphycus alvarezii (Doty) Doty. The study tested the efficacy of Acadian Marine Plant Extract Powder (AMPEP) on the growth rate and occurrence of macro-epiphytes from August to November, representing the wet season of the Philippines. The optimum concentration and dipping time were obtained at 0.1 g L−1 and 30 min, respectively. These optimum parameters were then further verified in a commercial nursery using the yellowish brown morphotype. In another experiment, K. alvarezii (tambalang purple morphotype) and Kappaphycus striatum (Schmitz) Doty (sacol green morphotype) with, and without, AMPEP dippings were tested for their total phenolic content, free radical scavenging and iron chelating activities. Seaweed dipped in AMPEP demonstrated higher growth rates than the control. Lower concentrations (i.e., 0.01-0.1 g L−1) and shorter dipping time (e.g., 30 min) produced higher growth rates than the highest concentration (1.0 g L−1) and longer (60 min) dipping time. The presence of macro-epiphytes such as filamentous Ulva did not adversely affect the robust growth of the three color morphotypes of K. alvarezii. The lowest and highest growth rates obtained in a commercial seaweed nursery using the optimum concentration and dipping time of AMPEP were observed in July and January with 0.8% and 6.7% day−1, respectively. The antioxidant content of K. alvarezii (tambalang purple) and K. striatum (sacol green) responded differently to AMPEP dipping. The changes in total antioxidant activity followed almost the same trend as in phenolic content, in both K. alvarezii (tambalang purple) and K. striatum (sacol green), whereas, the iron chelating ability of both seaweeds with and without AMPEP dipping varied monthly. The results obtained for the use of AMPEP dips for commercial Kappaphycus cultivation demonstrated an effective management tool for improved farming protocols.

Extract powder from the brown alga Ascophyllum nodosum (Linnaeus) Le Jolis (AMPEP): a “vaccine-like” effect on Kappaphycus alvarezii (Doty) Doty ex P.C. Silva

The benefits of using Acadian Marine Plant Extract Powder (AMPEP), obtained from the brown algae Ascophyllum nodosum, for improving growth of the red alga Kappaphycus alvarezii has been demonstrated by authors in the Philippines and Brazil, particularly for increasing daily growth rate and mitigation of epiphytes. However, the processes which occur have not been discussed. This study examined in vitro the relationship between those red algal defense mechanisms and K. alvarezii responses using AMPEP treatments. The administration of the extract reduced the effects of the oxidative burst (production of hydrogen peroxide) which may be extremely aggressive for an individual and its epiphyte. The bleaching of the non-corticated portions of Polysiphonia subtilissima thalli that were cultivated as simulated epiphytes with AMPEP samples confirmed that the reaction was evident in which AMPEP protected K. alvarezii from the hydrogen peroxide effects. It is proposed that the use of the brown seaweed powder AMPEP acts as a potential vaccine, eliciting activation of the red seaweed K. alvarezii natural defenses against pathogens and ameliorating the negative effects of long-term exposure to oxidative bursts.

Impact of AMPEP on the growth and occurrence of epiphytic Neosiphonia infestation on two varieties of commercially cultivated Kappaphycus alvarezii grown at different depths in the Philippines

Two varieties of the carrageenophyte Kappaphycus alvarezii (Tungawan, TUNG; and Giant tambalang, GTAM) from Zamboanga Sibugay, Philippines were used to test the efficacy of Acadian Marine Plant Extract Powder (AMPEP) as source of nutrients for growth, and to determine if applications had any effect on the percent occurrence of an epiphytic infestation of the red alga Neosiphonia sp. at four different depths in the sea. Results showed that the use of AMPEP significantly (P < 0.05) increased the growth rate of both Kappaphycus varieties tested but decreased the percent occurrence of Neosiphonia sp. The percent occurrence of Neosiphonia sp. infection (6–50% at all depths) of both Kappaphycus varieties with AMPEP treatment was significantly lower than the controls (i.e., 10–75% at all depths). Both the growth rate of the cultivated seaweed and the percent occurrence of the epiphytes decreased as the cultivation depth increased. Plants dipped in AMPEP and suspended at the surface had the highest growth rates (i.e., 4.1%, TUNG; 3.1%, GTAM) after 45 days; those without AMPEP dipping had the highest percent occurrence of Neosiphonia infection (viz. 70–75%). The occurrence of Neosiphonia infestation was found to be correlated with changes in irradiance and salinity at the depths observed. The results suggested that both varieties of K. alvarezii used in this study have the fastest growth rate when grown immediately at the water surface. However, in order to minimize damage caused by the occurrence of epiphytic Neosiphonia, K. alvarezii should be grown within a depth range of 50–100 cm. These observations are important for the improved management of Kappaphycus for commercial farming. Furthermore, the use of AMPEP treatments for enhancement of growth and reduction deleterious Neosiphonia sp. infections is encouraging.

Optimization of culture conditions for tissue culture production of young plantlets of carrageenophyte Kappaphycus

To improve the production of Kappaphycus plantlets in tissue culture, optimum media concentrations of an Ascophyllum nodosum extract (Acadian Marine Plant Extract Powder, AMPEP), plant growth regulators (PGR), pH–temperature combinations, and explant density were determined. Kappaphycus alvarezii var. tambalang purple (PUR), kapilaran brown (KAP), vanguard brown (VAN), adik-adik (AA), tungawan green (TGR), and K. striatum var. sacol green (GS) were used as explants. Based on the shortest period for shoot emergence and the economical use of AMPEP, the optimum enriched media was 3.0 mg L−1 AMPEP and 0.1 mg L−1 AMPEP + PGR 1 mg L−1 each phenylacetic acid (PAA) and zeatin for PUR, 1.0 mg L−1 AMPEP + PGR for KAP and GS, 0.1 mg L−1 AMPEP + PGR for VAN, and 3.0 mg L−1 AMPEP and 0.001 mg L−1 AMPEP + PGR for AA and TGR. Results showed that the addition of PGR to low concentrations of AMPEP hastened shoot formation. pH–temperature combinations for the most rapid shoot formation were determined for the brown (KAP) and purple (PUR) color morphotypes of K. alvarezii var. tambalang and the green morphotype of K. striatum var. sacol (GS) cultured in 1.0 mg L−1 AMPEP + PGR. The brown morphotype produced the most number of shoots at pH 7.7 at 20°C after as little as 20 days. Purple K. alvarezii showed an increased shoot formation at pH 6.7 at 25°C and the green K. striatum morphotype at pH 8.7 at 25°C. The optimum number of explants added to the culture media was also determined for tungawan green (TGR), brown (KAP), and tambalang purple (PUR) varieties of K. alvarezii in 1.0 mg L−1 AMPEP + PGR. The number of explants and the volume of the culture media combination were also tested. The highest average number of shoots formed occurred in two explants:1 mL culture media (2:1) for KAP and PUR (35.00% and 16.67%, respectively) and 1 explant: 2 mL culture media for the TGR (100.00%) with a range of 0.5–3.0 mm shoot length after 40 days in culture. The earliest shoot formation was observed after 21 days for the brown and 9 days for both the green and purple color morphotypes of Kappaphycus, in all densities investigated. This indicated that within the range tested, the density of explants did not have a significant effect on the rate of shoot formation but did influence the average number generated from the culture. The rate of production of new and improved Kappaphycus explants for a commercial nursery stock was improved through the use of AMPEP with optimized culture media pH, temperature, and density conditions.

Use of Acadian marine plant extract powder from Ascophyllum nodosum in tissue culture of Kappaphycus varieties

Three varieties of Kappaphycus alvarezii (Kapilaran, KAP), Tambalang purple (PUR), Adik-adik (AA), and one variety of Kappaphycus striatum var. sacol (green sacol (GS) were used to determine the efficiency of Acadian marine plant extract powder (AMPEP) as a culture medium at different concentrations, for the regeneration of young plants of Kappaphycus varieties, using tissue culture techniques for the production of seed stock for nursery and outplanting purposes for the commercial cultivation of carrageenophytes. A shorter duration for shoot formation was observed when the explant was treated with AMPEP + Plant Growth Regulator (PGR = PAA + zeatin at 1 mg L−1) compared to AMPEP when used singly. However, four explants responded differently to the number of days required for shoot formation. The KAP variety took 46 days to form shoots at 3–4 mg L−1 AMPEP + PGR; while PUR required 21 days at 3–5 mg L−1 AMPEP and 3–4 mg L−1 AMPEP + PGR. AA required 17 days at 3–5 mg L−1 AMPEP and AMPEP + PGR; and GS 25 days at 1 mg L−1 AMPEP + PGR. It was observed that among the four explants used, PUR and AA initiated shoot formation with the use of AMPEP only at higher concentrations (3–5 mg L−1) after a shorter period. Only PUR responded positively to ESS/2 for shoot initiation. The use of AMPEP alone and/or in combination with PGR as a culture medium in the propagation of microplantlets using tissue culture technique is highly encouraging.