Optimization of culture conditions for tissue culture production of young plantlets of carrageenophyte <Emphasis Type="Italic">Kappaphycus</Emphasis>

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.     

Factors Influencing the Growth Rates of Three Commercial Eucheumoids at Coastal Sites in Southern Kenya

As a possible means of improving the livelihoods of local villagers, off-bottom rope cultivation of commercial eucheumoids was investigated on the southern Kenyan coast at three sites, representative of the variety of environments. The morphotypes used were brown Eucheuma denticulatum and green and brown Kappaphycus alvarezii. The study was carried out over a 15 month period from August 2001 until October 2002. Relative growth rates were highest at a sandy flat in a mangrove system (Gazi; 5.6% d⁻¹), and lowest in an intertidal reef flat (Kibuyuni; 3.2% d⁻¹) with a lagoon being intermediate (Mkwiro; 4.8% d⁻¹). The brown E. denticulatum had the highest growth rate of 4.7% d⁻¹ compared to the green and brown K. alvarezii which were 4.3% d⁻¹ and 4.2% d⁻¹, respectively. Growth was more variable at Kibuyuni and Mkwiro. The growth was higher during the southeast monsoon (4.7% d⁻¹) than during the northeast monsoon (4.0% d⁻¹). This is part of a larger study and the effects of water motion, salinity, temperature, thallus nitrogen, and ‘ice-ice’ syndrome on growth of morphotypes is discussed. The water motion was observed to increase thallus nitrogen and hence the growth of eucheumoids. The ‘ice-ice’ condition affected both brown E. denticulatum and brown K. alvarezii but not green K. alvarezii. The results suggest that commercial cultivation of eucheumoids in Kenya will be feasible.     

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.     

Growth and carrageenan quality of <Emphasis Type="Italic">Kappaphycus striatum</Emphasis> var. <Emphasis Type="Italic">sacol</Emphasis> grown at different stocking densities,duration of culture and depth

Kappaphycus striatum var. sacol was grown in two separate studies: (1) at two stocking densities, and (2) at four different depths, each for three different durations of culture (30, 45 and 60 days) in order to determine the growth rate of the seaweed and evaluate the carrageenan content and its molecular weight. The results demonstrated that stocking density, duration of culture and depth significantly (P < 0.01) affected the growth rate, carrageenan content and molecular weight of K. striatum var. sacol. Decreasing growth rate was observed at both stocking densities and at four depths as duration of culture increased. A lower stocking density (500 g m⁻¹line⁻¹) showed a higher growth rate for the shortest durations, i.e. 30 days, as compared to those grown at a higher density. Likewise, decreasing growth rate was observed as depth increased, except at 50 cm after 60 days of culture. A 45-day culture period produced the highest molecular weight at both stocking densities (500 g m⁻¹line⁻¹ = 1,079.5 ± 31.8 kDa, 1,000 g m⁻¹line⁻¹ = 1,167 ± 270.6 kDa). ‘Sacol’ grown for 30 days at 50 cm (1,178 kDa) to 100 cm (1,200 kDa) depth showed the highest values of molecular weight of carrageenan extracted. The results suggested that K. striatum var. sacol is best grown at a stocking density of 500 g m⁻¹line⁻¹, at a depth of 50–100 cm, and for a duration of 30 days in order to provide the highest growth rate, carrageenan content and molecular weight.     

Callus induction and micropropagation improved by colchicine and phytoregulators in Kappaphycus alvarezii (Rhodophyta, Solieriaceae)

Tissue culture techniques were applied for micropropagation of the red alga Kappaphycus alvarezii in order to select the best strain and experimental system for in vitro culture. Five strains were tested: brown (BR), green (GR) and red (RD) tetrasporophytes, brown female gametophyte (BFG), and a strain originating from tetraspore germination (“Edison de Paula”, EP). The effects of three culture media were tested on callus formation, regeneration from explants and from callus in the three tetrasporophytic and EP strains: seawater enriched with half-strength of von Stosch’s (VS 50) and Guillard & Ryther’s (F/2 50) solutions, plus synthetic ASP 12-NTA medium, with or without gelling agent. Explants of the EP strain were treated with glycerol and the phytoregulators indole-3-acetic acid (IAA); 2,4-diclorophenoxyacetic acid (2,4-D); and benzylaminopurine (BA), alone or in combination. The effects of colchicine (0.01%) during 24, 48, 72 hours and 14 days were analyzed in the BFG and EP strains. The EP strain showed the highest percentage of explants forming callus and regeneration from explants in VS 50, indicating its high potential for micropropagation in comparison to the other strains. Regeneration from callus was very rare. Treatments with glycerol and IAA:BA (5:1 mg L⁻¹) stimulated the regeneration from explants. Significant differences were observed in the percentages of regeneration of EP strain explants treated with colchicine for 14 days. Our results indicate that IAA and BA stimulated the regeneration process, and that colchicine produced explants with high potential for regeneration, being useful for improving the micropropagation of K. alvarezii.     

Distribution and symptoms of epiphyte infection in major carrageenophyte-producing farms

High density commercial farming of carrageenophyte Kappaphycus alvarezii is often plagued with “ice-ice” disease and epiphyte infection, which eventually leads to reduced production and in some cases collapse of crop. Epiphyte outbreak has been occurring regularly in major carrageenophyte farms in the Philippines, Indonesia, Malaysia and Tanzania. Infected materials from these countries were studied to establish baseline information on the epiphyte’s identity, density, symptoms and secondary infection on the host seaweed. The causative organism was identified as Neosiphonia apiculata (Hollenberg) Masuda et Kogame, based on its morphological features. Epiphyte density on host seaweed materials decreased in the following order: the Philippines (88.5 epi cm⁻²), Tanzania (69.0 epi cm⁻²), Indonesia (56.5 epi cm⁻²) and Malaysia (42.0 epi cm⁻²). Initial symptoms were the presence of tiny black spots, indicating the embedded tetrasporeling in seaweed cortex layer. Vegetative form emerged after 2 weeks measuring less than 0.5 mm in length with a density of less than 25.0 epi cm⁻². Upon maturation, infected seaweed takes on a “hairy” appearance with “goose-bumps” like cortical swellings. The epiphyte appears as a solitary plant with multiple secondary rhizoids or as multiple epiphytes appearing from a single cortical opening. At the end of infection, the epiphytes left dark pits on the cortical swelling, and the carrageenophytes are infected by opportunistic bacteria. Bacterial enumeration of healthy and infected seaweed materials showed an increase of more than 300% in total bacterial count on infected materials dominated by Alteromonas sp., Flavobacterium sp. and Vibrio sp.     

Use of plant growth regulators in micropropagation of Kappaphycus alvarezii (Doty) in airlift bioreactors

The effects of plant growth regulators on callus induction rate and regeneration of K. alvarezii explants was evaluated. K. alvarezii calluses were induced in vitro with kinetin (K), 6-benzylaminopurine (B), 1-naphtalene acetic acid (N) and spermine (S). After 30 days, K. alvarezii explants produced filamentous calluses and isolated crystalline filaments growing from the medullar region and from cortical cells at the cut edge. The plant growth regulators 1-naphtalene acetic acid (1 mg L⁻¹) and 6-benzylaminopurine (1 mg L⁻¹) and the 1-naphtalene acetic acid + kinetin + spermine (1, 1, 0.018 mg L⁻¹ respectively) combination produced 85 to 129% more calluses, with significant differences versus the control (p<0.05). Spermine at 0.018 mg L⁻¹ produced calluses in the apical, intercalary and basal regions of explants. Spermine also reduced callus induction time to 7 days, which is faster than previously reported induction times with other plant growth regulators. An airlift bioreactor was designed and characterized to micropropagate K. alvarezii calluses. The bioreactor had mixing times ranging from 4.6–10.3 s at T ₉₀ and T ₉₅, which is shorter than those for the Fernbach (5.2–13.4 s) and balloon flasks (6.3–17.3 s). Mixing time standard deviations were smaller for the bioreactor (1.1–4.6) than for the Fernbach (9.3–13.6) and balloon flasks (5.5–15.8), suggesting an adequate flow regime within the bioreactor. The results are useful for improving callus induction in K. alvarezii and propagating microplantlets in an airlift bioreactor, and provide baseline data for macroalgal bioreactor culture.     

Use of lysozyme for treatment of bacterial contamination in <Emphasis Type="Italic">in vitro</Emphasis> shoot cultures of fruit plants

Summary Use of lysozyme was tested for treatment of bacterial contaminations in in vitro shoot cultures of quince (Cydonia oblonga) ‘BA 29’ and the hybrid (Prunus persica × P. amygdalus) rootstock ‘GF 677’. Shoots which had been contaminated for about 1 yr by Bacillus circulans and Sphingomonas paucimobilis were treated in liquid culture, at pH 4.5, with 9–36 mg ml⁻¹ egg white lysozyme (EWL), and compared to each other and to untreated cultures for their growth, proliferation, and number of bacterial colony-forming units in the tissues. EWL did not negatively affect shoot growth up to 18 mg ml⁻¹; furthermore, the proliferation rates of EWL-treated shoots were sometimes higher than those of controls. In contrast, the concentration of 36 mg ml⁻¹ had some deleterious effect on the regrowth capacity and shoot production of ‘GF 677’ at the first subculture to solid medium after EWL, treatments. EWL had a simple bacteriostatic effect against Sphingomonas paucimobilis; in contrast, it was effective at 18 mg ml⁻¹ in eliminating Bacillus circulans in both ‘BA 29’ and ‘GF 677’ cultures, after optimal treatment duration.     

Occurrence of Polysiphonia Epiphytes in Kappaphycus Farms at Calaguas Is., Camarines Norte, Phillippines

This paper describes the occurrence of an epiphyte infestation of Kappaphycus farms in Calaguas Is. Camarines Norte, Philippines. In particular, percentage cover of ‘goose bump’-Polysiphonia and ‘ice-ice’ disease, and some environmental parameters that influence the thallus condition of Kappaphycus alvarezii in Calaguas Is. were assessed during 3 separate visits and are discussed.Commercial cultivation of Kappaphycus at Calaguas Is. began in the early 1990s. After five years of farming, the stock was destroyed by a strong typhoon. The area was re-planted the following year and production increased annually and reached its peak in 1998–1999. However, the following year, the first occurrence of a Polysiphonia epiphyte infestation occurred concurrently with an ‘ice-ice’ disease. Consequently, annual production and the number of seaweed planters declined rapidly, and this situation persists to the present time. This paper highlights the etiological factors and their consequences.Results show that farm-site selection is critical for the success of Kappaphycus production. Characteristics of water movement and light intensity in farming areas contributed to the occurrence and detrimental effect of the phenomenon described as ‘goose bumps’: a morphological distortion of the host seaweed due to the presence of a Polysiphonia sp. epiphyte. A strong inverse correlation was observed between the occurrence of Polysiphonia and water movement: areas with low water motion registered a higher % cover (65%) of Polysiphonia than those in more exposed areas (17%). Although ‘goose bump’-Polysiphonia infestation and ‘ice-ice’ disease pose a tremendous problem to the seaweed farmers, the results of this limited assessment provide a useful baseline for future work.     

Chemical and physical-chemical characteristics of dietary fibres from Ulva lactuca (L.) Thuret and Enteromorpha compressa (L.) Grev.

Dietary fibres from Ulva lactuca (L.) Thuret (sea lettuce) and Enteromorpha compressa (L.) Grev. (A.O. nori) were measured according to a ‘standard’ method and a ‘physiological’ protocol simulating the gastric and intestinal environments. U. lactuca contained 15.8–8.0% soluble and 24.2–32.6% insoluble fibres according to the ‘standard’ and ‘physiological’ methods, respectively. For E. compressa, these values were 14.9–15.9 and 21.6–28.7%, respectively. For both algae, the composition suggests that the soluble fibres were xylorhamnoglycuronans sulphates and insoluble fibres were essentially composed of glucans. No marked chemical compositional variation was observed between soluble fractions extracted under the simulated gastric and intestinal conditions. Fibres in both algae are hydrophilic but the water holding capacities were higher after extraction of soluble fibres (5.5–9.5 g g⁻¹ for the dry algae; 14.0–16.0 g g⁻¹ for the standard insoluble fibres). Water soluble fibres demonstrated low intrinsic viscosities at 37 °C in buffers, particularly those from E. compressa (36.0–36.5 ml g⁻¹), and was affected by pH for those of U. lactuca (147.5 ml g⁻¹ at pH 3.0 and 175.0 ml g⁻¹ at pH 7.3).     

Cage culture ofKappaphycus alvarezii var.tambalang (Gigartinales,Rhodophyceae)

Kappaphycus alvarezii var.tambalang was cultured in a 3 × 3 m bamboo raft installed inside a 4 × 4 m floating net cage ofLates calcarifer (sea bass) broodstock at SEAFDEC Sub-station Igang, Guimaras, Philippines, from December 1989 to May 1990. Growth and production ofK. alvarezii var.tambalang was influenced by the culture months. The highest growth rate and production were recorded in January and May, respectively, while the lowest growth rate and production were observed in March. Average growth rate (wet weight) ranged from 3.72 to 7.17% day^-1, while production ranged from 575.5 to 2377 g m^-1 line^-1. A total production of approximately 123 t (fresh) or 37 t (dried) ha^-1 in the 5-month harvest season was produced from this culture system. Cultivation ofK. alvarezii var.tambalang in cages is possible, which indicates that seaweeds can be grown with carnivorous finfish, a practice which is still untapped.     

Economics of cultivatingKappaphycus alvarezii using the fixed-bottom line and hanging-long line methods in Panagatan Cays,Caluya, Antique,Philippines

A socio-economic survey was conducted among theKappaphycus alvarezii planters of Panagatan Cay, Caluya, Antique, Philippines to determine some social information, farming practices and cost and returns of farming the seaweed. Cultivation is dominated by brown and green morphotypes using the fixed-bottom and hanging-long line methods. Approximately 9.3 t d. wt ha^–1 and 7.2 t d. wt ha^–1 is produced from fixed-bottom and hanging-long lines methods, respectively, after 60–90 days of culture. The former method requires a working capital and total investment of P7490 and P1870, respectively, compared to the hanging-long line which requires P8455 and P25464, respectively (US$ 1 = P26). A higher total revenue (P139500), net income ((P187895), and return of investment 1002%), but a shorter pay back period (0.10 years) were obtained in fixed-bottom than in hanging-long line. A lower total expenses were incurred in fixed-bottom (P21354) than in hanging-long line (P24566). The farming ofK. alvarezii in this area has brought tremendous economic impact to the marginal fishermen.Author for correspondence     

Effects of salinity on the growth rate,carrageenan yield,and cellular structure of <Emphasis Type="Italic">Kappaphycus alvarezii</Emphasis> (Rhodophyta,Gigartinales) cultured in vitro

Kappaphycus alvarezii was cultured in vitro under salinities ranging from 15 to 55 psu for 35 days to determine the differential effect on growth rate, carrageenan yield, and cellular structure. Plants kept in 15 psu died after 3 days, while plants cultured in 55 psu presented low growth rates during the entire experimental period (0.28% day⁻¹). Plants cultured in 25, 35, and 45 psu showed growth rates normally associated with this species (between 3% and 4% day⁻¹) and similar cellular morphology. Carrageenan yield was significantly higher in plants cultured in 25 psu in relation to the other treatments. As observed by light microscopy, plants cultured in 15 psu showed cellular turgidity and increased cell wall thickness, both consequences of hyposalinity. Chloroplasts and other membranous organelles underwent rupture and considerable disorganization in ultrastructure. Although branches from the 55 psu samples showed plasmolysis, cells were able to maintain chloroplast integrity, despite their rudimentary features. In high salinities, great concentrations of floridean starch grains were observed in subcortical cells, indicating their probable participation in osmoregulation. Based on these results, we defined the range of 25 to 45 psu as the limits of saline tolerance for K. alvarezii. While new field studies are required to confirm these results, it can be concluded that new sites, such as inactive or abandoned shrimp tanks with salinities up to 25 psu, could be considered for commercial farming.     

The effects of selected cultivation conditions on the carrageenan characteristics of <Emphasis Type="Italic">Kappaphycus alvarezii</Emphasis> (Rhodophyta,Solieriaceae) in Ubatuba Bay,São Paulo,Brazil

The yield and the quality of carrageenan depend, among other things, on the cultivar or strain and on the cultivation and processing techniques. This work presents carrageenan yields and some properties of Kappaphycus alvarezii under selected cultivation conditions i.e. cultivation period, depth and planting density. Growth rates (GR) ranged from 5.2–7.2% day⁻¹, with the highest GR at 28 days, at 0–0.5 m depth, and planting density of 12 and 8.4 plants m⁻². Highest productivity was observed in samples after 44 and 59 day cultivation period, which were grown at 0–0.5 m depth, and a planting density of 24 plants m⁻². Carrageenan yields, iota fraction, viscosity, molecular weight and gel strength were measured. A cultivation period of 28 days during the winter had a significant higher carrageenan yield, while samples from 59 days showed a significantly higher iota fraction. Carrageenan also presented an increasing molecular weight under longer cultivation periods. A similar trend was observed for viscosity and gel strength. All samples cultivated in Brazil gave higher values when compared to a K. alvarezii commercial reference sample, with the exception of carrageenan yield values, which were lower in this study. Taking into account all parameters, the culture condition which provided the best carrageenan from a commercial perspective were 45 days of cultivation, growing at the surface, with a planting density of 12 plants m⁻². Considering that this study was performed in the least favorable season (winter), these results indicate that the site is suitable for the implementation of commercial cultivation.     

Effects of carbon source and concentration on development of lingonberry (<Emphasis Type="Italic">Vaccinium vitis-idaea</Emphasis> L.) shoots cultivated <Emphasis Type="Italic">in vitro</Emphasis> from nodal explants

Summary Carbohydrate type and concentration and their interactive effects on in vitro shoot proliferation of three lingonberry (Vaccinium vitis-idaea ssp. vitis-idaea L.) cultivars (‘Regal’, ‘Splendor’, and ‘Erntedank’) and two V. vitis-idaea ssp. minus (Lodd) clones (‘NL1’ and ‘NL2’) were studied. Nodal explants were grown in vitro on medium with 2 μM zeatin and either glucose, sorbitol, or sucrose at a concentration of 0, 10, 20, or 30 gl⁻¹. The interactive effects of carbohydrate type and concentration and genotype were important for shoot proliferation. The best response was afforded by sucrose at 20 gl⁻¹ both in terms of explant response and shoot developing potential, although glucose supported shoot growth equally well, and in ‘NL1’ at 10 gl⁻¹ it resulted in better in vitro growth than sucrose. Carbohydrate concentration had little effect on shoot vigor. The genotypes differed in terms of shoots per explant, length, and vigor, leaves per shoot, and callus formation at the base of explants; this was manifested with various types and concentrations of carbohydrate. Changing the positioning of explants on the medium from vertically upright to horizontal increased the shoot and callus size, but decreased shoot height and leaves per shoot. Proliferated shoots were rooted on a peat:perlite (1∶1, v/v) medium and the plantlets were acclimatized and eventually established in the greenhouse.     

Seasonal changes in growth rate, carrageenan yield and lectin content in the red alga Kappaphycus alvarezii cultivated in Camranh Bay, Vietnam

The three color morphotypes of the red alga Kappaphycus alvarezii (brown, red and green) were cultured in Camranh Bay, Vietnam, using the fixed off-bottom monoline culture method to evaluate the growth rate, carrageenan yield, 3,6-anhydrogalactose, gel strength and lectin content. The brown morphotype was cultivated over a 12-month period; the red and green morphotypes were over a 6-month period. At the 60-day culture timepoint, the brown morphotype showed a higher growth rate (3.5–4.6% day⁻¹) from September to February, and lower growth rate (1.6–2.8% day⁻¹) from March to August. Significant (P < 0.05) differences in growth rate between culture months were found with the brown morphotype. High growth rates for the red (3.6–4.4% day⁻¹) and green (3.7–4.2% day⁻¹) morphotypes were obtained from September to February. The carrageenan yield, 3,6-anhydrogalactose and gel strength of the three morphotypes showed little variation, with the highest values obtained in November–December. At the 30-day sampling point, the brown morphotype had a higher lectin content (167–302 μg g⁻¹ dry alga) from August to March and a lower lectin content (23–104 μg g⁻¹ dry alga) from April to July. High lectin contents were recorded for the red (139–338 μg g⁻¹ dry alga) and green (124–259 μg g⁻¹ dry alga) morphotypes from September to February. This study shows that the different morphotypes of K. alvarezii can be grown in the tropical waters of the Camranh during the northeast monsoon, and part of the southwest monsoon, especially the brown morphotype, which can be grown during any season.     

Highly efficient embryogenesis and plant regeneration of tall fescue (Festuca arundinacea Schreb.) from mature seed-derived calli

Summary We report a protocol for efficient plant regeneration of four tall fescue (Festuca arundinacea Schreb.) cultivars (‘Surpro’, ‘Coronado’, ‘Summer Lawn’, and ‘Fawn’) via somatic embryogenesis. Calli were initiated from mature seeds grown on modified Murashige and Skoog (MMS) medium supplemented with 7.0mgl⁻¹ (31.7μM) 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.05 mgl⁻¹ (0.23 μM) kinetin (Kin). Calli were maintained and proliferated by subculture at monthly intervals on MMS medium containing 4.5 mgl⁻¹ (20.4 μM) 2,4-D and 0.2mgl⁻¹ (0.9 μM) Kin. Somatic embryos (SE) were induced from seed-derived calli on SE-induction medium (MMS supplemented with 2.0 mgl⁻¹ 2,4-D and 0.2mgl⁻¹ Kin). Plantlets were regenerated from somatic embryogenic calli grown on modified SH medium supplemented with 2 mgl⁻¹ Kin. Using this optimized protocol, 78.6–82.3% of mature seeds of all four cultivars produced SE clusters, of which 93.5–95.3% regenerated into plants within 10 wk. The regenerants showed no phenotypic abnormalities.     

Iota-carrageenan from a newly farmed, rare variety of eucheumoid seaweed—“endong”

A seaweed with the local name “endong” is a rare eucheumoid variety that has recently been farmed in Tawi-Tawi, southern Philippines. Local farmers describe it as “eel-like” because of its long, slender and smooth thalli. Being somewhat similar in appearance to the dominantly farmed seaweed “tambalang” (Kappaphycus alvarezii var. tambalangii), farmers mix their harvests of “endong” with those of “tambalang”. We analyzed the hydrocolloid extract from “endong” using spectroscopic and chemical techniques. The extract’s infrared and nuclear magnetic resonance (¹H and ¹³C) spectra, and sulfate and 3,6-anhydrogalactose contents revealed carrageenan of the iota-type, similar to extracts from another farmed species, Eucheuma denticulatum. This result implies that “endong” carrageenan exhibits physico-chemical properties different from those of kappa-carrageenan of “tambalang”. The findings of this study are of significance to the seaweed industry, as carrageenan quality problems could potentially arise from a mixed harvest. Seaweed farmers are advised to separate their harvests of “endong” and “tambalang”. Presented at the 19th International Seaweed Symposium, Kobe, Japan     

Heavy metal chelation by non-living biomass of three color forms of Kappaphycus alvarezii (Doty) Doty

Water pollution by toxic heavy metals is a burning environmental problem and has presented a challenge to humans. Removal of heavy metals using non-living biomass of seaweeds could be a potential solution to this problem. In the present investigation, biomass of three color forms of Kappaphycus alvarezii, viz. brown, green and pale yellow, were studied in the laboratory for their heavy metal chelating capacity using cadmium, cobalt, chromium and copper. Amongst the four concentrations used (25, 50, 75 and 100 mg L⁻¹) maximum chelation of Cd, Co and Cu was recorded at 25 mg L⁻¹ concentration. The highest amount of Cr was chelated at 100 mg L⁻¹ by all the three color forms. The pale yellow form showed maximum chelation for all four metals studied. Further, chelation in all the color forms was found to be: Cd 5.37 ± 0.59–15.84 ± 0.32 %, Co 21.19 ± 0.13–32.32 ± 0.62 %, Cr 65.38 ± 0.27–88.09 ± 0.51 % and Cu 59.53 ± 0.37–90.28 ± 0.89 %. All the three color forms of K. alvarezii serve as an excellent biodetoxifier as they all chelated considerable amounts of heavy metals.     

Efficient and simple plant regeneration via organogenesis from leaf segment cultures of persimmon (Diospyros kaki thunb.)

Summary An efficient and simple plant regeneration system via organogenesis from leaf segments of persimmon (Diospyros kaki Thunb.) cultivars ‘Fuyu’ and ‘Nishimurawase’ has been developed. The regeneration capacity was influenced by the culture vessels, gelling agents, plant growth regulators, and light conditions. Leaf explants taken from in vitro shoots were cultured on a modified Murashige and Skoog medium (MS1/2N), for 16 wk without transfer to fresh medium. Adventious shoots appeared after 4 and 8 wk in culture of ‘Nishimurawase’ and ‘Fuyu’ tissues, respectively. The culture of leaf explants in Erlenmeyer flasks with medium containing 4 g l⁻¹ agar enhanced shoot formation in comparison to media with increased agar concentrations. Optimal shoot regeneration was obtained with 5 mg l⁻¹ (22.8 μM) zeatin and 0.1 mg l⁻¹ (0.05 μM) indole-3-butyric acid (IBA) for ‘Nishimurawase’, and 10 mg l⁻¹ (45.6 μM) zeatin and 0.1 mg l⁻¹ (0.05 μM) IBA for ‘Fuyn’. Shoot regeneration frequencies in both cultivars were 100%, and shoot numbers per explant reached up to 9.2 for ‘Nishimurawase’ and 2.2 for ‘Fuyu’. Dark incubation during the first 4–5 wk was the most effective condition to successfully influence shoot regeneration in both cultivars. While dark incubation was essential for adventitious shoot formation by ‘Fuyu’, it was only slightly beneficial to ‘Nishimurawase’. More than 80% of the regenerated shoots rooted within 4 wk on hormone-free MS1/2N demium after having been dipped for 30 s in 250 mg l⁻¹ (1.1. mM) IBA solution.