Growth,photosynthesis and agar in wild-type strains of Gracilaria verrucosa and G. conferta (Gracilariales,Rhodophyta), as a strain selection experiment

The local species Gracilaria conferta and the foreign G. verrucosa were grown together under a wide range of photon flux density and temperature conditions. Gracilaria verrucosa showed a higher growth rate, especially under low temperatures, and higher photosynthetic performances as well as higher ribulose-1,5-bisphosphate carboxylase activity as compared with G. conferta. Gracilaria verrucosa also showed a better quality and yield of agar, suggesting that this species could be more suitable than G. conferta for outdoor cultivation in Israel and may improve winter growth in ponds. Growth rate and agar quality (gel strength) were rated as the most suitable characteristics influencing the preference of strains for outdoor cultivation.     

Seasonal growth activity of local and foreign gracilarioid strains in Israel

Six gracilarioid strains originating from different climatical environments were cultured in two cultivation systems: a short-term indoor one with a cross gradiant table, and a long-term outdoor one. Seasonal growth performances of the different strains were determined. The growth results in the two culture systems showed similar trends. The tropical species.Gracilaria cornea andG. cornea mutant, showed highest growth rates during summer and no growth at all during winter. The temperate species,Gracilaria verrucosa andGracilariopsis lemaneiformis, showed best growth performances during winter with small fluctuations between seasons. The subtropical speciesGracilaria conferta (local) showed seasonal growth fluctuations all over the year. The foreign species definitely did not acclimate under local conditions, but successfully preserved their original response to temperature. Regression equations confirmed that temperature was the dominant environmental variable in most of the gracilarioid strains. The growth rate results obtained showed encouraging prospects for high algal productivity as compared to other cultivation systems. Seasonal cultivation strategy ofGracilaria spp. in Israel is discussed.     

Farming of agarophytes in India—a long-time sustainability for the industry and preserving wild stocks

In India, food-grade agar is produced from Gracilaria edulis while bacteriological-grade agar is extracted from Gelidiella acerosa. Agarose is directly obtained from Gracilaria dura. Seaweeds for agar production mainly come from wild stocks located in reefs of the Gulf of Mannar, southeast coast. Landings of Gr. edulis were peak (982 dry t) during 1990 while landings of Ge. acerosa reached maximum (665 dry t) during 2002–2003. Overexploitation is leading to a decrease in biomass of these two algae at an alarming rate for the last decade. Commercial cultivation of agarophytes is yet to increase in India in spite of growing pressure on wild stocks triggered by increase in demand of raw material by the industry. Taking this into consideration, a number of efforts have been initiated to develop feasible cultivation methods for agarophytes. The present paper summarizes various methods developed for commercial cultivation of Gr. edulis, Gr. dura, Gracilaria verrucosa and Ge. acerosa in the open sea. We also discuss the challenges in the development of commercial farming of agarophytes in India.     

Recent approaches to the taxonomy of the Gracilariaceae (Gracilariales,Rhodophyta) and the Gracilaria verrucosa problem

Recognition of species in the Gracilariaceae, often notoriously difficult, is being aided by a combination of classical and modern techniques. We review some recent findings and present new results that may lead to redefinition of Gracilaria verrucosa, the type species of its genus. Plastid DNA restriction profiles (patterns of banding obtained by electrophoresis of DNA after restriction endonuclease digestion) from eleven strains ascribed to G. verrucosa indicated that the concept of this species in northern Europe includes possibly three taxa, one of which is known now to be a species of Gracilariopsis. In contrast, restriction profiles from Argentinian and Japanese strains were closely similar to the predominant pattern for European G. verrucosa. Profiles of several other strains, from the western Atlantic and eastern Pacific, were dissimilar to the European group and to each other. A chromosome number of n = 24 was determined for a representative of the predominant European group, and preliminary results of hybridization trials suggest that these strains, and others with approximately the same plastid DNA restriction pattern, are interfertile.NRCC 30559     

Effects of temperature and salinity on the growth of Gracilaria verrucosa and G. chorda, with the Potential for mariculture in Korea

Effects of temperature and salinity on the growth of the two agarophytes, Gracilaria verrucosa (Hudson) Papenfuss and Gracilaria chorda Holmes were examined in Korea. Both species grew over a wide range of temperatures (10–30 ^∘C) and salinities (5–35‰), and grew well at 17–30 ^∘C and a salinity of 15–30‰. In culture, G. verrucosa grew faster than G. chorda and their maximum growth rates were 4.95% day⁻¹ (30 ^∘C, 25‰) and 4.47% day⁻¹ (at 25 ^∘C, 25‰), respectively. In the field population the maximum growth and fertility of G. chorda were observed in summer. The growth rate of G. verrucosa was slightly higher than that of G. chorda for 2 weeks on the cultivation rope and in culture but it was much lower after being contaminated with epiphytes. The biomass of the epiphytes was 0.82 g dry wt. per host plant in G. verrucosa and 0.001 g in G. chorda. G. chorda exhibited resistance to epiphytism and grew 7 times in length and the dry weight increased 15 times after 55 days. In conclusion, G. chorda appears to be a good agarophyte with a fast growth rate and resistance to epiphytesm, and compared with G. verrucosa, has good potential for commercial cultivation.     

Forest and laboratory evaluations of hybridizedApanteles melanoscelus [Hym.: Braconidae], a parasitoid ofPorthetria dispar [Lep.: Lymantriidae]

Hybrids ofApanteles melanoscelus (Ratzeburg) were produced from colonies originating from France, Yugoslavia, and Connecticut. All strains, as well as freshly collected “wild” Connecticut parasitoids of the same species were evaluated in the laboratory for developmental rate, host attack rate, and sex ratios. Development was significantly slower in all the laboratory strains compared to the progeny of forest collected Connecticut females. Progeny production was greater (almost 2X) for the “wild” females and the French-Yugoslavian-Connecticut hybrid than for the laboratory Connecticut strain. The proportion of females collected from the “wild” (Connecticut) strain was higher than that observed in any laboratory strain. A field test was conducted using the triple hybrid in 3 release plots with ca. 6000A. melanoscelus cocoons released per plot in central Connecticut, U.S.A. Weekly collections of gypsy moth larvae showed that the % parasitism was significantly higher in release plots than in the 3 check plots. These results suggest the value of inundative releases ofA. melanoscelus for reduction of sparse gypsy moth populations, but they did not show that hybridization of these strains produced a more effective parasitoid under forest conditions.     

Mutagenic effects of heavy-ion beam irradiation on in vitro nodal segments of Artemisia annua L.

Artemisia annua L. is a commercial source of artemisinin. Nevertheless, artemisinin content within the plant is relatively low and varies depending on genotype and environment. To broaden the genetic variability, the mutation effect of ¹²C-ion beam irradiation on A. annua was examined. Irradiation at 2.5 Gy had a slight lethal effect to nodal segments while a noticeable lethal effect was observed at 5 and 10 Gy. Furthermore, at higher doses (20 and 50 Gy), a severe lethal effect was observed. Mutations at the DNA level of axillary bud-derived shoots were performed by RAPD. The mutation frequency at 10 Gy was about 1.7 and 2.1 times higher than that at 2.5 and 5 Gy, respectively. After growth and artemisinin production observation of 72 irradiated mutants, around 14 and 7 % of them showed higher artemisinin content and artemisinin yield compared to the controls, respectively. The highest artemisinin content in a mutant was 1.43 % DW, which was 3.2-fold higher than the original wild type. Additionally, the highest artemisinin yield in mutants was 3.68 mg/plant, which was around 1.4-fold higher than in the wild type. Moreover, irradiated mutants exhibited antibacterial activity against S. aureus, but the wild types did not. This study presents an effective application of heavy ion beam irradiation to create variations and improve artemisinin production in A. annua.     

The diversity of rhizobia nodulating the <Emphasis Type="Italic">Medicago</Emphasis>, <Emphasis Type="Italic">Melilotus</Emphasis> and <Emphasis Type="Italic">Trigonella</Emphasis> inoculation group in Egypt is marked by the dominance of two genetic types

Twenty four rhizobial strains were isolated from root nodules of Melilotus, Medicago and Trigonella plants growing wild in soils throughout Egypt. The nearly complete 16S rRNA gene sequence from each strain showed that 12 strains (50 %) were closely related to the Ensifer meliloti LMG6133^T type strain with identity values higher than 99.0 %, that 9 (37.5 %) strains were more than 99 % identical to the E. medicae WSM419^T type strain, and that 3 (12.5 %) strains showed 100 % identity with the type strain of N. huautlense S02^T. Accordingly, the diversity of rhizobial strains nodulating wild Melilotus, Medicago and Trigonella species in Egypt is marked by predominance of two genetic types, E. meliloti and E. medicae, although the frequency of isolation was slightly higher in E. meliloti. Sequencing of the symbiotic nodC gene from selected Medicago and Melilotus strains revealed that they were all similar to those of the E. meliloti LMG6133^T and E. medicae WSM419^T type strains, respectively. Similarly, nodC sequences of strains identified as members of the genus Neorhizobium were more than 99 % identical to that of N. galegae symbiovar officinalis HAMBI 114.     

Multiple factors control the level of resistance induced in tomato by Fusarium oxysporum f. sp. cucumerinum

Fourteen wild type and three UV-irradiated isolates of Fusarium oxysporum f. sp. cucumerinum (Foc) were evaluated as to the level of resistance they could induce in tomato to late blight caused by Phytophthora infestans. Tomato plants were induced by applying a suspension of Foc microconidia directly to the surface of the potting media without disturbing the tomato roots. Upper leaves of tomato plants were inoculated with P. infestans, and a reduction in lesion expansion was used as an index of induced resistance. All fourteen wild type isolates of Foc significantly reduced expansion of late blight lesions. One of the wild type isolates produced a significantly weaker resistance response than the other isolates. None of the UV-irradiated isolates induced significant resistance. The same Foc isolates were compared as to their virulence and their pigment production in culture, and considerable variation among them was revealed for both characteristics. Positive correlations existed both between the level of induced resistance and virulence, and between the level of induced resistance and pigmentation. The gradual increment in the level of induced resistance and the exceptions to the correlations between induced resistance and the two characteristics investigated suggest that multiple factors contribute to the induction of resistance by Foc.     

Evaluation of ethanol production and bioadsorption of heavy metals by various red seaweeds

The objective of this study was to evaluate ethanol production and bioadsorption with four red seaweeds, Gelidium amansii, Gracilaria verrucosa, Kappaphycus alvarezii and Eucheuma denticulatum. To produce ethanol, thermal acid hydrolysis, enzymatic saccharification and fermentation was carried out. After pretreatment, 38.5, 39.9, 31.0 and 27.5 g/L of monosaccharides were obtained from G. amansii, G. verrucosa, K. alvarezii and E. denticulatum, respectively. Ethanol fermentation was performed with Saccharomyces cerevisiae KCCM 1129 adapted to 80 g/L galactose. The ethanol productions by G. amansii, G. verrucosa, K. alvarezii and E. denticulatum were 18.8 g/L with Y _EtOH = 0.49, 19.1 g/L with Y _EtOH = 0.48, 14.5 g/L with Y _EtOH = 0.47 and 13.0 g/L with Y _EtOH = 0.47, respectively. The waste seaweed slurries after the ethanol fermentation were reused to adsorb Cd(II), Pb(II) and Cu(II). Using langmuir isotherm model, Cu(II) had the highest affinity for waste seaweeds with the highest q _max and electronegativity values among three heavy metals.     

Detection and characterization of phosphatidylcholine in various strains of the genus Chlamydomonas (Volvocales,Chlorophyceae)

The laboratory strains of Chlamydomonas reinhardtii have been reported to contain no phosphatidylcholine (PC), which is considered to be replaced by another zwitterionic lipid, diacylglyceryl-N,N,N-trimethylhomoserine (DGTS). According to the recently published classification, the strains belonged to the subgroup Reinhardtinia. Screening for PC in 13 selected strains of Chlamydomonas in the NIES Algal Collection, which are different in habitats and belong to different phylogenetic subgroups in the genus, revealed the presence of PC in four strains: a strain in the subgroup Polytominia, and three strains in Reinhardtinia. PC was not detected in three other strains of Reinhardtinia analyzed. The presence/absence of PC was not related to the phylogenetic relationship based on 18S rRNA. DGTS was detected in all the strains analyzed. The rare isomer of linolenic acid, 18:3(5,9,12), which has been found in the DGTS of C. reinhardtii, was found in the PC of the two strains and in the DGTS of the five strains. The occurrence of this fatty acid seems limited to a branch of Reinhardtinia. Acquisition and loss of PC in various strains of Chlamydomonas are discussed from the viewpoint of evolution of PC biosynthetic pathway.     

Changes in fruit shape inproboscidea parviftora ssp.parviflora (martyniaceae) with domestication

Aboriginal cultivators selected a phylogenetically novel fruit shape during the domestication ofProboscidea parviflora ssp.parviflora var.Hohokamiana. The extremely long rostrum of the domesticated var.Hohokamiana is unique for the Martyniaceae. Rostrum length and width are significantly correlated in the wild var.parviflora, but in var.Hohokamiana aboriginal cultivators apparently selected for a weaker correlation between these variables. In both varieties there is an allometric relationship between rostrum and “capsule” lengths, with the former increasing proportionately faster than the latter. The above allometric relationship and the decrease in correlation between rostrum length and width are advantageous to aboriginal cultivators because the proportionately longer and thinner rostra are better for basketmaking. Comparisons of the data presented herein with data from other plants suggest that similar growth factors have been responsible for changes in fruit shape resulting from domestication.     

Reinstatement of the genus Hymenaea (Leguminosae: Caesalpinioideae) in Africa

The monotypic genusTrachylobium [T. verrucosum (Gaertn.) Oliv.] from East Africa and adjacent islands is returned toHymenaea verrucosa Gaertn. on the basis of reevaluation of inflorescence, flower, and fruit characters together with new evidence from seedlings, pollen, leaf venation, chromosome number, and chemistry of the resins.Hymenaea verrucosa appears more similar to an Amazonian and Cuban group of species, which may have been derived from a common African ancestral stock in the Tertiary period, than to another group of species that probably evolved from the evergreen forest species and then radiated into an expanding area of drier habitats during the later Tertiary and dry intervals of the Pleistocene. Originally the genusTrachylobium was distinguished fromHymenaea by a morphological comparison with the probably more recently evolved species. A possible explanation is presented for the disjunct amphi-Atlantic distribution ofH. verrucosa and the neotropical species ofHymenaea.     

DISSECTION OF TWO DISTINCT DEFENSE‐RELATED RESPONSES TO AGAR OLIGOSACCHARIDES IN GRACILARIA CHILENSIS (RHODOPHYTA) AND GRACILARIA CONFERTA (RHODOPHYTA)1

The two agar‐producing red algae, Gracilaria chilensis C. J. Bird, McLachlan & E. C. Oliveira and Gracilaria conferta (Schousboe ex Montagne) Montagne, responded with hydrogen peroxide (H₂O₂) release when agar oligosaccharides were added to the medium. In G. conferta, a transient release was observed, followed by a refractory state of 6 h. This response was sensitive to chemical inhibitors of NADPH oxidase, protein kinases, protein phosphatases, and calcium translocation in the cell, whereas it was insensitive to inhibitors of metalloenzymes. Transmission electron microscopic observations of the H₂O₂‐dependent formation of cerium peroxide from cerium chloride indicated oxygen activation at the plasma membrane of G. conferta. A putative system, consisting of a receptor specific to agar oligosaccharides and a plasma membrane‐located NADPH oxidase, appears to be responsible for the release of H₂O₂ in G. conferta. Subcellular examination of G. chilensis showed that the H₂O₂ release was located in the cell wall. It was sensitive to inhibitors of metalloenzymes and flavoenzymes, and no refractory state was observed. The release was correlated with accumulation of an aldehyde in the algal medium, suggesting that an agar oligosaccharide oxidase is present in the apoplast of G. chilensis. The presence of this enzyme could also be demonstrated by polyacrylamide electrophoresis under nondenaturating conditions and proven to be variable. Cultivation of G. chilensis at 16 to 17°C resulted in significantly stronger expression of agar oligosaccharide oxidase than cultivation at 12°C, which indicates that the enzyme is used under conditions that generally favor microbial agar macerating activity.     

Enhancement of Festuca rubra L. germination and seedling growth by seed treatment with pathogenic Agrobacterium rhizogenes

The study concerned effects of two methods of red fescue seeds treating with wild type Agrobacterium rhizogenes strains (15834 and LBA 1334): soaking and matriconditioning with Micro-Cel E solid carrier. The effects were assessed from germination and seedlings growth rates. The bacterial indole-3-acetic acid (IAA) production and ACC deaminase activity were also determined. Both strains are able to produce the IAA and showed ACC deaminase activity in various amounts. The strains accelerated seeds germination, seedling emergence and development. The beneficial effect on those processes was visible when seeds were soaked for 1 h in bacterial suspension and, especially, when the bacteria were present during Micro-Cel E conditioning. The main effect observed upon inoculation of seeds during priming was increased growth of lateral roots and more complex architecture of the branching root system. Treating seeds with A. rhizogenes simultaneously with Micro-Cel E priming as a carrier is a promising method of enhancing grass germination and seedlings growth, particularly by improving the root system development.