Genetic Map Construction and Quantitative Trait Locus (QTL) Detection of Six Economic Traits Using an F2 Population of the Hybrid from Saccharina longissima and Saccharina japonica

Saccharina (Laminaria) is one of the most important economic seaweeds. Previously, four genetic linkage maps of Saccharina have been constructed and five QTLs have been identified. However, they were not enough for its breeding. In this work, Saccharina longissima (♀) and Saccharina japonica (♂), which showed obvious differences in morphology and genetics, were applied in hybridization to yield the F₂ mapping population with 102 individuals. Using these 102 F₂ hybrids, the genetic linkage map of Saccharina was constructed by MapMaker software based on 37 amplified fragment length polymorphisms (AFLPs), 22 sequence-related amplified polymorphisms (SRAPs) and 139 simple sequence repeats (SSRs) markers. Meanwhile, QTL analysis was performed for six economic traits. The linkage map constructed in this research consisted of 422 marker loci (137 AFLPs, 57 SRAPs and 228 SSRs), which formed 45 linkage groups (LGs) with an average marker space of 7.92 cM; they spanned a total length of 2233.1 cM, covering the whole estimated genome size. A total of 29 QTLs were identified for six economic traits, which explained 1.06 to 64.00% of phenotypic variation, including three QTLs for frond length (FL) and raw weight (RW), five QTLs for frond width (FW), two QTLs for frond fascia width (FFW) and frond thickness (FT), and fourteen QTLs for base shape (BS). The results of this research will improve the breeding efficiency and be beneficial for marker-assisted selection (MAS) schemes in Saccharina breeding.     

Plant response to overcrowding – Lemna minor example

Plants adopt various strategies in response to increasing density. We tested that response in two populations of Lemna minor L. – a free floating aquatic plant that frequently experiences intraspecific competition for space. Surface area of fronds and colonies, colony size (the number of fronds per colony), the rate of reproduction (based on the number of produced fronds) and growth rate (enlargement of surface area of all colonies) were the analysed factors presumably affected by density. The study was performed in natural stands and in experimental conditions with the use of two contrasting plant densities. Plants growing in natural conditions produced fronds of smaller and less variable surface area as a response to overcrowding but the number of fronds per colony was unrelated to plant density. Stable experimental conditions facilitated formation of fronds and colonies larger than in the field but frond detachment decreasing colony size was more intensive at high than at low density. This strategy allowed plants to more efficiently occupy limited available space. No self-thinning was observed during experimental cultures. Due to increasing frond area in cultures, growth rate was always higher than the rate of plant reproduction. Both were strongly negatively affected by high density. Performed calculations indicate that density-dependent growth inhibition starts when L. minor colonies cover the available water surface with a mono-layer mat. Some types of responses were found to significantly differ between analysed populations, which was also shown by genetic differences tested with he ISSR-PCR technique. Possible causal relationship between plant strategies and their genomic structure needs, however, further studies.     

Translocation of C in Macrocystis pyrifera (Giant Kelp)

The pattern of import and export of ¹⁴C-labeled assimilates in Macrocystis pyrifera (L.) C. A. Agardh in southern California was studied by labeling single blades on fronds, in situ, with [¹⁴C]NaHCO₃ for 24 hours. The pattern was found to be similar to that known in dicotyledons: actively growing tissue imported and did not export. As a blade reached maturity it began to export, at first only acropetally to the apex which formed it, later also down the frond to sporophylls and frond initials at the base of the frond, and into the apical regions of juvenile fronds; finally there was a phase of declining export, late in the life of the blade, when transport was only downward. Young fronds imported from older fronds until they were approximately 3 meters long, by which time they had developed mature, upward exporting blades. No translocation was found from a younger frond to an older frond, nor was there transport upward from a blade on a frond lacking the apical region.     

Identification of SCAR marker linking to longer frond length of <Emphasis Type="Italic">Saccharina japonica</Emphasis> (Laminariales,Phaeophyta) using bulked-segregant analysis

To construct a molecular-marker-assisted selection (MAS) system, research was done on identifying molecular markers linking to longer frond length, a crucial selection index in the breeding of the commercially important seaweed Saccharina japonica. An F₂-segregant population of 92 individuals was obtained by crossing two prominent S. japonica strains. Genomic DNA from ten individuals with the longest frond and ten individuals with the shortest frond in the F₂-segregant population were mixed to create two DNA pools for screening polymorphic markers. In bulked-segregant analysis (BSA), out of 100 random amplified polymorphic DNA (RAPD) primers only two produced three polymorphic RAPD markers between the two DNA pools. In conversion of the three RAPD markers into sequence-characterized amplified region (SCAR) markers, only one was successfully converted into a SCAR marker FL-569 linking to the trait of longer frond. Test of the marker FL-569 showed that 80% of the individuals with longest fronds in a wild population and 87.5% of individuals with the longest fronds in an inbred line “Zhongke No. 2” could be detected by FL-569. Additionally, genetic linkage analysis showed that the SCAR marker could be integrated into the reported genetic map and QTL mapping showed that FL-569 linking to qL1-1. The obtained marker FL-569 will be beneficial to MAS in S. japonica breeding.     

DEMOGRAPHY OF FRONDS OF SARGASSUM LAPAZEANUM FROM SOUTHERN BAJA CALIFORNIA, MEXICO, DURING EARLY STAGES

In terms of biomass, Sargassum lapazeanum (Phaeophyceae, Fucales) is one of the most important seaweeds in La Paz Bay, on the southeastern coast of the Baja California Peninsula, Mexico. This species can be found year-round in shallow subtidal sites. Standing biomass is lowest between fall and winter and highest between spring and summer. We are currently studying the annual demography of fronds as a necessary step to understanding the mechanisms of population regulation. Work is being done on a bed of about 90 m in length and three m in width. At this stage of abstract submission, we can report results for winter only. Random samples were collected in February and in March 2000 to estimate stand biomass, frond density, and size (frond length) structure. In addition, we labeled fronds with numbered plastic tags to estimate their rates of growth and of mortality during this period. Rates of recruitment were estimated from a combined analysis of the above. In February, fronds were all shorter than 6.5 cm. Between February and March, mean total frond density increased from 122 to 776 fronds m⁻² (n = 30 quadrats). The mean recruitment rate was 667 fronds m⁻², the mean growth rate was 0.5 cm day⁻¹ (n = 60 fronds), and the mean mortality rate was 43 fronds m⁻². From last year's preliminary observations, we expect to observe peaks of biomass and of reproduction in late spring, followed by negative growth rates and high mortality rates during summer.     

Characterization of Senescence in Lemna gibba G3: A Determinate Growth System

Frond senescence in Lemna gibba G3 was characterized, and itscontrol by light, ABA and kinetin investigated. The plant exhibitsa determinate growth pattern with a frond producing a set numberof daughter fronds before undergoing senescence and death regardlessof whether or not it flowers. When a frond was cut in half,the distal half (half frond) which lacks any meristem underwentrapid senescence as compared with intact fronds. In both intactand half fronds, the onset of senescence was accelerated byABA and retarded by kinetin. Continuous white light acceleratedsenescence in both intact and half fronds over the dark controls.Under different photoperiodic light regime, the pace of daughterfrond production is accelerated in proportion to the lengthof light period. In half fronds, however, very short photoperiodiclight treatments (e.g. 1L: 23D or 3L: 21D) rather delayed senescenceover the dark controls. Two separate light control systems operatingin opposite directions in Lemana senescence appear to exist. ¹Present address: Department of Biology, Yonsei University,Seoul 120-749, Korea ²Present address: U.S. Department of Agriculture, Aero SpaceBuilding, Rm. 323, 901 D Street, S.W., Washington, D.C. 20251-2200, U.S.A. (Received July 13, 1989; Accepted May 8, 1990)     

Differential Response in the Water Status of Immature and Mature Fronds of the Ostrich Fern (Matteuccia struthiopteris [L.] Todaro) to a Mild Water Stress

Experiments were conducted in growth chambers to examine the effect of a mild water stress (−200 kilopascals polyethylene glycol) on frond elongation and water status of the ostrich fern (Matteuccia struthiopteris [L.] Todaro). Measurements were taken for two days, starting one day after the application of polyethylene glycol. Total water potential in control (well-watered) plants was always significantly higher in immature fronds than in mature fronds. The osmotic potential in mature fronds was always significantly lower (about 800 kilopascals) than in immature fronds in both control and stressed plants. In immature fronds, the stress decreased elongation and total water and pressure potentials, while in mature fronds it increased total water and pressure potentials. The decreases in total and pressure potentials in immature fronds were approximately equal to the increases in mature fronds.     

STRUCTURE AND DYNAMICS OF A POPULATION OF PALMARIA PALMATA (RHODOPHYTA) IN NORTHERN SPAIN1

Palmaria palmata (Linnaeus) O. Kuntze (Rhodophyta, Palmariaceae) is a seaweed commercially harvested for human consumption. Its population density, size structure, and frond dynamics were investigated from May 1999 to May 2001 at one intertidal locality in the northern coast of Spain, which is within the southern distributional boundary of the species in the eastern Atlantic coasts. The effect of size, age, and the life‐history phase (haploid vs. diploid) on frond growth and mortality were also evaluated. The study was carried out by mapping and monitoring fronds in the field. New fronds (macroscopic recruits or sprouts) appeared in spring, but subsequent mortality of these young fronds and detachment of the host plant led to lower density values in January. Palmaria palmata exhibited a distinctive seasonal growth cycle, with positive net growth from March to August and breakage from August to March. Interannual differences were also detected, with higher net growth in 2000 than in 1999. Net growth was apparently independent of age, reproductive status (fertile vs. reproductive), and life‐history phase (haploid vs. diploid) but was dependent on size, as longer fronds showed minor growth or greater breakage than small ones. Mortality, on the other hand, was more dependent on age than on size in the period analyzed (March–May 2000). Results of the study indicate that both size and age should be included as state variables and temporal changes in transition probabilities considered in the development of demographic models of the species.     

TRANSLOCATION OF 14C IN MACROCYSTIS INTEGRIFOLIA (PHAEOPHYCEAE)1,2

Translocation patterns in the giant kelp, Macrocystis integrifolia Bory, were investigated in situ using ¹⁴C tracer; sources and sinks were identified. Export was first detected after 4 h of labeling; experiments were routinely 24 h continuous ¹⁴C application. Mature blades exported ¹⁴C to young blades on the same frond and on younger fronds, as well as to sporophylls and frond initials at the bases of the fronds. Blades <0.3 m from the apex imported and did not export; this distance did not change seasonally. In spring export from blades 0.3–1.25 m from the apex was exclusively upwards; older blades also exported downwards. In fall downward export began 0.5 m from the apex, and blades >2 m from the apex exported exclusively downwards. Carbon imported by frond initials, young fronds, and sporophylls in fall may partly be stored for growth in early spring. No translocation was seen in very young plants until one blade (secondary frond initial) bad been freed from the apical blade; this blade exported to the apical blade for a time, but imported when it began to develop into a frond. The second and third formed blades on the primary fronds (sporophylls also exported when <0.3 m from the apex, and later stopped. Frond initials and sporophylls on later-formed fronds did not export at all. The translocation pattern in M. integrifolia differs from that previously reported in M. pyrifera in seasonal change and in distances from the apex at which the changes take place.     

Reproduction of the colonial hydroid Obelia geniculata (L., 1758) (Cnidaria,Hydrozoa) in the White Sea

Populations of the colonial hydroid Obelia geniculata in the White Sea reproduce asexually by frustule formation. Young medusae appear in the plankton during July and August. The number of medusae rarely exceeds 36 per m³, and the average number varies every year from 0.4 to 10 per m³. The size of medusae is smaller than reported from other regions. The umbrella of the largest recorded medusa was only 0.57 mm in diameter and the specimen had just 35 tentacles. Only a few mature medusae were found during the study. The colonies in the White Sea are epiphytic and grow only on laminarian thalli. At the beginning of July there are no colonies on thalli from the upper subtidal zone. By the end of August, colonies of O.␣geniculata had increased in density to 30 per m². Hydroid recruitment was attributed to active frustule production by colonies living below that zone. The frustules detach from the stems of the hydroids and are found in plankton. Production of frustules on branches occurs continuously during colony growth until water temperatures climb above 0 °C. We found that water temperature in this Arctic environment is generally too low for medusa maturation and planula development in the species. Propagation by frustule formation is the principal means of reproduction in Obelia geniculata within the White Sea, and this phenomenon accounts for the species being a dominant epiphyte on laminarian thalli there.     

Differential sensitivity of four Lemnaceae species to zinc sulphate

Lemnaceae are currently the only freshwater plants required for regulatory toxicity testing of pesticides and other chemicals. Toxicological protocols allow for the use of different Lemnaceae species in tests. However, few studies have compared the relative sensitivity of individual duckweed species. Zinc is an essential plant nutrient but is also a common pollutant in aquatic environments and elevated levels are phytotoxic. This study shows that four species of Lemnaceae differ in their relative sensitivities to zinc sulphate, a commonly used reference chemical. Comparative zinc sensitivity, in order, from most tolerant to most sensitive was: Landoltia punctata > Lemna minor > Wolffia brasiliensis > Lemna gibba. Zinc sensitivity was also endpoint dependant. EC₅₀ values typically increased in order of: specific biomass growth rate < specific frond number growth rate < chlorophyll absorbance. However, specific frond number growth rate was the most sensitive endpoint for L. punctata. Unlike the other species, L. punctata displayed no significant colony disintegration. Lemna species and L. punctata appear to be employing distinct response strategies when exposed to zinc. L. gibba and L. minor produce and release young, single fronds which are severely affected by zinc. In contrast, L. punctata produces fewer fronds, which are not released and form large colonies of high biomass that are relatively zinc tolerant.     

ABSCISIC ACID EFFECTS ON FRONDS AND ROOTS OF LEMNA MINOR L.

Plants of Lemna minor L. were grown in axenic culture in order to investigate the direct effect of abscisic acid (ABA) on individual fronds and roots. Both frond and root growth rates were inhibited by 60%, and total growth of both organs was reduced by 30% with ABA concentrations of 2 mg/l. Abscisic acid inhibited frond reproduction, and treated fronds tended to remain attached to parents. Reduction of root elongation and frond expansion occurred within 1–4 hr. Prolonged exposure to ABA inhibited both cell enlargement and cell division in the roots. The results are discussed in the light of current views on the control of growth with endogenous levels of promoter and inhibitor hormones.     

Effects of medium components and light on callus induction, growth, and frond regeneration in Lemna gibba (Duckweed)

Summary Basal media, plant growth regulator type and concentration, sucrose, and light were examined for their effects on duckweed (Lemna gibba) frond proliferation, callus induction and growth, and frond regeneration. Murashige and Skoog medium proved best for callus induction and growth, while Schenk and Hildebrandt medium proved best for frond proliferation. The ability of auxin to induce callus was associated with the relative strength of the four auxins tested, with 20 or 50 μM 2,4-dichlorophenoxyacetic acid giving the highest frequency (10%) of fronds producing callus. Auxin combinations did not improve callus induction frequency. Auxin in combination with other plant growth regulators was needed for long-term callus growth; the two superior plant growth regulator combinations were 10 μM naphthaleneacetic acid, 10 μM gibberellic acid, and 2 μM benzyladenine with either 1 or 20 μM 2,4-dichlorophenoxyacetic acid. Three percent sucrose was best for callus induction and growth. Callus induction and growth required light. Callus that proliferated from each frond’s meristematic zone contained a mixture of dedifferentiated and somewhat organized cell masses. Continual callus selection was required to produce mostly dedifferentiated, slow-growing callus cell lines. Frond regeneration occurred on Schenk and Hildebrandt medium without plant growth regulators but was promoted by 1 μM benzyladenine. Callus maintained its ability to regenerate fronds for at least 10 mo. Regenerated fronds showed a slower growth rate than normal fronds and a low percentage of abnormal morphologies that reverted to normal after one or two subcultures.     

Effects of wave exposure on population and reproductive phenology of an algal turf, Gelidium pusillum (Gelidales,Rhodophyta), Songkhla,Thailand

The effect of wave exposure on the population and reproductive phenology of the common red alga, Gelidium pusillum (Stackhouse) Le Jolis, was investigated between July 2003 and June 2004, at Suan Song Tha Le, Songkla Province, Thailand. Lengths of thalli, percentage cover, percentage of reproductive fronds and the number of reproductive structures were examined monthly in relation to different degrees of wave exposure (sheltered vs. exposed), temperature, rainfall, day length and tidal cycles. Frond length and percentage cover of G. pusillum were different among sites and seasons. Shorter fronds were found on the exposed shore which had a greater percentage cover. Fronds bleached and died off during the summer months (April and May), which resulted in shorter fronds and reduced percentage cover in June. The thalli reproduced throughout the year, with a predominance of tetrasporophytes. The highest percentage of tetrasporic fronds was 33% in February 2004 and 13% of cystocarpic fronds in April 2004, but male gametophytic fronds were never observed. Rainfall showed a strongly negative influence on reproduction since no reproductive fronds were observed during the rainy season (R² = 0.49, P = 0.01). The dominance of G. pusillum at this study site and throughout elsewhere in turf habitats might be a function of persistent vegetative growth, densely clumped, and the ability to reproduce almost throughout the entire year.     

Early-spring gas exchange and uptake of deuterium-labelled water in the poikilohydric fernPolypodium virginianum

This study was carried out to determine if the desiccation-tolerant fernPolypodium virgimanum L. ecologically resembles lower plants by absorbing atmospheric water through its fronds and actively growing in early spring when the soil along cliff edges is still frozen. Three times between February and April, 1991,P. virginianum clonal mats were treated with deuterium-labelled water. Following each application, fronds were collected over several days and analyzed for the presence of deuterium. Two treatment groups plus a control were used: fronds were sprayed directly while covering the soil, or the roots were watered while protecting the fronds. The control mats were left untreated. Soil, air, and frond temperatures, plus photosynthesis and frond conductance were monitored throughout the study period. At subfreezing temperatures in February, no labelled water was taken up from the soil and no photosynthesis took place. Small amounts of label were absorbed from the soil in March during freeze-thaw cycles when rates of photosynthesis and stomatal conductance were both low. Large amounts of label were taken up from the soil in April when the soil was fully thawed and gas exchange was at normal seasonal levels. Label was not absorbed directly through the fronds when the plants were actively growing. Despite the desiccation tolerance ofP. virginianum, the timing and patterns of its water uptake and gas exchange in early spring resemble those found in higher vascular plants, not poikilohydric lower plants.     

Crowding in clonal seaweeds: Does self-thinning occur in Mastocarpus papillatus shortly before stand biomass peaks?

Fronds from crowded stands of clonal seaweeds, particularly those in which holdfasts are mostly perennial and are the major source of new fronds every year, are thought not to undergo self-thinning during the growth season, unlike those from crowded stands of unitary seaweeds. For clonal seaweeds, it is not known, however, what happens at the very end of the growth season, when crowding is highest for the year. By sampling twice more frequently than previously done for similar species, the possible occurrence of frond self-thinning was tested for Mastocarpus papillatus (Rhodophyta, Gigartinales, Petrocelidaceae) from western Canada during the growth season (spring) of 2003. Initially, stand biomass increased together with frond density, as found previously for similar clonal seaweeds. Shortly before stand biomass peaked for the year (June), frond density remained statistically unchanged. Thus, the increased sampling precision of this study confirms that fronds of these clonal seaweeds do not undergo self-thinning, not even shortly before crowding is highest. Frond size inequality for M. papillatus remained statistically similar during the growth season, which is also consistent with a model of no self-thinning. There are similarities in biomass–density dynamics and in size inequality dynamics between clonal seaweeds and clonal vascular plants.     

The influences of temperature and irradiance on thallus length of <Emphasis Type="Italic">Saccharina japonica</Emphasis> (Phaeophyta) during the early stages of cultivation

Korean Saccharina japonica is highly valued, both for human consumption and abalone feed. For the stable production of abalone feed, fresh seaweed biomass is required throughout the year. However, currently, the production of farmed Saccharina is limited by environmental conditions such as temperature, irradiance, and nutrient availability between August and November. Due to shortages experienced in supply, the production of early-season biomass can be highly profitable and, therefore, some famers attempt to start their cultivation activities before prevailing, surface seawater temperatures (SST) are optimal. However, attempting to cultivate too early, can lead to total crop failure. Young kelp sporophytes are easily destroyed between 18 and 22 °C SST, which can occur during the early nursery period when the materials are confined to tanks. This study investigated the growth of S. japonica thalli and photosynthetic quantum yield (F_v/F_m) under five temperatures (i.e., 18–26 °C, at 2° increments) and five irradiances (i.e., 5, 10, 20, 40, and 80 μmol photons m^?2 s^?1). This was undertaken for four different size groups of sporophyte thalli (i.e., 0.25, 1, 5, 10 mm). There were different responses of the initial groups of S. japonica showing different tolerances to temperature and irradiance. In general, the smaller plants (1 mm) were more tolerant of sub-optimal conditions than their larger cohorts. These results indicated the optimum temperature and irradiance ranges for different size groups of S. japonica thalli which, if adopted in management protocols, could contribute to enhanced profitability and a more stable and evenly distributed production of Saccharina raw materials over an entire annual basis.     

Protein and nitrate content of lemna sp. As a function of developmental stage and incubation temperature

Lemna protein per frond and per root increases with developmental stage until plants are at least two generations old. Protein per frond, per root, and per unit dry weight is greater in plants grown at 23.9 C than at 18.3 C. More protein is found in fronds than in roots, and more nitrate occurs in roots than in fronds. Nitrate per root increases with developmental stage and is higher (per root) in plants grown at 23.9 C than in those grown at 18.3 C. The distribution of generations within a growing population is constant for at least eight doubling times. Whether populations multiply slowly at 15.6 C or more rapidly at 23.9 C, fronds which have not yet produced progeny form 62% of the population; fronds which are one generation old form 24% of the population; and fronds which are two generations old form 9% of the population.