镇海棘螈和大凉疣螈年龄结构研究

通过趾骨切片可以准确鉴定年龄，了解一个物种的最长寿命，也为我们研究确定一个物种的生长特点、性成熟期,以及一个地区一个物种的年龄结构、种群生态(Marnell，1998)和群落生态提供重要信息(Morrison，et a1.，2004)。 本论文使用骨骼鉴龄法对中国浙江省宁波市北仑瑞岩寺林场的镇海棘螈（Echinotriton chinhaiensis）雌性繁群进行了年龄结构研究。结果显示：第一次参加繁殖的年龄为3龄；繁群中数量占优势的是5龄、6龄。而在6龄以后参加繁殖的雌性个体数便开始随着年龄的增大而逐渐减少。参加繁殖的雌性年龄最大个体为8龄。平均年龄为5.13龄。同时对其年龄和头体长、体全长的相关性检验,发现其年龄与头体长和体全长不相关,镇海棘螈雌性的生长方式表现为性成熟后能量主要用于繁殖。 另外,对李子坪大凉疣螈(Tylototriton taliangensis) 雄性繁群进行了年龄结构研究。结果显示：大凉疣螈雄性第一次参加繁殖的年龄为4龄；繁群中数量占优势的是5龄、6龄、7龄。而在7龄以后参加繁殖的雄性个体数便开始随着年龄的增大而逐渐减少。参加繁殖的雄性中年龄最大的个体为10龄。平均年龄为6.7龄。对其年龄和头体长、体全长的相关性检验,发现其年龄与头体长和体全长不相关,大凉疣螈雄性生长特点也表现为性成熟后生长缓慢的特点。 研究材料方面，本文采用野外采样与标本馆标本相结合的方式获得了中国蝾螈科2个重要保护物种繁殖群体的剪（指）趾材料，使得建立于其上的年龄结构工作更加可靠、更加具有代表性。 此外,本论文讨论了镇海棘螈瑞岩寺种群繁殖总量年度间的差异及其产生原因。将1998、1999、2000、2008、2009年镇海棘螈（Echinotriton chinhaiensis） 瑞岩寺种群的繁殖量进行比较，发现虽然雌性平均窝卵数比较稳定，但繁殖总量小于1998、1999、2000年任何一年总产卵量的50%。对2008年镇海棘螈繁殖量大幅下降的原因分析发现, 2007年9、10月影响严重台风的两次强台风、瑞岩寺景区开发等因素可能是造成近年该种群繁殖量大幅下降的原因。而2008年初50年不遇的低温是否影响镇海棘螈的繁殖值得进一步追踪研究。2009年繁殖量较2008年没有明显的增长，可能是由于2007年的台风影响了其繁殖营养的积累。台风的影响可能存在滞后现象,对此有待进一步监测证明。 本研究首次对中国蝾螈科物种进行的年龄结构鉴定，为进一步了解中国蝾螈科动物的种群生态打下了坚实的基础。 Using skeletochronology, we can know the life span of a species, age of reaching sexual mature, and of course age structure, which are vital(Morrison，et a1.，2004). Skeletochronology was performed on Echinotriton chinhaiensis Ruiyansi female population. The result shows that: The oldest individuals were 8 years old and the youngest ones were 3 years old. Individuals of age class 5(39.13%) and 6(21.74%) were most numerous. The number of individuals participated in reproduction decreased with the increase of age after the sixth year. Average age is 5.13 years. There is no correlation between age and body size (SVL and TL). For female chinhai salamander, energy is devoted to reproduction after reaching sexual maturation. While using skeletochronology to study Tylototriton taliangensis Liziping male population, the oldest individuals is 10 years old, and the youngest ones is 4 years old. Individuals of the age class 5, 6, and 7 dominat this population. The number of individuals decrease with the increase of age also after the seventh year. Average age is 6.7 years old in this population. there is also no correlation between age and body size (SVL and TL).It turned out that T. taliangensis tend to grow slowly after reaching sexual maturation. In this thesis, specimens from both wild and museum were used to gain enough toe clipping samples. A big sample size guarantees the reliability of this study. In the meantime, E. chinhaiensis’s annual reproduction of the year 1998, 1999, 2000 ,2008,and 2009 was compared. The result shows there is a huge decline in E. chinhaiensis’s annual reproduction in 2008,even the egg clutch is very stable. After analyzing, it turned out the huge decline in 2008 was probably caused by typhoon in 2007, besides the effect of tourism development and cash crop planting. While the impact of extreme weather of 2008 on reproduction needs further investigation. In the year 2009, there is no obvious increase in annual reproduction. It maybe due to lasting impact of typhoon in 2007. It is the first age-structure study on these two Chinese salamanders. A solid foundation was laid for further population ecology study of these two species.

镇海棘螈繁殖群体生态和遗传结构研究

genetics, such as: population size, reproduction, mating system, growth, development,genetic structure and systematics status; The main results are presented below: The seasonal variation of the operational sexual ratio of this animal was found in the field and the ration always bias the female in the breeding season. Aiming at this character and considering the distance of time and space of both sexual habitat in breeding season, we census female population first by toe-clipping mark-recapture method, then estimated the population size with the definitive sexual ratio. Up to now, this species was found only at the Beilun district of the Ningbo City. The population size of the Ruiyan Temple Forest Park approximates to 369. The status of this population is extremely endangered, so besides protecting this population at the original locality, we also suggested to breed the salamander in fenced locality and to hatch embryos artificially, and send metamorphosed juveniles back to nature. We can transfer some individuals to other similar habitats or breed them under artificial conditions for saving this species from extinction. The early developmental stage of the Chinhai salamander is the same as its relative species, E. andersoni. Their balanceres are poorly developed and disappear very early. Temperature and moisture significantly influence the embryonic development of the Chinhai salamander. The embryonic stage is approx. 29 days under room temperature. The hatchling grows in a logarithmic curve. The larvae stage in water is approx. 58- 88 days. Many factors influence the nomal development, including two aspects of internal and external. Due to these factors, the effective protected measures were presented in detail. The breeding migration of E. chinhaiensis takes place at late March～late April every year. This salamander's hatching rate is high, but the rate of hatchling migrating into water is low. The average effectiveness of all the nest sites is 36.7%. The maternal self-conservation was contrary to the reproductive success of the egg-laying strategy. In the strategy of egg-laying behavior, the first factor selected by the female was its self-conservation, the second is embryonic survival rate, and the last is rate of hatchling survival rate. The oviposition selection is significant for the survival of the larvae. Based on the analysis of the evolutionary process of reproductive behaviors nad egg-laying site selections of all genera of the family Salamandridae, we deduced that perhaps Echinotriton is a transitional type in the evolutionary process from water to land. Due to its location in the adaptive stage in the terrestrial evolution, Echinotriton chinhaiensis's terrestrial nest may be one of important reason that causes this species to be endangered. The genetic deversity analysis shows that although the population size of the Chinhai salamander is quite small compared to other Chinese salamandrid species, the genetic diversity of this population is not reduce remarkably. We explain this phenomena with the polygamy mating system of this species. The result of 4 families' parenthood determinations shows that the parenhood determination can be taken without any paternal information. The "children" of every female include rich genetic information from at least two "fathers". It implies that female Chinhai salamander mates more than once with different males in a breeding season. The molecular evidence, the behavioral observation evidences and the sperm evidence in the female cloaca proved that this species has a polygamy mating system. The kin recognition in the mating of adult salamander was first discussed. The taxonomic status and phylogenetic relationships of 12 species representing 6 genera in the family Salamandridae were studied using DNA fingerprinting. The results showed that the DNA fingerprinting. The results showed that the DNA fingerprinting patterns demonstrated rich genetic diversity and species diversity, and also revealed the taxonomic status and phylogenetic relationshipes of higher taxa to a certain extent. The results are highly consistent with those obtained from the studies based on the morphology, ecology, cytology and molecular biology. The compreshensive analysis indicate that Tylototrition hainanensis and T. wenxianensis should be valid species; Echinotriton should be a valid genus;Tylotortriton is a natural cluster; Tylotortriton asperrimus should be put in Tylototrition rather than in Echinotriton, Hypselotriton and Allomestriton are synonyms of Cynops and Paramesotriton, respectively. There are three main groups in Chinese salamandride: Cynops, Paramesotriton and Pachytrition from the first group, the species of the Tylototriton from the second, and E. chinhaiensis composes the third.

同源四倍体水稻雌配子体发育特性及变异胚囊研究

通过秋水仙素诱导获得同源四倍体水稻10个株系，包括6个恢复系、3个保持系和1个不育系，这些株系具有加倍的染色体组。田间观察表明10个株系具有特殊的农艺性状：茎杆变粗壮、植株颜色加深、叶片变厚、叶宽适度增加、分蘖数减少、有效分蘖的比率下降等。根尖有丝分裂鉴定表明，同源四倍体水稻10个株系具有正常的有丝分裂，观察细胞的染色体数目皆为2n=48。花粉母细胞减数分裂鉴定表明10个株系具有比较理想的减数分裂行为，后期I染色体滞后、末期I微核生成和末期II异常小孢子比率较低，能较好的完成减数分裂过程，其中后期I染色体滞后比率约为10%-20%，末期I微核生成比率约为1%-6%，末期II异常小孢子比率约为1%-8%。这提示，染色体联合和分离不规则导致三价体、单价体 和落后染色体等产生，并进一步导致在后期和末期不均横分离产生异常小孢子，这可能是同源四倍体株系结实率不高的原因之一。 同源四倍体水稻正常胚囊为蓼型，变异胚囊具有多种类型，其比率显著高于二倍体对照，变化范围为39.62%－69.85%。按变异胚囊的结构特点和形成方式，分为四种类型：退化型，结构变异型，无融合生殖型和反足细胞增殖型。退化型胚囊的平均比率为29.17%，包括小胚囊（15.04%）和完全退化胚囊（14.13%），前者仍有较小胚囊腔而后者胚囊腔缺失。结构变异胚囊包括结构缺失、结构重复和位置异常，反映了蓼型胚囊八核七细胞结构的变异，其在各株系的平均比率为18.96%。无融合生殖胚囊发生比率极低，平均比率为1.77%，类型包括反足胚和卵细胞胚。反足细胞增殖胚囊是反足细胞团频繁增殖形成，伴随上述三种变异发生使异常胚囊的多样性进一步增加，其在各株系的平均比率为10.62%。相关分析表明，同源四倍体水稻结实可能主要来自三部分：正常胚囊、正常型小胚囊和反足细胞增殖型胚囊。这三种胚囊具有相对完整的蓼型结构，可能具有较好的育性，其对结实率的贡献程度估计值分别为72.44%、15.12%、12.44%。此外，完全退化型胚囊和位置异常型胚囊对结实率分别表现出显著（-0.66）和极显著（-0.92）的负相关，这表明二者可能是结实性的抑制因素。 Ten autotetraploid strains, which include six restoring lines, three maintaining lines and a sterile line, are derived from artificial induction by colchicine treatments. Variations of agronomical traits are observed which include large organs, sturdy plants, long panicle length and deep leaf color and so on. It has been confirmed that autotetraploid strains exhibit normal chromosome behaviors in mitosis and the chromosome numbers are all 48. Moreover, abnormal chromosome behaviors are investigated in meiosis including univalent, trivalent, quatrivalent, chromosome lagging and microkernel and so on. It evaluates that the percentage of chromosome lagging in anaphase I is about 10%-20%, the percentage of microkernel in telophase I is about 1%-6% and the percentage of abnormal microspore in telophase II is about 1%-8%. In all, abnormal behaviors of chromosomes could induce univalent, trivalent and et al. and subsequently induce infertile microspore. That may be one of the causes of low seed sets in autotetraploid strains. Embryo sacs of autotetraploid strains are formed according to the Polygonum type. However, these strains exhibit variations of abnormal embryo sacs at high frequencies (39.62% - 69.85%). The variations are frequently involved in the spikelets of the main panicles and the first tillers, leading to obvious decreases of the percentages of normal embryo sacs among the strains. Four types of abnormal embryo sacs are classified basing on their different structures and origins: degenerated embryo sac (DES), structure variation (SV), apomixis (Apo) and antipodal cell proliferation (ACP). Embryo sacs of DES (29.17%) exhibit small embryo sacs (15.04%) or no embryo sac (14.13%), the former showing embryo sacs with decreased size and the latter showing no sac. Embryo sacs of AS (18.96%) include three subtypes: structure deletion, structure duplication and location variation, which suggests abnormalities of the eight nuclei, seven celled pattern of the Polygonum type. Embryo sacs of Apo (only 1.77%) include two origins of apomictic embryos: antipodal embryo and egg embryo. Embryo sacs of ACP are observed frequently (10.62%) in autotetraploid strains which subsequently increase the variations of abnormal embryo sacs. It evaluates by the Pearson’s correlation analysis that seed set is probably contributed by three origins of embryo sacs: normal embryo sacs, small embryo sacs (normal pattern) and embryo sacs of ACP. These three origins exhibit comparatively good structure of the Polygonum type and could account for seed set at a percentage of 72.44%, 15.12%, 12.44%, respectively. Moreover, the subtype of no embryo sac (NES) negatively related to seed set at the P>0.01 level (-0.92) and the subtype of location variation (LV) negatively related to seed set at the P>0.05 level (-0.66). Which suggest the two subtypes may have strong stress on seed set.

Electrochemical studies of lipophilic ion transport through BLM. The influence of sterols on its transport

Cyclic voltammetry was employed to study the influence of sterols on the lipophilic ion transport through the BLM. The mole fraction of the sterols (cholesterol, oxidized cholesterol). as referred to total lipid, was varied in a range of 0-0.8. Data demonstrate that a thin-layer model is suitable to this BLM system. By this model, the number of charges transported per lipophilic ion, the concentration of the ion in the membrane bulk phase and the aqueous/membrane phase partition coefficient could be calculated. These parameters proved that sterols had an obvious influence on the lipophilic ion transport. Cholesterol had a stronger influence on the ion transport than oxidized cholesterol. Its incorporation into egg lecithin membranes increased the partition coefficient beta of the ion up to more than 3-fold. Yet, oxidized cholesterol incorporated into egg lecithin membranes only increased the beta up to less than 2-fold, and the beta had no great variation at different oxidized cholesterol mole fractions. The higher beta obtained was partly due to the trace amount of solvent existing in the core of the lipid bilayers. At the different sterol mole fractions, combining the change of beta with the change of peak current, we also concluded that sterols had somewhat inhibiting effect on the ion transport at the higher sterols mole fraction (>0.4). These results are explained in terms of the possible change of dipole potential of the membrane produced by sterols and the decrease of the membrane fluidity caused by the condensation effect of sterols and the thinning effect caused by sterols. The substituting group (in the oxidized cholesterol) had some inhibiting effects on the ion transport at higher mole fractions (oxidized cholesterol mole fraction >0.4).

Effect of lanthanum ions on the lipid polymorphism of phosphatidylethanolamines

The influence of lanthanum ions on the polymorphic phase of egg phosphatidylethanolamine and dielaidoylphosphatidylethanolamine (PE and DEPE) has been investigated by means of P-31-nuclear magnetic resonance (P-31-NMR) and high sensitivity differential scanning calorimetry (DSC) techniques. P-31-NMR experiments show that lanthanum ions promote the formation of the hexagonal II phase at temperatures lower than those of the pure egg PE, DSC results also show that lanthanum ions induce the formation of hexagonal II phase in DEPE liposomes even al very low ion concentration, The effect of lanthanum ions on the polymorphism of PE liposomes is much greater than that of calcium.

Cultivation of the brown alga Hizikia fusiformis (Harvey) Okamura: enhanced seedling production in tumbled culture

The reuse of holdfasts for regeneration of young seedlings or using wild juvenile plants as the seedling source has played the major role in commercial cultivation of the brown alga Hizikia fusiformis in East Asia over the past 20 years. The possibility of employing zygote-derived germlings for producing seedlings has been discussed in the literature, but has not yet become a reality. Three main obstacles have limited the use of zygotes as a main source of seedlings, (1) the dioecious nature of the algal life cycle which may lead to asynchronous male and female receptacle development and thus different timing of egg and spermatozoa expulsion, (2) the low attachment rate when using zygote-derived germlings with developed rhizoids from wild parental plants for seeding production, and (3) the problem of culturing young germlings in regions where water temperature is high in summer. In this investigation, shifting the timing of receptacle formation earlier than in nature was performed by tumbling the algae in a long-day tank (16-h light per day). Synchronization of egg and spermatozoa expulsion and thereafter fertilization were conducted in indoor tanks. Receptacle formation in constant long days could be shifted by 20 days earlier than in plants cultured on long lines in the open sea, or I month earlier than in plants growing on intertidal rocks. Synchronized expulsion of eggs and spermatozoon led to a high rate of fertilization. This was achieved by tumbling the male and female receptacle-bearing branchlets in the same tank at low density in high irradiance. In two independent trials, a total of 1,400,000 zygote-derived germlings were obtained from 620 g (fresh weight) female sporophytes. The germlings shed from the receptacles were at an identical developmental stage indicating high synchronization of expulsion of eggs and spermatozoon followed by fertilization. Approximately 63% ( +/-9.6%) of the germlings were shed from the receptacle between 16 and 24 It after fertilization and 20% ( +/-11.9%) remained on the receptacle for 3 days after fertilization. Germlings were seeded on string collectors before rhizoids started to elongate and the attachment efficiency was enhanced. Young seedlings reached 800 ( +/-50) mum in length in 25 days at 25 degreesC before they were transferred to open sea cultivation. These results provide the basis of a practical way of seedling production by use of zygote-derived germlings in the commercial cultivation of Hizikia fusiformis. (C) 2004 Elsevier B.V All rights reserved.

Optimization of triploid induction by heat shock in Chinese shrimp Fenneropenaeus chinensis

Chromosome manipulation for commercially valuable marine animals plays an important role in aquaculture. The special reproductive characteristics of shrimp make it difficult to control fertilization and synchronize egg development, so research on chromosome manipulation in shrimp has proceeded very slowly. In the present study, triploid shrimp Fenneropenaeus chinensis were induced by heat shocks and the optimal-inducing condition was screened at different spawning temperatures. Level of triploid induction for each treatment was evaluated by flow cytometry at nauplius stage. The highest level of triploid induction reached to more than 90%. Starting time for each treatment was very crucial for triploid induction in shrimp. One optimal treatment condition for triploid induction was heat shock (29-32 degreesC), starting at 18-20 min for duration of 10 min. These conditions varied depending on the temperature at spawning. Triploid level at embryo stage and nauplius stage was not different, suggesting the same hatching rate between diploids and triploids. Heat shock is a very effective way to induce triploids in this species, and can be easily used on large scale without any harmful effect on the environment as compared with chemical treatment. (C) 2003 Elsevier Science B.V. All rights reserved.

beta-catenin in early development of the lancelet embryo indicates specific determination of embryonic polarity

The lancelet (amphioxus) embryo develops from a miolecithal egg and starts gastrulation when it is approximately 400 cells in size, in a fashion similar to that of some non-chordate deuterostomes. Throughout this type of gastrulation, the embryo develops characteristics such as the notochord and hollow nerve cord that commonly appear in chordates. beta-Catenin is an important factor in initiating body patterning. The behavior and developmental pattern of this protein in early lancelet development was examined in this study. Cytoplasmic beta-catenin was localized to the animal pole after fertilization and then was incorporated asymmetrically into the blastomeres during the first cleavage. Asymmetric distribution was observed at least until the 32-cell stage. The first nuclear localization was at the 64-cell stage, and involved all of the cells. At the initial gastrula stage, however, concentrated beta-catenin was found on the dorsal side. LiCl treatment affected the asymmetric pattern of beta-catenin during the first cleavage. LiCl also changed distribution of nuclear beta-catenin at the initial gastrula stage: distribution extended to cells on the animal side. Apparently associated with this change, expression domains of goosecoid, lhx3 and otx also changed to a radially symmetric pattern centered at the animal pole. However, LiCl-treated embryos were able to establish embryonic polarity. The present study suggests that in the lancelet embryo, polarity determination is independent of dorsal morphogenesis.

Size and time of day affect the response of postlarvae and early juvenile grooved tiger prawns Penaeus semisulcatus De Haan (Decapoda:Penaeidae) to natural and artificial seagrass in the laboratory

Juvenile tiger prawns (Penaeus semisulcatus De Haan and P. esculentus Haswell) show a strong association with vegetated habitats and are rarely caught on non-vegetated areas. This pattern of distribution may be caused by postlarvae selecting vegetation when they settle, or to differences in post-settlement mortality in different habitats. In this study, we examined whether the postlarvae and early juvenile stages of P. semisulcatus would distinguish between seagrass (Zostera capricorni Aschers) without epiphytes, artificial seagrass and bare substratum in the laboratory. The responses of prawns reared from the egg to different stages of postlarval and juvenile development were tested to determine whether, and when, each size class showed a response to a particular habitat. Five size classes of postlarvae (average carapace lengths [CL] of 1.2, 1.4, 1.6, 1.7 and 2.0 mm) were offered a choice between Z. capricorni and bare sand. Small size classes of postlarvae either did not respond to Z. capricorni (1.2 and 1.6 mm CL), or were more abundant on bare substratum than Z. capricorni. In contrast, the largest size classes of postlarvae (1.7 and 2.0 mm CL) were more abundant on Z. capricorni during the day but not at night. The behaviour of postlarvae changed markedly at a size of 1.7 mm CL (22 days from the first nauplius): smaller postlarvae frequently swam in the water column; 1.7 and 2.0 mm CL postlarvae spent much more. time resting on the substrate and perched on seagrass leaves. This size at which postlarvae first respond to seagrass during the day, and show mainly benthic behaviour, is similar to the size at which they are found on shallow seagrass beds in northern Australia. Large postlarvae (2.7 mm CL) and juveniles (4.1 mm CL) both were more abundant on artificial seagrass than bare sand during the day but not at night, indicating that they respond to structured habitats. When large postlarvae (2.4 mm CL) and juveniles (3.5 mm CL) were offered a choice between Z. capricorni without epiphytes and artificial seagrass, they were more abundant on the Z. capricorni, which suggests that chemical cues from seagrass may explain some of the responses of P. semisulcatus to seagrass. (C) 1997 Elsevier Science B.V.

Influence of albumen on aggregation and O-2 evolution of protoplasm from Bryopsis hypnoides Lamourou

The extruded protoplasm from the coenocytic green alga, Bryopsis hypnoides Lamouroux, was able to reform a cell wall and develop further into a mature alga in seawater. In this paper, the influence of albumen on the ability of aggregation and on the photosynthesis of protoplasm was examined. Results show that the protoplasm of B. hypnoides could aggregate in either albumen or chicken egg, which is similar to that in seawater. However unlike in seawater, the aggregation from B. hypnoides in albumen and chicken egg failed to develop into a mature individual. Interestingly, the protoplasm of B. hypnoides could maintain its photosynthetic O-2 evolution in albumen and chicken egg, while the time in chicken egg was longer than that in albumen.

Induction of diploid gynogenesis in turbot Scophthalmus maximus with left-eyed flounder Paralichthys olivaceus sperm

Turbot Scophthalmus maximus exhibits sexually dimorphic growth, with females growing faster and reaching larger adult sizes than males. Thus, development of techniques for preferentially producing females is necessary to optimize production of these species. In this paper, gynogenetic diploids of turbot were induced by activating egg development with ultraviolet (UV)-irradiated left-eyed flounder Paralichthys olivaceus sperm combined with cold shock to prevent extrusion of the second polar body. The results of UV irradiation experiments showed that survival, motility, and duration of activity of P. olivaceus sperm generally decreased with increase in UV dose. The typical Hertwig's effect was observed after fertilized turbot eggs with UV-irradiated P. olivaceus sperm and the optimal UV dose for gynogenetic haploid production was 36,000 erg mm(-2). At 15 degrees C, appropriate timing of cold shock for retention of the second polar body in turbot eggs was at 6 min after fertilization. Results of different combinations of two shock temperatures (1 or 3 degrees C) and four shock durations (15, 25, 35 or 45 min) at 6 min after fertilization demonstrated that shock of 25 min at 1 degrees C gave the highest production of diploid gynogens (39.58% relative to its diploid control). The results of this study reveal that the use of UV-irradiated P. olivaceus sperm for activation of turbot eggs and cold shock for polar body retention is an effective method to produce gynogenetic offspring.

Circadian rhythms in the growth and reproduction of the brown alga Undaria pinnatifida and gametogenesis under different photoperiods

Circadian growth rhythm of the juvenile sporophyte of the brown alga Undaria pirznatifida was measured with the computer-aided image analysis system in constant florescent white light under constant temperature (10 degrees C). The growth rhythm persisted for 4 d in constant light with a free-running period of 25. 6 h. Egg release from filamentous gametophytes pre-cultured in the light - dark regime was evaluated for six consecutive days at fixed time intervals in constant white light and 12 h light per day. Egg release rhythm persisted for 3 d in both regimes, indicating the endogenous nature. Temporal scale of egg release and gametogenesis in 18, 16, 12 and 8 h light per day were evaluated respectively using vegetatively propagated filamentous gametophytes. Egg release occurred 2 h after the onset of dark phase and peaked at midnight. Evaluation of the rates of oogonium formation, egg release or fertilization revealed no significant differences in four light-dark regimes, indicating; the great plasticity of sexual reproduction. No photoperiodic effect in gametogenesis in terms of oogonium formation and egg release was found, but fertilization in short days was significantly higher than in long days. Results of this investigation further confirmed the general occurrence of circadian rhythms in inter-tidal seaweed species.

Early development of germlings of Sargassum thunbergii (Fucales, Phaeophyta) under laboratory conditions

Morphology and culture studies on germlings of Sargassum thunbergii (Mertens et Roth) Kuntze were carried out under controlled laboratory conditions. Growth characteristics of these germlings grown under different temperatures (from 10 to 25A degrees C), irradiances (from 9 to 88 mu mol photons m(-2) s(-1)), and under blue and white light conditions are described. The development of embryonic germlings follows the classic "8 nuclei 1 egg" type described for Sargassaceae. Fertilized eggs spent 5-6 h developing into multicellular germlings with abundant rhizoids after fertilization. Under conditions of 20A degrees C, 44 mu mol photons m(-2) s(-1) and photoperiod of 12 h, young germlings with one or two leaflets reached 2-3 mm in length after 8 weeks. Temperature variations (10, 15, 20, 25A degrees C) under 88 mu mol photons m(-2) s(-1) significantly influenced the growth rate within the first week, although this effect became less obvious after 8 weeks, especially at 15 and 20A degrees C. Variation in germling growth was highly significant under different irradiances (9, 18, 44, 88 mu mol photons m(-2) s(-1)) at 25A degrees C. Low temperature (10A degrees C) reduced germling growth. Growth of germlings cultured under blue light was lower than in white light. Optimal growth of these germlings occurred at 25A degrees C and 44 mu mol photons m(-2) s(-1).

A Dicer-1 gene from white shrimp Litopenaeus vannamei: Expression pattern in the processes of immune response and larval development

Dicer is a member of the RNAase III family which catalyzes the cleavage of double-stranded RNA to small interfering RNAs and micro RNAs, and then directs sequence-specific gene silencing. In this paper, the full-length cDNA of Dicer-1 was cloned from white shrimp Litopenaeus vannamei (designated as LvDcr1). It was of 7636 bp, including a poly A tail, a 5' UTR of 136 bp, a 3' UTR of 78 bp, and an open reading frame (ORF) of 7422 bp encoding a putative protein of 2473 amino acids. The predicted amino acid sequence comprised all recognized functional domains found in other Dicer-1 homologues and showed the highest (97.7%) similarity to the Dicer-1 from tiger shrimp Penaeus mondon. Quantitative real-time PCR was employed to investigate the tissue distribution of LvDcr1 mRNA, and its expression in shrimps under virus challenge and larvae at different developmental stages. The LvDcr1 mRNA could be detected in all examined tissues with the highest expression level in hemocyte, and was up-regulated in hemocytes and gills after virus injection. These results indicated that LvDcr1 was involved in antiviral defense in adult shrimp. During the developmental stages from fertilized egg to postlarva VII, LvDcr1 was constitutively expressed at all examined development stages, but the expression level varied significantly. The highest expression level was observed in fertilized eggs and followed a decrease from fertilized egg to nauplius I stage. Then, the higher levels of expression were detected at nauplius V and postlarva stages. LvDcr1 expression regularly increased at the upper phase of nauplius, zoea and mysis stages than their prophase. The different expression of LvDcr1 in the larval stages could provide clues for understanding the early innate immunity in the process of shrimp larval development. (C) 2010 Elsevier Ltd. All rights reserved.