Cultivation of the intertidal brown alga Hizikia fusiformis (Harvey) Okamura: mass production of zygote-derived seedlings under commercial cultivation conditions, a case study experience

Mariculture of the brown alga Hizikia fusiformis (Harvey) Okamura as an export-oriented human food has been there more for than 20 years in China. It is now one of the five major farmed algal species along the Chinese coast. Stable and sufficient supply of young seedlings for scaling up the cultivation has been a problem throughout the farming history of this species due to the unique dioecious life cycle and relatively short time window of sexual reproduction in nature. These two factors led to a practical difficulty in obtaining zygotes at identical developmental stage in viable amounts for seedling production. A key solution to this problem is to control the synchronization of the receptacle development and to realize the simultaneous discharge of male and female gametes, such that the fertilization rate could be greatly enhanced. Focusing on one of the farmed populations in this report, we present our results on mass production of seedlings using the synchronization technique on a large scale performed in 2007. Totally 5.5 hundred million embryos were obtained from 100 kg female sporophytes. The seedlings were raised up to 3.5 mm in length in greenhouse tanks over a month and were further grown in open sea for over 3 months at two experimental sites. The success of mass production of seedlings in this alga helped to lay the basis for future trials in other species in the genus of Sargassum that have identical life cycle.

A preliminary genetic map of Zhikong scallop (Chlamys farreri Jones et Preston 1904)

Zhikong scallop (Chlamys farreri Jones et Preston 1904) is one of the most important aquaculture species in China. The development of a genetic linkage map would provide a powerful tool for the genetic improvement of this species. Amplified fragment length polymorphism (AFLP) is a PCR-based technique that has proven to be powerful in genome fingerprinting and mapping, and population analysis. Genetic maps of C. farreri were constructed using AFLP markers and a full-sib family with 60 progeny. A total of 503 segregating AFLP markers were obtained, with 472 following the Mendelian segregation ratio of 1:1 and 31 markers showing significant (P< 0.05) segregation distortion. The male map contained 166 informative AFLP markers in 23 linkage groups covering 2468 cM. The average distance between markers was 14.9 cM. The female genetic map consisted of 198 markers in 25 linkage groups spanning 3130 cM with an average inter-marker spacing of 15.8 cM. DNA polymorphisms that segregated in a 3:1 ratio as well as the AFLP markers that were heterozygous in both parents were included to construct combined linkage genetic map. Five shared linkage groups, ranging from 61.1 to 162.5 cM, were identified between the male and female maps, covering 431 cM. Amplified fragment length polymorphism markers appeared to be evenly distributed within the linkage groups. Although preliminary, these maps provide a starting point for the mapping of the functional genes and quantitative trait loci in C. farreri.

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.

Assessment of genetic diversities of selected Laminaria (Laminariales, Phaeophyta) gametophytes by inter-simple sequence repeat analysis

Inter-simple sequence repeat (ISSR) analysis was used to assess genetic diversity among 10 pairs of male and female Laminaria gametophytes. A total of 58 amplification loci was obtained from 10 selected ISSR primers, of which 34 revealed polymorphism among the gametophytes. Genetic distances were calculated with the Dice coefficient ranging from 0.006 to 0.223. A dendrogram based on the unweighted pair-group method arithmetic (UPGMA) average showed that most male and female gametophytes of the same species were clustered together and that 10 pairs of gametophytes were divided into four groups. This was generally consistent with the taxonomic categories. The main group consisted of six pairs of gametophytes, which were selected from Laminaria japonica Aresch. by intensive inbreeding through artificial hybridization. One specific marker was cloned, but was not converted successfully into a sequence characterized amplified region (SCAR) marker. Our results demonstrate the feasibility of applying ISSR markers to evaluate Laminaria germplasm diversities.

Growth promotion of vegetative gametophytes of Undaria pinnatifida by blue light

Through an acclimation period of 10 days, compared to white light, the maximal net photosynthetic rates were significantly higher for gametophytes of Undaria pinnatifida cultivated under blue light (400-500 nm), and were lower under red light (600-700 nm). Chlorophyll c and the carotenoid content of gametophytes were similar under blue light and red light but were much lower under white light. The growth rate of female gametophytes under blue light was higher than that under other lights, and the growth rate of male gametophytes showed little variation with respect to blue and white light. Male and female gametophytes were mixed together to form sporophytes under white, blue and red light. After approximately 5 days, 50% gametophytes became fertile under blue and white light, but remained vegetative under red light after 10 days.

New life cycles of Porphyra katadae var. hemiphylla in culture

Porphyra katadae Miura var. hemiphylla Tseng et T. J. Chang, a species distributed around the Liaodong and Shandong Peninsulas of China, produces gametophytes from late winter to early spring. These are monoecious with male and female reproductive tissues in distinct halves or sectors. Vegetative tissues from sectors expected to differentiate into sexual tissue were cultured in the laboratory. Male and female reproductive organs, as well as conchocelis and blades, were differentiated from these tissues. The male and female reproductive tissues were in patches and mixed on the cultured tissue pieces. This was quite different from the wild-type sectored individuals. The F-1 conchospore germlings also produced monospores, carposporangia, spermatangia and conchocelis. These carposporangia and spermatangia were in patches and were mixed on the F-1 fronds. The results imply that P. katadae var. hemiphylla is possibly sex-differentiated rather than sex-determined. This is the first report of such a dimorphic life history in the genus Porphyra.

DNA fingerprinting of selected Laminaria (Phaeophyta) gametophytes by RAPD markers

Random amplified polymorphism DNA (RAPD) analysis was applied to germplasm characterization in 33 different selected Laminaria male and female gametophytes. The positional homology of the RAPD analysis using sequence characterized applied region (SCAR) method was successfully conducted. A total of 233 polymorphic loci were obtained from 18 selected primers after screening, of which 27 stable and clear bands were selected to construct a fingerprint map for discrimination of each gametophyte. Seven RAPD markers from five primers were finally determined by a computer program to construct the fingerprint map. Three specific markers closely related with gametophytes were obtained and were converted to gametophytic SCAR markers, the first SCAR marker report on Laminaria germplasm and applicable to cultivars identification. These results demonstrated the feasibility of applying RAPD markers to germplasm characterization in selected Laminaria gametophytes, and can provide a molecular basis for breeding new Laminaria strains. (C) 2004 Elsevier B.V. All rights reserved.

Genetic analysis of the gametophytes of Undaria pinnatifida (Phaeophyceae) with ISSR method

Inter-simple sequence repeat (ISSR) analysis was used to assess eleven pairs of Undaria pinnatifida (Harv.) Suringar male and female gametophytes. After screening fifty primers, 18 ISSR primers were selected for final analysis. A total of 104 loci were obtained, of which 77 were polymorphic, among the gametophytes studied. Genetic relationships were analyzed with simple matching (S), Jaccard's (J) and Dice's (D) distance coefficients. Little genetic variations were found among the selected Undaria gametophytes, for instance, the genetic distances ranging from 0.010 to 0.125 with Dice coefficients. UPGMA dendrograms showed that 11 pairs of Undaria gametophytes were distributed into five groups. Most Undaria strains cultivated in China exhibited closely genetic relationships with the strains from Japan. However, gametophytes from Qingdao appeared as distinct clades from other Undaria strains with all three distance coefficients used. Mantel test showed that the three distance measurements generated congruent clustering patterns on the same data. Our results demonstrated the feasibility of applying ISSR markers for genetic analysis of Undaria gametophytes. (c) 2006 Elsevier B.V. All rights reserved.

高原鼢鼠的挖掘行为及其与土壤硬度的关系(英文)

Burrowing behaviors of the plateau zokors ( Myospalax bailely) in relation to soil hardness were investigated by using radio2telemetry at Haibei Alpine Meadow Ecosystem Research Station ,Qinghai , China. The results showed that there were no differences of burrowing behaviors and the efficiencies of the excavating segment between male and female , except the digging duration of the male was longer than that of the female in the same soil. With increase of soil hardness , digging duration in incisors significantly increased , but dried soil mass dug out by the zokors in each bout reduced. Mean-while the field investigation also showed that soil hardness affected the distribution of the zokor.

工科大学生心理应激、应对与自我概念的关系研究

In the period of college, an individual matures rapidly in all aspects. College engineering students are the important parts of undergraduates. The state of an individual’s mental health may affect and even decide his future life and work. The level of the student’s self-concept and the kind of coping styles the students adopt are directly related to their mental health. So, it is significant to study the psychological stress, coping and self-concept of college engineering students for the mental health education and research of college engineering students. Based on overviews of former research, with the China College Student Psychological Stress Scale, the Coping Styles Scale and the Tennessee Self-Concept Scale, 559 college engineering students were investigated to explore the characteristics of and the relationship between the psychological stress, coping styles and self-concept of college engineering students. The results showed: 1. The stresses of learning, living and daily hassles were the main psychological stresses of college engineering students. There were significant differences in psychological stress between students from the countryside and those from urban areas, between needy students and non-needy students, between single-parent students and non-single-parent students, among students from different grades, with different academic achievements and of different postgraduate targets, between student party members and non-party members, between student cadres and non-cadres. However, there were no significant differences between male and female, between those from single-child families and from multiple-child families. 2. The coping styles of solving problem, seeking help and rationalization were the main coping styles of college engineering students. There were significant differences in the coping styles between needy students and non-needy students, among students from different grades, with different academic achievements and of different postgraduate targets, between student party members and non-party members, between student cadres and non-cadres. However, there were no significant differences between students from the countryside and from urban areas, between male and female, between single-parent students and non-single-parent students, between those from single-child families and from multiple-child families. 3. The self-concept of college engineering students was positive in general. There were significant differences in self-concept between students from the countryside and those from urban areas, between male and female, between needy students and non-needy students, between single-parent students and non-single-parent students, among students from different grades, with different academic achievements and of different postgraduate targets, between student party members and non-party members, between student cadres and non-cadres. However, there were no significant differences between those from single-child families and from multiple-child families. 4. The psychological stress had significantly negative correlation to the immature coping styles, and had partial correlation to the mature coping styles. Coping style has significant predictability on psychological stress. 5. The positive factors of the self-concept had significantly negative correlation to psychological stress, but self-criticism had positive correlation to psychological stress. There are significant differences between high self-concept students and low self-concept students for psychological stress. Self-concept has significant predictability on psychological stress. 6. The positive factors of the self-concept had significantly negative correlation to the coping styles of self-blame, illusion, avoidance, and rationalization, but had significantly positive correlation to the coping style of solving problem and seeking help. Self-criticism had significantly negative correlation to the coping styles of self-blame, illusion, avoidance, and rationalization. There are significant differences between high self-concept students and low self-concept students for coping styles. Self-concept has significant predictability on coping styles. 7. The self-concept of college engineering students had an effect on psychological stress by coping styles. However, the effect by the immature coping styles was higher than that to the mental health directly, and the effect by the mature and mixed coping styles was slighter than that to the mental health directly. According to the results, improving the college engineering students’ self-concept level and establishing right self-concept, developing the middle school student’ active coping styles and overcoming the negative coping styles are essential and important to the college engineering students’ mental health and provide useful clues for the psychological education of the college engineering students.

攻击性、情绪调节评价对青少年攻击行为的作用

The aggressive behavior and the violent crime of the Adolenscents have aroused social widespread attention. The Social Implicit Cognition Method could overcome the insufficiency of explicit research methods such as Self-Report Scale, and carry on the thorough discussion to the intrinsic mechanism of the Adolenscent’s aggressive behavior. In this research, Implicit Association Test and Explicit Questionnaire are synthetically applied to inspect the relations among the aggression, Evaluation of Emotion Regulation and the aggressive behavior of the normal middle-school students and the Adolenscent violent criminals under the explicit and implicit conditions, and have carried the supplement to the internal condition of the General aggression model (GAM). The result indicates, 1. The concept of implicit aggression exists independently and separates from explicit aggression. However the relations between implicit aggression and the groups are remarkable, i.e. both the implicit and explicit aggressions of the Adolenscent violent criminals are remarkably higher than those of the normal middle-school students, but not influenced by sex factors. 2. Implicit Evaluation of Emotion Regulation is suitable to Chinese Adolenscents. The correlation of groups and sex factors exist to Implicit Evaluation of Emotion Regulation, i.e. in the group of criminals, the female inclines to the mood control compared with the male in the aspect of Implicit Evaluation of Emotion Regulation; however, in the group of normal persons, the difference between male and female is not remarkable in the aspect of Implicit Evaluation of Emotion Regulation. 3. Under the explicit conditions, it is discovered that the remarkable explicit aggression can predict violent criminals. But it is adjusted by Explicit Evaluation of Emotion Regulation, i.e. the predicting function is not remarkable when the Explicit Evaluation inclines to expression. Meanwhile, when checking the functions and the effects of the implicit variables after the explicit conditions are under control, it is discovered that the predicting function of implicit aggression is still remarkable, and the predicting function of Implicit Evaluation of Emotion Regulation is remarkable at the edge and also adjusted by sex factors. These findings help us not only understanding thoroughly the intrinsic mechanism of the Adolenscent’s aggressive behavior, discovering the moderating role of Evaluation of Emotion Regulation on aggression, but also providing an academic foundation on the prevention and intervention of the Adolenscent’s aggressive behavior.