59 resultados para Sexual reproduction


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As a large conspicuous intertidal brown alga, individuals of Sargassum horneri can reach a length of more than 7 m with a fresh weight of 3 kg along the coasts of the Eastern China Sea. The biomass of this alga as a vital component in coastal water ecology has been well documented. In recent years, a steady disappearance of the algal biomass along the once densely populated coastal areas of the Eastern China Sea has drawn attention in China. Efforts have been made to reconstruct the subtidal algal flora or even to grow the alga by use of long-lines. As part of the efforts to establish an efficient technique for producing seedlings of S. horneri, in this investigation a series of culture experiments were carried out in indoor raceway and rectangular tanks under reduced solar irradiance at ambient temperature in 2007-2008. The investigation demonstrated that: (1) sexual reproduction of S. horneri could be accelerated in elevated temperature and light climates, at least 3 months earlier than in the wild; (2) eggs of S. horneri had the potential to be fertilized up to 48 h, much longer than that of known related species; (3) suspension and fixed culture methods were both effective in growing the seedlings to the long-line cultivation stage; and (4) the life cycle of S. horneri in culture could be shortened to 4.5 months, thus establishing this alga as an appropriate model for investigating sexual reproduction in dieocious species of this genus.

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The maturation pattern of sexual reproduction in Hizikia fusiformis (Harvey) Okamura (Sargassaceae, Phaeaophyta) was examined in 2003 at Yunao Bay, Nanao Island, Shantou, China. Maturation began in mid-April (seawater temperature 19-21 degrees C), reached the peak in mid-May (maturation rate ca. 70%, and seawater temperature 23.5-25 degrees C) and finished in late-June (seawater temperature 27.5-30 degrees C). The Hizikia plants continued to gain the length from the beginning of maturation season to reach a maximum mean length of 34.8 cm in mid-May, after which the mean length was reduced drastically due to the senescence and rupture of the larger plants in size. The major portion of the mature plants belonged to the larger plants between April and May, but to the smaller ones in June. It is suggested that the plant must achieve a critical size before reproductive maturation occurred. There was a positive relationship between the number of receptacles (NR), as well as the reproductive allocation (RA), and the plant size of Hizikia population, with the recorded maximum values of NR and RA being 1220 and 64.3% respectively, for a single plant.

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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.

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羊草 (Leymus chinensis (Trin.) Tzvel. ) ,隶属禾本科赖草属,是欧亚大陆草原区东部的重要草原建群种之一。羊草是牧草之王,属于我国有比较优势的战略性生物资源,对我国北方畜牧业发展以及保护生态环境均具有重要作用。 羊草较弱的有性生殖特性限制了其应用,本文从实验生物学角度,研究了羊草有性生殖的基本特点,并试图通过现代分子生物学手段探讨自交不亲和性的有关机理。本论文的主要结果如下: 1. 实验发现羊草具有自交不亲和性。以6 个羊草居群为材料,测定得知开放授粉时的结实率在6.5% - 56.7%之间,自交时结实率为0.6% - 4.3%,差异极其显著; FDA 染色法检测结果显示羊草成熟花药中有活性的花粉达到92.2% 以上;在发育时间顺序和空间结构上,羊草雌蕊、雄蕊适于异花和自花授粉;花粉柱头亲和性实验表明,自交花粉只有5.5%-11.7% 是亲和的,杂交花粉亲和率达到了60.0%-84.8%,说明自交花粉在柱头上萌发受到抑制,其次,荧光显微镜还观察到“不亲和花粉”在进入柱头后生长缓慢,或停止延伸。 2. 初步确定羊草自交不亲和性具有配子体型遗传特点。以不同居群羊草杂交后的姊妹系作为实验材料,观察到自交组合的亲和率变幅为0 % - 6.9 %,杂交组合的亲和率具有连续性变异和变幅较宽的特点(47.5% - 96.0 %),且正反交结果具有一定的一致性(88.2%),表现出配子体遗传特性。 3. 羊草居群内结实率存在一定变异。以羊草单株为单位分别进行自交、随机互交和开放授粉,结果显示三者的平均结实率分别为4.6%,18.1% 和35.7%,株间的变异系数分别为33.4%,21.2%和17.1%,这些株间的变异均达到统计上的显著差异;同时羊草自交、杂交和开放授粉之间具有一定的相关性,显示羊草的这种株间差异与株系本身的生理特性相关。 4. 分离了羊草硫氧化还原蛋白 H 基因(ThioLc)并对其功能进行了分析。克隆了ThioLc全长和cDNA序列。序列分析结果显示,DNA全长2257 bp,包括3 个内含子和4 个外显子,与水稻Thio h 的cDNA 序列相比,具有 32.0% 的同源性;Southern 杂交显示 ThioLc 在羊草基因组中是单拷贝;Northern 杂交显示 ThioLc 在羊草根、茎、叶和幼小的雌蕊中没有表达, 在成熟雌蕊和幼小的花粉中微量表达, 在成熟花粉中大量表达,说明分离的羊草硫氧化还原蛋白H 基因具有花粉特异表达特点。 5. 原核表达的ThioLc 蛋白具有较高的催化活性。构建了ThioLc 基因的原核表达载体,检测证明ThioLc基因在大肠杆菌中正常表达;提取表达蛋白,纯化,用胰岛素和二硫苏糖醇反应体系进行硫氧化还原蛋白的催化作用反应,结果表明表达的蛋白具有催化活性。这一结果为进一步搜寻靶向蛋白和研究该蛋白的结构、功能和作用方式奠定了基础。

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预计到本世纪末,大气CO2浓度将会增加到540~970ppm,大气CO2浓度升高所引起的全球气候变化已经受到广泛的关注。植物生长依赖CO2,并且对大气CO2浓度升高在结构和生理上产生响应。目前已有大量报道,从生态系统、群落、种群、个体、器官、组织、生理以及生化等水平上研究高浓度CO2所对植物产生的影响。但是有关高浓度CO2对植物有性生殖影响的报道却很少,同时多数实验均建立在短期的生殖响应,忽视了植物在长期高CO2浓度下具有的反馈作用和CO2浓度变化对植物的驯化作用。植物有性生殖与其生态适应性和农作物籽粒产量的关系极为密切;同时,植物有性生殖特性的变化,也可作为预测植物对全球气候变化响应的重要指标之一。为此,利用高浓度CO2对植物进行长期选择实验将很有必要。研究结果将为预测未来大气CO2浓度增加的条件下陆地生态系统的演变趋势、全球变化对植物有性生殖响应的方式和机制提供新的思路和有效方法。   在本研究中,我们以模式植物拟南芥(Arabidopsis thaliana)作为实验材料,利用370和700ppm CO2对其进行连续8个世代处理,首先研究高浓度CO2对每一个世代的拟南芥有性生殖特性的影响,然后比较各个世代中各种生殖特性指标变化的规律,从细胞、组织和个体尺度上揭示拟南芥有性生殖对全球变化的响应模式。此外,在700ppm CO2处理下,我们对拟南芥叶片生理、生化以及结构的变化进行了相关研究。两部分研究结果及主要结论如下:   首先,在每一个世代中,与370ppm CO2相比较,700ppm CO2处理显著促进了拟南芥开花,缩短生长周期,增加花、角果及种子等生殖的产量,降低种子N含量,提高种子C/N比、种子千粒重以及生殖生物量所占总生物量的比例等,而对种子萌发率、角果所含种子数目以及角果长度则无显著影响。但是, 通过对相同CO2浓度处理条件下,不同世代之间的研究结果比较发现,不同世代之间相关的生殖生物学指标并无显著差异。   其次,高浓度CO2显著降低叶片气孔密度、气孔指数、气孔导度以及蒸腾速率。在高浓度CO2处理下,叶肉细胞中叶绿体数目、叶绿体宽度和表观面积、淀粉粒大小和数量、叶片和细胞壁厚度等都显著增加,但是基粒内囊体膜的数量却显著下降。叶片中碳水化合物如可溶性总糖、淀粉以及纤维素含量在高浓度CO2下分别显著增加71.9%、78.7% 和 22.3%。此外,在高浓度CO2处理下,叶片中多数激素如如吲哚乙酸(indole-3-acetic acid, IAA)、赤霉素(gibberellin, GA)、玉米素核苷(zeatin riboside, ZR)、二氢玉米素核苷(dihydrozeatin riboside, DHZR)和异戊烯基腺苷(isopentenyl adenosine, iPA)均都显著地增加,而脱落酸(abscisic acid, ABA)含量却有所下降。最后,叶片中各种矿物质元素含量如N、P、K、Ca和Mg等含量在高浓度CO2处理下也都显著下降,而C/N比增加24.8%。   以上结果表明:   (1) 在每一个世代中,700ppm CO2处理对拟南芥各种有性生殖特性具有显著的影响,但是高浓度CO2处理对植物所引起的效应在多个世代以内并不能够传递给后代,所以在多个有性生殖世代内,高浓度CO2处理对植物生长、生殖没有驯化作用。   (2) 在高浓度CO2处理下,拟南芥叶片中叶绿体超微结构的变化,可能主要是由于叶绿体中淀粉粒数量和体积大小显著增加而引起。   (3) 在高浓度CO2处理下,由于拟南芥叶片内与促进细胞分裂与伸长的激素含量显著增加,从而对拟南芥植株生长发育速率的提高起了重要的作用。   (4) 拟南芥生长在高浓度CO2条件下,其叶片中各种矿质元素含量(如N、P、K、Ca和Mg)均显著降低,究其原因可能是,第一由于叶片中碳水化合物含量的显著增加而对矿物质元素具有稀释作用;第二由于蒸腾速率下降,引起矿质元素从根部随着蒸腾流运输到地上部分的含量相应减少。   

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To investigate germline development and germ cell specification, we identified a Dazl homolog (CagDazl) from gynogenetic gibel carp (Carassius auratus gibelio). Its cDNA sequence and BAC clone sequence analyses revealed the genomic organization conservation and conserved synteny of the Dazl family members and their neighborhood genes among vertebrates, especially in fish. Moreover, a polyclonal antibody specific to CagDazl was produced and used to examine its expression and distribution throughout germline development at protein level. Firstly, ovary-specific expression pattern of CagDazl was confirmed in adult tissues by RT-PCR and Western blot. In addition, in situ hybridization and immunofluorescence localization demonstrated its specific expression in germ cells, and both its transcript and protein were localized to germ plasm. Then, co-localization of CagDazl and mitochondrial cloud was found, confirming that CagDazl transcript and its protein are germ plasm component and move via METRO pathway during oogenesis. Furthermore, the CagDazl is abundant and continuous throughout germline development and germ cell specification including primordial germ cell (PGC) formation, oogonium differentiation, oocyte development, and embryogenesis, and the dynamic distribution occurs at different development stages. The data suggest that maternal CagDazl might play an important role in gibel carp PGC formation. Therefore, CagDazl is a useful and specific marker for tracing germ plasm and germ cell development in the gynogenetic gibel carp. In addition, in comparison with previous studies in sexual reproduction species, the continuous and dynamic distribution of CagDazl protein in the germ plasm throughout the life cycle seems to have significant implication in sex evolution of vertebrates. J. Exp. Zool. (Mol. Deu. Euol.) 312B:855-871, 2009. (C) 2009 Wiley-Liss, Inc.

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Although the peritrichous ciliate Carchesium polypinum is common in freshwater, its population genetic structure is largely unknown. We used inter-simple sequence repeat (ISSR) fingerprinting to analyze the genetic structure of 48 different isolates of the species from four lakes in Wuhan, central China. Using eight polymorphic primers, 81 discernible DNA fragments were detected, among which 76 (93.83%) were polymorphic, indicating high genetic diversity at the isolate level. Further, Nei's gene diversity (h) and Shannon's Information index (I) between the different isolates both revealed a remarkable genetic diversity, higher than previously indicated by their morphology. At the same time, substantial gene flow was found. So the main factors responsible for the high level of diversity within populations are probably due to conjugation (sexual reproduction) and wide distribution of swarmers. Analysis of molecular variance (AMOVA) showed that there was low genetic differentiation among the four populations probably due to common ancestry and flooding events. The cluster analysis and principal component analysis (PCA) suggested that genotypes isolated from the same lake displayed a higher genetic similarity than those from different lakes. Both analyses separated C. polypinum isolates into subgroups according to the geographical locations. However, there is only a weak positive correlation between the genetic distance and geographical distance, suggesting a minor effect of geographical distance on the distribution of genetic diversity between populations of C. polypinum at the local level. In conclusion, our studies clearly demonstrated that a single morphospecies may harbor high levels of genetic diversity, and that the degree of resolution offered by morphology as a marker for measuring distribution patterns of genetically distinct entities is too low.

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有性繁殖是植物适应干扰的重要机制。目前,人们对于植物如何从有性繁殖方面适应风沙环境还知之甚少。本论文以半干旱草原区的科尔沁沙地为对象,采用实验室分析、模拟试验、野外调查等方法,研究了种子形态及其功能意义、种子传播与植物沙生适应性的关系、埋藏种子萌发与沙丘植物更新的关系,探讨了沙丘土壤种子库时空格局、土壤种子库持久性与沙丘植被恢复的关系,揭示了生殖物候对植物沙生适应性的影响,探索了有性繁殖在多年生沙生植物适应风蚀中的作用。 研究结果表明:(1)沙生植物通过调节种子重量和形状降低种子位移,增大幼苗补充几率;(2)沙生植物通过将种子脱落推迟至“风季结束、雨季来临”时期,或者通过种子遇水溶出粘液粘沙增大重量,分摊干旱和风沙干扰给幼苗补充带来的风险,维持种群繁衍;(3)埋藏能调节沙生植物种子萌发时间和萌发率;(4)沙生植物通过生殖物候可塑性和延长结实期增强沙生适应性;(5)沙生植物具有持久种子库,能提供稳定种源;(6)多年生沙生植物可通过有性繁殖适应风蚀,入侵流动沙丘区。本研究为植物有性繁殖研究提供了方法论借鉴。根据研究结果我们提出了沙丘植被恢复和管理建议。

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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.

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Nets in traditional Porphyra mariculture are seeded with conchospores derived from the conchocelis phase, and spend a nursery period in culture tanks or calm coastal waters until they reach several centimeters in length. Some species of Porphyra can regenerate the foliose phase directly through asexual reproduction, which suggests that the time, infrastructure, and costs associated with conchocelis culture might be avoided by seeding nets with asexual spores. Here, we present work from a short-term mariculture study using nets seeded with asexual spores (neutral spores) of a native Maine species of Porphyra. Porphyra umbilicalis (L.) Kutzing was selected for this proof of concept research because of its reproductive biology, abundance across seasons in Maine, and evidence of its promise as a mariculture crop. We studied the maturation, release, and germination of the neutral spores to develop an appropriate seeding protocol for nets, followed by development of a nursery raceway to provide an easily manipulated environment for the seeded nets. Neutral spores were produced throughout the year on the central Maine coast,however, there was a temporal variability in the number and survival of released neutral spores, depending upon thallus position in the intertidal zone. Small thalli were strictly vegetative, but most thalli reproduced by neutral spores- sexual reproduction was absent. Neutral spores germinated quickly at 10 and 15 'C, but germination was delayed at 5 degrees C. Unlike some algal zygotes and spores, neutral spores of R umbilicalis required light to germinate; however, irradiances of 25 and 100 mu mol photons M-2 S-1 were equally sufficient for germination. Rafts of seeded nets were deployed in Cobscook Bay, Maine, at two distances from salmon aquaculture pens and at a control site on a nearby, fallow aquaculture site (no salmon). There was no difference in nitrogen content of harvested thalli; however, both the density and the surface area of harvested thalli were different among the sites. The possible causes of these differences are discussed in the context of potential use of P umbilicalis in IMTA. (C) 2007 Elsevier B.V. All rights reserved.

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Commercial farming of the intertidal brown alga Hizikia fusiformis (Harvey) Okamura in China and South Korea in the sea depends on three sources of seedlings: holdfast-derived regenerated seedlings, young plants from wild population and zygote-derived seedlings. Like many successfully farmed seaweed species, the sustainable development of Hizikia farming will rely on a stable supply of artificial seedlings via sexual reproduction under controlled conditions. However, the high rate of detachment of seedlings after transfer to open sea is one of the main obstacles, and has limited large-scale application of zygote-derived seedlings. To seek the optimal condition for growing seedlings on substratum in land-based tanks for avoidance of detachment in this investigation, young seedlings were grown in both outdoor tanks exposed directly to sunlight and in indoor raceway tanks in reduced, filtered sunlight. Results showed that young seedlings, immediately after fertilization, could withstand a daily fluctuation of direct solar irradiance up to a level of 1800 mu mol photons m(-1)s(-1), and maintained a faster growth rate than seedlings grown in indoor tanks. Detailed experiments by use of chlorophyll fluorescence measurements further demonstrated that the overnight (12 h) recovery of optimal fluorescence quantum yield (F-v/F-m) of seedlings after 1 h treatment at 40 degrees C was 98%, and the 48 h recovery of F-v/F-m of seedlings after 1 h exposure to 1800 mu mol m(-2)s(-1) was 92%. Forty-one-day-old seedlings showed no significant decrease of optimal fluorescence quantum yield at salinity ranging from 30 to 5 ppt for a treatment up to 17 h. Six-hour desiccation treatment did not have any influence on the optimal fluorescence quantum yield. Exposure to 18 mmol L-1 sodium hypochlorite for 10 min did not damage the PSII efficiency, and thus could be used to remove epiphytic algae. The strong tolerance of young seedlings to high temperature, high irradiance, low salinity and desiccation found in this investigation supports the view that mass production of Hizikia seedlings should be performed in ambient light and temperature instead of in shaded greenhouse tanks.

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Bangia fuscopurpurea (Rhodophyta) was cultivated in Putian (Fujian province, China). The characteristics of the life history concerned with cultivation were investigated and the cultivation procedure was presented. The gametophytic phase (thallus) and the sporophytic phase (conchocelis) occurred alternately in the life history of B. fuscopurpurea. Young thalli produced archeospores, and the number depended on the environmental factors. Temperature affected the number of archeospore release and percent of germination, and photo flux density (PFD) mainly affected the time of spore release and germination. Thalli matured from December to February and developed into the conchocelis phase through sexual reproduction. The conchocelis grown in shells had three developmental stages: vegetative conchocelis, conchosporangiall formation and conchospore formation. Pit-connections were present in the first 2 stages but absent after conchospore formation. Vegetative conchocelis and conchosporangial. branches can transform into each other. However, conchospores only developed into the gametophytic phase. Cultivation of B. fuscopurpurea was based on characterization of the life history, consisting of 3 steps: zygotospores collection, indoor cultivation of conchocelis and outdoor cultivation of thalli. Young thalli that developed from conchospores produced numerous archeospores before December. Over 90% of the crop was from the development of archeospores. The results indicated that conchosporelings were a good source of archeospores, and the development of the large quantity of archeospores acted as a more prevailing means to increase the population size. (C) 2008 Elsevier B.V. All rights reserved.

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Commercial cultivation of the dioecious brown macroalga Hizikia fusiformis (Harvey) Okamura in East Asia depends on the supply of young seedlings from regenerated holdfasts or from wild population. Recent development of synchronized release of male and female gametes in tumble culture provides a possibility of mass production of young seedlings via sexual reproduction. In this paper, we demonstrate that controlled fertilization can be efficiently realized in ambient light and temperature in a specially designed raceway tank in which the sperm-containing water has been recirculated. The effective fertilization time of eggs by sperm was found to be within six hours. Fast growth and development of the young seedlings relied on the presence of water currents. Velocity tests demonstrated that young seedlings of 2-3 mm in length could withstand a water current of 190 cm s(-1) stop without detachment. Culture experiments at 24 h postfertilization showed that elongation of both the seedlings and their rhizoids were not hampered by high irradiance up to 600 mu mol photons m(-2) stop s(-1) stop. However, growth was slightly retarded if cultured at a temperature of 16 degrees C compared to other culture temperatures of 22, 25 and 29 degrees C. No seedling detachment was observed after transfer of the young seedlings to raft cultivation in the sea after one and 1.5 months post-fertilization, indicating the feasibility of obtaining large quantity of seedlings in such a system.

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红毛菜(Bangia Lyngb.)属于红藻门,与紫菜属同属红毛菜科,其味道和营养都优于紫菜。目前红毛菜栽培产业已在我国福建莆田展开,但栽培技术还有待提高。海藻栽培技术的发展和成熟依赖于对其生长发育过程的认识。本研究针对红毛菜发育过程及相关光合生理展开,并初步探讨了一采自山西娘子关泉淡水红毛菜群体(FWB)的系统地位。 色素突变标记的壳孢子萌发特征表明最初两次分裂产生的4细胞决定了完整植株的形态建成。成熟植株,为雌雄异体。雌性生殖器果胞的标志性分化结构为原始受精丝,环境因子是促发原始受精丝发展的外部因素,其膨大程度随受精的延迟而增大。原孢子是主要的无性生殖孢子类型,在不良环境中,藻体也会形成内生孢子或休眠包囊,或者藻体断裂后重新形成完整的植株。 红毛菜的生长发育很大程度上受环境因子的控制。高温不利于配子体的发育,15-20 ºC比较适宜。红毛菜无性繁殖的最适温度-光照组合为20 ºC-8 h,有性繁殖为15 ºC-12h。 不同发育阶段,PSII实际光合效率(Y(II))与细胞的健康状况以及光合器官完整性及其在细胞内的分布有关,而与细胞的类型关系不大。健康的假根细胞、已分化未成熟的精子以及果孢子细胞均具有很高的Y(II)。色素体由中间位变为围周位,中央大液泡(营养藻丝)和大小纤维囊泡(成熟孢子与精子)的产生,使得细胞Y(II)降低。刚放散的壳孢子Y(II)很低,说明在壳孢子由贝壳基质释放到自由水体过程,光合作用受到一定程度抑制;而2h后,Y(II)开始恢复,rbcL的转录水平非常高,为孢子的萌发储备物质和能量需求。 在失水和低盐胁迫下,藻体均维持较高的Y(II)。干出处理至藻体重量不再变化,复水后Y(II)可回复初始水平。海生红毛菜在100%淡水培养基中(约20ºC)培养7天后,部分雄性藻体依然活着。从而体现了红毛菜位居高潮带的生理优势。 FWB终生行无性繁殖,藻体形态与发生以及染色体数目(4条)与海生群体没有区别。而rbcL-rbcS Spacer序列显示,红毛菜海生群体(无性和有性)具有完全相同的序列,而FWB与它们有5bp差异,但是与欧洲、北美地区的淡水群体仅1bp不同,初步说明所有淡水红毛菜群体具有共同的原始起源。