靖海湾条斑紫菜的营养及鲜味评价

以不同收割期的靖海湾条斑紫菜(Pyropia yezoensis)为研究对象,对比分析营养价值并评价其鲜味。通过测定基本营养成分、无机元素组成和氨基酸组成,分析其营养学特征,通过测定游离氨基酸与呈味核苷酸组成,进而计算滋味活性值(Taste active value, TAV)与味精当量(Equivalent umami concentration, EUC)。结果显示,靖海湾条斑紫菜蛋白质含量丰富,头水和二水紫菜的蛋白含量均在49%以上,显著高于四水和六水紫菜(P<0.05)。不同收割期条斑紫菜的脂肪含量较低,均小于1.1%。常量元素中P含量最低,K含量最高,约为2.59×104~5.52×104 mg/kg;微量元素以Fe含量最高,Zn次之,Co最低;Pb含量明显低于GB 2762-2017的限量要求。头水、二水紫菜与四水、六水紫菜的必需氨基酸及氨基酸总量间存在显著差异(P<0.05)。游离谷氨酸和次黄嘌呤核苷酸对条斑紫菜的鲜味起主要贡献作用,头水、二水、四水和六水紫菜的EUC(以干基计)依次降低,分别为239.71、190.03、108.05和51.56 g MSG/100 g。靖海湾条斑紫菜具有较高的营养价值,且收割前期的靖海湾条斑紫菜鲜味度较高,可作为开发调味品的原料。     

坛紫菜雌雄叶状体的细胞分化比较

以室内培养20～90 d的坛紫菜雌雄叶状体为研究材料,用酶解法分别获取单离细胞进行再生培养。在雌雄叶状体的体细胞再生体中,都出现9种不同发育类型。再生体发育类型的数目和比例与种藻日龄密切相关等结果,证实了离体培养的单离细胞发育成不同形态的再生体是基于其离体前处于不同分化时期所致;由壳孢子分化成性母细胞大致可划分成8个不同阶段。雌雄叶状体的细胞分化途经大致相同,但也有一定差异,雌性叶状体的细胞最终分化成雌性性母细胞,并产生大量的丝状体;而雄性叶状体的细胞最终分化成雄性性母细胞,绝大部分生成精子,但极少数产生丝状体。在雌雄叶状体的细胞再生体中,均产生 “类单孢子”并长成正常叶状体。雄性叶状体成熟较雌性早,与其细胞分化速度较快有关。成熟期不同的雌性品系观察结果表明,叶状体成熟越早、生长期越短,其体细胞分化速度也越快。     

条斑紫菜(Pyropia yezoensis)绿斑病病原菌的分离鉴定

绿斑病(Green spot disease)是一种常见的海区栽培条斑紫菜(Pyropia yezoensis)病害,在整个紫菜栽培期间都可能发生,以每年11~12月份最为严重,主要出现在幼叶期和成叶期。首先,在叶状体上出现红色或淡红色小斑,而后逐渐转变为绿色,病斑继续扩展,在叶状体表面形成若干孔洞,后期几乎整个藻体变绿。本研究对日照地区患绿斑病的条斑紫菜进行病原菌分离纯化,得到5株优势菌(编号为Y1~Y5),人工回感实验结果显示,Y1可以引起健康条斑紫菜发生绿斑病。对Y1进行了生理生化检测、16S rRNA、dnaA和dnaN基因序列分析,确定病原菌为海洋假交替单胞菌(Pseudoalteromonas marina)。对绿斑病的发病进程进行了观察,并检测了培养温度、海水比重和养殖密度等环境因子对绿斑病发生的影响,结果显示,高温和高密度养殖会加速绿斑病病情的发展,海水比重为1.022时,绿斑病发病较严重。本研究确定了一株引起条斑紫菜绿斑病的病原菌,并分析了部分理化因子对感染的影响,为条斑紫菜绿斑病的防控提供了基础数据。     

Dietary supplementation of red alga Pyropia spheroplasts on growth,feed utilization and body composition of sea cucumber,Apostichopus japonicus (Selenka)

A feeding experiment was conducted in a closed recirculating system to evaluate the effects of freeze‐dried spheroplasts prepared from Pyropia yezoensis (Ueda) on feed intake, growth and biochemical composition of sea cucumber, Apostichopus japonicus (Selenka). Pyropea spheroplasts (PS) were prepared through enzymatic treatment to break down the complex mixture of polysaccharides cell walls that might be easier for growth energy partitioning. Sea cucumbers were fed‐formulated diets with 10 (Diet 1), 30 (Diet 2) and 50 g/kg (Diet 3) inclusion level of PS. A diet without PS was used as a control (Diet 4). The experiment was conducted for 6 weeks maintaining water temperature 15 ± 1°C, photoperiod 18:06 hours (D:L). Feed was supplied ad‐libitum at 16.00 h once in a day, and the remaining feed and faeces were removed in the next day. Results showed that the highest growth was observed in the 50 g/kg PS diet compared to other treatments. Total weight gain, mean weight gain, net yield, protein efficiency ratio (PER) and protein gain (%) were significantly higher in the 50 g/kg PS diet (p < .05). A significantly higher percentage of energy was allocated for growth in the 50 g/kg PS diet. The highest specific growth rate and feed conversion efficiency (p < .05) were observed in the higher percentage of PS diet. Both the growth performance and biochemical analysis showed that superior growth was observed with increasing levels of PS in the diet. We infer that PS can be used as a new, cheaper feed ingredient in the formulated diet of A. japonicus.     

坛紫菜叶状体的细菌性红烂病研究

本实验对发生在福建省平埠岛自然海区的野生坛紫菜（Porphyra haitanensis）叶状体上的红烂病进行了研究。患病叶状体上存在大小不等、肉眼可见的圆形或亚圆形病斑,镜检发现病斑内存在大量铁锈红色的死细胞和少量已解离的发绿或发白死细胞。将患病叶状体与健康坛紫菜叶状体共培养3d后,后者也出现了相同的红烂病,表明该病是由传染性病原侵入引起的。从患病叶状体中分离到一种病原菌,能感染健康叶状体使其出现相同的病症。该病原菌具有分泌毒素和微弱的消化琼胶能力,经高压灭菌或煮沸后的病原菌液,其杀死紫菜细胞的能力比未处理的病原菌液增加了数倍,这说明该菌可能通过释放内毒素来杀死叶状体细胞。鉴于此病是由于病原菌侵入叶状体并释放内毒素杀死紫菜细胞,且死亡细胞呈铁锈红色,故将其命名为“坛紫菜细菌性红烂病”。[中国水产科学,2008,15（2）：313—322]     

Anti-Aging Effects of R-Phycocyanin from Porphyra haitanensis on HUVEC Cells and Drosophila melanogaster

Aging has become a global public health challenge. Many studies have revealed that the excessive generation of ROS and oxidative stress could be the major causative factors contributing to aging. In this study, R-phycocyanin (R-PC) was isolated from Porphyra haitanensis, and its anti-aging ability was explored by natural aging Drosophila melanogaster and H₂O₂-induced HUVEC cells as the aging model. Results showed that R-PC α and β subunits expressed have antioxidant activity and can inhibit the generation of radicals, exhibiting a protective effect against H₂O₂-induced apoptotic HUVEC cells death. R-PC prevented the H₂O₂-induced HUVEC cell cycle phase arrest by regulating cell cycle-related protein. Furthermore, R-PC prevented the H₂O₂-induced HUVEC cell cycle phase arrest by regulating cell-cycle-related protein expression. In vivo study also indicated that R-PC significantly increased the survival time and alleviated the oxidative stress of Drosophila melanogaster. Moreover, R-PC notably decreased levels of ROS in natural aging flies and inhibited lipid peroxidation by enhancing the expressions of the endogenous stress marker genes (SOD1, SOD2, CAT of Drosophila melanogaster). Taken together, a study on the antioxidation extract from Porphyra haitanensis, such as R-PC, may open a new window for the prevention of anti-aging.     

紫菜养殖对养殖水体中细菌多样性分布及环境因子的影响

采用基于16S rDNA的高通量测序技术对山东长岛紫菜养殖区的细菌多样性分布进行了研究,并与环境因子进行关联分析,以探究紫菜养殖对环境的影响。结果显示,长岛海区水环境细菌多样性较丰富,呈现由近岸区(CDCNS)、养殖区(CDPF)、外海区(CDCOS)递减的趋势。分别基于所有细菌组成以及相对丰度最高的前20个属/种的聚类结果均显示,CDCNS和CDCOS区聚在一起,明显区别于CDPF区。CDPF区的特异优势菌包括鼠尾菌属(Muricauda)、假单胞菌属(Pseudomonas)、盐单胞菌属(Halomonas)、赤杆菌属(Erythrobacter)、海杆菌属(Marinobacter)。3个区域均未检测到河豚毒素假交替单胞菌(Pseudoalteromonas tetraodonis)、柠檬假交替单胞菌(Pseudoalteromonas citrea)等疑似紫菜致病菌。溶解氧(DO)和pH呈现由近岸向外海逐渐降低的趋势,总颗粒悬浮物(TSS)反之。CDPF区氨氮(NH4+-N)含量最低,盐度最高。关联分析发现,环境因子与环境微生物之间具有一定的相关性,如嗜氨菌属(Ammoniphilus)的丰度与NH4+-N 浓度具显著正相关性,CDPF区最低,CDCOS区最高;盐单胞菌属的分布与盐度具显著正相关性,CDPF区盐单胞菌属含量显著高于其他2个海区。研究表明,紫菜养殖海区细菌群落结构特征与环境因子具有关联性,其间的互作机制尚需进一步深入研究。     

坛紫菜转录因子NhbZIP1克隆和功能验证

高温是制约坛紫菜(Neoporphyra haitanensis)产业发展的主要因素之一,阐明坛紫菜高温胁迫应答机理对耐高温品种选育至关重要。人们已分离多个坛紫菜抗逆相关基因,但尚不清楚这些基因的表达调控机制。本研究通过分子生物学和生物信息学技术分离了坛紫菜转录因子NhbZIP1基因。该基因开放阅读框长825 bp,编码274个氨基酸。从开放阅读框推导的氨基酸序列有5个低复杂度区域和1个BRLZ结构。其中,BRLZ是bZIP家族的保守结构域,含有一个α卷曲螺旋结构(121～171aa)。实时荧光定量PCR(qRT-PCR)检测发现,NhbZIP1受高温胁迫显著诱导。为进一步阐明NhbZIP1的功能,将其转入莱茵衣藻(Chlamydomonas reinhardtii)中。结果显示,高温胁迫下转基因藻株生物量始终高于野生型,且随处理时间增加差异越来越显著。转基因藻株中热激蛋白家族和抗氧化系统相关基因的表达量显著高于野生型。研究表明,NhbZIP1激活下游抗逆基因表达,在坛紫菜应答高温胁迫中发挥重要作用。研究结果有助于阐明bZIP调控坛紫菜响应高温胁迫的分子机制,为耐高温新品种选育提供了基...     

Determination of the Extraction,Physicochemical Characterization,and Digestibility of Sulfated Polysaccharides in Seaweed—Porphyra haitanensis

The aim of the study was to extract Porphyra haitanensis polysaccharides (PHPs) using the water extraction and alcohol precipitation methods and explore their antioxidant activity and physicochemical properties. The single-factor and Box-Behnken response surface methodologies were used to optimize the extraction of polysaccharides from Porphyra haitanensis. Our results showed that the polysaccharide yield was as high as 20.48% with a raw material to water ratio of 0.04, and extraction time of 3 h at 80 °C. The extraction rate observed was similar to the actual extraction rate, thus proving the reliability of the optimization model. The extracted polysaccharides primarily consisted of galactose, glucose, and fucose in the molar ratio 76.2:2.1:1, respectively. The high performance gel permeation chromatography (HPGPC) results showed that the molecular weight of the PHPs obtained was 6.3 × 10⁵ Da, and the sulfate content was 2.7 mg/mL. Fourier infrared spectroscopy was used to analyze the functional groups and structures of the polysaccharides. The effect of concentration, temperature, and pH on the apparent viscosity of the PHPs solution were studied using rheology experiments, which revealed that PHPs were a “non-Newtonian fluid” with shear-thinning behavior. The viscosity of the PHPs gradually increased with increasing sugar concentration, and decreased with increasing temperature, acidity, and alkalinity. Detection of the antioxidant activity of OH*, DPPH*, and ABTS* revealed that the scavenging activity of ABTS* was higher than that of OH* and DPPH* in the concentration range of 1–5 mg/mL. In the experiments of simulating gastric juice and alpha amylase in vitro, it was found that PHPs can better resist digestion of alpha amylase, and have better resistance than fructooligosaccharide (FOS), so PHPs have potential prebiotic activity. These findings demonstrate the potential of PHPs for use in the food and cosmetic industries.     

用人工海水培养坛紫菜的初步研究

用天然海水与海水晶配置成的人工海水按10∶0,8∶2,6∶4,4∶6,2∶8,0∶10的体积比配成6种不同的培养液,用它们培养野生型坛紫菜叶状体。培养30 d后,叶状体的生长情况如下:在100%天然海水中培养的坛紫菜叶状体生长最快,其它组的生长快慢依次为:含20%>40%>60%>80%的海水晶人工海水组。在100%海水晶人工海水组培养的叶状体5 d后死亡。叶状体叶绿素a(Chl.a)含量在20%海水晶人工海水组中最高,其余各组的含量相差不大,但藻红蛋白(PE)和藻蓝蛋白(PC)含量均比100%天然海水组稍低。用上述6种培养液培养坛紫菜丝状体,结果表明在60%海水晶人工海水组中培养的自由丝状体鲜重增加最明显,生长情况最好,100%天然海水组增重最少,其余各组增重差异不明显。坛紫菜丝状体在100%海水晶人工海水中生长良好,这说明可以用海水晶人工海水来培养坛紫菜丝状体。