大型藻酶解单细胞用于贝类育苗的研究

用海藻工具酶将紫菜和裙带菜解离成单细胞作为饵料对海湾扇贝亲贝和幼体、魁蚶亲贝和皱纹盘鲍稚鲍进行了研究。结果表明：(1)海湾扇贝亲贝全喂紫菜细胞其效果与喂全微藻的效果相近，亲贝都能成熟，排放精卵和正常胚胎发育。微藻与紫菜细胞混合投喂可加速性腺发育，性腺指数较高。全喂紫菜细胞能够满足幼体的生长发育和变态。但变态率、生长率较全喂微藻和混合投喂的低。(2)紫菜细胞和微藻分别或混合投喂魁蚶亲贝，都能满足亲贝的性腺发育，进行正常受精、孵化和变为D形幼体。(3)用裙带菜细胞投喂稚鲍与人工配合饵料比较，能显著提高稚鲍的存活率。研究表明。由大型藻生产单细胞饵料，在海产动物育苗中具有广阔的应用前景。     

条斑紫菜叶状体细胞的抗生素敏感性研究

为了进行条斑紫菜基因工程研究和培育转基因紫菜 ,首先必须确定适合的阳性选择标记基因。条斑紫菜对 8种抗生素的敏感性试验表明 ,它对各种抗生素的敏感性不同 ,其中 zeocin、腐草霉素和氯霉素半致死剂量都很低。因此 Cat(氯霉素乙酰转移酶 )基因和 Sh.ble(sh.bleomycin)基因均有望成为条斑紫菜基因工程研究中理想的选择标记基因。     

几种红藻和蓝藻的光合作用色素

分离纯化出几种海产红藻和一种蓝藻的光合作用色素,并测定了它们的化学性质和光谱学性质。这些藻类是3种红藻:多管藻(Polysiphonia urceolata)、橡叶藻(Phycodrys sp.)和条斑紫菜(Porphyra yezoensis);蓝藻:钝顶螺旋藻(Spirulina platensis)。用羟基磷灰石柱层析法从上述藻类中分离到几种不同的藻胆蛋白。经SDS-PAGE及光谱测定,发现条斑紫菜中的藻红蛋白不同于其它两种红藻。而橡叶藻中存在的两条藻红蛋白也有差异,条斑紫菜和钝顶螺旋藻中的两种别藻蓝蛋白之间也有区别。叶绿素分析表明,钝顶螺旋藻中叶绿素a的含量高于红藻中叶绿素a的含量。     

紫菜体细胞对抗生素的敏感性实验

近几年来栽培紫菜常发生大面积的病烂,给养殖生产造成巨大的损失,因此,培育出抗逆、高产、优质的紫菜新品种成为亟需解决的问题。利用细胞工程、尤其基因工程等生物技术研究紫菜的生物学特性、培育紫菜的新品种成为必然的发展方向。秦松等对海带等大型经济褐藻类的研究取得了一些进展[1,2]并发现氯霉素是海带基因工程的有效选择压力。上海水产大学王素娟在紫菜领域进行了这方面的研究工作,探索外源基因导入紫菜细胞的方法。本文实验表明,氯霉素在紫菜基因工程中可作为有效选择压力,另外四环素也有一定的效果。1　材料与方法1.1　材料、体细…     

福建海区条斑紫菜的栽培试验

本文报告了对采集自福建自然海区的条斑紫菜进行的2a栽培试验。结果表明，条斑紫菜壳孢子的放散时间比坛紫菜迟1－1．5个月；壳孢子放散延续时间比坛紫菜长；放散高峰期间出现在11月中旬；骤然降温可以促进壳孢子集中放散；单孢子放散以水温20℃左右较佳。莆田及其以北海区可能比厦门更适宜栽培条斑紫菜。     

五种紫菜同工酶标记的遗传分析

用垂直凝胶电泳来研究采自中国的（条斑紫菜×坛紫菜）杂交种、坛紫菜、条斑紫菜、少精紫菜、半叶紫菜的同工酶多态性。然后分析最小可能位点数和观察同位基因数、遗传差异性和采用平均连接聚类法。选取23种酶带中的六个酶（MDH，ME，LDH，GDH，IDH和G-6-PDH）进行分析。最小可能等位基因位点数显示五种紫菜在ME等位基因位点都只有一个。LDH和GDH等位基因位点半叶紫菜和少精紫菜和另外的三种紫菜不同，而在分析MDH时半叶紫菜比较独特。在IDH位点分析时少精紫菜和坛紫菜和其他三种不同，而条斑紫菜和坛紫菜和其他三种不同在G-6-PDH位点不一致。总的来看，最小可能等位基因位点数分析半叶紫菜是最分离的。遗传多样性结果分析表明当遗传相似度为0．7550时，五种紫菜中的遗传变异研究受到限制。条斑紫菜和坛紫菜的杂交种非常接近少精紫菜。当遗传相似度达到0．8937时，出乎意料的是条斑紫菜和坛紫菜遗传同一。性很低。条斑紫菜4个株系的平均遗传相似度仅为0．7428，差异比较大。在所有紫菜中半叶紫菜是最分化的分支，它与最小可能等位基因位点数分析一致。     

Physiological Responses of Cultured Seaweed Pyropia yezoensis to Phosphorous Limitation in the Nakdong River Estuary,Korea

Ocean Science Journal - Pyropia yezoensis cultured in the estuary of the Nakdong River in Korea is an important fisheries resource for the local economy. Over the past few decades, the...     

Extraction of plasmid-like DNA and high-quality total DNA from Porphyra yezoensis

I~ODWrIONExtraction of DNA from Porphyra po~nsis is very difficult due tO its high content ofpolysaccharides. TO avoid the interference Of POlysaccharides, HOng et al. (1992) used LiCI tosoften Porphyra thalli cells to release DNA directly without lysing the thalli, hoWever, the procedure is problematic in the isolation of high-quality DNA. Kitade et al. (1996) established a complicated procedure which includes the grindiflg of material in liquid nitrogen, lysing of material inextra…     

外界因素对条斑紫菜体细胞分化发育的影响——Ⅰ. 温度、光强实验

本实验利用酶解技术得到单个体细胞,较详细地研究了不同温度、光照强度对体细胞分化发育的影响,从而确定出体细胞培养的最佳条件,为进一步利用条斑紫菜体细胞育苗这一新技术打下了基础。     

Fe2+对条斑紫菜细胞生长发育的影响

对条斑紫菜(Porphyra yezoen sis)离体细胞在含Fe2+培养条件下的生长发育进行了研究。在消毒海水培养基中添 加不同浓度的Fe2+,培养经酶解分离而获得的条斑紫菜单细胞。结果表明,低浓度的 Fe2+能促进其生长发育;而高浓度的Fe2+可以导致苗大量畸变,细胞变小,苗 反复崩解成单孢子。当Fe2+浓度达到50mg/L以上时,细胞完全致死。     

条斑紫菜杂交重组品系(A-18)的筛选与特性分析

为筛选出藻体生长快且颜色与野生型色泽相近的条斑紫菜新品系，本研究从条斑紫菜绿色突变体和红色突变体种内杂交产生的后代中，分离出了优良品系A-18。日龄60 d时的叶状体平均长度、长宽比和单株湿质量，A-18品系分别为84.95 cm、49.46和0.52 g，分别是条斑紫菜野生型品系（WT）的3.12、7.01和1.36倍。日龄60 d时叶状体的叶绿素a和总藻胆蛋白含量，A-18品系分别为8.37和53.81 mg/g，均与WT品系较接近。日龄60 d时的叶状体平均厚度，A-18品系仅为20.22 μm，比WT品系降低了29%。另外，A-18品系的壳孢子放散总量为916.01万个/贝壳，是WT品系的1.55倍。综上所述，A-18品系具有生长快、长宽比值大、藻体薄、壳孢子放散量大等优良特性，且藻体颜色与野生型色泽相近，有望在生产中运用。     

几种红藻和蓝藻藻胆体的分离和光谱特性

本文分析了高等红藻龙须菜(Gracilaria lemaneiformis)藻胆体的吸收光谱和荧光光谱,并与低等红藻条斑紫菜(Porphyra yezoensis)及蓝藻钝顶螺旋藻(Spirulina platensis)作了比较研究。龙须菜藻胆体的吸收峰位于498nm、541nm、565nm、620nm和650nm。荧光发射峰位于576nm和672nm。从光谱学性质和能量传递分析,龙须菜和条斑紫菜胆体的组成、结构相似,但有细微差别。而螺旋藻藻胆体的组成、结构则与两种红藻差别较大。     

条斑紫菜高纯度总DNA及其质粒状DNA的提取

提取条斑紫菜高纯度总DNA及其质粒状DNA的新方法。先用海螺酶处理紫菜叶状体制备细胞,然后用SDS-蛋白酶K裂解细胞提取总DNA,再用玻璃粉浆(glassmilk)对其纯化,经纯化后的总DNA能被EcoRI,Dral与HaeⅢ等限制酶完全酶切,并在酶切图谱上形成明显的DNA带型。当用异硫氰酸胍一十二烷基肌氨酸钠裂解紫菜细胞时,在总DNA提取物中直接发现有一条质粒状DNA带(2.3Kb),即建立了一种极简便的质粒状DNA提取方法。     

Genetic variation of Porphyra yezoensis by using AFLP1

Genetic variation of 11 lines of Porphyra yezoensis from the coastline of Kagoshima ofJapan, Qingdao, Nantong, Putuo and Nanji Islands of China were studied by using amplified fragment length polymorphism (AFLP).778 bands were obtained with AFLP analysis of 16 primer combinations, among which 15 were unique, about 98.07% were polymorphic. The AFLP data showed that the closest genetic distance was 0.180 between two Kagoshima samples, and the farthest one was 0.397 between Kagoshima No.1 and Nantong No.9 line. The genetic distance showed that the variation was within the inner species scope. Neighbor-joining cluster and UPGMA cluster indicated that samples from Kagoshima and Qingdao were with high similarity and either with the samples of Nantong, Putuo and Nanji Islands. P. yezoensis in China shared high genetic diversity, and the genetic distance showed positive correlation with the geographic distance.     

利用AFLP技术研究条斑紫菜的遗传变异

利用AFLP技术对11个条斑紫菜品系进行了研究.由16对引物共扩增出778条谱带,其中仅15条为共有谱带,多态位点的比例高达98.07%.日本鹿儿岛,我国的青岛、江苏、浙江等不同地点的野生或栽培条斑紫菜品系之间最近的遗传距离为0.180(两个日本样本之间),最远为0.397(日本样本与江苏样本之间),属于物种内种群间的遗传变异.聚类结果显示日本的样本与青岛的野生样本间的遗传相似系数较高而聚合在一起;浙江野生条斑紫菜与江苏的样本遗传距离相近,聚合在一起.结果表明,我国条斑紫菜的遗传多样性程度较高而且各品系间的遗传距离与其地理间距呈正相关关系.     

Induction Effect and Genetic Analysis of NG to Conchospores of Porphyra

The new spreading conchospores ot Porphyra yezoensis, P. haitanensis and P. katadai var. hemiphylla are induced by N-Methy-N‘-Nitro-N-Nitrosoguanidine (NG), a strong mutagen to induce the mutant of chromatophore. It is showed that (1) no mutants have been investigated in all control group. (2)The conchospores of all the three species of Porphyra are easy to be induced by NG for they are in the stage of breeding, In the lowest dose (10 μ g/mL) of this experiment, the mutant rate of P. yezoensis is up to 57.9%, that of P. haitanensis 30.7% and that of P. katadai var. hemiphylla 51.7%. (3)The mutant rate can not increase obviously by increasing the reagent concentration or prolonging the induction time. Within the scope of experiment, the inducement effects of three species of Porphyra ar e much similar, and the mutant rate is near 50%. The optimum induction concentration of NG is 10 μ g/mL, and the optimum induction time is 30 minutes.     

超声波对条斑紫菜硅藻附着症的作用

为使条斑紫菜(Porphyra yezoensis)上附着的硅藻脱落,从超声波的生物学效应出发提出了新的思路。选择生长状态较好的附着硅藻的紫菜分别在不同的输出功率和时间下用超声波处理。结果表明,当超声波的输出功率为300 W、处理时间25 s时,可以成功地去除附着的硅藻及黏质柄,而对条斑紫菜未产生不良影响。该研究结果在条斑紫菜的加工生产上有应用与推广价值。     

条斑紫菜藻胆蛋白性质的研究Ⅲ.不同环境条件下R-藻红蛋白吸收光谱类型的转变

条斑紫菜的R-藻红蛋白在pH:5.8—8.4的几种缓冲液中保存,经过大约4个星期,其吸收光谱由原有的双峰型转变为三峰型;在pH=8.4的聚丙烯酰胺梯度凝胶电泳后或在微量四甲基乙二胺作用下发生了同样的变化。但在变性剂SDS或尿素存在下,吸收光谱中540nm和565nm的峰强度比逐渐改变,直至565nm吸收峰消失,始终没有从双峰型转变为三峰型的迹象。显然,条斑紫菜R-藻红蛋白吸收光谱类型的转变不是变性引起的。     

紫菜(Porphyra遗传差异的ISSR分析

采用ISSR标记技术对来自不同产地的6个坛紫菜(Porphyra haitanensis)无性系和两个条斑紫菜(P.yezoensis)无性系进行了遗传差异的分析。结果表明,7条ISSR引物在8个紫菜系中共扩增出59条片段,全部表现出多态性。根据Nei等的相似性系数得出8个紫菜系间的遗传距离在0.274—0.746之间。用UPGMA方法作出的系统树中各紫菜基本上是按照产地进行聚类,从而推测紫菜的遗传差异可能与地域分布相关。本文结果也证明了ISSR与RAPD、AFLP等标记技术一样适用于紫菜的遗传多样性分析。     

Induction Effect and Genetic Analysis of NG to Thallus of Porphyra

The young thalli of Porphyra yezoensis and P. haitanensis are induced by N-Methy-N‘-Nitro-N-Nitrosoguanidine (NG), a strong mutagen to induce the mutant of chromatophore. It is showed that no mutants have been investigated in all control groups,while in the induced groups, it is easy to find mutant cells or mutant cell masses in the thallus.The pigmentation mutants may be the result of NG inducement. The mutant rate increases obviously with the mutagen concentration or by prolonging the induction time. Within the scope of experiment, the inducement effects of Porphyra yezoensis are raised from 11.2% to 28.7%, and those of P. haitanensis from 10.1% to 20.2%. According to our experiments, the amount of mutant cells in the same area does not increase with the induction intensity The amount of mutant cells in every group of inducement of both species of Porphyra changes very little with the fixed fields check method. The lowest dose (25μg/mL) of this experiment has a high mutant rate. The mutant rate shows the percentage of mutant cells in all the survived cells, while the amount of mutant cells reflects the effect of inducement. Thus the optimum induction concentration of NG is 25μg/mL, and the optimum induction time is 30minutes.