营养盐可得性对坛紫菜氮磷吸收、生长及藻红蛋白含量的影响

实验室条件下,研究了N、P浓度、不同化合态N及N∶P比值对坛紫菜(Porphyra haitanensis)N、P吸收速率,以及P浓度对坛紫菜生长速率和藻红蛋白含量的影响。结果表明,随着营养盐浓度的升高,坛紫菜对N、P的吸收速率也随之增高,当无机氮浓度达到100μmol/L时,坛紫菜对N、P的吸收速率趋向接近最大值;当NO3--N∶NH4+-N比值为1∶5时,坛紫菜对N的吸收达到最大值;坛紫菜对P的吸收速率随NO3--N∶NH4+-N比值的减小而略有增大;坛紫菜对N的吸收速率随着N∶P比值的增大而增大,而对P的吸收速率随着N∶P比值的增大而减小;在磷浓度低于12μmol/L的情况下,坛紫菜的生长速率和藻红蛋白含量随着P浓度的升高而增加,高于12μmol/L时,则不再增加。     

不同采收期坛紫菜的风味比较

坛紫菜的采收有"分茬"的特点,为科学评价不同采收期的坛紫菜在风味方面的差异,采用电子舌、电子鼻和气相离子迁移谱仪对一水、二水、三水、四水和五水坛紫菜的滋味和挥发性成分进行检测。结果表明:在滋味方面,前期采收的坛紫菜鲜味和鲜味回味等令人愉悦的滋味更为强烈,随着采收期的延后,苦味、涩味等不愉快的滋味强度逐渐增强;在挥发性成分方面,不同采收期的坛紫菜差别明显,采收期邻近的样品挥发性成分的种类和含量也较为接近,一水坛紫菜的挥发性成分以2-乙基呋喃和阈值相对较高的酮类、醇类为主,二水和三水坛紫菜中丁内酯、2-己烯-1-醇、1-辛烯-3-醇含量相对较高,四水和五水坛紫菜中壬醛(青草味)和苯甲醛(苦杏仁味)含量较高。采用电子舌和电子鼻可以快速、准确区别不同采收期坛紫菜的风味。     

条斑紫菜壳孢子的抗生素敏感性研究

以往研究紫菜细胞对抗生素的敏感性时,常以叶状体的酶解单离体细胞为材料,但由于来自同一个叶状体的体细胞处于不同的分化阶段,离体培养时,体细胞的成活、再生和发育途径均不相同,从而无法准确地判断抗生素对紫菜离体细胞的生长和发育影响。以成活、萌发和发育途径等几乎一致的条斑紫菜壳孢子为研究材料,较系统地观察了3种抗生素对壳孢子的成活、生长和发育的影响。结果表明：卡那霉素能促进条斑紫菜壳孢子的萌发、生长和发育。加卡那霉素培养7 d,26%的壳孢子萌发体,其细胞数多于20个,比对照组多2倍,培养11 d和18 d,萌发体的平均长度分别为对照组的1.6倍和2.1倍。氨苄青霉素能显著地抑制条斑紫菜的壳孢子萌发、生长和发育,造成细胞的分裂速度减缓、叶状体发育异常。加氨苄青霉素培养7 d,80%的壳孢子萌发体的细胞数少于15个,培养11 d和18 d,各试验组的壳孢子萌发体的平均长度均显著低于对照组,并且随着氨苄青霉素浓度的提高,萌发体的平均长度下降。氯霉素对条斑紫菜的壳孢子具有强烈的致死作用。加氯霉素培养7 d,浓度大于100 μg/mL的各试验组的壳孢子萌发体全部死亡;培养11 d,低浓度组（50 μg/mL）的萌发体也全部死亡,90%以上的萌发体的细胞数小于5个。由此得出,氯霉素是开展条斑紫菜基因工程的理想抗性选择压力。     

紫菜种内原生质体的融合和融合体再生

利用酶解法分离出野生型叶状体和红色突变体的坛紫菜原生质体以及条斑紫菜野生型叶状体的原生质体，再用聚乙二醇（PEG）法进行种内原生质体融合，当加入原生质体液内的PEG被稀释除去百，原生质体发生融合并形成许多杂合体，种内原生质体融合率坛紫菜为9.7%-12.4%，条斑紫菜为10%-11.5%。PEG分子量在1540-6000之间，随着分子量的提高，原生质体融合率增高，原生质体融合体被挑出来进行单个培养，经过30天培养坛紫菜的一个杂合体（含一个红色原生质体和一个进驻生型原生质体）再生成一个呈红色和野生色的相嵌叶状体，鸸 个含两个红色原生质体的融合体再生成一个在基部和头部均长假根的叶状体，但来自这个再生体的原生质体只长成具有一个假根的叶状体，从原生质体融保体的再生体以及它们无性繁殖体的结果来看，坛紫菜的种内原生质体融合体可能只发生了细胞质融合，而真正的核融合并没有发生，培养15-20天，条斑紫菜的原生质体融合体先再生成细胞团，然后由细胞团释放出许胞子并萌发成正常野生型叶状体。     

3种植物生长调节剂对坛紫菜自由丝状体生长与光合色素含量的影响

采用单因素分析法,比较研究了添加3种外源植物生长调节剂(吲哚乙酸IAA、2,4-二氯苯氧乙酸2,4-D、激动素KT)对野生型坛紫菜(Pyropia haitanensis)自由丝状体生长及光合色素含量的影响。结果表明:与对照组相比,低浓度的IAA(0.5~4.0 mg·L~(-1))和KT(0.5~2.0 mg·L~(-1))对藻体的特定生长率有显著影响,以2.0 mg·L~(-1)IAA和1.0 mg·L~(-1)KT对藻体的促生长效应最大,培养25 d时藻体鲜重增加最大,分别是对照组的1.90倍和1.52倍(P0.05)。高浓度的IAA(6.0~10.0 mg·L~(-1))和KT(4.0~10.0 mg·L~(-1))却会抑制藻体生长(P0.05)。培养25 d时,与对照组相比,2.0 mg·L~(-1)IAA和1.0 mg·L~(-1)KT均有利于叶绿素a(Chl a)和类胡萝卜素(Car)的合成,Chl a含量分别增加了23.7%和23.8%;Car含量分别增加了31.0%和28.6%(P0.05)。然而,高浓度的IAA和KT(6.0、10.0 mg·L~(-1))不利于Chl a的合成,其处理浓度越高,Chl a含量下降越多。与对照组相比,较低浓度的IAA(1.0~2.0 mg·L~(-1))明显促进藻红蛋白(PE)和藻蓝蛋白(PC)合成,其中2.0 mg·L~(-1)IAA处理组PE、PC含量最高,分别增加了59.1%和45.7%(P0.05)。较低浓度的KT(0.5~2.0 mg·L~(-1))均有利于PE、PC合成,其中0.5 mg·L~(-1)KT处理组的PE、PC含量增加最多,分别提高了48.4%和50.5%(P0.05)。高浓度的IAA(6.0、10.0 mg·L~(-1))和KT(10.0 mg·L~(-1))则明显降低了PE、PC合成(P0.05)。2,4-D对藻体的特定生长率和光合色素合成无显著影响(P0.05)。     

Seeding nets with neutral spores of the red alga Porphyra umbilicalis (L.) Kützing for use in integrated multi-trophic aquaculture (IMTA)

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.) Kützing 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 °C. Unlike some algal zygotes and spores, neutral spores of P. umbilicalis required light to germinate; however, irradiances of 25 and 100 μmol 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.     

条斑紫菜蛋白酶解液脱色脱腥技术研究

以条斑紫菜为原料采用木瓜蛋白酶制备紫菜蛋白酶解液,以活性回收率和脱色率为评价指标,研究酶解液的脱色脱腥技术。试验结果表明,紫菜盐酸提取液色素最大吸收峰为550nm;在单因素试验基础上,通过正交试验优化后的脱色工艺条件为活性炭用量10g/L、脱色温度60℃、脱色时间40min、pH 6.5,脱色后的酶解液呈浅黄色,活性回收率逾90%。     

生态因子对条斑紫菜自由丝状体生长的影响

报道了生态因子对条斑紫菜自由丝状体生长的影响,结果表明,条斑紫菜自由丝状体生长的最适宜盐度范围为２０￣３０,低于１０的盐度不能生长,最适宜温度范围为１５℃￣２５℃,高限为３０℃,适宜光辐射照度１５μＥｍ％－２Ｓ＾－１￣６０μＥｍ＾２Ｓ＾－１,添加的适宜Ｎ浓度２０ｇ／ｍ＾３￣６０ｇ／ｍ＾３,Ｐ为１．５ｇ／ｍ＾３￣５ｇ／ｍ＾３。     

坛紫菜耐高温品系的选育与海区中试

利用人工诱变和体细胞再生技术,选育出可在30℃下生长的坛紫菜耐高温品(Q-1),实验结果表明:(1)在高温(28℃和30℃)下培养15d,Q-1品系的壳孢子成活率分别为76.8%和60.1%,分裂率分别为100%和83%;而对照组野生型(WT)品系的壳孢子成活率分别为15.9%和6.7%,分裂率分别为90.4%和63.8%。Q-1品系的壳孢子成活率和分裂率均远高于WT品系。(2)在常温(24℃)下培养35d的Q-1和WT品系壳孢子苗,在24℃、28℃和30℃组中再分别培养25d,Q-1品系的苗平均体长分别增加了19.4、10.8和2.8倍,而WT品系的苗平均体长分别只增加了7.3、1.7和0.9倍;Q-1品系的苗平均体长分别为WT品系的2.5、5.8和1.9倍。另外,在28℃和30℃中培养15d的WT品系幼苗发生了大面积腐烂;而Q-1品系幼苗没有出现任何烂苗迹象,表现出较好的生长。Q-1品系在海区中试中也表现出很好的耐高温特性。2008年秋季,坛紫菜采苗后遇到了长时间的高温天气,当地传统养殖的坛紫菜野生种发生了大规模腐烂与脱苗,产量大幅度减少;而Q-1品系的苗仍然维持良好的生长势头,没有出现烂苗,取得了高产。由此证实,Q-1品系是既能较快生长又耐高温的品系,有很大的生产应用价值。     

坛紫菜自由丝状体对无机碳的利用

Free-living Porphyra haitanensis conchocelis was cultured under continual light,then it was treated with Van,DIDS and SITS,the inhibitors of inorganic carbon utilization,to study the mechanism of inorganic carbon utilization in the conchocelis.The results indicate that Van inhibits the utilization of inorganic carbon most,with a rate of 71.3%,and the rates of DIDS and SITS are relatively lower,42% and 35.5% respectively.The inhibition rate of Az is 25.3%,which indicates the external CA is not an important part of its inorganic carbon uptake.A complementary conclusion is that the main part of inorganic carbon absorption in P.haitanensis conchocelis is through active transport of HCO3^- and CO2.