Experimental evaluation of heterobeltiosis and heterosis between two populations of Pacific abalone Haliotis discus hannai Ino

A2 × 2 factorial cross between two populations of Pacific abalone Haliotis discus hannai Ino, collected separately from Dalian （D） in China and Miyagi （M） in Japan, was conducted to compare performances in fertilization rate, hatching rate, metamorphosis rate and growth at days 20, 43, 160 and 330 between purebreds （DD and MM） and crossbreds （DM and MD） and investigate the magnitude of heterobeltiosis （better parent） and heterosis （mid-parent）. Heterobeltiosis and heterosis for all the traits analyzed were evidently different between crossbreds DM and MD. Heterobeltiosis in the crossbred DM varied among traits, with values of 2.5% for the fertilization rate, 2.2% for the hatching rate, - 1.9% for the metamorphosis rate and 7.4% for the growth at the day 330. The crossbred DM displayed positive heterotic values for fertilization rate （5.4%）, hatching rate （7.4%）, metamorphosis rate （7.6%） and growth （ 12.0% ） at the day 330. However, both heterobeltiosis and heterosis for all the traits in the crossbred MD were negative except those for the growth at days 20 and 43. The results indicate the importance of selecting superior hybrid varieties if the exploitation of hybrid vigor is considered in the Pacific abalone breeding program.     

Induction of larval settlement and metamorphosis ofHaliotis discus hannai Ino (Gastropoda,Mollusca)

Conspecific foot mucus, excessive [K⁺] and gamma-aminobutyric acid (GABA) showed different metamorphosis-inductive effect on the veliger ofHaliotis discus hannai. The inductive effect of excessive [K⁺] and GABA was developmental stage-dependent and dose-dependent, while that of conspecific foot mucus was only developmental stage-dependent. At 20°C the veliger larvae became competent within 4 days after fertilization.H. discus hannai larvae showed gregarious settlement pattern on the conspecific foot mucus under the conditions of either presence or absence of KCl or GABA. The present studies showed that the effect of conspecific foot mucus on abalone larvae metamorphosis could be dose-independent.     

DIETARY PHOSPHORUS REQUIREMENT OF YOUNG ABALONE, HALIOTIS DISCUS HANNAI INO

An experiment was performed to determine the dietary phosphorus requirement of the young abalone,Haliotis discus hannai. Five semi-purified diets were formulated to provide a series of graded levels of dietary total phosphorus (0.23%–1.98) from monobasic potassium phosphate (KH2P04). The brown alga,Laminaria japonica, was used as a control diet. Similar size abalone were distributed in a single-pass, flow-through system using a completely randomized design with six treatments and three replicates each treatment. The abalone were hand-fed to satiation with appropriate diets in excess, once daily at 17:00. The feeding trial was run for 120-d. Survival rate and soft-body to shell ratio (SB/S) were constantly maintained regardless of dietary treatment. However, the weight gain rate (WGR), daily increment in shell length (DISL), muscle RNA to DNA ratio (RNA/DNA), carcass levels of lipid and protein, soft-body alkaline phosphatase (SBAKP), and phosphorus concentrations of whole body (WB) and soft body (SB) were significantly (ANOVA, P<0.05) affected by the dietary phosphorus level. The dietary phosphorus requirements of the abalone were evaluated from the WGR, DISL, and RNA/DNA ratio respectively, by using second-order polynomial regression analysis. Based on these criteria, about 1.0%–1.2% total dietary phosphorus, i.e. 0.9%–1.1% dietary available phosphorus is recommended for the maximum growth of the abalone. Project 39670572 supported by the NSFC.     

几种环境因子对皱纹盘鲍的胁迫作用及其阈值

针对福建等海域养殖皱纹盘鲍度夏死亡率高的现象,试验研究了不同盐度、温度和溶解氧条件,对皱纹盘鲍幼鲍和1龄鲍死亡率的影响,并根据试验结果设定了皱纹盘鲍海上养殖盐度、温度和溶解氧预警阈值。结果表明:低盐胁迫对皱纹盘鲍的死亡率影响显著,在水温24～27℃时,盐度20是幼鲍和1龄鲍试验组和对照组死亡率有显著差异的临界点,盐度预警阈值为22;高温胁迫对皱纹盘鲍的死亡率影响显著,温度28℃是幼鲍和1龄鲍试验组和对照组死亡率有显著差异的临界点,温度预警阈值为27℃;皱纹盘鲍有较强的耐低氧能力,在水温26℃,盐度为30,溶解氧低于2.0mg/L时,开始大量死亡,溶解氧预警阈值为3.5mg/L;高温胁迫和低盐胁迫对皱纹盘鲍的死亡率有显著的交互作用,温度升高,低盐耐受能力下降。     

杂色鲍碱性磷酸酶在醇、醛和酮溶液中的失活作用

报道了杂色鲍碱性磷酸酶(ALP)在甲醇、乙醇、丙醇、甲醛、丙酮等溶液中的失 活动力学．结果表明:酶的剩余活力随着有机溶剂浓度的增大而迅速下降．测得上述 5种有机溶剂对该酶的半抑制率(IC50)分别为5．41、3．07、1．80、31．0×10-3、2．30 mol／dm3．在这些有机溶剂溶液中酶的失活过程都是可逆反应．检测醇、醛、酮对该酶 的失活作用机理,结果表明甲醇、乙醇、丙醇对杂色鲍ALP的失活作用均为非竞争性 机制,其抑制常数分别为5．36、3．02、1．80mol／dm3．甲醛、丙酮对杂色鲍ALP的失活 作用都呈现为混合型抑制,其对游离酶的抑制常数(K1)分别为24．7×10-3、1．66 mol／dm3;对酶-底物络合物的抑制常数(KIS)分别为56．2×10-3、5．40mol／dm3．     

AVAILABILITY OF PHOSPHORUS FROM SELECTED INORGANIC PHOSPHATES TO JUVENILE ABALONE, HALIOTIS DISCUS HANNAI INO

The availability of phosphorus to juvenile abalone,Haliotis discus hannai from primary, secondary and tertiary calcium phosphate, primary sodium or potassium phosphate separately or in combination was determined in a 120-day feeding trial. Seven semi-purified diets were formulated to contain relatively constant dietary phosphorus, ranging from 0.78%–0.82%. The concentrations of dietary phosphorus were presumed to be slightly below the requirement for abalone. A brown alga,Laminaria japonica, was used as a control diet. Abalone juveniles of silimar size (average weight 1.18 g; average shell length 18.74 mm) were distributed in a single-pass, flow-through system using a completely randomized design with eight treatments and three replicates each treatment. The survival ranged from 71.1% to 81.1%, and was not significantly (P>0.05) related to dietary treatment. However, the weight gain rate (41.72%–65.02%), daily increment in shell length (36.87–55.07 μm) and muscle RNA-DNA ratio (3.44–4.69) were significantly (P<0.05) affected by dietary treatment. Soft body alkaline phosphatase activity (10.9–19.8 U/g wet tissue) and carcass levels of lipid (7.71%–9.33%) and protein (46.68%–49.35%) were significantly (P<0.05) responsive to available phosphorus of the diets. Dietary treatment had significant effect (P<0.05) on concentrations of phosphorus in the whole body (WB) and soft body (SB). Apparent digestibility coefficients (45%–97%) of phosphorus were also significantly (P<0.05) different between dietary treatments. The results indicated that among these inorganic compounds, primary calcium, potassium and sodium phosphates separately or in combination could be utilized effectively by juvenile abalone as dietary phosphorus sources (availability ranging from 87%–97%). However, secondary and tertiary calcium phosphates were found to be low in availability, the values being 45% and 77%, repectively. Project 39670572 supported by the Natural Science Foundation of China (NSFC).     

AVAILABILITY OF PHOSPHORUS FROM SELECTED INORGANIC PHOSPHATES TO JUVENILE ABALONE,HALIOTIS DISCUS HANNAI INO.

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笼养密度对九孔鲍生长的影响

在室内水池内以五层笼摞叠养殖方式,试验了入笼时平均壳长21.2 mm(平均体重1.1 7 g)、37.0 mm(平均体重5.43 g)、51.8 mm(平均体重16.08 g)小、中、大3种规格的九孔鲍的不同放养密度对其生长的影响.笼子大小为40 cm×30 cm×12 cm.小规格设每笼放养量为25、30、35、40、45与50只6种密度;中为25、30、35、40、与45只5种密度;大为20、25、30与40只4种密度.经60 d试验,结果表明(1)小个体组以30只/笼生长最快,平均160μm/d,33 mg/d;(2)中个体组以25只/笼生长最快,平均123um/d,62 mg/d;(3)大个体组以20只/笼生长最快,平均95 μm/d,116 mg/d.     

皱纹盘鲍(Haliotis discushannai Ino)胚胎RNA分离和cDNA文库构建

从皱纹盘鲍胚胎样本中分离RNA并构建了cDNA文库。分别收集孵化8、10和12h的皱纹盘鲍胚胎,用抽滤除菌的海水反复悬浮胚胎,将这3个发育阶段的胚胎样本等量混合后用TRIZOL试剂提取总RNA,将提取物用酚氯仿异戊醇再抽提2次,分离获得高质量的总RNA。从总RNA中分离mRNA,构建了皱纹盘鲍混合胚胎的cDNA文库,未扩增胚胎文库的滴度为5.0×106pfu/ml,重组率为94.12%,插入片段均大于400bp,70.6%克隆的插入片段分布在1000—1500bp之间,扩增后的文库滴度为3.06×109pfu/ml。     

Characterization of defensin gene from abalone Haliotis discus hannai and its deduced protein

Defensin is one of preserved ancient host defensive materials formed in biological evolution. As a regulator and effector molecule, it is very important in animals' acquired immune system. This paper reports the defensin gene from the mixed liver and kidney cDNA library of abalone Haliotis discus hannai Ino. Sequence analysis shows that the gene sequence of full-length cDNA encodes 42 mature peptides (including six Cys), molecular weight of 4 323 Da, and pI of 8.02. Amino acid sequence homology analysis shows that the peptides are highly similar (70% in common) to other insects defensin. Because of a typical insect-defensin structural character of mature peptide in the secondary structure, the polypeptide named Haliotis discus defensin (hd-def), a novel of antimicrobial peptides, belongs to insects defensin subfamily. The RT-PCR result of Haliotis discus defensin shows that the gene can be expressed only in the hepatopancreas by Gram-negative and positive bacteria stimulation, which is ascribed to inducible expression. Therefore, it is revealed that the Haliotis discus defensin gene expression was related to the antibacterial infection of Haliotis discus hannai Ino.