Japanese scallop is one of the most important cultured seashells in the world. Following about 20 years’ artificial culture, we need DNA markers for genetic research. Microsatellites recently have become popular mark...Japanese scallop is one of the most important cultured seashells in the world. Following about 20 years’ artificial culture, we need DNA markers for genetic research. Microsatellites recently have become popular markers in a variety of genetic investigations. In this research, we isolated microsatellite DNAs from the Japanese scallop Patinopecten yessoensis genome with magnetic beads. As a result, 136 positive clones were identified from 192 clones,and 179 microsatellites were found. Among the 179 microsatellites, 50.8% were perfect, 43.0% were imperfect and the rest were compound type (6.1%). Finally, 85 microsatellite primers were designed by software Primer Premier 5.0, and 40 pairs were screened; 15 loci showed polymorphism. In these microsatellite loci, genetic diversity was analysed in 2 cultured populations. The average number of allele (A) ranged from 3.27 to 3.40; the average heterozygosity (H_o) in 2 populations was 0.447 to 0.540; average excepted heterozygosity (H_e) was 0.468 to 0.493; and average polymorphism information content (PIC) was 0.443 to 0.463. The genetic distance was 0.1155 and genetic identity was 0.8909. Thirteen microsatellite loci except HLJX-028 and HJLX-031 could be used for genetic展开更多
采用半静水式毒性试验,研究了6种苯系物(苯、甲苯、乙基苯、邻–二甲苯、间–二甲苯、对–二甲苯)对虾夷扇贝的生殖毒性作用。在水温(16±0.5)℃、盐度30.0、p H 8.0条件下,用0.5、2.5、12.5 mg·L-1的苯、甲苯、乙基苯、邻–...采用半静水式毒性试验,研究了6种苯系物(苯、甲苯、乙基苯、邻–二甲苯、间–二甲苯、对–二甲苯)对虾夷扇贝的生殖毒性作用。在水温(16±0.5)℃、盐度30.0、p H 8.0条件下,用0.5、2.5、12.5 mg·L-1的苯、甲苯、乙基苯、邻–二甲苯、间–二甲苯、对–二甲苯处理虾夷扇贝的精子、胚胎和幼体,观察6种苯系物对虾夷扇贝的精子活力、卵子受精率、胚胎延滞率、胚胎畸形率、孵化率、幼体畸形率的影响。同时通过透射电镜观察6种苯系物(2.5 mg·L-1)对虾夷扇贝精子超微结构的损伤作用。结果发现:不同浓度苯系物处理组与对照组(0.0 mg·L-1)相比,虾夷扇贝精子的运动时间、卵子受精率及胚胎孵化率显著降低;胚胎发育延滞率、胚胎畸形率和幼体畸形率显著增加为并与处理浓度之间存在显著的剂量—效应关系。以上结果表明实验浓度下,6种苯系物对虾夷扇贝具有较强的胚胎毒性和生殖毒性作用。通过电镜切片发现,2.5 mg·L-1的苯、甲苯、乙基苯、邻–二甲苯、间–二甲苯、对–二甲苯导致虾夷扇贝精子超微结构损伤,表现为:精子质膜断裂、部分溶解,线粒体质膜和内嵴断裂、部分溶解。苯系物对虾夷扇贝精子超微结构的损伤可能是影响其精子活力和降低卵子受精率的主要原因之一。上述结果为苯系物对海洋贝类的生殖毒性评价提供基础数据。展开更多
Triploid shellfish are useful for aquaculture because of their sterility,superior growth and improved meat quality.Tetraploid are also valuable for 100% producing triploids through mating with diploid.We tested polypl...Triploid shellfish are useful for aquaculture because of their sterility,superior growth and improved meat quality.Tetraploid are also valuable for 100% producing triploids through mating with diploid.We tested polyploid induction in Japanese scallop,Patinopecten yessoensis,by inhibiting polar body Ⅰ (PB group) and both polar bodyⅠandⅡ (PPB group) in newly fertilized eggs.Cytochalasin B (0.6 mg/L) was applied at 11～22 min post fertilization (PF),and terminated in PB group when polar body Ⅰ was released about 70% in untreated eggs,in PPB group when polar lobe was observed in control group.The treatment and its control were repeated 5～7 times using different pairs of parents.The ploidy was determined by counting chromosome number at embryo stage,and then was detected by flow cytometry (FCM) at larvae stage and juvenile stage.\;In PB group,aneuploid (31.13%),triploid (3.96%),tetraploid (17.46%) and pentaploid (46.65%) embryos were produced,and in PPB groups,pentaploid embryos became higher (56.2%),triploid and tetraploid were 2.42% and 9.11%.At day 3 PF,the larvae showed tetraploid,pentaploid and aneuploid peaks through checking with FCM in PB group,and showed mainly higher pentaploidy peak in PPB groups.However,at day 14 PF diploids were mainly left,sometimes with small triploid peak.It suggested that most tetraploid,aneuploid and pentaploid larvae were died within the first two weeks PF.At three months PF,a few diploid juveniles were harvested in three control groups.Only 12 juvenile scallops were harvested in one of treated group (PB-7),and 11 of them died accidentally,the alive one in treated group was triploid through checking with FCM.展开更多
The glutathione S-transferases(GSTs)are a superfamily of enzymes that function in cellular protection against toxic substances and oxidative stress.Bivalves could accumulate high concentration of paralytic shellfish t...The glutathione S-transferases(GSTs)are a superfamily of enzymes that function in cellular protection against toxic substances and oxidative stress.Bivalves could accumulate high concentration of paralytic shellfish toxins(PSTs)from harmful algae.To understand the possible involvement of GSTs in protecting bivalves during PST accumulation and metabolism,the GST genes were systemically analyzed in two cultured scallops,Azumapecten farreri and Mizuhopecten yessoensis,which were reported for PST deposition during harmful algae bloom.A total of 35 and 37 GSTs were identified in A.farreri(AfGSTs)and M.yessoensis(MyGSTs)genome,respectively,and the expansion of the sigma class from the cytosolic subfamily was observed.In both scallop species,sigma class GSTs showed higher expression than other members.The high GSTs expression was detected mainly during/after larvae stages and in the two most toxic organs,hepatopancreas and kidney.After ingesting PST-producing dinoflagellates,all the regulated AfGSTs in the hepatopancrcas were from the sigma class,but with opposite regulation pattern between Alexandrium catenella and A.minutum exposure.In scallop kidneys,where PSTs transformed into higher toxicity,more AfGSTs were regulated than in the hepatopancreas,and most of them were from the sigma class,with similar regulation pattern between A.catenella and A.minutum exposure.In M.yessoensis exposed to A.catenella,MyGST-σ2 was the only up-regulated MyGST in both hepatopancreas and kidney.Our results suggested the possible diverse function of scallop GSTs and the importance of sigma class in the defense against PSTs,which would contribute to the adaptive evolution of scallops in marine environments.展开更多
文摘Japanese scallop is one of the most important cultured seashells in the world. Following about 20 years’ artificial culture, we need DNA markers for genetic research. Microsatellites recently have become popular markers in a variety of genetic investigations. In this research, we isolated microsatellite DNAs from the Japanese scallop Patinopecten yessoensis genome with magnetic beads. As a result, 136 positive clones were identified from 192 clones,and 179 microsatellites were found. Among the 179 microsatellites, 50.8% were perfect, 43.0% were imperfect and the rest were compound type (6.1%). Finally, 85 microsatellite primers were designed by software Primer Premier 5.0, and 40 pairs were screened; 15 loci showed polymorphism. In these microsatellite loci, genetic diversity was analysed in 2 cultured populations. The average number of allele (A) ranged from 3.27 to 3.40; the average heterozygosity (H_o) in 2 populations was 0.447 to 0.540; average excepted heterozygosity (H_e) was 0.468 to 0.493; and average polymorphism information content (PIC) was 0.443 to 0.463. The genetic distance was 0.1155 and genetic identity was 0.8909. Thirteen microsatellite loci except HLJX-028 and HJLX-031 could be used for genetic
文摘Triploid shellfish are useful for aquaculture because of their sterility,superior growth and improved meat quality.Tetraploid are also valuable for 100% producing triploids through mating with diploid.We tested polyploid induction in Japanese scallop,Patinopecten yessoensis,by inhibiting polar body Ⅰ (PB group) and both polar bodyⅠandⅡ (PPB group) in newly fertilized eggs.Cytochalasin B (0.6 mg/L) was applied at 11～22 min post fertilization (PF),and terminated in PB group when polar body Ⅰ was released about 70% in untreated eggs,in PPB group when polar lobe was observed in control group.The treatment and its control were repeated 5～7 times using different pairs of parents.The ploidy was determined by counting chromosome number at embryo stage,and then was detected by flow cytometry (FCM) at larvae stage and juvenile stage.\;In PB group,aneuploid (31.13%),triploid (3.96%),tetraploid (17.46%) and pentaploid (46.65%) embryos were produced,and in PPB groups,pentaploid embryos became higher (56.2%),triploid and tetraploid were 2.42% and 9.11%.At day 3 PF,the larvae showed tetraploid,pentaploid and aneuploid peaks through checking with FCM in PB group,and showed mainly higher pentaploidy peak in PPB groups.However,at day 14 PF diploids were mainly left,sometimes with small triploid peak.It suggested that most tetraploid,aneuploid and pentaploid larvae were died within the first two weeks PF.At three months PF,a few diploid juveniles were harvested in three control groups.Only 12 juvenile scallops were harvested in one of treated group (PB-7),and 11 of them died accidentally,the alive one in treated group was triploid through checking with FCM.
基金This work was funded by the National Key R&D Program of China(2019YFC1605704)the National Natural Science Foundation of China(31630081 and 31802292)Taishan Industry Leading Talent Project.
文摘The glutathione S-transferases(GSTs)are a superfamily of enzymes that function in cellular protection against toxic substances and oxidative stress.Bivalves could accumulate high concentration of paralytic shellfish toxins(PSTs)from harmful algae.To understand the possible involvement of GSTs in protecting bivalves during PST accumulation and metabolism,the GST genes were systemically analyzed in two cultured scallops,Azumapecten farreri and Mizuhopecten yessoensis,which were reported for PST deposition during harmful algae bloom.A total of 35 and 37 GSTs were identified in A.farreri(AfGSTs)and M.yessoensis(MyGSTs)genome,respectively,and the expansion of the sigma class from the cytosolic subfamily was observed.In both scallop species,sigma class GSTs showed higher expression than other members.The high GSTs expression was detected mainly during/after larvae stages and in the two most toxic organs,hepatopancreas and kidney.After ingesting PST-producing dinoflagellates,all the regulated AfGSTs in the hepatopancrcas were from the sigma class,but with opposite regulation pattern between Alexandrium catenella and A.minutum exposure.In scallop kidneys,where PSTs transformed into higher toxicity,more AfGSTs were regulated than in the hepatopancreas,and most of them were from the sigma class,with similar regulation pattern between A.catenella and A.minutum exposure.In M.yessoensis exposed to A.catenella,MyGST-σ2 was the only up-regulated MyGST in both hepatopancreas and kidney.Our results suggested the possible diverse function of scallop GSTs and the importance of sigma class in the defense against PSTs,which would contribute to the adaptive evolution of scallops in marine environments.
