Isozyme Analysis of Spotted Halibut Verasper variegatus Temminck et Schlegel

To investigate the tissue-specificities of isozymes and the genetic structure of wild spotted halibut ( Verasper variegatus) population, horizontal starch gel electrophoresis was performed on 45 individuals collected in part of the Yellow Sea. The performances of 17 isozymes in 8 kinds of tissues or organs were screened preliminarily in a TC-7.0 buffer system. The results showed that the screened isozymes displayed remarkable tissue-specificities. Finally, 14 enzymes (AAT, ADH, EST, GPI, G3PDH, IDHP, LAP, LDH, MDH, MPI, PGDH, PGM, SDH and SOD) and 4 kinds of tissues (eye, skeleton muscle, liver and heart) were selected for genetic analysis. Fourteen isozymes are encoded by 20 loci, and 9 of them are poly-morphic. The polymorphic loci are AAT-1*, GPI-2*, G3PDH*, IDHP-1*, LDH*, MPI*, PGM-1*, PGM-2* and SDH*, and the proportion of polymorphic loci is 0.450 0 (P0.99). The mean values of observed and expected heterozygosities are 0.027 8 and 0.026 5, respectively and the average effective number of alleles is 1.067 5.     

褐菖鲉同工酶的组织特异性及群体遗传结构

用聚丙烯酰胺垂直板不连续凝胶电泳法对湛江海域褐菖鲉(Sebastiscus marmoratus)心、肝、肾、性腺、肌肉、鳃、眼、脑等8种组织的12种同工酶(ADH、a-AMY、CAT、EST、GDH、LDH、MDH、ME、POD、PPO、SDH、SOD)进行分析。结果表明,12种同工酶均有明显的组织特异性。经筛选,对肝脏和眼的8种同工酶(ADH、EST、LDH、MDH、ME、POD、SDH、SOD)进行群体遗传结构分析,共记录了18个基因位点,其中呈多态性的基因位点有5个(Adh-1、Est-2、m-Pod-1、s-Pod-1、s-Sod-1),多态位点比例(P)为27.78%,平均有效等位基因数(Ae)为1.238 8,平均观测杂合度(Ho)值为0.1401。     

Isozymes Analysis of the Golden Cuttlefish Sepia esculenta（Cephalopoda ： Sepiidae）

Thirty-nine isozymes in four tissues (mantle muscle, buccal bulb muscle, eye and liver) of Sepia esculenta were screened for enzymatic analysis using starch gel electrophoretic technique. Eighteen enzymes (G3PDH, LDH, MDH, MEP, IDHP, PGDH, GRS, NP, AAT, CK, AK, EST, ALP, ACP, FBP, MPI, GPI and PGM) show strong activities and good convergence in zymogram. They are proved to be suitable genetic markers in Sepia esculenta. Among the tissues used, mantle muscle is the best for electrophoretic analysis of isozymes. Eye and liver are fairly good for some special enzymes, such as LDH, EST, MPI, etc. Twenty-six loci are detected. The proportion of polymorphic loci is 0.115 in the Qingdao sample and 0.153 in the Rizhao sample (P<0.99). The mean values of the observed and expected heterozygosity per locus of Qingdao sample are 0.016 and 0.017, while those of the Rizhao sample are 0.023 and 0.025 respectively.     

光裸星虫不同组织同工酶谱分析

采用聚丙烯酰胺梯度凝胶垂直平板电泳对湛江地区的光裸星虫(Sipunculus nudus)3种组织(肠组织、体腔液和体壁肌肉)12种同工酶(SOD、EST、MDH、LDH、ALP、AMY、ME、ACP、G6PDH、SDH、ADH、GPI)进行检测分析。结果表明:检测的12种同工酶中的7种酶(SOD、EST、MDH、LDH、ALP、AMY、ME)具有明显的组织特异性;GPI在组织中表达不稳定,未检测到其他4种同工酶(ACP、G6PDH、SDH、ADH)的活性;光裸星虫酶谱带数目少,酶谱构型单一。并说明了对其种群内遗传生化特性进行系统研究的必要性。     

Isozymes Analysis of the Golden Cuttlefish Sepia esculenta (Cephalopoda: Sepiidae)

1　Introduction　ThegoldencuttlefishSepiaesculenta,aneriticanddemersalspecies,livesinthedepthof 1 0 - 1 0 0meters,andsometimesinseabottomsand .Itiswidelydistrib utedfromcentralHonshutoVietnamandthePhilip pines .Thegoldencuttlefishisalsoanimportantobjectof…     

BIOCHEMICAL GENETIC STUDIES ON CUTTLEFISH SEPIELLA MAINDRONI (CEPHALOPODA: SEPIIDAE)──ACTIVE LOCI SCREENING OF ISOZYME

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Genetic analysis of four populations of redlip mullet (Chelon haematocheilus) collected in China seas

Horizontal starch gel electrophoresis was used to investigate genetic structures of four populations (Dandong, Rizhao, Ningbo and Guangzhou) of redlip mullet (Chelon haematocheilus). Fourteen loci of ten enzymes (G3PDH, IDHP, LAP, LDH, MDH, MPI, PGDH, PGM, SDH, and SOD) were identified. The proportions of polymorphic loci varied from 0.07 to 0.14. The average observed and expected heterozygosities ranged from 0.02 to 0.05 and from 0.02 to 0.04 respectively among the 4 populations. The average number of efficient alleles of each locus varied from 1.02 to 1.06. The genetic distance among populations was from 0.0004 to 0.0021. The genetic distances between Dandong, Rizhao and Ningbo populations were small, while those between Guangzhou population and other three populations were relatively high. The result indicated the possible divergence of redlip mullet between the South China Sea and the other sea areas of China due to geographic isolation.     

BIOCHEMICAL GENETIC STUDIES ON CUTYLEFISH SEPIELLA MAINDRONI （CEPHALOPODA： SEPIIDAE）——ACTIVE LOCI SCREENING OF ISOZYME

Screening of 46 putative enzyme-coding loci and 4 different kinds of tissues of Sepiella maindroni de Rochebrone, 1884 for enzymatic activities using starch gel electrophoretic technique proved that the 21 enzymes such as AAT, AK, ALP, AP, CK, DIA, ES, FBP, G3PDH, GPI, GRS,IDH, LDH, MDH, MEP, MPI, NP, PGDH, PGM, SOD and XO* , were active to Sepiella maindroni after being stained. The tissue exhibiting stable and clear bands was also determined. Among tissues tested, mantle muscle tissue was the best for electrophoretic survey of isozymes. Buccal bulb muscle, eye and liver were fairly good for some special enzymes, such as DIA, ES, MPI, NT, etc.     

A Comparative Study of Genetic Variation betwccn Chinese Mitten Crab Eriocheir Sinensis and Hepu Mitten Crab E. Hepuensis

Horizontal starch gel electrophoresis was used to investigate the biochemical genetic structure of Chinese mittencrab Eriocheir sinensis and Hepu mitten crab E. hepuensis. Sixteen putative enzyme-coding loci were examined for forty-nineChinese mitten crabs and thirty-eight Hepu mitten crabs. Nine loci, AAT-1 *, AAT-2 *, G3PDH*, GPI*, IDHP-1 *, IDHP-2 *,MDH-1 *, MDH-2 * and PGM * , are polymorphic in Chinese mitten crab, and seven, AAT-1 *, AAT-2 *, GPI *, IDHP-1 *,MDH-1 *, MDH-2 * and PGM * , are polymorphic in Hepu mitten crab. The proportion of polymorphic loci and the expectedheterozygosity are 0.562 5 and 0.080 3 for Chinese mitten crab, and 0.437 5 and 0.075 4 for Hepu mitten crab. The Nei' s ge-netic distance between two species is 0.002 4.     

Genetic Analysis of Four Populations of Redlip Mullet(Chelon haematocheilus) Collected in China Seas

1 Introduction The redlip mullet, (Chelon haematocheilus) (Tem-minck et Schlegel, 1845), belongs to the family Mugili-dae. It is mainly distributed in the coastal areas of Korea, Japan and China (Wang et al., 2000). Because of the good adaptation to envir…     

Ontogenic study of isozymes in giant tiger prawn,Penaeus monodon (Crustacea: Decapoda)

The patterns of isozyme variation throughout the life cycle of the giant tiger prawn,Penaeus monodon were described for ten enzyme systems (G3PD, MDH, LGG, GPI, MPI, FBP, PGM, LDH, PGD, and PROT). The zymograms showed little variation in the number of isozymic loci expressed during larval development compared with results reported for other penaeid prawns. There were specific ontogenic patterns at some loci. As larvae developed beyond the embryonic stage, there was a tendency for intensity of staining and complexity of banding patterns to increase.     

月鳢不同组织中乳酸脱氢酶和酯酶同工酶的比较

采用垂直板聚丙烯酰胺凝胶电泳技术，对月鳢十种组织中乳酸脱氢酶（ＬＤＨ）和酯酶（ＥＳＴ）同工酶进行了比较研究，结果表明：月鳢组织内ＬＤＨ和ＥＳＴ具有明显的组织特异性，ＥＳＴ由２个座位编码，ＬＤＨ则由Ｌｄｈ－ａ、Ｌｄｈ－ｂ两个基因编码。     

Sequence comparison of partial cytochrome b genes of two Coilia species

Sequence variation of partial cytochrome b genes between two Coilia species, C. ectenes and C. mystus, was investigated. Of the 402 nucleotides, twenty-seven (6.72%) are polymorphic and all are synonymous substitutions. At the third positions of genetic condon of cytochrome b gene, the two species show an extreme anti-G bias (<4%) and a pronounced bias towards A and C (>68%). There is no amino acid sequence divergence between the partial cytochrome b genes of the two species, indicating a close genetic relationship between them. The k-2p genetic distance of partial cytochrome b segment of the two species is 0.072, suggesting that the species were separated 3.6 Ma ago, in the middle Pliocene. Our result reveals that the cytochrome b gene is an appropriate marker for studies of population genetic structures and phylogeographic patterns of the two species.     

Tissue Expression and Stock Variation of Isozymes of Stone Flounder （Kareius bicoloratus）

Tissue expression and stock variation of isozymes of stone flounder (Kareius bicoloratus) were analyzed with horizontal starch gel electrophoresis. For the fourteen enzymes assayed, 31 loci were recorded. The results indicated that all the isozymes ex- amined were obviously tissue-specific. The expressions of SOD*, GDH*, G3PDH-2* and ADH-2* were detected only in liver, SDH-1*, MDH-1* and ADH-1* only in muscle, and LDH-B* and LDH-C* only in eyes. In comparison, MDH-2*, GPI-3* and SDH-2* were detected in all tissues examined. Other loci examined were detected in a variety of tissues. Muscle and liver were se- lected to detect the isozyme variation of the two geographic stocks of Qingdao and Weihai, Shandong Province, China. The percent- ages of polymorphic loci (P0.99) were 29.17% and 25.00%, the observed heterozygosities (H0) were 0.028 ± 0.014 and 0.040 ± 0.019, and the expected heterozygosities (He) were 0.039 ± 0.017 and 0.052 ± 0.022 in Qingdao and Weihai stock, respectively. The coeffi- cient of gene differentiation (Fst) and genetic distance (D) between the two stocks was 0.012 and 0.0011, respectively, indicating that the genetic differentiation is low between them. Compared with other species of Pleuronectiformes, both the percentage of polymor- phic loci and the mean heterozygosity of K. bicoloratus were at a middle level.     

A comparative study of genetic variation between Chinese mitten crab <Emphasis Type="Italic">ericheir sinensis</Emphasis> and Hepu mitten crab <Emphasis Type="Italic">E. hepuensis</Emphasis>

Horizonal starch gel electrophoresis was used to investigate the biochemical genetic structure of Chinese mitten crab Eriocheir sinensis and Hepu mitten crab E. hepuensis. Sixteen putative enzyme-coding loci were examined for forty-nine chinese mitten crabs and thirty-eight Hepu mitten crabs. Nine loci, AAT-1 ^*, AAT-2 ^*, G3PDH ^*, GPI ^*, IDHP-1 ^*, IDHP-2 ^*, MDH-1 ^*, MDH-2 ^* and PGM ^*, are polymorphic in Chinese mitten crab, and seven, AAT-1 ^*, AAT-2 ^*, GPI ^*, IDHP-1 ^*, MDH-1 ^*, MDH-2 ^* and PGM ^*, are polymorphic in Hepu mitten crab. The proportion of polymorphic loci and the expected heterozygosity are 0.562 5 and 0.080 3 for Chinese mitten crab, and 0.437 5 and 0.075 4 for Hepu mitten crab. The Nei’s genetic distance between two species is 0.002 4.     

Isozyme expression of Chinese and Japanese populations of Chlamys farreri and their reciprocal hybrids

Chinese and Japanese population ofChlamys farreri and their reciprocal hybrids were surveyed in isozyme variability at 13 loci by polyacrytamide gel electrophoresis (PAGE). Isozyme banding patterns indicated these hybrids were diploid. Loci that were observed as being monomorphic in inbred populations ofC. farreri were also found to be monomorphic in filial progeny; loci that observed to be polymorphic in parental type generations were also polymorphic in hybrid generations. Differences existed among allelic frequency of the four types of cross. Within the reciprocal hybrids the expression of malic enzyme (ME) isozyme was sufficient to distinguishing individual hybrids because of the band, Rf=0.38. However, there were no noticeable variations among all the samples to differentiate one from another. Inbreeding was likely to be the main problem in aquaculture. The introduction of new broodstock can improve the genetic diversity. Hybrid vigor has manifested to a certain extent in the present study. Supported by the National “973” Program (G1999012008) and “863” Program (2002AA626020).     

Invasion and spreading of Cabomba caroliniana revealed by RAPD markers

Applying randomly amplified polymorphic DNA (RAPD), the genetic variation ofCabomba caroliniana Gray (cabomba or fanwort), a new alien plant in China, was analyzed in this paper. Total 143 bands, including 47 polymorphic bands, were amplified from 23 primers in 20 samples. The sampling distance was large, but its genetic diversity was low. The main results were that: (1) Cabomba, which grew and dispersed mainly in fragment, was an abundant and dominant species in freshwater, and its main dispersal mechanism was vegetative reproduction (2) Cabomba was originally introduced into China as an aquarium submerged plant. Somehow, those discarded cabomba became invasive species in the areas of Hangzhou, Shanghai, and Meicheng, and other places. (3) Although the level of genetic diversity in cabomba was low, their rapid dispersion and propagation could seriously harm to local aquatic community. Therefore, specific measure should be used to control cabomba from uncontrolled spreading and damage to local vegetation communities. Supported by National Natural Science Foundation of China (No. 39970145) and Major State Basic Research Development Program of China (No. G2000046803).     

Genetic differentiation withinEriochoir sinensis (milne, edwards) revealed by allozyme and RAPD

We analyzed 17 allozymes, and 20 primers in order to detect the genetic differentiation between commercial populations (Changjiang River, Liaohe River) ofEriochoir sinensis. Ten allozymes (LDH, MDH, ME, IDH, EST, ALP, AAT, CTL, POD, SOD) showed 21 loci by vertically discontinuos buffer system polyacrylamide gel electrophoresis. RAPD profiles generated by 12 ten-base primers showed 63 loci. The percentage of polymorphic loci and the expected heterozygosity obtained by using allozyme analysis were lower than those obtained by RAPD. The index of similarity between these two populations were 0.955 and 0.932 as revealed by allozyme analysis and RAPD technology. There was gene flow between the above populations. Contribution No. 4025 from the Institute of Oceanology and Limnology, Chinese Academy of Sciences. Project 39470141 supported by NSFC. Work also supported by Climbing Program B (PDB 6-5-3) and No. 863 High-technology Program 863-819-01-05.     

Application of AFLP markers for population genetic study on half-smooth tongue sole Cynoglossus semilaevis

The genetic diversity of wild and hatchery populations of half-smooth tongue sole Cynoglossus semilaevis, based on observation of amplified fragment length polymorphism (AFLP) was described. Two hundred individuals from four wild populations, Laizhou (LZ), Weihai (WH), Qingdao (QD), Rizhao (RZ), and one hatchery population, Mingbo (MB), were screened using eight different AFLP primer combinations. A total of 384 loci were screened in the five studied populations. 48.4%, 51.3%, 50.7%, 49.3% and 45.8% of these loci were polymorphic among the individuals tested in the LZ, WH, QD, RZ and MB populations, respectively. The number of polymorphic loci detected by single primer combinations ranged from 17 to 35. The average heterozygosity of the LZ, WH, QD, RZ and MB populations was 0.072, 0.093, 0.092, 0.090 and 0.063, respectively. The WH population showed the highest genetic diversity in terms of total number of AFLP bands, total number of polymorphic bands, average heterozygosity and percentage of low frequency (0-0.2) polymorphic loci among all the populations, while the LZ population was the lowest among the wild populations. Compared with the wild populations, the hatchery population showed a low genetic viability.     

Genetic differentiation of Japanese sardinella (Sardinella zunasi) populations in the Northwest Pacific revealed by ISSR analysis

Knowledge of population genetic structure plays an important role in fisheries management.In this research,In-ter-Simple-Sequence-Repeat(ISSR)markers were employed to evaluate the genetic structure of Japanese sardinella(Sardinella zun-asi)populations in the Northwest Pacific.Eighty seven individuals from 5 locations were screened using 4 highly polymorphic prim-ers.A total of 173 polymorphic loci were detected out of 191 loci amplified.Small but significant genetic differentiation was de-tected between the Chinese and Japanese populations by both AMOVA and pairwise FST analyses,which was further supported by cluster analysis.We consider that climate change during glaciations should be responsible for the genetic differentiation.Isolation by geographic distance among populations was observed,indicating that the distance might also lead to the genetic differentiation.However,no genetic structure was found within the populations off both the Chinese and Japanese coasts,indicating a high-level along-coast gene flow,which might result from ocean current transport and common ground for over-wintering.