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1.
Chinese shrimp(Fenneropenaeus chinensis)is an economically important aquaculture species in China.However,cytogenetic and genomic data is limited in the organism partly because the chromosomes are difficult to isolate and analyze.In this study,fluorescence in-situ hybridization(FISH) was used to identify the chromosomes of F.chinensis.The 5S ribosomal RNA gene(rDNA)of F. chinensis was isolated,cloned and then used as a hybridization probe.The results show that the 5S rDNA was located on one pair of homologous chromosomes in F.chinensis.In addition,triploid shrimp were used to evaluate the feasibility of chromosome identification using FISH and to validate the method.It was confirmed that 5S rDNA can be used as a chromosome-specific probe for chromosome identification in F.chinensis.The successful application of FISH in F.chinensis shows that chromosome-specific probes can be developed and this finding will facilitate further research on the chromosomes of penaeid shrimps. 相似文献
2.
The family Sciaenidae is remarkable for its species richness and economic importance. However, the cytogenetic data available in this fish group are still limited, especially those obtained using fl uorescence in situ hybridization(FISH). In the present study, the chromosome characteristics of a sciaenid species, Argyrosomus amoyensis, were examined with several cytogenetic methods, including dual-FISH with 18 S and 5 S rDNA probes, and a self-genomic in situ hybridization procedure(Self-GISH). The karyotype of A. amoyensis comprised 2 n=48 acrocentric chromosomes. A single pair of nucleolar organizer regions(NORs) was located at the proximal position of chromosome 1, which was positive for silver nitrate impregnation(AgNO_3) staining and denaturation-propidium iodide(DPI) staining but negative for Giemsa staining and 4',6-diamidino-2-phenylindole(DAPI) staining, and was confi rmed by FISH with 18 S rDNA probes. The 5 S rDNA sites were located at the centromeric region of chromosome 3. Telomeric FISH signals were detected at all chromosome ends with dif ferent intensities, but internal telomeric sequences(ITSs) were not found. Self-GISH resulted in strong signals distributed at the centromeric regions of all chromosomes. C-banding revealed not only centromeric heterochromatin, but also heterochromatin that located on NORs, in interstitial and distal telomeric regions of specifi c chromosomes. These results suggest that the karyotype of Amoy croaker was relatively conserved and primitive. By comparison with the reported cytogenetic data of other sciaenids, it can be deduced that although the karyotypic macrostructure and chromosomal localization of 18 S rDNA are conserved, the distribution of 5 S rDNA varies dynamically among sciaenid species. Thus, the 5 S rDNA sites may have different evolutionary dynamics in relation to other chromosomal regions, and have the potential to be ef fective cytotaxonomic markers in Sciaenidae. 相似文献
3.
《中国海洋湖沼学报》2021,(2)
It has been reported that there was a linkage of 5 S rRNA gene to 18 S-5.8 S-25 S rRNA gene in a few of species in Ochrophyta.In regard to the usual two positions of linked 5 S rDNA to the 3' end of 25 S rDNA,two pairs of primers were designed for amplification to verify this linkage of two genes in a kelp cultivar of Saccharina japonica,one of species in Ochrophyta.This result supplemented the previous report that 5 S rDNA was unlinked to 25 S rDNA in this kelp.In order to simultaneously visualize this unlinkage of two genes,dual-color fluorescence in situ hybridization(FISH) technique was applied to the cytogenetics of S.japonica.Dual-color FISH images showed that two and four hybridization signals were present in the kelp gametophyte and sporophyte,respectively,metaphase nuclei hybridized simultaneously with the labeled probes of 18 S rDNA and 5 S rDNA.Both haploid and diploid karyotypes in decreasing length of chromosomes showed that 18 S-5.8 S-25 S rDNA was localized at the interstitial region of Chromosome 23,whereas 5 S rDNA resided at the sub-telomeric region of Chromosome 27.These karyotypes suggested that the kelp nuclear genome had only one locus of each rRNA gene,and their loci on different chromosomes indicated the physical unlinkage of 5 S rDNA to 18 S-5.8 S-25 S rDNA in this kelp.Therefore,dual-color FISH seems to be a powerful technique for the discrimination and pairing of chromosomes featured in both small size and nearly identical shape in S.japonica. 相似文献
4.
The pathological changes of hemocytes in the haemolymph and hepatopancreas were examined in experimentally and naturally WSSV
(white spot syndrome virus) infected Fenneropenaeus chinensis. The results showed that the pathological manifestations of hemocytes were similar among moribund shrimps infected via injection,
feeding and by nature. Firstly, the total hemocyte counts (THCs) in WSSV-infected shrimp were significantly lower than those
in healthy shrimp. Secondly, necrotic, broken and disintegrated cells were often observed, and a typical hematolysis was present
in the haemolymph smear of WSSV-infected shrimp. Thirdly, necrosis and typical apoptosis of hemocytes were detected with TEM
in the peripheral haemolymph of WSSV-infected shrimp. Hyalinocytes and semi-granulocytes with masses of WSSVs in their nuclei
often appeared, whereas no granular hemocytes with WSSV were found in the hepatopancreas of moribund infected shrimps. All
our results supported that hemocytes were the main target cells of WSSV, and hyalinocytes and semigranular hemocytes seemed
to be more favorable for WSSV infection in F. chinensis. 相似文献
5.
A muscle cDNA library of Chinese shrimp (Fenneropenaeus chinensis) was constructed with the SMART™ cDNA Library Construction Kit. The titer of optimal primary library was 7.7×105 pfu mL−1 and that of the amplified library was 3.0×109 pfu mL−1. The percentages of the recombinant clones of primary and amplified libraries were over 98%. The insert sizes were longer
than 400 bp with an average of 1000 bp. A positive clone containing a 794 bp insert was sequenced and identified encoding
fast skeletal troponin I gene. This library provided a useful resource for the functional genomic research of F. chinensis. 相似文献
6.
YU Shanshan KONG Xiaoyu LI Yulong XU Hui 《中国海洋大学学报(英文版)》2007,6(2):187-192
The complete mitochondrial cytochrome oxidase subunit Ⅱ （COⅡ） gene of Penaeinae shrimp Fenneropenaeus chinensis was cloned and sequenced. The gene is 688 bp in length and codes for 229 amino acids. It shows 83.2%, 87.0% and 83.8% sequence similarity to Marsupenaeus Japonicus, Penaeus monodon and Farfantepenaeus notialis, respectively. The A＋T content of the whole gene and that at the third position of codons are 64.7% and 78.2%, respectively. The phylogenetic relationship between F. chinensis and three other species representing genera Farfanatepenaeus, Marsupenaeus and Penaeus was analyzed. Results showed that the genetic distances among the four taxa ranged from 0.144 0 to 0.200 5, exceeding those estimated with COⅠ and partial 16S rRNA gene sequences among Marsupenaeus, Litopenaeus and Melicertus, and being therefore larger than the value among subgenera. It has been suggested that the COⅡ gene has a faster evolutionary rate than that of the COⅠ gene and partial 16S rRNA gene and could be used for phylogenetic analysis at genus or species level. The results of the present study indicated that Farfantepenaeus, Fenneropenaeus, Marsupenaeus and Penaeus are at a higher phylogenetic level than subgenus, which supports the opinion of the elevation of phylogenetic status of the four subgenera to genus level. 相似文献
7.
Stable isotope methods can be used to determine the food sources and prey items of aquatic organisms accurately and reliably.
This study examined the relative contribution of artificial foods (the formulated feed and Artemia) and natural foods to shrimp growth in an intensive Fenneropenaeus chinensis pond by using carbon and nitrogen stable isotopes. The results showed that the nutrition utilization efficiency of the harvested
shrimp was low, only 33.18% of feed nitrogen and 21.73% of feed carbon being converted to shrimp flesh. Our stable isotope
results showed that the shrimp obtained nutrition for maximum growth from artificial foods, whose contribution was 93.5%,
with the remaining attributed to the natural foods. However, there was 0.94 t harvested shrimp derived from natural foods
(the rest of 13.56 t harvested shrimp derived from artificial foods) in 1ha intensive pond with a shrimp production of 14.50
t ha−1. Therefore, unit area shrimp production can be increased by increasing the contribution proportion of natural foods in intensive
shrimp farming. 相似文献
8.
In this study, the intestinal microbiota of kuruma shrimp (Marsupenaeus japonicus) was examined by molecular analysis of the 16S rDNA to identify the dominant intestinal bacteria and to investigate the effects
of Bacillus spp. on intestinal microbial diversity. Samples of the intestines of kuruma shrimp fed normal feed and Bacillus spp. amended feed. PCR and denaturing gradient gel electrophoresis (DGGE) analyses were then performed on DNA extracted directly
from the guts. Population fingerprints of the predominant organisms were generated by DGGE analysis of the universal V3 16S
rDNA amplicons, and distinct bands in the gels were sequenced. The results suggested that the gut of kuruma shrimp was dominated
by Vibrio sp. and uncultured gamma proteobacterium. Overall, the results of this study suggest that PCR-DGGE is a possible method of studying the intestinal microbial diversity
of shrimp. 相似文献
9.
Eight hundred and thirty eight base pair fragment of mitochondrial COI gene of wild and cultured populations (CP1, CP4, CP5
and CP6) of Fenneropenaeus chinensis was amplified and sequenced. The A, T, G and C contents of the sequence were 235 bp (28.0%), 307 bp (36.6%), 138 bp (16.5%)
and 158 bp (18.9%), respectively. Furthermore, 556 bp fragment of the sequence was used to discuss the phylogenetic relationship
among 14 Penaeidae species using Alpheus armillatus as the outgroup. From the molecular phylogenetic tree constructed by neighbor-joining method, we obtained three large shrimp
groups: Farfantepenaeus, Litopenaeus and Fenneropenaeus group. The results also indicated that there were a closer genetic relationships between F. aztecus and F. paulensis, L. schmitti and L. setiferus, F. indicus and F. merguiensis, and the genus Farfantepenaeus was closer to Litopenaeus. 相似文献
10.
The Chinese shrimp Penaeus (Fenneropaeneus) chinensis is an important species in marine fishery and aquaculture in China. A female Chinese shrimp Penaeus (Fenneropaeneus) chinensis was captured from west coast of the Korean peninsula and mated with a “Yellow Sea No. 1” male to produce the first filial
generation (F1) 100 F2 full-sib progeny from brother-sister crosses between F1 families was used for the mapping study. A genetic linkage map of the Chinese shrimp was constructed, based on 354 markers,
including 300 amplified fragment length polymorphism (AFLP) markers, 42 microsatellite (SSR) markers, and 12 randomly amplified
polymorphism (RAPD) markers. Forty-seven linkage groups (LGs) were identified. The total map length was 4 580.5 cM, with an
average spacing of 11.3 cM, covering 75.8% of the estimated genome size. The construction of this genetic linkage map was
part of a genetic breeding program. This linkage map will contribute to the discovery of genes and quantitative trait loci
(QTLs) in Chinese shrimp. 相似文献
11.
12.
A harmful algae bloom (HAB) is a dense aggregation of algae in a marine or aquatic environment that can result in significant
environmental problems. To forecast the occurrence of HAB, development of a rapid and precise detection method is urgently
required. In this study, two Skeletonema costatum-like diatoms (SK-1 and SK-2), were identified morphologically under a light microscope, and detected using fluorescent in situ hybridization (FISH). Strain SK-1 was isolated from a frequently HAB affected area of the East China Sea, and strain SK-2
from an aquatic farm in Qingdao, China. Fluorescent DNA probes were designed that were complementary to the ITS sequence (including
5.8S rDNA) of strain SK-1. After hybridization, strong green fluorescence was observed in cells of strain SK-1 under an epifluorescence
microscope; however, no such fluorescence was observed with strain SK-2, which indicates that probes hybridized only the DNA
of the target strain, SK-1, in species-specific manner, and that the two strains do not belong to a same species. This finding
was confirmed by ITS sequence analysis. The FISH technique used in this study was sensitive, simple, and rapid, and is a promising
tool for detecting target HAB species in natural environments. 相似文献
13.
The wild Porphyra yezoensis collected from the Qingdao coast was used to prepare protoplasts by enzyme digestion. The pure line was constructed by cultivating the protoplasts. The 18S rDNA of the P. yezoensis pure line was cloned and sequenced. Sequence analysis was executed for this sequence and other 22 sequences retrieved from GenBank. A phylogenetic tree was constructed using the neighbor-joining method. The results revealed a high diversity of 18S rDNA sequences in genus Porphyra and the considerable variation of 18S rDNA sequences in different strains of the same species P. yezoensis and P. tenera. Significant difference of 18S rDNA sequence was observed between P. yezoensis from Qingdao, China, and the two strains of P. yezoensis from Japan, but the three strains of P. yezoensis formed a stable clade in the phylogenetic tree. These results indicate the possibility of interspecies and intraspecies discrimination of Porphyra using the 18S rDNA sequences. 相似文献
14.
This study examined the distribution and elimination of Norfloxacin(NFLX) in Fenneropenaeus chinensis ovary and egg and newly hatched larvae.Mature parental shrimp were exposed to 4 or 10 mg L 1NFLX for 2 or 5 d.Ovary and eggs of the shrimp were sampled after spawning in order to detect NFLX residue using high-performance liquid chromatography(HPLC).Results showed that NFLX residue accumulated in F.chinensis eggs after the parental exposure,with the highest residue detected in ovary.To examine the fate of NFLX residue in larvae,we further determined the concentration of NFLX residue in F.chinensis eggs and larvae at 4 different developmental stages after 24-h exposure.From the newly metamorphosed larvae(0 h post-metamorphosis,h.p.m),samples were taken at different time intervals to 72 h.p.m.HPLC assay showed that the concentrations of NFLX residue in zoea exposed to 4 and 10 mg L 1NFLX were the highest at 1.5 h,i.e.,0.332 and 0.454 μg g 1,respectively.At the two NFLX exposure levels,the elimination time of half NFLX(half life) in nauplius was 45.36 and 49.85 h,respectively,followed by that in zoea(31.68 and 33.13 h),mysis larvae(42.24 and 47.28 h) and postlarvae(24.48 and 30.96 h).Both NFLX exposure levels had a germicidal effect.The distribution and elimination of NFLX residue in F.chinensis tissue,eggs and larvae correlated well with the drug exposure level.The disappearance of NFLX residue coincided with the larval growth,and the half-life of NFLX decreased with the larval development. 相似文献
15.
The marine green alga Chaetomorpha valida fouls aquaculture ponds along the coastal cities of Dalian and Rongcheng, China. Unialgal cultures were observed under a microscope to determine the developmental morphological characters of C. valida. Results reveal that gametophytic filaments often produce lateral branches under laboratory culture conditions, suggesting an atypical heteromorphic life cycle of C. valida between unbranched sporophytes and branched gametophytes, which differs from typical isomorphic alternation of Chaetomorpha species. The shape of the basal attachment cell, an important taxonomic character within the genus, was found variable depending on environmental conditions. The 18S rDNA and 28S rDNA regions were used to explore the phylogenetic affinity of the taxa. Inferred trees from 18S rDNA sequences revealed a close relationship between C. valida and Chaetomorpha moniligera. These results would enrich information in general biology and morphological plasticity of C. valida and provided a basis for future identification of green tide forming algae. 相似文献
16.
Potassium, sodium, magnesium and iodine requirements of shrimp,Penaeus chinensis were studied. Orthogonal design was employed in this experiment. The composition of the basal diet consisted of fish meal,
peanut cake, corn meal, soybean cake, wheat bran, vitamin mix and mineral mix, and supplementations of potassium, sodium,
magnesium and iodine in the basal diet were made according to the L9(34) orthogonal table. The results indicated that iodine supplementation improved growth of the shrimp significantly and raised
survival very significantly, iodine requirement of the shrimp was 0.003%; sodium requirement was 0.87% or less; and that 1.1–1.3%
potassium and 0.18–0.38% magnesium in the diet were proper nutrition supplements for the shrimp.
Contribution No. 2342 from the Institute of Oceanology, Chinese Academy of Sciences. 相似文献
17.
18.
HUANGGuoqiang DONGShuanglin WANGFang 《中国海洋大学学报(英文版)》2005,4(1):56-60
Attractabilities of different diets and dietary selectivity of Chinese shrimp, Fenneropenaeus chinensis were studied through behavior observation and feeding experiment, respectively. The five diets used in the experiment are: Fish Flesh (FF), Shrimp Flesh (SF), Clam Foot (CF), Polychaete Worm (PW), and Formulated Diet (FD). No significant differences of attractability exist between any two different diets when every two natural diets or all five diets are provided simultaneously. On the other hand, significant differences of attractability exist between FD and every single natural diet when they are provided simultaneously. Results of behavioral observation indicate that natural diets are more attractive than FD. In feeding experiment, Chinese shrimp has distinct selectivity on different diets. It positively selects CF and PW, negatively selects FF and SF, and excludes FD absolutely. The results of the present studies indicate that the dietary selectivity of shrimp was based not only on the attractabilities of the diets, but also on the responses such as growth and food conversion. 相似文献
19.
We collected nine Enteromorpha specimens from the coast of Yantai and evaluated their diversity based on analyses of their ITS （internal transcribed spacer） and 5S rDNA NTS （non-transcribed spacer） sequences. The ITS sequences showed slight nucleotide divergences between Enteromorpha linza and Enteromorpha prolifera. In contrast, multiple highly variable regions were found in the ITS region of Enteromorpha flexuosa. In general, there were more variable sites in the NTS region than in the ITS region in the three species. The variations in 5S rDNA NTS sequences indicated that the molecular diversity of Enteromorpha from the coast of Yantai is very high. However, a phylogenetic tree constructed using 5S rDNA NTS sequence data indicated that genetic differences were not directly related to geographical distribution. 相似文献
20.
The dinoflagellate Alexandrium minutum is often associated with harmful algal blooms (HABs). This species consists of many strains that differ in their ability to produce toxins but have similar morphology, making identification difficult. In this study, species-specific rRNA probes were designed for whole-cell fluorescence in situ hybridization (FISH) to distinguish A. minutum from two phylogenetic clades. We acquired the complete SSU to LSU rDNA sequences (GenBank accession numbers JF906989-JF906999) of 11 Alexandrium strains and used these to design rRNA targeted oligonucleotide probes. Three ribotype-specific probes, M-GC-1, M-PC-2, and M-PC-3, were designed. The former is specific for the GC clade ("Global clade") of A. minutum, the majority of which have been found non-toxic, and the latter two are specific for the PSP (paralytic shellfish poisoning)-producing PC clade ("Pacific clade"). The specificity of these three probes was confirmed by FISH. All cells in observed fields of view were fluorescently labeled when probes and target species were incubated under optimized FISH conditions. However, the accessibility of rRNA molecules in ribosomes varied among the probe binding positions. Thus, there was variation in the distribution of positive signals in labeled cells within nucleolus and cytosol (M-GC-1, M-PC-3), or just nucleolus (M-PC-2). Our results provide a methodological basis for studying the biogeography and population dynamics of A. minutum, and providing an early warning of toxic HABs. 相似文献