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In this study, bulked segregant analysis (BSA) was used on Larix leptolepis × Larix olgensis hybrids to identify a random amplified polymorphic DNA (RAPD) marker associated with high rooting ability in larch. Two DNA bulks: H (high rooting ability) bulk and L (low rooting ability) bulk were constructed according to the rooting percentages of the stock plants. Among the 328 primers, only S356 could amplify a specific band, named S356445, which only existed in the H bulk and was further confirmed following selective genotyping of individual hybrids. Grounded on the border sequences, S356445 was converted to a sequence characterized amplified region (SCAR) marker, HRL445, which can be useful in marker-assisted selection (MAS) to screen for larch with high rooting ability. All the results strongly indicated that S356445 and HRL445 were closely associated with high rooting ability in larch.

In this study, seven cDNA fragments of genes, differentially expressed in ovaries after pollination in Cymbidium hybridium, were identified and characterized by mRNA differential display reverse transcription polymerase chain reaction (DDRT-PCR). Four (CDD-313, CDD-272, CDD-265, CDD-243) among these seven cDNA fragments showed no significant homology with ESTs or genes in the databases of NCBI; another three (CDD-193, CDD-218, CDD-470) showed significant homologies with sequences encoding components of an ABC-type transporter, a GTPase and 40S ribosomal S3 proteins (RPS3), respectively. The differential expression patterns of them were confirmed by reverse Northern dot blot analysis. More interestingly, CDD-470 appeared to be present and highly expressed in the pollinated ovaries and encoded a new factor of RPS3 participating in cell growth and proliferation. We deduced that this 40s ribosomal S3 like protein was involved in ovary development of orchids.

The positive surgical margins are associated with postsurgical recurrence in hepatocellular carcinoma patients, and molecular margin analysis is considered more sensitive in detecting preneoplastic lesions than conventional histological margin examination. To evaluate the feasibility of methylation-based molecular margin analysis in HCC and explore its clinical application, we investigated CDKN2A methylation status in the surgical margins of 20 HCC patients using a nested BS-MSP protocol and compared the methylation patterns in resection margins with those in the corresponding tumor and adjacent nonmalignant tissues. The results showed that a considerable frequency (35%, 7 of 20) of CDKN2A methylation was present in histologically negative margins, and methylation pattern analysis might be valuable for studying the cellular origin of recurrent carcinoma. Therefore, methylation-based molecular surgical margin analysis offers a promising tool in prognosis for HCC patients who underwent hepatectomy.

Amplified Fragment Length Polymorphism (AFLP) markers were successfully employed to analyze 15Artemia species and strains for genetic diversity. AFLP markers are extremely sensitive to even a small sequence variation. They are stable and more polymorphic than RAPD. Twelve pairs of primer combinations were used to detect AFLP bands, of which 384 were polymorphic, and DNA fingerprintings were obtained by using silver staining. The polymorphism analysis leads us to the following conclusions: 1.Artemia tibetiana seems to differentiate fromA. sinica. 2. The parthenogenetic populations from inland salt lakes could follow an evolutionary path that is different from that of the coastal parthenogenetic populations.

A simple method to create a chromosome-specific DNA library of rice, including microdissection, amplification, characterization and cloning, is described. Rice chromosome 4 from a metaphase cell has been isolated and amplified by the Linker Adapter PCR (LA-PCR). The PCR products were labeled as probes with DIG-11-dUTP using the random priming method. Southern blot analysis with rice genomic DNA and specific RFLP markers demonstrated that the PCR products were derived from rice chromosome 4. A large library comprising over 100,000 recombinant plasmid microclones from rice chromosome 4 was constructed. Colony hybridization showed that 58% of the clones contained single or low-copy sequences and 42% contained repetitive sequences. The size of inserts generated by PCR ranged from 140bp to 500bp. This method will facilitate cloning of the specific chromosome DNA markers and important genes of rice.

To investigate the prevalence and time of Dickkopf (DKK) family methylation and its clinical significance in hepatocarcinogenesis.Methylation of DKK family genes was quantitatively analyzed in 115 liver tissue samples, including 50 pairs of primary hepatocellular carcinoma (HCC) and matched noncancerous cirrhotic tissue samples, as well as 15 liver cirrhosis biopsy samples.The methylation level of DKK3 was significantly higher in HCC tissue samples than in matched noncancerous cirrhotic tissue samples (P0.0001) or in liver cirrhosis biopsy samples (P = 0.0139). Receiver operator characteristic curve analysis confirmed that the percent of methylated reference (PMR) values of DKK3 could effectively discriminate HCC tissue samples from noncancerous tissue samples (AUC = 0.8146) or liver cirrhosis biopsy samples (AUC = 0.7093). Kaplan-Meier survival curves revealed that the progression-free survival time of patients with a higher DKK3 methylation level (PMR1%) was significantly shorter than that of those with a lower DKK3 methylation level (PMRor = 1%) (P = 0.0255). Multivariate Cox analysis indicated that methylated DKK3 was significantly and independently related with a shorter survival time (relative risk = 2.527, 95% CI: 1.063-6.008, P = 0.036) of HCC patients.Methylation of DKK3 is an important event in early malignant transformation and HCC progression, and therefore might be a prognostic indicator for risk assessment of HCC.

Black rot, caused by Xanthomonas campestris pv. campestris (Pammel) Dowson (Xcc), is one of the most damaging diseases of cauliflower and other crucifers. In order to investigate the molecular resistance mechanisms and to find the genes related to black rot resistance in cauliflower, a suppression subtractive hybridization (SSH) cDNA library was constructed using resistant line C712 and its susceptible near-isogenic line C731 as tester and driver, respectively. A total of 280 clones were obtained from the library by reverse northern blotting. Sequencing analysis and homology searching showed that these clones represent 202 unique sequences. The library included many defense/disease-resistant related genes, such as plant defensin gene PDF1.2, lipid transfer protein, thioredoxin h. Gene expression profiles of 12 genes corresponding to different functional categories were monitored by real-time RT-PCR. The results showed that the expression induction of these genes in the susceptible line C712 in response to Xcc was quicker and more intense, while in C731 the reaction was delayed and limited. Our results imply that these up-regulated genes might be involved in cauliflower responses against Xcc infection. Information obtained from this study could be used to understand the molecular mechanisms of disease response in cauliflower under Xcc stress.

To screen and determine the regions of copy number variation (CNV) associated with hepatocellular carcinoma (HCC) using SNP array and fluorescence quantitative PCR.The CNV from HCC cell line TJ3ZX-01 was analyzed using GeneChip Human Mapping 500K SNP array. According to the data obtained by SNP array analysis, four candidate amplification regions were verified in 41 primary HCC samples by fluorescence quantitative PCR.Four regions of copy number amplification at 1q21.2, 1q22 approximately 23.1, 7p22.1 and 22q13.1 were detected by SNP array analysis. The four candidate amplicons occurred in 56.1% (23/41) of HCC samples at 1q21.2; 80.5% (33/41) at 1q22 approximately 23.1; 75.6% (31/41) at 7p22.1 and 31.7% (13/41) at 22q13.1 analyzed with sequence tagged site (STS) markers by quantitative PCR.In four candidate amplification regions selected by SNP array analysis and detected by fluorescence quantitative PCR, three amplification regions show increased copy number in more than 50.0% HCC tissues. This result indicates that these amplification regions are associated with pathogenesis of hepatocellular carcinoma.

DNA methylation is one of the major epigenetic modifications. It is very important to the regulation of gene expression. In present study, an autoploidy series (2x, 3x and 4x) in watermelon (Citrullus lanatus) was constructed and MSAP (Methylation-Sensitive Amplification Polymorphism) analysis was conducted to elucidate the level and pattern of DNA methylation at CCGG sites in different ploidy watermelons. Totally, 1883 genetic loci were produced by 23 pairs of selective primers, of which 647, 655 and 581 sites were detected in diploid, autotriploid and autotetraploid, respectively. The methylation sites were 181, 150 and 159, and the corresponding total methylation ratios were 28.0%, 22.9% and 27.4% in 2x, 3x and 4x, respectively, of which the fully methylation sites were 121, 80 and 82, and the corresponding fully methylation ratios were 18.7%, 12.2% and 14.1%. Further analysis of the pattern of DNA methylation suggested that compared 4x with 2x, about half of detected sites (54.4%) shown changes of DNA methylation patterns. Similarly, compared 4x with 3x, 45.4% sites also shown changes of DNA methylation patterns. Moreover, the trend of DNA methylation adjustment mainly involved increase of DNA methylation levels in 4x. However, compared 3x with 2x or 4x, although the changes of DNA methylation pattern also widely occurred, which involved 41.6% (compared 3x with 2x) and 45.4% (compared 3x with 4x) sites, respectively, the trend of DNA methylation adjustment mainly involved decrease of DNA methylation levels in 3x. All these results indicated that DNA methylation events were widely existed in different ploidy watermelons. However, not only based on the total DNA methylation ratio or fully DNA methylation ratio, the results both implied that the DNA methylation levels were not closely associated with the autopolyploidy level in watermelon. Autotriploid watermelon shows obvious low level of DNA methylation. Analysis of DNA methylation patterns also suggested that the adjustment of DNA methylation patterns in autotriploid mainly involved demethylation events, implying the unusual characteristic of DNA methylation status in 3x watermelon. The present results are valuable to further explore the nature of triploid vigor and autopolyploidizaion in watermelon from the view of epigenetics.

Genome copy number variation (CNV) is one of the mechanisms to regulate the expression level of genes which contributes to the development and progression of cancer. In order to investigate the regions of high-level amplification and potential target genes within these amplicons in hepatocellular carcinoma (HCC), we analyzed HCC cell line (TJ3ZX-01) for CNV regions at the whole genome level using GeneChip Human Mapping 500K array, and also examined the relative copy number and expression levels of the related genes at candidate amplicons in 41 HCC tissues via real-time fluorescence quantitative PCR methods. Through analysis of sequence tag site (STS) markers by quantitative PCR, The two candidate amplicons at 1q found by SNP array were shown to occur in 56.1% (23/41) HCC samples at 1q21 and 80.5% (33/41) at 1q22–23.1. Wilcoxon signed rank test showed expression of CD1d, which located at amplicon of 1q22–23.1 increased significantly within tumor tissues compared with paired nontumor tissues. Our study provides evidences that a novel, high-level amplicon at 1q22–23.1 occurs in both HCC cell line and tissues. CD1d is a potential target for this amplicon in HCC. The up-regulation of CD1d may be used as a novel molecular signature for diagnosis and prognosis of HCC.

Analysis of ISSR (Inter-Simple Sequence Repeat) and DDRT-PCR (Differential Display Reverse Transcriptase Polymerase Chain Reaction) was performed between cytoplasmic male sterility cauliflower ogura-A and its corresponding maintainer line ogura-B. Totally, 306 detectable bands were obtained by ISSR using thirty oligonucleotide primers. Commonly, six to twelve bands were produced per primer. Among all these primers only the amplification of primer ISSR3 was polymorphic, an 1100 bp specific band was only detected in maintainer line, named ISSR3(1100). Analysis of this sequence indicated that ISSR3(1100) was high homologous with the corresponding sequences of mitochondrial genome in Brassica napus and Arabidopsis thaliana,which suggested that ISSR3(1100) may derive from mitochondrial genome in cauliflower. To carry out DDRT-PCR analysis, three anchor primers and fifteen random primers were selected to combine. Totally, 1122 bands from 1 000 bp to 50 bp were detected. However, only four bands, named ogura-A 205, ogura-A383, ogura-B307 and ogura-B352, were confirmed to be different display in both lines. This result was further identified by reverse Northern dot blotting analysis. Among these four bands, ogura-A205 and ogura-A383 only express in cytoplasmic male sterility line, while ogura-B307 and ogura-B352 were only detected in maintainer line. Analysis of these sequences indicated that it was the first time that these four sequences were reported in cauliflower. Interestingly, ogura-A205 and ogura-B307 did not exhibit any similarities to other reported sequences in other species, more investigations were required to obtain further information. ogura-A383 and ogura-B352 were also two new sequences, they showed high similarities to corresponding chloroplast sequences of Arabidopsis thaliana and Brassica rapa subsp. pekinensis. So we speculated that these two sequences may derive from chloroplast genome. All these results obtained in this study offer new and significant information to investigate the molecular mechanism of cytoplasmic male sterility and fertile maintenance in cauliflower.

Nucleotide binding site (NBS) profiling, a new method was used to map resistance gene analogues (RGAs) in cauliflower (Brassica oleracea var. botrytis). This method allows amplification and the mapping of genetic markers anchored in the conserved NBS encoding domain of plant disease resistance genes. AFLP was also performed to construct the cauliflower intervarietal genetic map. The aim of constructing genetic map was to identify potential molecular markers linked to important agronomic traits that would be particularly useful for development and improving the species. Using 17 AFLP primer combinations and two degeneration primer/enzyme combinations, a total of 234 AFLP markers and 21 NBS markers were mapped in the F2 population derived from self-pollinating a single F1 plant of the cross AD White Flower x C-8. The markers were mapped in 9 of major linkage groups spanning 668.4 cM, with an average distance of 2.9 cM between adjacent mapped markers. The AFLP markers were well distributed throughout the linkage groups. The linkage groups contained from 12 to 47 loci each and the distance between two consecutive loci ranged from 0 to 14.9 cM. NBS markers were mapped on 8 of the 9 linkage groups of the genetic map. Most of these markers were organized in clusters. This result demonstrates the feasibility of the NBS-profiling method for generating NBS markers for resistance loci in cauliflower. The clustering of the markers mapped in this study adds to the evidence that most of them could be real RGAs.

In order to clarify the genetic background of Pinellia ternata germplasm resources in China, the chromosomal constitution and cytogeographical distribution of P. ternata were investigated in 27 different populations among 16 provinces and regions in China systematically.Cytological and cytogeographical methods were used in the study.P. ternata in China is a polyploid complex, which contains septuploid (2n = 7x = 91) , octoploid (2n = 8x = 104) , nonuploid (2n = 9x = 117) and decaploid (2n = 10x = 130). Meanwhile the aneuploid series (2n = 92, 103, 105, 115) of a minority of P. ternata were also found.The genetic differentiation and the phenomenon of ploidy miscellany commonly exist in the species of P. ternata in China, both for natural populations and cultivated populations. Toxicity and chemical components of different ploidy P. ternata should be clarified before the superior multiploid is selected for normalized plantation of the plant.

To explore the methylation status of 5' CpG island of fragile histidine triad (FHIT) gene in plasma and the expression of FHIT protein in cancer tissue of cervical cancer patients.Methylation-specific PCR (MS-PCR) was employed to examine methylation of FHIT gene in 151 plasma samples before treatment. The immunohistochemistry was used to the expression of FHIT protein in cancer tissues.CpG island methylation of FHIT was detected in 31.13% of plasma samples. The expression of FHIT protein was decreased or discarded in 59.60% of cervical cancer tissues. Among them 47.78% was included in methylation positive samples.CpG island methylation of FHIT gene in plasma plays an important role on cervical cancer, which results in decreased expression of FHIT protein. It can be used to diagnose and evaluate the effect of treatment to cervical cancers.

Recent evidence suggests that tumor cells may release DNA into the serum and plasma of afflicted cancer patients. However, no report existed regarding the methylation status of the fragile histidine triad (FHIT) and E-cadherin genes in plasma samples of cervical cancer patients. Methylation-specific PCR (MSP) was employed to examine CpG island methylation of the FHIT and E-cadherin genes in 151 pretreatment plasma samples and 30 tumor tissue samples from cervical cancer patients. MSP products were cloned and sequenced. CpG island methylation of the FHIT and E-cadherin genes was detected in 30.46% and 39.74% of plasma samples, respectively, and in 53.33% and 60.0% of tissue samples, respectively. The total concordance rate of methylation between plasma samples and tissue samples in FHIT gene was 80.00% and that in E-cadherin gene was 76.66%. At least one of the two methylated genes was detected in 56.29% of plasma samples and 76.7% of tissue samples. The presence of both methylated genes was detected in 13.9% of plasma samples and 36.67% of tissue samples. We found that the higher the clinical stage and histologic grade, the higher the rate of methylation in both genes in plasma samples. CpG island methylation of the FHIT and E-cadherin genes is present in plasma of cervical cancer patients. Using the two genes as markers simultaneously may allow clinicians to diagnose and evaluate the effect of treatment earlier and using fewer invasive procedures.

To evaluate the methylation status of the 5' CpG islands in FHIT gene using plasma and tissue samples from cervical cancer patients and find a novel marker for non-invasive diagnosis of cervical cancer, methylation-specific PCR (MSP) was employed to examine CpG island methylation in FHIT gene in 151 pretreatment plasma samples and 30 tumor tissue samples obtained from cervical cancer patients. MSP product was cloned and sequenced directly. CpG island methylation of FHIT was detected in 31.13% of the plasma samples, and in 53.33% of the tissue samples. The total concordant rate of methylation status between plasma and tissue samples in FHIT gene was 80.00%. We found a strong positive correlation between FHIT methylation in the plasma and the clinical stage and histological grade of the tumor. The data showed that CpG island methylation of the FHIT gene is prevalent in the plasma and tissue samples from cervical cancer patients. FHIT detection may be used as a non-invasive marker for diagnosis of cervical cancer and prognostic treatment evaluation.

Analysis of RAPD (Randomly Amplification Polymorphic DNA) was performed between cytoplasmic male sterility line and its maintainer line of cauliflower. Totally 2160 detectable bands were obtained by RAPD using 406 10-bp random primers. Averagely, 5 to 10 bands were produced per primer. Among all the primers only the amplification of primer S2121 was polymorphic in two lines. A 934-bp specific band was only detected in maintainer line. After cloning and sequencing, specific primers were designed to transform the RAPD marker to PCR marker, which was named S2121(900). To identify the specificity of S2121(900), southern dot blotting was performed. To further identify its specificity, individual plant and candidate materials testing were also performed. All these results indicated that the S2121(900) was specific. It can be used to screen the maintainer lines of cauliflower in early stage. Analysis of the sequence suggested that this fragment was high homologous with the part sequences of mitochondrial genome in Brassica napus and Arabidopsis thaliana. So we supposed the S2121(900) may also derive from mitochondrial genome. Our results here offer new clues for explaining the molecular mechanism of cytoplasic male sterility of cauliflower in other way.

Since single nucleotide polymorphisms (SNPs) can serve as gene markers, polymorphism profiles may help scientists to identify the full collection of genes that contribute to the development of complex diseases such as cancer. The distribution of interleukin-10 (IL-10) promoter polymorphisms in Chinese Han ethnic patients with hepatitis B virus (HBV) infection and hepatocellular carcinoma (HCC) was investigated in this study.The polymorphisms of IL-10 promoter region were detected by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and sequencing. Sixty-six health controls, 42 patients with HBV infection, 30 HCC patients, and cell line SMMC-7721 were examined this way.Polymorphisms of T/C or T/N on -872 site occurred frequently in Han ethnic population. Polymorphisms were detected in HBV and HCC patients and cell line SMMC-7721. The hotspot among the polymorphisms was inserting base A between -1058 and -1057.Polymorphisms of IL-10 promoter in HBV and HCC patients may be associated with HBV infection and HCC development.

The understanding of cSNPs of cancer-related genes harboring in high frequency loss regions of tumor chromosomes can advance the disclosure of genetic and variant mechanisms of tumorigenesis, and the investigation of cancer susceptibility. In preparing a gene chip for detecting polymorphisms on coding region of genes in hepatocellular carcinoma tissues, some cSNPs are of interest for their potential links with phenotype.The genes harboring in loss regions with high frequency of hepatocellular carcinoma (HCC) were selected, the related information of cSNP sequences was obtained from the SNP database (dbSNP) of the National Center for Biotechnology Information (NCBI). Then appropriate primers and oligonucleotide probes were designed according to the SNP sites, and a gene chip for the detection of SNPs was constructed. The chip included 48 cSNPs of 25 hepatocellular carcinoma-related genes. The PCR products labeled by Dig-dUTP were hybridized with the cSNP chip.The sensitivity, influence by probe concentration, and reiteration of the chip were detected, with a high sensitivity of 6X10(-3) ng/mul. The signal of hybridization was reduced with a lower concentration of probe. Seven polymorphisms of caspase 9 (rs2308941)C--T and DOK2(rs2242241) T--G, 6 of polymorphisms of EGFL3 (rs947345)A --G, caspase 9 ( rs2308938) C--G and PHGDH(rs1801955)T--A, 5 of polymorphisms of E2F2(rs3218170) G--A,4 of polymorphisms of MUTYH(rs1140507)T--C and BNIP3L(rs1055806)G--T, and 1 of polymorphism of TNFRSF1B (rs1061622)T--G were detected by the chip in the tissues of 10 HCC. Samples of caspase 9 (rs2308941G) and (rs2308941A) were verified by PCR-SSCP and sequencing.The cSNP chip of hepatocellular carcinoma-related genes can accelerate the discovery of polymorphic markers on hepatocellular carcinoma.

The homology-based candidate gene method was used to identified the specific PCR markers linked to cytoplasmic male sterility (CMS) in cauliflower( Brassica oleracea var botrytis.). Searching the DNA and protein data-base of NCBI , correlative genes or open reading frames were identified . Analysis of biosoft, based on the conservative regions ,five primers were designed . Among them, only primer P9/P10 produced a 313- bp specific fragment. Identified by individual plant testing , analysis of RT-PCR and dot blot ,this fragment was only existed in CMS cauliflower knxd612. Analysis of the sequence indicated it was high homologous(98%) with orf138 of Ogura CMS radish. Primary result suggested that the cytoplasmic type of CMS cauliflower knxd612 may belong to Ogura type. This research offered a good foundation to further investigate the CMS mechanism of cauliflower in molecular level.

Sex-determining mechanisms are highly variable between phyla. However, there is an apparent exception in which structurally and functionally related genes control sex determination in different phyla: the sexual regulators DSX of Drosophila melanogaster and MAB-3 of Caenorhabditis elegans both containing a DNA-binding motif, DM domain. Proteins containing the domain may also play a role in sexual development of vertebrates. For examples, both the human DMRT1 (doublesex and mab-3 related transcription factor 1) gene and mouse Dmrt1 gene are necessary for male development. In this paper, through the degenerated PCR, a DNA fragment ADM was amplified out from genomic DNA of brine shrimp, Artemia sinica from YunCheng Salt Lake, Shanxi, China and Artemia parthenogenetica from GaHai, Qinghai, China, respectively. ADM encodes 47 amino acids and is highly homologous to amino acid sequence of the known DM domains. By comparing total of 27 DM domains in distant related species, a phylogenic tree of DM domain was constructed. In the tree, these DM domains were divided into different branches according to their subtypes. Among the DM domains that were compared, ADM is most homologous to the DM domain contained in human DMRT3 and mouse Dmrt3, which shares 83% identity between them. In addition, the same length of ADM could also be amplified out from cDNA of Artemia sinica and Artemia parthenogenetica, which indicated that ADM was expressed and located in one exon. The DM domain in brine shrimp reported here would make it possible for cloning the full-length cDNA containing the DM domain and further elucidating their functions.

The origin and evolution of B chromosomes could be explained by the specific DNA sequence on them. But the specific sequences known were quite limited. To investigate maize B chromosome sqicific DNA sequeces, maize genomes with and without B chromosomes were analyzed by AFLP. Only 5 markers were found specific to genomes with B chromosomes among about 2000 AFLP markers. Southern hybridization and sequence analysis revealed that only the sequence of M8-2D was a B chromosome specific sequence. This sequence contained the telomeric repeat unit AGGGTTT conserved in plant chromosome telomeres. In addition, the sequence of M8-2D shared low homology to clones from maize chromosome 4 centromere as well. M8-2D were localized to B chromosome centromeric and telomeric regions.

To gain a preliminary understanding of cytoplasmic male sterility (CMS) in cauliflower NKC-A, seven primer pairs were designed to amplify atpa, atp6, atp9, coxI, coxII, orfB and orf224, which have been reported in other CMS systems. The amplification pattern of orfB is different in the CMS cauliflower and its maintainer line NKC-B. Results suggest that orfB only expresses in the CMS line, in both flowers and leaves. Sequence analysis indicated that orfB is highly homologous with orf138, a chimeric gene believed to closely associate with Ogura cytoplasmic male sterility in radish. Full-length sequence was obtained to elucidate the molecular basis of orfB. The sequence includes three open reading frames, trnfM, orf138 and orf158. Study of transcriptional patterns and RNA editing of this sequence revealed that orf138 and orf158 co-transcribe, similar to orf138 in Ogura CMS radish. Based on this, we concluded that the molecular basis of cytoplasmic male sterility in cauliflower NKC-A may be similar to that of the Ogura CMS in radish. However, the nuclear backgrounds and RNA editing of corresponding sequences in radish and cauliflower are different. The results of this study also provide important information for breeders selecting new CMS lines in cauliflower. A specific molecular marker was developed according to the CMS specific fragment in NKC-A.

We have applied the technique of random amplification of polymorphic DNA (RAPD) to the analysis of the relationships among four species of brine shrimp: Artemia franciscana, A. urmiana, A. sinica, and A. parthenogenetica. Seventy ten-base synthetic oligonucleotides were used to amplify a total of 458 distinct fragments. DNA polymorphisms were found in all the species examined; the highest percentage of polymorphic bands was found in A. parthenogenetica, with 28.8 per cent. Each species was scored for the presence or absence of every amplification product and the data entered into a binary data matrix. Cluster analysis was then performed to create a dendrogram using UPGMA by the NTSYS program. There are significant differences between bisexual species and parthenogenetic populations. A. parthenogenetica provided 94 specific molecular markers, while bisexual species gave 27 specific molecular markers. A. sinica is a species distinct from the other Old World bisexual species.