Fine mapping and candidate gene analysis of Brtri1, a gene controlling trichome development in Chinese cabbage (Brassica rapa L. ssp pekinensis).

Trichomes are derived from the epidermis and constitute an ideal system for studying cell division in plants. Here, a Chinese cabbage doubled haploid (DH) line (FT) without trichomes was crossed with another DH line (PurDH-1) with trichomes to develop an F2 population for fine mapping of trichome control genes. Genetic analysis showed that the trichome phenotype was controlled by a single dominant gene, Brtri1. Using 1226 glabrous individuals in the F2 segregation population, Brtri1 was localized to a 16.84 kb region between markers Pur6-31 and Pur6-39 on chromosome A06. One of the four complete open reading frames within the mapping region, Bra025311, encodes a MYB transcription factor and is highly homologous to the trichome regulatory gene GL1 in Arabidopsis thaliana. It was thus regarded as a candidate gene for Brtri1. Comparative sequencing showed a 5-bp deletion in the third exon of Bra025311 in FT, resulting in a frame-shift mutation. No expression of Bra025311 was detected in FT. A co-dominant indel marker close to this mutation site was developed for marker-assisted selection in Chinese cabbage breeding.

Fine mapping and characterization of the or gene in Chinese cabbage (Brassica rapa L. ssp pekinensis).

Orange inner leaves/heads is a qualitative trait in Chinese cabbage that is controlled by a single recessive gene. Orange head Chinese cabbage contain more carotenoids than its white head counterpart; hence, this trait is of interest to both researchers and consumers. In this study, we selected the orange head Chinese cabbage line 07A163 and the white head Chinese cabbage line Chiifu as test materials. We localized the target gene controlling the orange head trait to the A09 linkage group, with a physical distance of approximately 19.9 kb between the two markers, syau26 and syau28. This region contains six candidate genes, including Bra031539, which was predicted to encode CRTISO, a carotenoid isomerase specifically required for carotenoid biosynthesis. A comparison of the nucleic acid sequences of the two test materials revealed 88 and 7-bp deletions and 88 SNPs in the promoter region of Bra031539 in line 07A163, along with a 6-bp deletion in the first exon and early termination at the 3' end of this gene. BLAST analysis revealed that 22 amino acids were altered and 17 amino acids were lost in Bra031539 in the orange head line 07A163. We developed the BrPro1 molecular marker in the promoter region of Bra031539 that can be used for early identification of orange head materials, thereby accelerating the breeding process of orange head Chinese cabbage.

Correlation analysis between single nucleotide polymorphism of DRD1 gene and stereotyped behavior of blue fox.

This study was performed to investigate the correlation between stereotyped behavior of the blue fox and single nucleotide polymorphisms (SNPs) of the DRD1 gene. We choose the DRD1 gene as a major gene for investigating the correlation of gene polymorphism and self-biting disease by means of direct sequencing. Part of the DRD1 gene exon of the blue fox was cloned; the length of the whole sequence was 864 bp. Four SNPs were detected and analyzed by the chi-square analysis; the results showed that the gene polymorphism of T206C in the DRD1 gene had a significant correlation with self-biting (P < 0.01). Therefore, marker-assistant selection on self-biting of blue foxes using these SNPs can be applied to select healthy individuals.

Effects of p-chlorophenoxyisobutyric acid, arabinogalactan, and activated charcoal on microspore embryogenesis in kale.

To improve embryogenesis in microspore cultures of kale (Brassica oleracea L. var. acephala DC.), 6-benzylaminopurine (6-BA), naphthaleneacetic acid (NAA), arabinogalactan (AG), p-chlorophenoxyisobutyric acid (PCIB), and activated charcoal (AC) were added to the medium using four varieties of kale. The results showed that the addition of AG (0.1-0.2 g/L), AC (0.1-0.2 g/L) or a combination of 6-BA (0.1-0.2 mg/L) and NAA (0.1-0.2 mg/L) promoted embryo-genesis. Adding 40 µM PCIB or a combination of 40 µM PCIB and 0.2 g/L AC to NLN-13 medium at pH 5.8 effectively enhanced embryogenesis. Treatment with a combination of 40 µM PCIB and 10 mg/L AG gave the highest rate of embryonic induction, especially in genotype "Y007," which showed a twelve-fold increase in yield.

Association of regulator of G protein signaling (RGS5) gene variants and essential hypertension in Mongolian and Han populations.

Genetic variants of the RGS5 gene are believed to be risk factors for hypertension and cardiovascular diseases. In this study, we investigated the association between RGS5 gene variants and hypertension in the Mongolian and Han populations. Peripheral blood was obtained from 429 unrelated Mongolian herdsmen and 416 Han farmers [including essential hypertension (EH) patients and controls]. Nine tagSNPs within the RGS5 genes were retrieved from HapMap, and the samples were individually genotyped using the polymerase chain reaction/ligase detection reaction assay. The distribution of the allele frequency of rs12035879 differed significantly between hypertensive subjects and controls in the Han population, while the distribution of the allele and genotype frequencies of rs16849802 differed significantly between hypertensive subjects and controls in the Mongolian population. We observed an association between rs16849802 and EH in the Mongolian population. The frequency of haplotype GAA was significantly higher in the EH group than in controls in the Mongolian population. However, the EH group and controls did not differ significantly in all 6 haplotypes in the Han population. The rs16849802 and haplotype GAA independently increased the risk of EH in Mongolian patients, and may be used as a risk factor for the prediction of high blood pressure.

Association of NPRA and NPRC gene variants and hypertension in Mongolian population.

NPRA and NPRC are candidate susceptibility genes for essential hypertension (EH) and play a key role in the regulation of plasma levels and biological effects of natriuretic peptides. The aims of the present study were to find new genetic markers in the NPRA and NPRC genes and to assess relationships between variants and EH. A total of 797 unrelated Mongolian herdsmen were enrolled, including 389 EH patients and 408 normotensive controls. Genotyping was performed using the polymerase chain reaction/ligase detection reaction assay. The distribution of the T-allele frequency of rs1847018 in NPRC differed significantly between hypertensive subjects and controls. There was an association between rs1847018 and EH in the additive model in NPRC (P < 0.05). There were no significant differences in the genotype and allele frequency distributions for any of the 3 single nucleotide polymorphisms in NPRA between EH and normotensive individuals. In NPRA, the frequency of haplotype TCA in the EH group was significantly lower than in controls, while the frequency of haplotype TCG was significantly higher in the EH group than in controls; Individuals who possessed the TCA haplotype had a significantly lower risk of EH, whereas the presence of haplotype TCG was significantly associated with a higher risk of EH. However, there was no significant difference between the EH group and controls in any of the 8 haplotypes in NPRC. Rs1847018 is a genetic marker of EH in NPRC, and the frequency of haplotype TCA and TCG in NPRA is associated with EH in the Mongolian population.

Effect of CDH1 gene methylation on transforming growth factor (TGF-ß)-induced epithelial-mesenchymal transition in alveolar epithelial cell line A549.

The aim of this study was to investigate the specific molecular mechanism of the transforming growth factor ß (TGF-ß)-induced epithelial-mesenchymal transition in a lung cancer cell line, and to provide new ideas for targeting therapy of lung cancer. A549 cells were treated with different concentrations of TGF-ß and 5-aza-deoxycytidine (5-aza-dC). The morphological changes after the intervention were observed. The change in the expression of the epithelial marker E-cadherin (E-cad) was detected by Western blot. The proliferation of A549 cells was measured using the MTT assay. Cell movement and invasion capacity was evaluated with the cell scratch test and invasion test. TGF-ß induced A549 cells to transform to a mesenchymal cell morphology and downregulated the expression of E-cad, and 5-aza-dC inhibited this phenomenon. Compared with the control group, the number of transmembrane cells was higher and cell migration was markedly increased in the experimental group with continued culture in the presence of 10 ng/mL TGF-ß, showing significant differences (P < 0.05). CDH1 gene methylation is involved in TGF-ß-induced epithelial-mesenchymal transition in the alveolar epithelial cell line A549.

Relationship between serum neopterin levels and coronary heart disease.

The relationship between serum neopterin levels and coronary heart disease (CHD) was investigated. Eighty-six CHD patients were divided into an acute myocardial infarction (AMI) group (N = 21), an unstable angina pectoris (UAP) group (N = 35), and a stable angina pectoris (SAP) group (N = 30), based on coronary angiography (CAG), 30 subjects without CHD served as the control group. Serum neopterin levels were determined by enzyme linked immunosorbent assay (ELISA), and relationships between neopterin and the severity of stenosis, stenosis number, and the stability of coronary artery were analyzed. Serum neopterin levels were higher in the AMI and UAP groups than in the SAP and control groups (P < 0.01), but no significant differences were observed between the AMI and UAP groups or between the SAP and control groups (P > 0.05). Mean serum neopterin levels were higher in the single, double, and three vessel lesion groups than in the control group (P < 0.05), whereas there were no significant differences among the lesion groups (P > 0.05). Serum levels of neopterin were significantly higher in the type II than in the type I or type III, plaque groups (P < 0.01), the incidence of type II plaque was significantly higher in the AMI and UAP groups compared to the SAP group (P < 0.01). Neopterin likely plays a role in the occurrence and development of athermanous plaque and can serve as a useful biomarker of vulnerable plaques. Immunoreaction may be involved in the pathophysiological process of CHD.

Identification of sequence-related amplified polymorphism markers linked to the red leaf trait in ornamental kale (Brassica oleracea L. var. acephala).

Artistic diversiform leaf color is an important agronomic trait that affects the market value of ornamental kale. In the present study, genetic analysis showed that a single-dominant gene, Re (red leaf), determines the red leaf trait in ornamental kale. An F2 population consisting of 500 individuals from the cross of a red leaf double-haploid line 'D05' with a white leaf double-haploid line 'D10' was analyzed for the red leaf trait. By combining bulked segregant analysis and sequence-related amplified polymorphism technology, we identified 3 markers linked to the Re/re locus. A genetic map of the Re locus was constructed using these sequence-related amplified polymorphism markers. Two of the markers, Me8Em4 and Me8Em17, were located on one side of Re/re at distances of 2.2 and 6.4 cM, whereas the other marker, Me9Em11, was located on the other side of Re/re at a distance of 3.7 cM. These markers could be helpful for the subsequent cloning of the red trait gene and marker-assisted selection in ornamental kale breeding programs.