Characterization of the complete chloroplast genome of the Hongkong kumquat (Fortunella hindsii Swingle).

Hongkong qumquat (Fortunella hindsii Swingle) is a wild citrus species native to China. In this study, we firstly reporteded its complete chloroplast genome using BGISEQ-500 sequencing. The chloroplast genome is 160,145 bp in size, containing a large single copy region (87,467 bp), a small single copy region (18,730 bp), and a pair of IR regions (26,974 bp). The chloroplast genome contains 112 unique genes, including 79 protein-coding genes, 29 tRNAs, and 4 rRNAs. Phylogenetic maximum-likelihood analysis indicated that F. hindsii is closely related to Citrus species. The complete chloroplast genome would be subsequently used for citrus species researches.

Characterization of the complete chloroplast genome of Sanming wild banana (Musa itinerans) and phylogenetic relationships.

Sanming wild banana (Musa itinerans) is a cold resistant wild banana found in Fujian province of China. In this study, we characterized its complete chloroplast genome using BGISEQ-500 sequencing. The chloroplast genome is 171,815 bp in size, containing a pair of IR regions (35,142 bp), a large single copy region (89,995 bp), and a small single copy region (11,464 bp). The whole chloroplast genome contains 111 unique genes, including 78 protein-coding genes, 29 tRNAs, and 4 rRNAs. Phylogenetic maximum likelihood analysis revealed that Sanming wild banana showed the closest relationship with Musa itinerans that collected from Yunnan Province of China.

Characterization of the complete chloroplast genome of Gerbera jamesonii Bolus in China and phylogenetic relationships.

Gerbera (Gerbera jamesonii Bolus) is one of the most popular ornamental flowers and one of the top five cut flowers worldwide. In this study, we presented the complete chloroplast genome of it using BGISEQ-500 sequencing. Its chloroplast genome is 151,898 bp in size, containing a large single copy region (83,518 bp) and a small single copy region (18,244 bp), and a pair of IR regions (25,068 bp). The chloroplast genome contains 113 unique genes, including 80 protein-coding genes, 29 tRNAs, and 4 rRNAs. Phylogenetic maximum likelihood analysis based on chloroplast genomes of Gerbera and other 17 plant species indicated that the relationship between Gerbera and other plant species of family Compositae is not very close and its relationship with Atractylodes lancea is the closest.

Comparative Transcriptome and iTRAQ Proteome Analyses of Citrus Root Responses to Candidatus Liberibacter asiaticus Infection.

Root samples of 'Sanhu' red tangerine trees infected with and without Candidatus Liberibacter asiaticus (CLas) were collected at 50 days post inoculation and subjected to RNA-sequencing and isobaric tags for relative and absolute quantification (iTRAQ) to profile the differentially expressed genes (DEGs) and proteins (DEPs), respectively. Quantitative real-time PCR was subsequently used to confirm the expression of 16 selected DEGs. Results showed that a total of 3956 genes and 78 proteins were differentially regulated by HLB-infection. Among the most highly up-regulated DEPs were sperm specific protein 411, copper ion binding protein, germin-like proteins, subtilisin-like proteins and serine carboxypeptidase-like 40 proteins whose transcript levels were concomitantly up-regulated as shown by RNA-seq data. Comparison between our results and those of the previously reported showed that known HLB-modulated biological pathways including cell-wall modification, protease-involved protein degradation, carbohydrate metabolism, hormone synthesis and signaling, transcription activities, and stress responses were similarly regulated by HLB infection but different or root-specific changes did exist. The root unique changes included the down-regulation in genes of ubiquitin-dependent protein degradation pathway, secondary metabolism, cytochrome P450s, UDP-glucosyl transferases and pentatricopeptide repeat containing proteins. Notably, nutrient absorption was impaired by HLB-infection as the expression of the genes involved in Fe, Zn, N and P adsorption and transportation were significantly changed. HLB-infection induced some cellular defense responses but simultaneously reduced the biosynthesis of the three major classes of secondary metabolites, many of which are known to have anti-pathogen activities. Genes involved in callose deposition were up-regulated whereas those involved in callose degradation were also up-regulated, indicating that the sieve tube elements in roots were hanging on the balance of life and death at this stage. In addition, signs of carbohydrate starvation were already eminent in roots at this stage. Other interesting genes and pathways that were changed by HLB-infection were also discussed based on our findings.

[Effect of umbilicus moxibustion therapy on physical conditions of yang-deficiency volunteers].

OBJECTIVE: To observe the therapeutic effect of umbilicus moxibustion for sub-health people with yang-deficiency constitution. METHODS: Sixty sub-health volunteers with yang-deficiency constitution were equally and randomly divided into treatment and control groups. The subjects of the two groups were treated conventionally with Jingui Shenqi Pill (Goldern Chamber Pill for strengthening the Kidney-qi), 8 pills each time, 3 times a day. Additionally, those subjects of the treatment group were also treated by moxibustion over the umbilicus, twice a week, 3 months altogether. The integrative score was assessed before and after the treatment in accordance with the criteria of "Quantization Diagnosis of Chinese Medicine". RESULTS: After umbilicus moxibustion intervention for 3 months, the integrative scores of symptoms of mental fatigue, cold limbs and warmth preference, lassitude in loin and limbs, and loose stool were decreased significantly in the treatment group (P < 0.05), suggesting an improvement of these symptoms and being superior to those of the control group in the curative effect (P < 0.05). Simple Jingui Shenqi Pill treatment was also effective in reducing scores of cold limbs and warmth preference, and lassitude in loin and legs after the treatment (P < 0.05). Of the two 30 cases in the treatment and control groups, 18 (60.0%) and 7 (23.3%) experienced marked improvement in their symptoms, 10 (33.3%) and 12 (40.0%) had an improvement, 2 (6.7%) and 11 (36.7%) were failed, with the effective rates being 93.3% and 63.3%, respectively. CONCLUSION: Umbilicus moxibustion therapy can effectively improve clinical symptoms of sub-health subjects with yang-deficiency constitution.

Effects of arsenic exposure from drinking water on spatial memory, ultra-structures and NMDAR gene expression of hippocampus in rats.

Epidemiological investigations indicate that chronic arsenic exposure can damage neurobehavioral function in children. The present study was aimed to study the effects of arsenic exposure from drinking water on the spatial memory, and hippocampal ultra-structures and N-methyl-d-aspartate receptor (NMDAR) gene expression in rats. Sprague-Dawley rats were assigned to four groups: rats in control group drank regular water, rats in other groups drank water with final arsenic concentration of 2.72 mg/L (group A), 13.6 mg/L (group B) and 68 mg/L (group C), respectively, for 3 months. The levels of arsenic in blood serum and hippocampus were monitored. Rats were tested in Morris water maze (MWM) for memory status. Samples of hippocampus were collected from two rats in each group for transmission electron microscopic study and the detection of NMDAR expression by RT-PCR. The rats in group C showed a significant delay in hidden platform acquisition. Neurons and endothelial cells presented pathological changes and the expression of NR2A was down-regulated in hippocampus in arsenic exposed rats. Our data indicated that arsenic exposure of 68 mg/L caused spatial memory damage, of which the morphological and biochemical bases could be the ultra-structure changes and reduced NR2A expression in hippocampus.

Effects of different free fatty acids on insulin resistance in rats.

BACKGROUND: Much evidence demonstrates that elevated free fatty acids (FFAs) are associated with insulin resistance. However, it is not clear whether different FFAs can cause different degrees of peripheral insulin resistance. This study aimed to investigate the effects of short-term elevation of FFAs on hepatic and peripheral insulin action, and determine whether FFAs with different degrees of saturation have differential effects on hepatic insulin resistance. METHODS: Intralipid+heparin (IH, polyunsaturated fatty acids), oleate (OLE), lard oil+heparin (LOH), and saline (SAL) were separately infused intravenously for 7 hours in normal Wistar rats. During the last 2 hours of the fat/saline infusion, a hyperinsulinemic-euglycemic clamping was performed with [6-3H] glucose tracer. Plasma glucose was measured using the glucose oxygenase method. Plasma insulin and C-peptide were determined by radioimmunoassays. Plasma FFAs were measured using a colorimetric method. RESULTS: Compared with infusion of SAL, plasma FFA levels were significantly elevated by infusions of IH, OLE, and LOH (P<0.001). All three fat infusions caused remarkably higher hepatic glucose production (HGP) than SAL (P<0.001). OLE and LOH infusions induced much higher HGP than IH (P<0.01). Glucose utilization (GU) was decreased with all three fat infusions relative to SAL (P<0.001), but GU did not differ among the three types of fat infusions. CONCLUSIONS: Short-term elevation of FFAs can induce hepatic and peripheral insulin resistance. Polyunsaturated fatty acids induced less hepatic insulin resistance than monounsaturated or saturated fatty acids. However, IH, OLE, and LOH infusions induced similar peripheral insulin resistance.

[Effects of Huatan Xiezhuo Recipe on insulin resistance and leptin in rats with fatty liver].

OBJECTIVE: To observe the effects of Huatan Xiezhuo Recipe (HTXZR) on fatty liver of rats. METHODS: Forty-eight SD rats were randomly divided into five groups: normal control group, untreated group, Dongbao Gantai Tablet (DBGTT)-treated group, low-dose HTXZR-treated group and high-dose HTXZR-treated group. Fatty liver was induced in the rats by hyperlipid diet and intraperitoneal injection of tetracycline. The pathological changes of liver tissues in rats were observed, and the liver function, serum leptin, insulin resistance index, triglyceride (TG) and free fatty acid (FFA) in the liver were detected. RESULTS: The levels of serum leptin, insulin resistance index, FFA and TG in rats of the three treated groups were lower than those of the untreated group (P<0.01), and such effects in the HTXZR-treated groups were more significant than those in the DBGTT-treated group (P<0.05). CONCLUSION: The main action mechanisms of HTXZR in treating fatty liver are probably to promote the lipid metabolism, decrease the leptin and insulin resistance.

[Transgenic maize plants with low copy number of foreign genes were produced with maize Ubi-1 promoter].

Direct DNA delivery procedures (include biolistics method) often resulted in multiple copies of the transgenes in transformants and certain copies of them were rearranged. Integration of multiple copies of the introduced genes was the main reason of gene silencing which meant inhibition or loss of foreign gene expression in filial generations of transformants. In the present work, we compared the influences of maize Ubi-1 promoter and other promoters on copy number of transgenes in maize transgenic plants. Immature embryos from Zea mays L. plants of sib-pollinated of A188 x H99 genotype were used as initial materials. Type- I embryonic calluses derived from preculture of immature embryos were treated on N6 medium containing 0.6 mol/L sucrose for 3 approximately 5 hours and transformed via particle bombardment with PDS1000/He delivery system (Bio-Rad). Bombarded calluses were treated with hyperosmotic N6 medium for 16 approximately 20 hours continuously. Then the cultures were transferred onto normal N6 medium and incubated at 26 degrees C in dark for two weeks and subsequently selected on N6 medium supplemented with 2 or 5 mg/L phosphinothricin (PPT) but without casamino acid for another two weeks. The calluses after selective culture were transferred onto hormone-free MS medium containing 2 or 5 mg/L PPT but without casamino acid, and incubated at 24 degrees C under 16 h illumination for plant regeneration. Regenerated plantlets over 2 cm in height were transferred to Magenta box containing 1/2 hormone-free MS medium. Plantlets over 8 cm in height were transplanted to soil. After growing for one week in greenhouse, the plants were sprayed with 250 mg/L PPT solution. Fertile transgenic maize plants were regenerated and confirmed by Southern blotting and histochemical localization of beta-glucuronidase (GUS) activity. Relations between promoter and copy number of transgenes in transformants were analyzed. Maize transgenic plants possessing an intact copy and another incomplete copy of beta-glucuronidase gene (gus) were obtained in case gus gene under the control of maize Ubi-1 promoter (pUbi:GUS). Simultaneously the co-transformed phosphinothricin acetyltransferase gene (bar) controlled by CaMV 35S promoter in another plasmid (p35S:BAR) also existed with only one copy. When pDB1 and (pUbi:in2) were cobombarded, the regenerated transgenic maize plant exhibited with only one copy of in2 gene too. It suggested that the copy number of transgenes in maize transformants was low if the transgenes controlled by maize Ubi-1 promoter. The possible reason might be that the foreign genes were integrated site-specifically via homologous recombination between Ubi-1 promoter and its endogenous sequences in maize genome, and two cotransformed plasmids had reconstructed as one intact molecule before integrating into maize chromosome. On the contrary, if p35S:BAR was cobom-barded with plasmid pAct:In1 containing rice Act-1 promoter (without maize Ubi-1 promoter), the transgenic maize plants had 4 approximately 8 copies of bar gene. These results reflected that utilization of self gene promoter could reduce the copy number of the transgenes in transgenic plants of certain species itself and avoid the occurrence of gene silencing. T2 seeds have been harvested.