Three AFLP markers tightly linked to the genic male sterility <Emphasis Type="Italic">ms</Emphasis><Subscript><Emphasis Type="Italic">3</Emphasis></Subscript> gene in chili pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.) and conversion to a CAPS marker

Genic male sterility (GMS) has long been used as a tool for hybrid seed production in chili pepper (Capsicum annuum L.). We developed DNA markers linked to the GMS ms ₃ gene in a segregating population using bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) techniques. The segregating population was subjected to BSA-AFLP with 512 primer combinations. Three AFLP markers (Eagg/Mccc₂₇₆, Eagc/Mctt₁₇₈, and Ecag/Mtgc₂₀₄) were identified as tightly linked to the ms ₃ locus. Among them, we converted the AFLP marker Ecag/Mtgc₂₀₄ to the cleavage amplified polymorphic sequence (CAPS) marker, named GMS3-CAPS, based on sequencing analysis of internal and flanking regions for the markers between male-fertile and sterile plants. This marker will be useful for pepper breeding using the GMS system.     

Genetic, agronomic and quality comparisons of two 1AL.1RS. wheat-rye chromosomal translocations

The 1AL.1RS wheat-rye chromosomal translocation originally found in ‘Amigo’ wheat possesses resistance genes for stem rust, powdery mildew and greenbug biotypes B and C, but also has a negative effect on wheat processing quality. Recently, a second 1AL.1RS translocation carrying Gb6, a gene conferring resistance to greenbug biotypes B, C, E, G and I, was identified in the wheat germplasm line ‘GRS1201′. Protein analytical methods, and the DNA polymerase chain reaction were used to identify markers capable of differentiating the 1RS chromosome arms derived from ‘Amigo’ and ‘GRS1201′. The secalin proteins encoded by genes on 1RS chromosome arms differed in ‘Amigo’ and ‘GRS1201′. A 70 kDa secalin was found in the ‘Amigo’1AL.1RS, but did not occur in the ‘GRS1201’1AL.1RS. Polymorphisms detected by PCR primers derived from a family of moderately repetitive rye DNA sequences also differentiated the two translocations. When ‘GRS1201’was mated with a non-1RS wheat, no recombinants between 1RS markers were observed. In crosses between 1RS and non-1RS parents, both DNA markers and secalins would be useful as selectable markers for 1RS-derived greenbug resistance. Recombination between 1RS markers did occur when 1RS from ‘Amigo’ and 1RS from ‘GRS1201’were combined, but in such intermatings, the molecular markers described herein could still be used to develop a population enriched in lines carrying Gb6. No differences in grain yield or grain and flour quality characteristics were observed when lines carrying 1RS from ‘Amigo’ were compared with lines with 1RS from ‘GRS1201′. Hence, differences in secalin composition did not result in differential quality effects. When compared with sister lines with 1AL.1AS derived from the wheat cultivar ‘Redland’, lines with ‘GRS1201’had equal grain yield, but produced flours with significantly shorter mix times, weaker doughs, and lower sodium dodecyl sulphate sedimentation volumes.     

Dietary eicosapentaenoic acid requirement of juvenile rock bream,Oplegnathus fasciatus

A feeding trial was conducted to evaluate the optimum dietary level of eicosapentaenoic acid (EPA, 20:5n‐3) based on growth and non‐specific immune responses in juvenile rock bream. A basal diet without EPA supplementation was used as a control, and six other diets were prepared by supplementing with 4, 8, 12, 16, 20 or 40 g of EPA per kg diet. The actual EPA concentrations of the diets were 0.5, 4.3, 8.5, 13.0, 16.8, 21.0 and 41.2 g of EPA per kg diet, and the diets were abbreviated as EPA_0.5, EPA_4.3, EPA_8.5, EPA_13.0, EPA_16.8, EPA_21.0 and EPA_41.2, respectively. Triplicate groups of fish averaging 1.06 ± 0.01 g (mean ± SD) were fed one of the seven experimental diets at the apparent satiation for 8 weeks. At the end of the feeding trial, weight gain, specific growth rate and feed efficiency of fish fed EPA_16.8, EPA_21.0 and EPA_41.2 diets were significantly higher than those of fish fed EPA_0.5, EPA_4.3, EPA_8.5 and EPA_13.0 diets (p < .05). Superoxide dismutase activity of fish fed EPA_16.8, EPA_21.0 and EPA_41.2 diets were significantly higher than those of fish fed EPA_0.5, EPA_4.3 and EPA_8.5 diets. Fish fed EPA_21.0 and EPA_41.2 diets showed significantly higher lysozyme activity than did fish fed EPA_0.5, EPA_4.3, EPA_8.5 and EPA_13.0 diets. The broken‐line analysis of weight gain indicated that the optimum dietary EPA level was 16.7 g/kg diet. These results suggested that the optimum dietary EPA level in juvenile rock bream could be greater than 16.7 g/kg diet but less than or equal to 16.8 g/kg diet based on the broken‐line analysis and the ANOVA test of weight gain.     

山西猪场猪蓝耳病并发猪胸膜肺炎放线杆菌的病毒鉴定及细菌分离鉴定以及治疗方案

2020年2月份山西某大型规模猪场育肥猪突发呼吸困难,无过多症状表现,即突然死亡,发病率为2%,死亡率1.5%左右。实验室以病死猪为研究对象,综合死亡猪只脏器病变状况观察,无菌采集病死猪的肺脏、肝脏、脾脏病料,进行RT-PCR检测猪蓝耳病原;用含1%NAD和5%胎牛血清的TSA琼脂培养基进行细菌分离培养和纯化,经形态学鉴定、PCR鉴定、16Sr RNA基因序列进化树构建等方法,结果显示:该发病猪群为猪繁殖与呼吸障碍综合征并发猪胸膜肺炎放线杆菌感染。药敏试验结果显示:该猪胸膜肺炎放线杆菌对安普霉素、硫酸黏菌素、替米考星+氟苯尼考、替米考星+阿莫西林、氟苯尼考、头孢噻呋钠+硫酸黏菌素、头孢喹肟、头孢噻呋钠+新霉素、替米考星+头孢噻呋钠、多西环素+氟苯尼考、头孢噻呋钠+阿莫西林、阿莫西林+克拉维酸钾、头孢噻呋钠+恩诺沙星、新霉素、头孢噻呋钠、安普霉素+头孢噻呋钠敏感。对恩诺沙星耐药。本实验室根据临床及检测结果制定治疗方案,为该猪场的猪繁殖与呼吸障碍综合征并发猪胸膜肺炎的临床诊断及用药提供了理论依据。     

Output of methylmercury from a Catchment in northern Sweden

The runoff output of methylmercury (MeHg) from the 50 ha Svartberget Catchment in northern Sweden was studied during 1993. These outputs are compared to those from the Gårdsjön Catchment in the southwestern part of Sweden. Although the wet deposition of MeHg is several times higher in southern Sweden the output of MeHg per unit area from the two catchments was comparable (0.12 g /km_* ²yr). Furthermore, the concentration of MeHg (0.4 ng/L) in the Svartberget Catchment was more than twice that from the Gårdsjön Catchment. These results suggest that the contemporary wet deposition of MeHg in itself is not a good indicator of runoff concentration or output per unit area. DOC transport and catchment characteristics such as wetland area, or possibly other forms of contemporary atmospheric deposition may all be more important for explaining MeHg output from the terrestrial ecosystem.     

Metabolism of flupyrazofos in the isolated perfused rat liver

To investigate the hepatic metabolism of the new insecticide flupyrazofos [O,O-diethyl O-(1-phenyl-3-trifluoromethylpyrazol-5-yl) phosphorothioate], isolated rat liver was perfused with flupyrazofos under single-pass conditions. In outflow perfusate and bile, 1-phenyl-3-trifluoromethyl-5-hydroxyprazole (PTMHP), PTMHP-sulfate and PTMHP-glucuronide conjugates were identified as the metabolites of flupyrazofos. However, O,O-diethyl O-(1-phenyl-3-trifluoromethylpyrazol-5-yl) phosphate (flupyrazofos oxon) was not detected. A HPLC method with UV detection was used to investigate the hepatic disposition of flupyrazofos and its metabolite PTMHP. The concentrations of flupyrazofos, PTMHP and PTMHP conjugates in outflow perfusate reached steady-state levels within 20 min after commencing perfusion of 7.3 microM flupyrazofos. At steady state, the mean extraction ratio of flupyrazofos was 0.93 (+/- 0.01) and clearance was 26.1 (+/- 0.2) ml min-1 which nearly approached perfusate flow rate (28 ml min-1). PTMHP accounted for 55.7 (+/- 5.8)% of eliminated flupyrazofos and was recovered as unchanged PTMHP, PTMHP-sulfate and PTMHP-glucuronide in the bile as well as the outflow perfusate.     

Effect of catheter size and injection rate of contrast agent on enhancement and image quality for triple‐phase helical computed tomography of the liver in small dogs

Rapid contrast injection is recommended for triple‐phase helical computed tomography (CT) of the liver. However, a large‐gauge catheter is needed for faster contrast injection and this is not practical for small breed dogs or cats. The purpose of this crossover group study was to evaluate applicability of a lower injection rate with a small‐gauge (G) catheter for triple‐phase hepatic CT in small dogs. Triple‐phase CT images were acquired for six beagle dogs using three protocols: an injection rate of 1.5 ml/s with a 24 G catheter, 3.0 ml/s with a 22 G catheter, and 4.5 ml/s with a 20 G catheter. Enhancement of the aorta, portal vein, and hepatic parenchyma was measured in each phase (arterial, portal, and delayed) and image quality was scored subjectively by two observers. Injection duration, time to scan delay, and time to peak enhancement were also recorded. Contrast injection duration decreased with a higher injection rate (n = 6, P ≤ 0.01), but time to peak enhancement and time to scan delay were not significantly affected by injection rates and catheter sizes. Contrast injection rate did not significantly affect aortic, portal, and hepatic enhancement. In addition, separation between each phase and quality of images was subjectively scored as good regardless of injection rate. Findings from the current study supported using an injection rate of 1.5 ml/s with a catheter size of 24 G for triple‐phase hepatic CT in small dogs (weight < 12 kg).     

Effect of heat treatment on the antioxidant activity of extracts from citrus peels

The effect of heat treatment on the antioxidant activity of extracts from Citrus unshiu peels was evaluated. Citrus peels (CP) (5 g) were placed in Pyrex Petri dishes (8.0 cm diameter) and heat-treated at 50, 100, or 150 degrees C for 10, 20, 30, 40, 50, and 60 min in an electric muffle furnace. After heat treatment, 70% ethanol extract (EE) and water extract (WE) (0.1 g/10 mL) of CP were prepared, and total phenol contents (TPC), radical scavenging activity (RSA), and reducing power of the extracts were determined. The antioxidant activities of CP extracts increased as heating temperature increased. For example, heat treatment of CP at 150 degrees C for 60 min increased the TPC, RSA, and reducing power of EE from 71.8 to 171.0 microM, from 29.64 to 64.25%, and from 0.45 to 0.82, respectively, compared to non-heat-treated control. In the case of WE from CP heat-treated at the same conditions (150 degrees C for 60 min), the TPC, RSA, and reducing power also increased from 84.4 to 204.9 microM, from 15.81 to 58.26%, and from 0.27 to 0.96, respectively. Several low molecular weight phenolic compounds such as 2,3-diacetyl-1-phenylnaphthalene, ferulic acid, p-hydroxybenzaldoxime, 5-hydroxyvaleric acid, 2,3-diacetyl-1-phenylnaphthalene, and vanillic acid were newly formed in the CP heated at 150 degrees C for 30 min. These results indicated that the antioxidant activity of CP extracts was significantly affected by heating temperature and duration of treatment on CP and that the heating process can be used as a tool for increasing the antioxidant activity of CP.     

Sesame oleosin and prepro-2S albumin expressed as a fusion polypeptide in transgenic rice were split, processed and separately assembled into oil bodies and protein bodies

A recombinant polypeptide containing the precursor protein of a sesame storage protein, 2S albumin, fused to the C-terminus of a sesame oleosin was expressed in transgenic rice seeds under the control of a rice glutelin promoter. The recombinant polypeptide of 32 kDa, equivalent to the resultant molecular mass of sesame oleosin (15 kDa) and prepro-2S albumin (17 kDa), was detected in the endoplasmic reticulum fraction of maturing transgenic rice seeds, but not in the purified oil bodies or the soluble extract of transgenic seeds. However, sesame oleosin presumably fused with a 2 kDa C-terminal appendix originating from the signal sequence of prepro-2S albumin, was found in the purified oil bodies, and mature sesame 2S albumin apparently processed into two subunits (9 and 4 kDa) linked by disulfide bonds was detected in extracts of transgenic seeds. Immunogold labeling revealed that the sesame oleosin and 2S albumin were separately located in oil bodies and protein bodies of embryo cells of transgenic rice seeds. While sesame 2S albumin was also detected in protein bodies of endosperm cells of transgenic seeds, the co-expressed sesame oleosin, probably degraded due to the lack of oil bodies in this tissue, and was not detected. The results provide a new technique for introducing two recombinant polypeptides separately into rice oil bodies and protein bodies from one expression construct.