小麦条锈菌DNA提取方法比较及RAPD反应体系优化

分别采用CRAB法和CTAB/SDS法进行小麦条锈菌gDNA的提取,并对两种方法进行比较分析.结果表明:CTAB法具有较高的得率,其优化RAPD反应体系为10×Reaction Buffer 2.5 μL,MgCl<,2>(25 mmol/L)2 μL,dNTP(2.5 mmol/L each)1.5μL,模板DNA(...     

Restriction fragment length polymorphisms in Fusarium oxysporum f.sp. dianthi and other fusaria from Dianthus species

DNA restriction fragment length polymorphisms (RFLPs) among 46 isolates of Fusarium oxysporum from Dianthus spp., representing the known range of pathogenicity in carnation, were determined using total DNA digested with the restriction enzyme Hind III and a previously described probe, D4. Distinct multiple band RFLP patterns were found, which delineated RFLP groups as follows: (i) F. oxysporum f.sp. dianthi races I and 8; (ii) F. oxysporum f.sp. dianthi races 2, 5 and 6; (iii) F. oxysporum f.sp. dianthi race 4; (iv) a recently described race of F. oxysporum f.sp. dianthi (wilt-causing isolates from D. caryophyllus formerly classified as F. redolens); (v) wilt-causing isolates from D. barbatus formerly classified as F. redolens and (vi), (vii) and (viii), three further recently described races of F. oxysporum f.sp. dianthi. Isolate groups derived from analysis of RFLPs were consistent with existing and recently described vegetative compatibility groups (VCGs) in F. oxysporum f.sp. dianthi , but not in all cases with races. Isolates of F. oxysporum and F. proliferatum not associated with wilt disease had simpler RFLP patterns (with one exception) that were not associated with VCGs.     

Metastatic immune infiltrates correlate with those of the primary tumour in canine osteosarcoma

Our lack of understanding of the immune microenvironment in canine osteosarcoma (cOSA) has limited the identification of potential immunotherapeutic targets. In particular, our ability to utilize readily available tissue from a dog's primary tumour to predict the type and extent of immune response in their pulmonary metastatic lesions is unknown. We, therefore, collected 21 matched pairs of primary tumours and pulmonary metastatic lesions from dogs with OSA and performed immunohistochemistry to quantify T‐lymphocyte (CD3), FOXP3+ cell, B‐lymphocyte (Pax‐5), and CD204+ macrophage infiltration. We found that T‐lymphocytes and FOXP3+ infiltrates in primary tumours positively correlated with that of metastatic lesions (ρ = 0.512, P = 0.038 and ρ = 0.698, P = 0.007, respectively), while a strong trend existed for CD204+ infiltrates (ρ = 0.404, P = 0.087). We also observed T‐ and B‐lymphocytes, and CD204+ macrophages to be significantly higher in a dog's pulmonary metastasis compared to their primary tumour (P = 0.018, P = 0.018, P = 0.016, respectively), while FOXP3+ cells were only significantly higher in metastases when all primary tumour and metastasis lesions were compared without pairing (P = 0.036). Together, these findings suggest that the metastatic immune microenvironment may be influenced by that of the primary cOSA, and that primary tumour immune biomarkers could potentially be applied to predict immunotherapeutic responses in gross metastatic disease. We, therefore, provide a rationale for the treatment of cOSA pulmonary metastases with immunotherapeutics that enhance the anti‐tumour activity of these immune cells, particularly in dogs with moderate to high immune cell infiltration in their primary tumours.     

Influence of Apoptosis in Bovine Embryo's Development

The correlation between apoptosis and early bovine embryonic loss is still not fully elucidated. In the present study, the relationship between the arrest of bovine embryos at the different stages of development and apoptosis was evaluated. We used embryos 7 days after in vitro maturation and fertilization, and morphologic and biochemical apoptotic analyses were performed by using a phase contrast microscope and by the terminal transferase dUTP nick end‐labelling respectively. For the statistic, the apoptotic cell ratio (ACR) was determined as the percentage of apoptotic cells per embryo. To evaluate the relation between ACR and fragmentation pattern, embryos were divided into five groups, groups I–V. To assess the relation between ACR and cytoplasmatic fragmentation, embryos were divided into three groups, according to the fragmentation percentage (<5%; 5–15% and >15%). Of the total 139 embryos included, 65 arrested at 2–8 cells; 14 arrested at 9–16 cells; 18 compacted morula and 42 were non‐arrested blastocysts. The average number of embryonic fragmentation at different stages of the development, 2–8 cells, 9–16 cells, compacted morula and blastocyst, was 16.0 ± 1.5, 28.7 ± 4.4, 4.4 ± 2.4 and 1 ± 0.3 respectively. The embryos at the stage of arrested 9–16 cells and compacted morula had higher ACR than those at the blastocyst stage, excluding the stage of 2–8 cells (the genome is not yet active). The correlation detected between embryonic development and ACR was 0.92 (p < 0.01). It was observed that embryos possessing high fragmentation showed the higher ACR value (r = 0.98, p < 0.05). Comparing the results between fragmentation percentage and ACR, it was observed that the embryos with higher percentage of fragmentation corresponded to higher ACR (r = 0.97, p < 0.01). These results clearly demonstrated that bovine embryonic arrest at different stages of development is correlated with the apoptotic mechanisms.     

Rapid identification of virulence genes in enterotoxigenic Escherichia coli isolates associated with diarrhoea in Queensland piggeries

OBJECTIVE: To identify virulence genes in enterotoxigenic E coli (ETEC) isolates associated with diarrhoea in neonatal, 1 to 3 week-old and weaned pigs in southeast Queensland. DESIGN: Multiplex PCR and serotyping were applied to E coli isolates obtained over a 5-year period (1998-2002) from cases diagnosed at Toowoomba Veterinary Laboratory. PROCEDURE: A total of 126 isolates from 25 different Queensland piggeries were tested for haemolytic activity on 5% sheep blood agar and by multiplex PCR for the presence of five commonly recognised fimbrial (F4, F5, F6, F41 and F18) and three enterotoxin genes (STa, STb, LT). A subset of 62 representative isolates were serotyped by slide agglutination. For comparative purposes, multiplex PCR was also performed on the DNA of 31 ETEC isolates from 9 serotypes originating from piggeries in southern New South Wales. RESULTS: A total of 113 (89.7%) of the isolates from Queensland possessed ETEC virulence genes, including 14 of 15 isolates from neonatal pigs (93.3%), 18 of 23 isolates from 1 to 3 week old pigs (78.3%) and 81 of 88 isolates from weaned pigs (92.1%). F4:STa:STb:LT (serotype O149) was the most prevalent pathotype in neonatal and 1-3 week old pigs and F4:STa:STb:LT (serotype O149) and F18:STa:STb:LT (serotype O141) were most prevalent in weaned pigs. In comparison, isolates obtained from neonatal pigs from New South Wales belonged to a more diverse range of pathotypes and serotypes. CONCLUSION: Multiplex PCR was a rapid and specific method for detecting the presence of ETEC virulence genes in porcine E coli isolates. For isolates obtained from cases of suspected colibacillosis in Queensland, growth of a heavy pure culture of haemolytic E coli was a sensitive prognostic indicator of the presence of ETEC virulence genes in the isolate. ETEC pathotypes and serotypes remained stable in Queensland piggeries over the five-year study period and appear to have changed little over the last three decades.     

GAPB基因原核表达载体的构建

以拟南芥cDNA为模板,用PCR扩增出GAPB的基因全长,然后将GAPB基因片段连接到PET28a（＋）载体上,构建重组质粒并转化大肠杆菌DH5α,经菌落PCR和酶切鉴定筛选出阳性克隆,测序正确后,再将阳性克隆的质粒转化大肠杆菌BL21（DE3）。结果表明：成功构建了原核表达载体PET28a（＋）-GAPB,为后续的G...     

Paraphoma root rot of alfalfa (Medicago sativa) in Inner Mongolia,China

Root rot symptoms were observed in fields of alfalfa in Chifeng city, Inner Mongolia, China in 2016. Disease incidences of seven alfalfa varieties planted in 2014 ranged from 56% to 95%, while incidence of Gongnong No. 1 planted in 2016 was 8%, 31% and 76% in 2016, 2017 and 2018, respectively. Paraphoma isolates were consistently recovered from black necrotic root tissues of diseased plants with a frequency of 77.1%. Based on morphological characters and phylogenetic analysis of rDNA internal transcribed spacer (ITS), elongation factor 1-α (EF1-α) and β-tubulin (TUB), this fungus was identified as Paraphoma radicina. Glasshouse pathogenicity experiments showed that P. radicina significantly reduced above- and below-ground biomass of alfalfa plants 2 months after inoculation. Paraphoma radicina infected 70% of the plants inoculated with a root dip in conidia, and these symptoms were consistent with the symptoms in the field. Paraphoma radicina was successfully reisolated from disease roots of the inoculated alfalfa plants. This is the first report of P. radicina as the causal agent of alfalfa root rot in China.     

Short term soil priming effects and the mineralisation of biochar following its incorporation to soils of different pH

The aim of this work was to determine the magnitude of the priming effect, i.e. short-term changes in the rate (negative or positive) of mineralisation of native soil organic carbon (C), following addition of biochars. The biochars were made from Miscanthus giganteus, a C4 plant, naturally enriched with ¹³C. The biochars were produced at 350 °C (biochar350) and 700 °C (biochar700) and applied with and without ryegrass as a substrate to a clay-loam soil at pH 3.7 and 7.6. A secondary aim was to determine the effect of ryegrass addition on the mineralisation of the two biochars.After 87 days, biochar350 addition caused priming effects equivalent to 250 and 319 μg CO₂-C g⁻¹ soil, in the low and high pH soil, respectively. The largest priming effects occurred at the start of the incubations. The size of the priming effect was decreased at higher biochar pyrolysis temperatures, which may be a way of controlling priming effects following biochar incorporation to soil, if desired. The priming effect was probably induced by the water soluble components of the biochar. At 87 days of incubation, 0.14% and 0.18% of biochar700 and 0.61% and 0.84% of biochar350 were mineralized in the low and high pH soil, respectively. Ryegrass addition gave an increased biochar350 mineralisation of 33% and 40%, and increased biochar700 at 137% and 70%, in the low and high pH soils, respectively. Certainly, on the basis of our results, if biochar is used to sequester carbon a priming effect may occur, increasing CO₂-C evolved from soil and decreasing soil organic C. However, this will be more than compensated for by the increased soil C caused by biochar incorporation. A similar conclusion holds for accelerated mineralisation of biochar due to incorporation of fresh labile substrates. We consider that our results are the first to unequivocally demonstrate the initiation, progress and termination of a true positive priming effect by biochar on native soil organic C.     

美国乙醇市场和玉米消费

3月3日,美国乙醇工业协会(EIA)公布了12月美国乙醇产量报告,另外,近期报道称甲基叔丁基醚(MTBE)将停止使用,在此对乙醇产业市场和玉米消费以及今年剩余时间里可能会影响该产业的因素进行略为详尽的研究。图1显示出美国乙醇产业持续强劲增长的状况。3月3日EIA公布的2005年12月报告显示:美国每天乙醇产量为28万桶,再次创下月度高点。2005年美国乙醇产量为9300万桶(39亿加仑),全年平均日产量为25.5万桶。2005年乙醇产量高于2004年的8100万桶(34亿加仑),2004年日产量为22.1万桶。12月份数据显示当月用于乙醇生产的玉米用量约为1.29亿蒲式耳,…