三种DNA提取方法对检测草莓镶脉病毒稳定性的研究

草莓成熟叶片中含有大量的多糖、蛋白质、多酚和色素等物质,影响高质量DNA的提取和下游PCR反应的进行。为确定适用于PCR技术检测草莓镶脉病毒(Strawberry vein banding virus,SVBV)的DNA提取方法,从草莓叶片中提取DNA后,对样品中的SVBV进行特异性的PCR检测,比较确定不同DNA提取方法对PCR检测SVBV稳定性的影响。以感染SVBV的草莓叶片为材料,分别采用CTAB试剂盒法、高盐低pH法、改良SDS法提取草莓叶片的总DNA,将不同方法提取的DNA通过紫外分光光度法和琼脂糖凝胶电泳法进行测定,对DNA产量和质量进行分析。结果表明,3种方法均能获得较为完整的DNA片段,用CTAB试剂盒提取的DNA质量最高,改良SDS法提取的DNA质量较低。将不同方法提取的DNA经过PCR反应特异性检测SVBV后,将与目的片段大小一致的PCR扩增产物经序列测定及分析,证明与SVBV的序列相吻合,证明了方法的可靠性,其质量能够满足PCR特异性检测需要。     

毛乌素沙地杨树防护林内紫花苜蓿蒸散耗水规律的研究

本文利用验证后的Penman-Monteith模型,计算了内蒙古毛乌素沙地新疆杨防护林内紫花苜蓿的蒸散耗水,结果表明：(1)通过Penman-Monteith模型计算得到的蒸散量与涡度相关法实测值平均相对误差为14.73%(R²=0.84),说明利用Penman-Monteith模型计算林草复合模式内牧草蒸散量是可行的;(2)不同生育期内紫花苜蓿蒸散量差别较大,具体表现为：返青－分枝阶段、现蕾－开花阶段蒸散量较小,分别为5.03 mm和44.95 mm;分枝－现蕾、开花－结荚阶段蒸散量较大,分别为71.10 mm和101.74 mm;其中耗水最大的生育期为开花－结荚阶段,占整个生育期总耗水量的45.66%。在空间上表现为：随着距主防护林带距离增加,紫花苜蓿蒸散量逐渐增大,最大蒸散量出现在距离林带约115 m处(约7倍树高),为234.41 mm,之后,受另一侧防护林的影响,蒸散量逐渐减少。整个生育期防护林内紫花苜蓿总蒸散量为222.83 mm,对照地紫花苜蓿蒸散量为269.02 mm,与对照相比,防护林内紫花苜蓿比林外蒸散量降低了17.2%。防护林内和对照紫花苜蓿生物量干重分别为3287.28 kg/hm²和2959.93 kg/hm²,林内比对照增产11.1%。     

可溶性E2蛋白作为抗原的检测猪瘟病毒血清抗体间接ELISA方法的建立

以可溶性重组E2蛋白作为抗原,建立了猪瘟病毒(CSFV)血清抗体间接ELISA诊断方法(rE2-ELISA).将猪瘟疫苗毒株E2基因主要抗原区(A-D)基因克隆到表达栽体pGEX-6p-1上,转化E.coli,降低诱导温度至20℃,获得48 000大小E2融合蛋白,部分目的蛋白以可溶性形式表达.Western blotting试验证实,E2融合蛋白可以和CSFV阳性血清发生特异性结合.亲和层析纯化后的E2融合蛋白作为抗原,建立了检测CSFV血清抗体的间接rE2-ELISA方法.该方法的特异性试验结果表明,与PRRSV、PCV2、PPV和PRV阳性血清之间不存在交叉反应;用rE2-ELISA和国外同类试剂盒(CSF-Ab-Kit)检测142份田问血清样品,2种试剂盒的阳性检测率分别为83.81%和88.73%.因此,rE2-ELISA猪瘟抗体检测试剂盒具有良好的敏感性和特异性,适合应用在大规模的CSFV血清抗体的检测工作中.     

Antimicrobial resistance in Salmonella enterica serovars Enteritidis and Typhimurium isolated from animals in Korea: comparison of phenotypic and genotypic resistance characterization

Fourteen and 22 each of Salmonella Enteritidis and Salmonella Typhimurium (S. Typhimurium) were isolated from animals from 1983 to 1999 in Korea and tested for their antibiotic resistance patterns, phage types and resistance gene patterns. S. Typhimurium isolates were highly resistant to streptomycin, sulfisoxazole and tetracycline, 95, 95 and 86%, respectively. The incidence of multiple antibiotic resistance (resistant to more than two drugs tested) of S. Typhimurium isolates was extremely high (100%) comparing to S. Enteritidis isolates (21%). Two of the five ACSSuT (ampicillin, chloramphenicol, streptomycin, sulfisoxazole and tetracycline) resistant type S. Typhimurium isolates were phage type definitive type 104 (DT104).For the detection of resistance related genes in S. Enteritidis and S. Typhimurium isolates, particularly ACSSuT type S. Typhimurium, antibiotic resistance genes, cmlA/tetR, bla(PSE-1) and bla(TEM), and genus Salmonella specific gene, sipB/C, were amplified using four pairs of primers in a hot-start multiplex polymerase chain reaction (PCR). Two Korean isolates of S. Typhimurium DT104 showed bla(TEM) amplicons instead of bla(PSE-1) for the ampicillin resistance and they were susceptible to florfenicol. The multiplex PCR used in this study was useful in characterization of multiple drug resistant Salmonella isolates, especially ACSSuT type S. Typhimurium, and identification of beta-lactamase gene distribution among Salmonella isolates.     

半番鸭网上平养不同饲养密度试验

在网上平养条件下对育雏期和生长肥育期不同阶段的大型白羽半番鸭设定不同饲养密度,进行生产性能指标的对比试验,得出初步试验结果:育雏期(0～14日龄)饲养密度27羽/m2左右较为适宜,生长肥育期15～28日龄8～9羽/m2,29～42日龄5～6羽/m2,43 ～56日龄4～5羽/m2较为适宜.     

Role of TGF-β1/Smads and ERK expression in vascular remodeling induced by high-salt diet in Wistar rats

AIM: To investigate the role of transforming growth factor β₁ (TGF-β₁)/Smads and extracellular signal-regulated kinase(ERK) expression in vascular remodeling induced by high-salt diet in Wistar rats. METHODS: Wistar rats were randomly divided into 3 groups: normal control group (n=13), high salt (8%) model group and high salt+telmisartan group (n=13). Tail-cuff arterial pressure was determined every 2 weeks. After 24 weeks, the rats in high salt model group were divided into model animals with hypertension group (MH, n=12) and model animals without hypertension group (MN, n=12). The remodeling of aorta and mesenteric artery was observed by HE and Masson staining. In addition, the techniques of immunohistochemistry and real-time PCR were applied to detect the expression of proliferating cell nuclear antigen (PCNA), TGF-β₁, p-Smad2/3, p-ERK1/2 and Smad7 at both protein and mRNA levels. RESULTS: Compared with normal control group, blood pressure in MH group was much higher, and media thickness (MT) and collagen volume fraction (CVF) of arteries in MH and MN groups were higher.The mRNA expression of TGF-β₁, Smad2 and Smad7 in the aorta was significantly increased, and the protein levels of PCNA, p-ERK1/2, TGF-β₁ and p-Smad2/3 in the aorta and mesenteric artery media were elevated, but Smad7 decreased. After telmisartan treatment, MT and CVF were much lower,and the protein levels of PCNA, TGF-β₁, p-Smad2/3 and p-ERK1/2 were significantly reduced, whereas Smad7 was increased. CONCLUSION: The abnormal expression of TGF-β₁/Smads and ERK may be involved in the mechanism of remodeling of aorta and mesenteric artery induced by high-salt diet. Telmisartan prevents the vascular remodeling via regulating TGF-β₁/Smads and ERK signal pathways mediated by angiotensinⅡ type 1 (AT₁) receptor, at least in part.     

黄瓜地面覆盖对日光温室土壤温度变化的影响

以地膜和小麦秸秆为覆盖材料,研究了黄瓜地面覆盖对日光温室土壤温度变化的影响。结果表明,地面覆盖可以提高土壤温度,地膜覆盖的效果大于秸秆覆盖,覆盖效果随土层深度的增加而下降。地膜覆盖可以显著提高土壤平均温度,且对最低温度的提高作用大于最高温度;秸秆覆盖延缓了土壤温度变化的速度,对土壤增温作用小,且土壤最高温度低于对照而最低温度高于对照,温差最小,土壤温度保持相对稳定状态。因此,秸秆覆盖作用更为稳定,更具有推广价值。     

济南野生木本观赏植物资源研究

济南市共有观赏价值的野生木本植物148种及变种,现将其划分为绿化及风景植物、花卉植物、绿篱植物、垂直绿化植物及树桩盆景植物等五大类,并对济南野生木本观赏植物的保护及开发利用提出了建议。