Real‐time PCR detection of Erwinia amylovora on blossoms correlates with subsequent fire blight incidence

Fire blight is the most devastating bacterial disease of rosaceous plants. Forecasting fire blight infections is important to allow for countermeasures that reduce economic damage in pome fruit production. Current computerized forecasting models are solely based on physical factors such as temperature and moisture, but not on the actual presence of the pathogen Erwinia amylovora. Although the inoculum concentration is considered to be crucial for infection and disease outbreak, most current approaches used for identification of fire blight inoculum including morphological, biochemical, serological, and DNA‐based methods are nonquantitative. Based on a real‐time PCR approach previously published, an improved protocol to be used directly on whole bacteria in the field is described. The method allows for early detection and quantification of the pathogen prior to the occurrence of first symptoms. There is a clear correlation between bacterial abundance and subsequent disease development. However, in some cases, no disease symptoms could be observed despite a pathogen load of up to 3·4 × 10⁶ cells per blossom. Integration of the amount of pathogen detected into refined prediction algorithms may allow for the improvement of applied forecasting models, finally permitting a better abatement of fire blight.     

中药组方对黄曲霉毒素B1中毒雏鸭肝功能的影响

为观察黄曲霉毒素B1(AFB1)对试验雏鸭肝功能血清指标的影响及复方中药对AFB1的颉颃效应,本试验选用7日龄健康雏鸭90只,分为3组,每组30只。Ⅰ组为空白对照组,灌胃与试验组等体积二甲基亚砜;Ⅱ组、Ⅲ组为试验组。每天分别按0.1mg/kg剂量给Ⅱ组、Ⅲ组雏鸭灌胃AFB1一次,连续投药21d,试验期间给Ⅲ组雏鸭日粮中添加2%复方中药。分别在给雏鸭投药后7、14、21d,检测雏鸭肝功能部分血清指标。结果显示,Ⅱ组、Ⅲ组雏鸭丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)与Y-谷氨酰转肽酶(GGT)活性显著高于Ⅰ组(P<0.05),而血清总蛋白(TP)与白蛋白(ALB)含量显著下降(P<0.05);与Ⅱ组比较,在日粮中添加复方中药的Ⅲ组雏鸭各项血清指标均有显著改善(P<0.05)。说明黄曲霉毒素B1导致雏鸭肝功能发生显著的变化,而复方中药能明显改善其变化。     

甘蓝型波里马不育胞质向南方白菜型油菜转育及育性变化

白菜型油菜品种、自交系与甘蓝型油菜波里马细胞质雄性不育系杂交、回交至ＢＣ２Ｆ１，再用南方白菜型油菜自交系与ＢＣ２Ｆ１测交，花期研究测交后代育性变化规律及定株定时观察白菜型波里马细胞质雄性不育材料与２９Ａ、广丰Ａ、阿油Ａ三种白菜型细胞质雄性不育材料育性变化情况．结果表明，在２６个测交后代中，雄性不育的占２３．１％，恢复可育的占７．７％，介于两者之间或育性分离的占６９．２％．在南方白莱型油菜中发现能完全恢复育性的自交系。白菜型波里马细胞质雄性不育材料的不育性较其他三种不育材料稳定．可为油蔬两用的南方白菜型油菜杂种优势利用提供新的不育源。     

家蚕B型清道夫受体基因的鉴定及系统发生与表达分析

B型清道夫受体(SR-B)是清道夫受体超基因家族成员,参与机体的脂类代谢、动脉粥样硬化形成或保护、宿主免疫损害防御、凋亡细胞清除和细胞粘附等重要生命过程。基于家蚕全基因组数据,利用比较基因组学方法,鉴定获得了14个家蚕B型清道夫受体基因,这些基因分布在至少5条染色体上,其内含子的大小从几十到近万个碱基对。系统发生分析表明,14个家蚕B型清道夫受体基因分布在3个类群:第1类群中分布的9个基因和第3类群中分布的3个基因,分别与类群中其它昆虫的同源基因形成明显的直系同源关系;第2类群中分布有2个基因,此类群中不同昆虫的同源基因之间的进化关系较为复杂。EST数据和芯片数据分析表明,多数家蚕B型清道夫受体基因具有转录活性。RT-PCR检测结果显示,有8个家蚕B型清道夫受体基因在5龄第3天幼虫组织中表达,并具有不同的表达模式,推测这些基因可能行使不同的功能。研究结果为进一步诠释家蚕B型清道夫受体基因的功能奠定了基础。     

水稻三系保持系辐射M2代突变类型分析

用60Coγ射线直接辐射保持系嘉浙B诱发突变,M2代出现多种变异,按变异频率大小从高到底依次为叶色突变>早熟突变>颖尖不退花突变>高外露率突变>薄颖壳突变>花时早突变>紫稃尖突变>迟熟突变,上述8个选择项的总突变频率达到2.419%,说明辐射诱变改良保持系性状十分有效.     

枸杞的扦插繁殖

以细叶枸杞为试材,研究了枸杞的扦插繁殖。结果表明：绿色嫩枝不宜作扦插材料。木质化、半木质化插条用２００ｍｇ／ｋｇ ＮＡＡ或ＩＢＡ处理３０－６０ｍｉｎ或用５００ｍｇ／ｋｇＨ３ＢＯ３处理３０ｍｉｎ均可提高生根率和平均生根条数,但降低平均根长度。     

F18ac菌毛A亚单位基因的克隆与序列分析

根据已经发表的F18ab菌毛A亚单位(FedA/ab)的基因(fedA/ab)[1],设计一对引物,利用PCR技术从表达F18ac菌毛的大肠杆菌2134P株[2]、8199株[3]、8813株[3]中分别扩增到一段序列,并克隆至pGEM-T载体,获得重组质粒T2134PA、T8199A、T8813A.琼脂糖凝胶电泳、序列测定及分析表明,该3个序列大小均为516bp,与fedA/ab(513bp)具有较高的同源性,分别为96.3%、96.5%、95.9%,推导的Fed/ac氨基酸序列与FedA/ab同源性分别为93.0%、93.6%、92.4%.数据表明该实验所克隆的序列均为F18ac菌毛A亚单位(FedA/ac)的基因(fedA/ac).     

Inflammatory response in rat kidney with contrast-induced nephropathy

AIM：To observe the expression of tumor necrosis factor α (TNF-α) and nuclear factor κB (NF-κB) in the renal tissue of the rats with contrast-induced nephropathy (CIN). METHODS：Male Sprague-Dawley rats (n=96) were randomly divided into control group (n=48) and CIN group (n=48). The model rats in CIN group were intravenously injected with iodinated contrast media (76% compound diatrizoate injection,10 mL/kg), while the rats in control group were injected with the same volume of saline. Six rats in each group were sacrificed at 6 h, 12 h, 24 h, 48 h, 72 h, 5 d, 10 d and 15 d after intravenous injection, respectively, and the blood and kidney samples of the rats were obtained. The renal tubular injury was assessed by histological examination (HE staining). The expression of kidney injury molecule-1 (KIM-1), TNF-α and NF-κB at mRNA and protein levels in the renal tissues were semiquantitatively measured by the methods of RT-PCR and immunohistochemistry, respectively. The correlations between the expression of TNF-α, NF-κB and tubular injury score, KIM-1 expression in renal tissue of CIN group were analyzed. RESULTS：The levels of serum creatinine (SCr) and blood urea nitrogen (BUN) in control group were not changed between different time points (P>0.05). The levels of SCr and BUN in CIN group displayed significant increases at different time points (except 15 d) compared with control group (P<0.05). The renal tubular injury score in CIN group was significantly higher at all time points than that in control group (P<0.05). The expression of KIM-1, TNF-α and NF-κB at mRNA and protein levels up-regulated significantly at 6 h and the peaking of KIM-1 expression was at 24 h, while the peaking of TNF-α and NF-κB expression was at 48 h in CIN group. The expression of KIM-1,TNF-α and NF-κB was significantly increased in CIN group compared with control group except at 15 d (P<0.05). The expression of TNF-α and NF-κB at mRNA and protein levels showed close correlations with renal tubular injury score (r=0.843, 0.758, 0.743 and 0.707, P<0.05). The expression of TNF-α and NF-κB at mRNA and protein levels was also positively correlated with KIM-1 expression (r=0.863, 0.807, 0.839 and 0.855, P<0.05). CONCLUSION：The expression of TNF-α and NF-κB at mRNA and protein levels in the renal tissues of CIN group is up-regulated and is closely related with renal tubular injury, indicating that the inflammatory response is involved in the pathogenesis of CIN.     

猪带绦虫45W-4BX和TSOL18的高效表达及免疫效果研究

RT-PCR扩增45W-4B和TSOL18基因,PCR截去45W-4B基因的N端信号肽和C端疏水氨基酸序列形成45W-4BX。将45W-4BX和TSOL18 PCR产物分别亚克隆人pGEX-4T-1,用IPTG诱导表达,取产物进行SDS-PAGE和Western blot分析。纯化表达产物制成油佐剂疫苗分别免疫家猪,用25000枚猪带绦虫成熟虫卵进行攻击感染,ELISA检测各组的抗体水平,90d后剖检计算各组的减虫率。结果表明,45W-4BX和TSOL18基因在大肠杆菌中分别以可溶性和包涵体形式获得高效表达,并能被囊虫病人血清所识别。重组蛋白免疫猪15d血清抗体即为阳性,30d左右达到峰值。2种重组抗原的减虫率均在88％以上,与囊虫粗抗原免疫效果相当。这为进一步研制基于45W-4BX和TSOL18的猪囊虫重组基因工程疫苗奠定了基础。     

山西白羊草灌丛草地牧草产量动态研究

对山西白羊草灌丛草地牧草产量动态进行了研究，经３年的定位研究其结果表明，白羊草灌丛草地牧草产量的季节动态规律较稳定，各年度动态曲线均呈单峰型，高峰值出现在８月份，峰期后牧草产量下降较缓慢。牧草营养物质的变化，生长前期粗蛋白质含量高随生育期进展而逐渐减少，粗纤维含量则相反，粗蛋白质产量高峰期在８月，这与干草产量的峰值正好吻合，此时正是该类草地利用的最佳时期。