超表达OsPR1A增强了Xa21介导的水稻对白叶枯病的抗性反应

【背景】前期研究发现,水稻病程相关蛋白质OsPR1A的表达受上游抗病基因Xa21调控,接菌后早期启动Xa21介导的OsPR1A较高水平表达对水稻抵抗白叶枯病菌至关重要。同时OsPR1A也受到水稻白叶枯病菌（Xanthomonas oryzae pv. oryzae,Xoo）的诱导表达。对于OsPR1A的研究绝大部分是作为抗性反应发生的标志基因佐证其他基因或途径在抗性中的作用,缺乏直接的证据证实OsPR1A本身的生物学功能。【目的】通过获得OsPR1a-OX超表达转基因植株,调查其表型及农艺性状,并明确OsPR1A蛋白质表达与抗性的关系,为鉴定OsPR1A功能提供依据。【方法】通过农杆菌介导法,将构建的OsPR1a-OX转化载体转入到水稻受体4021中,利用PCR和免疫印迹（western blot,WB）技术分别在基因水平和蛋白质水平上筛选并鉴定OsPR1A超表达阳性纯合株系。在成熟期,调查OsPR1A超表达转基因植株的表型及农艺性状（株高、穗长、分蘖数、结实率和籽粒大小等）。在31℃条件下,将生长2周的水稻幼苗TP309、4021和OsPR1A超表达转基因植株接种水稻白叶枯病菌,并在接菌0、2、4、6、8、10和12 d时测量病斑长度。在接菌0、4和6 d时,收集TP309、4021和OsPR1A超表达转基因植株的水稻叶片,提取蛋白质,利用WB技术检测OsPR1A的表达特征。【结果】构建了OsPR1a-OX转化载体,并转入到受体4021中,筛选并鉴定到2个OsPR1A超表达转基因纯合株系（#704和#709）。调查了OsPR1A超表达转基因植株在成熟期的表型及农艺性状,与对照4021相比,#704和#709的株高较矮、穗长较短、分蘖数减少、结实率降低,但籽粒稍大,可能与结实率低有关。在31℃条件下,OsPR1A超表达转基因植株的病斑长度与对照4021相比明显缩短,结果具有显著性差异（P<0.05）。在接菌0、4和6 d的材料中,超表达转基因植株#704和#709中OsPR1A始终有较高水平的表达丰度,从而提高了对白叶枯病菌的抗性。【结论】采用农杆菌介导法,获得OsPR1A超表达转基因植株;超表达OsPR1A影响到水稻的正常发育过程;超表达OsPR1A后增强了Xa21介导的水稻对白叶枯病的抗性。     

鸡细菌性腹泻防治

我国从古至今都是农业大国,在经济体制改革的推动下,我国工业发展迅猛的情况下,农业依旧是我国的产业支柱,对促进我国经济发展有极大帮助。养殖业是农业的重要部分,而养殖业中,鸡养殖有占据极大比例,且其规模也在不断扩大。近年,鸡的一些疾病发病率逐渐增加,其中细菌性腹泻是常见的1种,会影响鸡群身体健康与生长发育,甚至会导致其死亡,危害极大,给养殖户乃至养殖产业造成极大的经济损失,做好鸡细菌性腹泻预防与治疗工作尤为重要。该文对鸡细菌性腹泻的特点进行探讨,并提出一些防治措施。     

稻曲病菌交配型基因MAT1-2-1和MAT1-2-8的特性分析

 有性生殖在真菌的生活史和进化过程中具有重要作用,而交配型基因是控制有性生殖的关键因子。前期研究发现稻曲病菌(Villosiclava virens)MAT1-2型菌株中包含MAT1-2-1和MAT1-2-8两个交配型基因,但是它们如何调控稻曲病菌有性生殖依然不清楚。本文研究了它们在不同侵染和生长发育时期的表达模式和编码的蛋白结构特性。研究表明MAT1-2-1在侵染不同阶段一直下调表达;而MAT1-2-8在侵染早期(5 dpi)上调表达,在侵染后期下调表达。与营养菌丝阶段比较,MAT1-2-1和MAT1-2-8在有性发育过程菌核形成、菌核萌发、子座原基形成和子座成熟4个阶段的表达量都是下降的,在菌核形成阶段表达量最低。生物信息学分析显示MAT1-2-1和MAT1-2-8具有磷酸化位点,为非分泌蛋白,无明显的跨膜结构域。蛋白同源比对分析表明MAT1-2-1与香柱菌(Epichloë typhina)的MAT1-2-1同源性最高,而MAT1-2-8与绿僵菌(Metarhizium)的MBR_08192蛋白同源性最高。进一步研究发现MAT1-2-1和MAT1-2-8能够互作,并分别主要定位在细胞核和细胞基质中。通过质谱技术鉴定到MAT1-2-1的一些候选互作蛋白,如假定Ran交换因子Prp20/Pim1(KDB12229.1)、假定rRNA处理蛋白Ebp2(KDB12923.1)及组蛋白H1(KDB12711.1)等。因此,以上结果为研究稻曲病菌交配型基因MAT1-2-1和MAT1-2-8调控有性生殖的生物学功能奠定了基础。     

马尾松叶表微生物多样性

为发掘对林木生长发育有利的优良微生物资源,并筛选适合叶表微生物的收集方法,以马尾松针叶为试验材料,分别用悬摇法和超声波法收集马尾松叶表微生物,用扩增子高通量测序技术、MUSCLE和Qiime软件研究马尾松叶表微生物的多样性。结果表明:扫描电镜观测结果显示,马尾松针叶表面定殖有大量微生物,包括真菌(菌丝及孢子)和细菌。扩增子高通量测序结果表明,马尾松叶表微生物物种丰富,包含细菌运算分类单位(OTUs)490个,真菌OTUs 1273个。马尾松叶表细菌以未分类的蓝细菌属(unidentified_Cyanobacteria)(36.53%)、未分类的拜叶林克氏菌属(unidentified_Beijerinckia)(28.60%)、鞘氨醇单胞菌属(Sphingomonas)(2.35%)为优势属;叶表真菌以枝孢属(Cladosporium)(2.45%)、拟盘多毛孢属(Pestalotiopsis)(0.92%)、无头孢菌属(Capnobotryella)(0.91%)为优势属。针对叶表细菌多样性的研究表明,悬摇法和超声波法均有较高的物种检出度;在叶表真菌多样性的研究中超声波法优于悬摇法,但超声波法样品间数据变异性较大,测定结果不稳定。     

Comparative effect of advanced soy products or corn protein concentrate with porcine meal on growth,body composition,and distal intestine histology of Florida pompano,Trachinotus carolinus

The present study was designed to investigate the effects of diets containing advanced soy products (enzyme‐treated soy and fermented soy) or corn protein concentrate (CPC) in combination with porcine meal (PM) to completely replace poultry byproduct meal (PBM) on growth performance, body composition, and distal intestine histology of Florida pompano, Trachinotus carolinus. Four experimental diets were formulated to be isonitrogenous and isolipidic, to contain 400 g/kg crude protein and 80 g/kg lipid. A reference diet (PBM diet [PBMD]) contained 150 g/kg PBM and 495 g/kg soybean meal (SBM), and three test diets were formulated replacing PBM with 15 g/kg of CPC (CPC diet [CPCD]) or replacing all SBM and PBM with 535 g/kg fermented soy (fermented soybean meal diet [FSBMD]) or 451.3 g/kg enzyme‐treated soy (enzyme‐treated soybean meal diet [ESBMD]). All three test diets were supplemented with 38 g/kg of PM. Diets were fed based on a percentage of bodyweight adjusted after sampling the fish every 2 weeks to triplicate groups of Florida pompano juveniles (mean weight 8.06 ± 0.22 g). After 8 weeks of feeding, fish fed CPCD and ESBMD performed equally well in terms of final body weight, thermal growth coefficient, and percentage weight gain in comparison to fish fed PBMD. In all cases, feeding FSBMD resulted in poor feed conversion and lower feed intake compared to other treatments. Protein retention efficiency, whole‐body proximate composition, phosphorus, sulfur, potassium, magnesium, calcium, sodium, and zinc contents were not significantly influenced by the dietary treatments. The results obtained in the present histological study showed no significant differences in the thickness of serous layer, muscular layer, and submucosal layer of the intestine among treatments. Fish fed CPCD showed a significant widening of the lamina propria with an increase of cellular infiltration and higher presence of goblet cells compared to other dietary treatment. Based on these results, 451 g/kg ESBM or combination of 150 g/kg of CPC and 495 g/kg SBM supplemented with 38 g/kg PM can be utilized to develop a practical diet for juvenile Florida pompano without impacting growth, nutritive parameters, and several distal intestine health parameters.     

Zerumbone attenuates MPP+-induced cytotoxicity in human neuroblastoma SH-SY5Y cells by inhibition of oxidative stress

AIM: To investigate the role of zerumbone (ZER) in 1-methyl-4-phenylpyridinium (MPP⁺)-induced cytotoxicity of human neuroblastoma SH-SY5Y cells. METHODS: Human neuroblastoma SH-SY5Y cells were cultured in vitro and the protective effect of ZER against MPP⁺-induced cytotoxicity was measured by CCK-8 assay. Flow cytometry was used to determine the apoptosis and reactive oxygen species (ROS). The expression of Parkinson disease protein 7 (PARK7) was knocked-down by using PARK7-specific short hairpin RNA (shRNA). The protein levels of PARK7, nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were determined by Western blot. RESULTS: MMP⁺ remarkably reduced the cell viability in a dose-dependent and time-dependent manner. The SH-SY5Y cell injury model was established by treatment with MPP⁺ at 600 μmol/L for 24 h. ZER up-regulated the protein levels of PARK7 and Nrf2 (P<0.05), alleviated apoptosis (P<0.05), and reduced ROS production (P<0.05) in the SH-SY5Y cell injury model. Meanwhile, N-acetyl-L-cysteine (NAC) had the similar functions. Moreover, significant reductions in the protein levels of Nrf2 and HO-1 (P<0.05), and obvious increases in apoptosis (P<0.05) and ROS level (P<0.05) were demonstrated in PARK7-knockdown cells. CONCLUSION: ZER protects SH-SY5Y cells against MPP⁺-induced cytotoxi-city, which may be related to activation of PARK7/Nrf2/HO-1 pathway, and subsequent attenuation of oxidative stress and apoptosis.     

品种间大豆脂肪氧化酶活性差异及β-胡萝卜素漂白性研究

为了解东北地区大豆品种间大豆脂肪氧化酶(LOX)活性差异规律,整理适于东北地区种植的大豆种质资源307份(6份半野生品种,17份地方品种及4份国外品种,其余为栽培品种),统一种植,秋季收获后,测定各品种成熟籽粒脂肪氧化酶总活性。分析品种间脂肪氧化酶总活性分布规律、大豆脂肪氧化酶总活性与β-胡萝卜素漂白性之间关系。结果表明,307份大豆品种间脂肪氧化酶总活性差异极显著,平均值为21.68 U·mL~(-1),最小值为2.51 U·mL~(-1),最大值为42.38 U·mL~(-1)。根据脂肪氧化酶总活性差异,通过聚类分析将307份大豆品种分为3类。低活性品种占总量43.65%,脂肪氧化酶总活性平均值为15.25 U·mL~(-1);中等活性品种占总量43.97%,脂肪氧化酶活性平均值为24.64 U·mL~(-1);高活性品种占总量12.38%,脂肪氧化酶总活性平均值为33.87 U·mL~(-1)。脂肪氧化酶总活性与油分含量呈正相关,与蛋白含量呈负相关,与漂白性呈正相关。     

异硫氰酸丙烯酯毒杀松材线虫作用机制初步研究

对异硫氰酸丙烯酯毒杀松材线虫作用机制进行了初步研究。测定了其对线虫虫体形态、运动行为、体内总糖和蛋白质含量的影响,以及处理前后线虫体内过氧化氢酶(CAT)、乙酰胆碱酯酶(AChE)和谷胱甘肽-S-转移酶(GST)活性变化。结果表明:经异硫氰酸丙烯酯处理后的松材线虫虫体形态严重扭曲,运动形态出现由正常的S形变化为多节弯曲和螺旋状卷曲等异常状况,线虫行动随时间的推移变得迟缓,最终呈C形、新月形或直接死亡。随着异硫氰酸丙烯酯处理质量浓度的增大,线虫体内总糖和蛋白质含量显著上升,糖和蛋白质的代谢发生紊乱;线虫体内CAT、AChE和GST活性显著降低,且均低于对照组,表明线虫体内氧化和抗氧化作用失衡;细胞受到极大的影响,神经系统遭到破坏,解毒能力明显降低。研究结果可为研究利用异硫氰酸丙烯酯作为植物源杀线虫剂提供依据。     

Effect of Heat Stress on Grain Growth, Starch Synthesis and Protein Synthesis in Grains of Wheat (Triticum aestivum L.) Varieties Differing in Grain Weight Stability

The effect of heat stress on the components of grain weight was analysed in wheat ( Triticum aestivum L.) varieties differing in grain weight stability. The wheat varieties PBW154, Sonalika and Hindi62 were raised in the field (New Delhi, India; 77°12' E, 28°40' N, 228.6 m a.s.l.) at three dates of sowing: 19 November 1992 (DOS I), 14 December 1992 (DOS II) and 18 January 1993 (DOS III). The late-sown crop (DOS III) experienced 6–8 °C warmer temperatures during grain development than the crop sown at the normal time (DOS I). The heat susceptibility index (S) revealed that grain weight was less susceptible to heat in Sonalika and PBW154 than in Hindi62. Heat stress reduced both the grain growth duration (GGD) and the grain growth rate (GGR). The grain weight reduction in PBW154 and Sonalika was mainly due to a reduction in GGR, while that of Hindi62 was due to a reduction in GGD. In vivo studies on starch and protein synthesis in excised endosperm at 15, 25 and 35 °C revealed that both processes were more thermotolerant in Hindi62 than in PBW154. The grain starch content was stable in Hindi62 while that in PBW154 was significantly reduced under heat stress. The grain nitrogen content at maturity increased in both varieties under heat stress. It was concluded that the heat susceptibility of grain weight in Hindi62 was mainly due to a reduction in GGD, although GGR, starch and protein synthesis were more thermotolerant in developing grains of Hindi62 than in those of PBW154.     

猪肺炎支原体及其生物学研究进展

猪肺炎支原体是引起猪支气管肺炎的主要病原 ,严重危害着养猪事业的发展。虽然猪肺炎支原体的结构简单 ,但由于它对营养要求较高 ,故培养难度大 ,也是人们不能对其深入研究的一个主要原因。近年来利用克隆技术 ,对猪肺炎支原体从基因水平上进行了多种研究 ,已经取得了一些可喜成果。文章从它的生物学特性、荚膜结构、膜蛋白研究以及基因水平的研究等方面进行了综合性论述 ,并简明地指出了目前研究中存在的问题以及未来展望