皂荚提取物对植物病原菌的抑制作用

 豆科植物皂荚是一种传统药用植物,为我国特有种。本文在室内测定了皂荚叶和棘刺乙醇提取物对枯草芽孢杆菌、胡萝卜软腐欧文氏杆菌、番茄疮痂病菌、番茄灰霉病菌、番茄叶霉病菌、棉花枯萎病菌、瓜果腐霉的抑制作用。叶提取物未检测出抗菌活性。棘刺提取物对枯草芽孢杆菌和番茄疮痂病菌都表现出一定的抗菌活性,最低抑菌浓度(MIC)分别为2.5 mg/mL和10 mg/mL。棘刺乙醇提取物在浓度为1 mg/mL时,对4种植物病原真菌的生长都表现出抑制活性,其中对瓜果腐霉生长的抑制率为28.4%。将棘刺乙醇提取物分为石油醚等5个极性不同的萃取部分,其中乙酸乙酯部分对枯草芽孢杆菌、番茄疮痂病菌、欧文氏杆菌都具有明显的抑制作用,且活性强于其它的极性段,对枯草芽孢杆菌和番茄疮痂病菌的MIC分别为1.25 mg/mL和5 mg/mL。乙酸乙酯部分对瓜果腐霉和棉花枯萎病菌的生长也有较强的抑制作用,在1mg/mL时抑制率分别为45.8%和23.9%。研究结果表明,皂英抗菌活性成分主要存在于棘刺中的乙酸乙酯极性段和正丁醇极性段中。     

Germination response of Orobanche seeds subjected to conditioning temperature, water potential and growth regulator treatments

Broomrapes (Orobanche spp.) are parasitic weeds that cause significant losses of crop yield. Experiments were conducted to investigate the seed response to the artificial germination stimulant GR₂₄ in three species of Orobanche subjected to preconditioning under various temperatures, water potentials and with plant growth regulators. The highest germination percentages were observed in Orobanche ramosa, Orobanche aegyptiaca and Orobanche minor seeds conditioned at 18°C for 7 days followed by germination stimulation at 18°C. With the increase of the conditioning period (7, 14, 21 and 28 days), the germination percentage of O. ramosa and O. aegyptiaca progressively decreased. When conditioned at −2 MPa, the germination percentage was lower than at 0 and −1 MPa, especially at 13 and 28°C. Orobanche minor seeds could retain relatively high germination if conditioned at 18, 23 or 28°C, even after significantly extended conditioning periods (up to 84 days). GA₃ (30–100 mg L⁻¹), norflurazon and fluridone (10–100 mg L⁻¹), and brassinolide (0.5–1.0 mg L⁻¹) increased seed germination, while 0.01 mg L⁻¹ uniconazole significantly reduced germination rates of all three Orobanche spp. The promotional effects of GA₃ and norflurazon and the inhibitory effect of uniconazole were evident, even when they were treated for 3 days. Germination of Orobanche seeds was much lower when the unconditioned seeds were directly exposed to GR₂₄ at 10⁻⁶ m . This early GR₂₄-induced inhibition was however alleviated or even eliminated by the inclusion of GA₃ or norflurazon (10–50 mg L⁻¹) in the conditioning medium. On the contrary, the inclusion of uniconazole increased the inhibitory effect of GR₂₄, particularly in the case of O. ramosa.     

小麦白粉病纯化菌种保存方法

利用经初选确定的5种具有较好保鲜能力的配方,采用离体叶段滤纸保存法、离体叶段琼脂保存法和试管苗保存法对小麦白粉病菌纯化菌系保存效果进行了研究.结果表明,对小麦幼苗第一叶叶段保鲜效果最好的配方是沈保6号,当使用40～60μg/mL最适浓度时,第25天叶段绿色仍可达到90%以上.保存菌种的最适环境条件研究结果表明,在4℃冰箱中,以离体叶片沈保6号琼脂保存和离体叶沈保6号浸润滤纸保存效果最佳,在第40天叶片绿色仍分别达到78.0%和68.0%,孢子萌发率可达26.7%和26.8%.     

华东地区部分猪源大肠杆菌K88黏附素的生物学特性

近年来，从华东地区患腹泻仔猪中分离到一些表达K88菌毛的大肠杆菌，这些菌株只与K88a因子单抗反应，而不与b、c、d因子单抗反应。通过K88常规血清交叉吸收试验、SDS-PAGE、Western印迹，表明这些菌株不仅与K88ac参考菌株C83907制备的c因子血清反应，而且与以分离株SEC586制备且经K88ab、K88ac、K88ad参考菌株吸收后的血清也反应。对分离株SEC586、SEC464的K88主要亚单位结构基因faeG的克隆、测序，发现该基因由846对核苷酸组成，编码菌毛主要亚单位的262个氨基酸及21个氨基酸的信号肽，比国外报道的K88ac FaeG亚单位（263个氨基酸）少了1个氨基酸，比K88ab、K88ad（265个氨基酸）少了3个氨基酸。SEC586、SEC464菌株的FaeG亚单位氨基酸序列的同源性为97．7％，它们与K88ac的同源性为94．7％和96．2％；与K88ab的同源性为90．1％和91．2％；与K88ad的同源性为87．0％和88，6％。结果表明，新分离的K88ac大肠杆菌黏附素主要亚单位已发生了部分变异。     

生长抑素基因疫苗质粒pEGS/2SS的构建及表达

将pcS／2SS中的乙肝表面抗原（HBsAg）S基因及插入S基因中的生长抑素（somatostatin，ss）基因亚克隆到pUC19中，构建成pUSS／S质粒；PCR扩增pcS／SS中SS基因，调整阅读框，融合到pUSS／S质柱S基因后，构建成pUS／2SSG质粒；最后将S／2SSG融合基因克隆到pEGFPN1中EGFP基因的5’端，构建S／2SS-EGFP融合表达质粒pEGS／2SS。酶切、测序鉴定后脂质体包裹法将pEGS／2SS转染HeLa细胞，荧光显微镜下检测荧光，ELISA检测表达产物的SS免疫反应原性。酶切和测序鉴定表明，重组质粒pEGS／2SS构建成功。pEGS／2SS转染24h后的HeLa细胞检测到绿色荧光，72h检测到强烈荧光，表明融合基因在HeLa细胞获得高表达；ELISA证实融合蛋白具有SS的抗原抗体反应原性。质粒pEGS／2SS的成功构建及表达为生长抑素基因免疫的机理及安全性研究奠定了基础。     

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).     

新城疫病毒F48E9感染CEF细胞特异性表达基因的筛选鉴定

分别以NDV F48E9和La Sota株感染鸡胚成纤维细胞,于感染后8 h提取NDV感染CEF细胞总RNA,通过mRNA差异显示技术筛选病毒感染诱导表达的特异性基因.主要方法是先以锚定引物经反转录后,然后以9～10 bp随机引物为上游引物,以锚定引物Oligod(T)18为下游引物,进行PCR扩增,PCR产物采用8%的尿素变性PAGE胶电泳进行分离鉴定.对于特征性差异带进行第二次PCR扩增,克隆后测定其核苷酸序列.结果我们发现数个差异条带,选取差异带在500 bp以内的条带,经测序后显示经NDV F48E9感染后,有一条差异条带特异性表达的基因所编码的蛋白与Receptor(TNFRSF)-interacting serine-threonine kinase 2有47 %同源,是一个新基因,其蛋白功能不明确,有可能和细胞的凋亡有关.     

O型口蹄疫病毒VP1 T细胞与B细胞表位基因双拷贝串联表达产物的免疫应答

以一株O型口蹄疫病毒(FMDV)外壳蛋白VP1基因为模板,合成与细胞免疫及体液免疫相关抗原表位肽基因:21-40肽(20AA)和141-160肽(20AA)基因序列,运用基因工程技术构建了含有串联结构21-40(20AA)～141-160(20AA)～21-40(20AA)～141-160(20AA)的2020-2020VP1融合基因表达载体r2020-2020,转化宿主菌BL21(DE3)RIL后诱导表达,表达产物经SDS-PAGE及Western Blot分析显示重组融合蛋白的分子量约为18Ku.动物实验表明,较小剂量的融合蛋白就能诱导豚鼠产生特异性T淋巴细胞增殖反应及抗FMDV中和抗体,证明该融合蛋白可同时激活细胞免疫及体液免疫反应,具有开发成为抗FMDV疫苗的应用价值.     

Effects of gastrin on expression of cyclooxygenase and growth factors in rat gastric mucosa

AIM: To clarify the effects of gastrin on the expression of cyclooxygenase (COX) and several growth factors in rat gastric mucosa. METHODS: Male Sprague Dawley rats were fasted for 24 hours and subcutaneously injected with saline or gastrin 17 at doses of 1 μg/kg, 10 μg/kg and 100 μg/kg, respectively. The expression of COX-1, COX-2, heparin-binding epidermal growth factor-like growth factor (HB-EGF) and hepatocyte growth factor (HGF) in the gastric mucosa were examined using Western blotting and immunohistochemical staining. Effects of a potent gastrin receptor antagonist YM022 on the expression of COX-1, COX-2, HB-EGF and HGF in gastric mucosa were also evaluated. RESULTS: Gastrin dose-dependently increased the expression of COX-2 and HB-EGF in rat gastric mucosa while the expression of COX-1 and HGF did not change significantly after treatment with gastrin. However, pretreatment with YM022 dose-dependently abolished the up-regulation of COX-2 and HB-EGF expression induced by gastrin. CONCLUSIONS: This study demonstrates that gastrin up-regulates COX-2 and HB-EGF expression in rat gastric mucosa, indicating that COX-2 and HB-EGF are involved in pathogenesis of the gastrin-related gastric mucosal hyperplasia and carcinoma of stomach.     

包膜控释尿素对大棚番茄的增产与品质提高效应

对施用包膜控释尿素和普通尿素的大棚番茄进行了田间生产条件下产量与品质对比分析试验。结果表明:与普通尿素相比施用控释尿素有显著的增产效果。D 60控释处理(控释期60 d)产量最高,为68.85 t/hm2,比对照增产19.39 t/hm2,增产率为39.2%;比尿素增产10.10 t/hm2,增产率为17.2%;D 90(控释期90 d)比对照增产15.37 t/hm2,增产率为31.0%;比尿素增产6.08 t/hm2。增产率为10.3%;D 90与D 60两种控释尿素之间产量差异不显著。控释尿素具有增加单株果数、单果重的效果。D 90与D 60氮素当季利用率比普通尿素分别高65.5%和108.8%。硝酸盐含量,以D 90为最低,仅比对照高4.4%,比普通尿素低56%;D 60比对照高15%,比尿素处理低51.6%;尿素处理比对照高137.3%。V c和氨基酸含量,D 90为最高,分别比对照高1.00倍和2.46倍;比普通尿素处理高22%和100%;D 60与尿素两处理V c含量差异不显著,但氨基酸总量前者却比后者高48%。可溶性糖含量以D 60较高,普通尿素为最低。综合品质指标为:D 90>D 60>U。说明控释尿素具有显著地增产效应和提高蔬菜品质的突出效应,特别在提高蔬菜的安全品质方面贡献最大。综合各项,在番茄生产中以选用控释期为60 d的包膜控释尿素为最好。