日本结缕草ZjNAC1的克隆、转录激活活性、亚细胞定位及表达分析

采用RACE技术从日本结缕草中克隆得到ZjNAC1转录因子(GenBank No. MH428376),其开放阅读框为945bp,编码314个氨基酸。ZjNAC1编码的蛋白在N端有1个典型的NAM保守结构域,属于NAC转录因子家族。通过染色体步移的方法,获得ZjNAC1基因ATG上游1635bp序列,分析发现其包含响应脱落酸(ABA)、茉莉酸甲酯(MeJA)及逆境胁迫的作用元件。构建pGBKT7-ZjNAC1载体,转化Y2HGold酵母感受态细胞,发现ZjNAC1具有转录激活活性。农杆菌介导的35S-ZjNAC1-YFP载体注射本生烟草叶片瞬时表达结果显示,ZjNAC1定位于细胞核。实时荧光定量结果表明,ZjNAC1在日本结缕草根中表达量最高;此外,ZjNAC1的表达受10μmol/L MeJA和20%PEG4000处理的诱导,但受200μmol/L乙烯(ET)、10μmol/L ABA和300mmol/L NaCl处理的抑制。     

影响荷斯坦牛产犊难易的原因分析

通过研究不同胎次、产犊季节和犊牛初生重、犊牛性别、产犊方式等因素对荷斯坦牛难产程度的影响,旨在降低难产率,帮助牛场制定合理有效的繁殖管理措施。试验收集上海金山奶牛场2018年1~12月产犊难易记录共2 812条,采用GLM模型对影响奶牛难产程度的因素进行分析。结果表明,胎次、产犊季节、犊牛初生重、犊牛性别、产犊方式对难产程度有显著影响(P<0.05),且第二胎奶牛自然分娩比例最高;春季和夏季产犊低难产度所占比例较大;随着犊牛初生重的增加,难产程度逐渐增大;生公犊难产程度越大;双胎生产方式难产度增大。     

放牧对羊草根际土壤线虫群落及区系的影响

为了探索放牧干扰对羊草（Leymus chinensis Tzvel）根际土壤小型动物-土壤线虫群落的影响规律,本试验以土壤线虫为生物指标对典型草原放牧区及围栏区羊草根际土壤线虫群落特征及区系进行了研究,结果表明：羊草根际共捕获34属线虫,放牧影响羊草根际线虫总数量及优势类群,不同土层土壤线虫总数量的分布规律为：围栏区>放牧区,放牧区优势及次优势类群中植物寄生线虫类所占比例明显高于围栏区。放牧区羊草根际土壤线虫Shannon多样性指数显著高于围栏区。放牧区瓦斯乐斯卡指数及自由生活线虫成熟度指数低于围栏区,说明放牧区土壤健康状况较差。放牧影响羊草根际土壤线虫区系的分布,放牧区0~10 cm,10~20 cm羊草根际土壤线虫区系分布在B区和D区,表明放牧区土壤线虫食物网相对于围栏区受干扰大、土壤线虫食物网不稳定,且退化。     

Study on Proliferation and Apoptosis of EBL Cells Induced by P48 Protein of Mycoplasma bovis

The aim of this study was to investigate the effect of P48 protein on proliferation and apoptosis of embryonic bovine lung (EBL) cells.In this study,samples which co-incubated with P48 protein and EBL cells in different concentrations at different time points were collected,the proliferation rate of the cells was detected by MTT method,and the changes in the nuclear morphology of EBL cells were observed by DAPI staining method.Meanwhile,flow cytometry was used to detect the apoptosis rate of EBL cells induced by P48 protein.Real-time quantitative PCR was used to detect the changes in mRNA level of apoptotic marker genes,and Western blotting tested the Bax and Beclin-1 protein expressions.The results showed that under the condition of 72 h and 10 μg/mL of protein concentration treatment,P48 extremely significantly inhibited EBL cells proliferation (P<0.01),while 0.1 and 0.5 μg/mL protein concentration had no inhibitory effect (P>0.05).The nuclear morphology showed no significant change after protein induction for 12 h,but wrinkled and condensed at 24 h.The nucleus was fragmented,and a sprouted apoptotic body was appeared at 48 and 72 h.Apoptosis related genes expression showed no obvious increase at 2 and 12 h at mRNA level,but gradually increased at 24,48 and 72 h,and it showed a time-dependent manner.Accordingly,the expression of apoptosis marker proteins Bax and Beclin-1 significantly increased.Flow cytometry analysis showed that the apoptosis rate of EBL cells induced by P48 protein was 48.44%.In conclusion,P48 recombinant protein of Mycoplasmas bovis inhibited the proliferation of EBL cells and promoted their apoptosis,which provided reference for revealing the pathogenic mechanism of Mycoplasmas bovis.     

16种中药体外抗猪繁殖与呼吸综合征病毒作用的研究

To observe the antiviral action of 16 kinds of Chinese medicinal herbs (Houttuynia cordata Thunb,Flos Chrysanthemi,Flos Lonicerae Confusae,Anemarrhena asphodeloides,CassiaTwig,Herba Ephedrae,Radix Paeoniae Rubra,Bupleurum chinense,Dryopteris setosa,Folium Isatidis, Gardenia jasminoides Ellis,Mntha haplocalyx Briq,Paeonia suffruticosa Andr,Cornu Bubali, Herba Senecionis Scandentis,Radix Gentianae) against porcine reproductive and respiratory syndrome virus （PRRSV）, the blocking, inhibiting and killing roles were detected in present study in vitro. The results showed that 13 kinds of Chinese medicinal herbs had antiviral actions against PPRSV except Paeonia suffruticosa Andr, Herba Senecionis Scandentis and Folium Isatidis, and Houttuynia cordata Thunb, Flos Chrysanthemi, Anemarrhena asphodeloides, CassiaTwig and Radix Paeoniae Rubra exhibited the best antiviral actions followed by Herba Ephedrae, Bupleurum chinense and Dryopteris setosa.     

昆明小鼠胃肠道钙离子跨膜吸收途径相关基因表达模式分析

本研究旨在分析钙离子(Ca2+)跨膜吸收途径相关基因在昆明小鼠胃肠道中的表达模式。选取12只8周龄、平均体重(30.71±2.93)g的雌性昆明小鼠,取胃、十二指肠、空肠、回肠、盲肠和结肠组织样品,利用实时定量PCR法检测维生素D依赖性钙结合蛋白(CaBP-D9k)、瞬时性受体电位通道香草酸受体6(TRPV6)和维生素D受体(VDR)mRNA表达量。结果表明:1)CaBP-D9k、TRPV6和VDR mRNA在胃内属低水平表达,而在盲肠内表达量较高;2)随着小肠的延伸,CaBP-D9k、VDR mRNA表达量逐渐降低,而TRPV6 mRNA则在回肠内高水平表达;3)CaBP-D9k(P<0.05)、VDR(P<0.05)、TRPV6 mRNA的表达量(P>0.05)随着大肠肠段的延伸而不同程度地下降。结果提示,CaBP-D9k、TRPV6和VDR mRNA表达量与胃肠道Ca2+跨膜吸收能力存在关联性。     

塞北兔种质特性研究与利用现状

从外貌特征和生产性能两个方面对塞北兔的种质特性进行了详细描述,并从数量遗传学、生化遗传学、被毛颜色和耳型遗传规律、生理生化指标、营养需要特点等方面对塞北兔的研究情况进行了概括总结。同时针对目前塞北兔在研究利用方面存在的问题提出了合理建议。     

花溪灰萝卜区域试验研究

为筛选出具有推广潜力、产量较高、抗逆性强并适应贵州气候条件的饲用灰萝卜新品种,对3个灰萝卜品种的生育期、生产性能、抗逆性等进行了对比分析,结果表明,花溪灰萝卜品种优势突出,表现为产量高、抗逆性强,具有较大的推广潜力.     

高分辨率熔解曲线分析法检测牦牛脂蛋白脂酶外显子7的多态性及与生长性状相关性分析

脂蛋白脂酶(lipoprotein lipase,LPL)是一个分子质量为54 kb的糖蛋白,其活性受营养因素、激素水平等调节,因牦牛生存环境的特殊性,试验拟从分子水平研究天峻县牦牛脂蛋白脂酶外显子7(LPL-Exon7)基因的多态性,并与生长发育性状进行关联分析,从而为牦牛营养调控、品种资源保护和遗传育种提供科学理论依据。试验采集青海省海西州天峻县牦牛抗凝血106份,提取血样DNA,并利用PCR技术和高分辨率熔解曲线分析法对候选基因LPL-Exon7进行多态性分析,并于采血的同时对牦牛体重、体斜长、体高、胸围和管围等生长性状指标进行了相关性和遗传效应分析。结果表明,高原牦牛LPL-Exon7基因在高原牦牛和野血牦牛中都表现为多态,存在3种基因型,即AA、AB和BB,由1对等位基因A和B控制,A等位基因均为优势等位基因。经χ²检验,符合哈迪—温伯格(Hardy-Weinberg)定律,处于平衡状态(P>0.05)。该位点在高原牦牛和野血牦牛群体中公、母牦牛的多态信息含量为0.3588和0.2103、0.3219和0.3343,高原牦牛母牛的多态信息含量PIC<0.25,处于低度多态,高原牦牛公牛和野血牦牛公、母牛的多态信息含量均在0.250.05)。因此,初步推断牦牛LPL-Exon7可以作为牦牛标记辅助选择的遗传标记之一。     

我国特殊用途家蚕品种选育的研究进展

综述了我国彩色茧品种、丝胶品种、细纤度品种、雄蚕品种、限性品种等特殊用途家蚕品种选育的研究进展,并分析了特殊用途家蚕品种选育的发展趋势。