鸡Ii结合Rab5a和Rab7b分子的分析

本研究旨在探明鸡恒定链（invariant chain，Ii）与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征。首先，用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii_{（Cyt-Tra）}]、Ii CLIP （class Ⅱ-associated invariant chain peptide）-三聚体区[Ii_{（CLIP-TRIM）}]和Ii突变体[Ii_{（M81-87aa）}、Ii_{（M91-99aa）}和Ii_{（M81-99aa）}]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒。其次，将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T，观察它们的共定位。将构建的原核重组质粒进行表达和纯化，最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域。结果表明，成功构建Ii结构域及Ii突变体的重组质粒。Ii_{（Cyt-Tra）}及Ii突变体均能与Rab5a和Rab7b在细胞内共定位，而Ii_{（CLIP-TRIM）}与空载体却不能。Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域，而不是CLIP与三聚体区。综上所述，鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区，而不是内质网腔区。这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制，为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径。     

OsHemA gene,encoding glutamyl-tRNA reductase(GluTR) is essential for chlorophyll biosynthesis in rice(Oryza sativa)

Chlorophyll(Chl) biosynthesis is essential for photosynthesis and plant growth. Glutamyl-tRNA reductase(GluTR) catalyzes glutamyl-tRNA into glutamate-1-semialdehyde(GSA) and initiates the chlorophyll biosynthesis. Even though the main role of GluTR has been established, the effects caused by natural variations in its corresponding gene remain largely unknown. Here, we characterized a spontaneous mutant in paddy field with Chl biosynthesis deficiency, designated as cbd1. With intact thylakoid lamellar structure, the cbd1 plant showed light green leaves and reduced Chl and carotenoids(Cars) content significantly compared to the wild type. By map-based gene cloning, the mutation was restricted within a 57-kb region on chromosome 10, in which an mPingA miniature inverted-repeat transposable element(MITE) inserted in the promoter region of OsHemA gene. Both leaf color and the pigment contents in cbd1 were recovered in a complementation test, confirming OsHemA was responsible for the mutant phenotype. OsHemA was uniquely predicted to encode GluTR and its expression level was dramatically repressed in cbd1. Transient transformation in protoplasts demonstrated that GluTR localized in chloroplasts and a signal peptide exists in its N-terminus. A majority of Chl biosynthesis genes, except for POR and CHLG, were down-regulated synchronously by the repression of OsHemA, suggesting that an attenuation occurred in the Chl biosynthesis pathway. Interestingly, we found major agronomic traits involved in rice yield were statistically unaffected, except for the number of full grains per panicle was increased in cbd1. Collectively, OsHemA plays an essential role in Chl biosynthesis in rice and its weak allele can adjust leaf color and Chls content without compromise to rice yield.     

牛星状病毒EvaGreen实时荧光定量PCR检测方法的建立及应用

牛星状病毒(BAstV)是我国新发的犊牛腹泻病原,本试验的目的是建立检测BAstV的Real-time PCR方法。根据BAstV流行株的ORF1a基因序列设计引物,通过优化反应条件和体系,成功建立基于EvaGreen检测BAstV的Real-time PCR方法。结果表明,该检测方法的Ct值与标准品模板在1.36×10¹~1.36×10⁸拷贝/μL线性关系良好,相关系数R²=0.999,扩增效率为93.79%;该方法可特异性检出BAstV,对犊牛腹泻其他相关病原呈阴性;最低检测下限为13.6拷贝/μL;批间和批内的变异系数均小于2%,重复性好。对2017年9月至2019年5月采自河南省的221份犊牛腹泻样本进行检测,BAstV的检出率为18.1%(40/221),采样场阳性率为100.0%(14/14)。本试验所建方法灵敏度高、特异性强、稳定性好,为BAstV的检测和流行病学调查提供了有力手段。     

Site‐specific management is crucial to managing Mikania micrantha

Increasingly, weeds have been taking on global distributions. With the proliferation of invasive weeds has come the challenge of managing these species over broad geographical regions, with diverse habitats and political jurisdictions. Here, we review the management of Mikania micrantha Kunth (Asteraceae; mile‐a‐minute) throughout its invaded range, extending through most of the Pacific islands and southern and south‐east Asia. Context matters when determining the best course of action for managing M. micrantha, as it has invaded a large variety of agricultural and natural systems. In Queensland, Australia and Florida, USA, M. micrantha has been targeted in relatively successful eradication campaigns, highlighting the importance of early detection and rapid response methods, while elsewhere in its invaded range, populations are either still increasing or showing limited signs of decline. An inter‐regional approach to research and management should incorporate successful management strategies employed throughout the invaded range including, but not limited to, chemical and cultural control practices, manual and mechanical control, classical biological control using the rust fungus Puccinia spegazzinii, plant–plant competition and integrated approaches utilising two or more control methods concurrently. Additional knowledge of M. micrantha genetics is required to determine if management approaches could be fine‐tuned for particular populations. Countries bordering the Mekong River formed a network in 2011 to co‐ordinate the management of invasive species such as M. micrantha. Expanding such a collaborative approach to other regions could further reduce populations of M. micrantha and limit its spread.     

长期施肥处理对双季晚稻叶绿素荧光特征及籽粒产量的影响

研究施肥模式对水稻叶绿素含量及荧光参数的调节作用,对提高水稻光能利用效率及籽粒产量具有重要意义。本研究以水稻品种赣929为试验材料,比较长期定位施磷钾肥(PK)、施氮磷钾肥(NPK)、等养分条件下70%氮磷钾肥配合施用30%有机肥(70F+30M)、50%氮磷钾肥配合施用50%有机肥(50F+50M)、30%氮磷钾肥配合施用70%有机肥(30F+70M)条件下水稻叶绿素荧光参数和产量变化特征。结果表明,水稻旗叶叶绿素含量从齐穗期到成熟期呈下降趋势,且表现为30F+70M50F+50M70F+30MNPKPK。水稻叶绿素荧光参数Fv/Fm随水稻生育期的推进呈降低趋势,其中PK处理Fv/Fm值最低,而施氮处理比不施氮处理Fv/Fm值增加了2.85%～4.18%。叶绿素荧光参数qL、ΦPSII在NPK处理中表现为齐穗期较高, 20 d之后显著降低;在50F+50M、30F+70M处理中表现为齐穗期较低, 20 d以后显著增加;在PK处理、70F+30M处理中一直处于较高水平。NPQ变化趋势与qL基本相反。ETR-PAR光响应曲线拟合结果表明, 70F+30M处理ETRmax、?和Ek值在齐穗期和20 d后值均最高。综合看来,等养分条件下配施30%有机肥具有最优的叶绿素荧光指标组合及籽粒产量;而配施超过50%有机肥由于前期热耗散增大使得用于光合作用的光能份额减少,而施用氮磷钾肥处理则由于后期的衰老使得光能利用效率下降。     

施氮量对柳枝稷叶片叶绿素荧光特性及干物质积累的影响

研究施氮量对柳枝稷叶绿素荧光特性及干物质积累的影响,对提高盐碱地柳枝稷光能利用效率及生物质产量具有重要意义。在宁夏银北地区采用大田试验,以Cave-in-Rock品种柳枝稷为供试材料,在无氮添加(0 kg·hm^-2,N₀)、施低氮(60 kg·hm^-2,N₆₀)、中氮(120 kg·hm^-2,N₁₂₀)和高氮(240 kg·hm^-2,N₂₄₀)共4个施肥水平下,比较了开花期和成熟期柳枝稷叶片叶绿素荧光参数和干物质积累动态变化,采用灰色关联度分析法对柳枝稷叶片叶绿素荧光参数与干物质积累量进行研究。结果表明:与N₀相比,在N₆₀、N₁₂₀和N₂₄₀处理下柳枝稷开花期和成熟期叶片PSⅡ最大光化学效率(F_v/F_m)、PSⅡ实际光化学量子效率(Φ_PSⅡ)、潜在活性(F_v/F_o)、光化学猝灭系数(qP)、非光化学猝灭系数(NPQ)和干物质积累量均显著性提高,在N₂₄₀处理下达到峰值,而热耗散量子比率(F_o/F_m)显著性降低。在柳枝稷开花期,随着施氮量的增加,柳枝稷叶片PSⅡ潜在活性(F_v/F_o)显现出先上升后逐渐下降的总趋势。在N₁₂₀处理下柳枝稷叶片PSⅡ潜在活性(F_v/F_o)最大,为3.13,较N₀处理显著提高了16.26%。柳枝稷干物质积累量随着生育时期的推进和施氮量的增加,均有不同程度的提高,其中在开花期至成熟期,干物质积累缓慢,在灌浆期达到最大,成熟后期略有降低。柳枝稷成熟期干物质积累量在N₂₄₀处理下最高,每穴干物质积累达378.13 g,较N₀、N₆₀和N₁₂₀处理显著提高了24.33%、20.09%和7.24%。灰色关联度分析结果表明,在本试验条件范围内,N₂₄₀处理下加权关联度与理想施肥水平的关联度最大,有利于促进PSⅡ的光化学活性,从而提高柳枝稷干物质积累量。     

Up-regulation of miR-181a attenuates releases of CSE-induced pro-inflammatory factors and expression of collagen IV,fibronectin and α-SMA in human bronchial epithelial cells

AIM: To investigate the effect and potential mechanism of microRNA-181a (miR-181a) on cigarette smoke extract (CSE)-induced the productions of pro-inflammatory factors and the expression of collagen IV, fibronectin and α-smooth muscle actin (α-SMA) in human bronchial epithelial cells (HBECs). METHODS: CSE-induced miR-181a expression was detected by RT-qPCR in the HBECs. After tansfected with miR-181a mimic, the releases of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6 and transforming growth factor-β1 (TGF-β1) were measured by ELISA, the protein expression of collagen IV, fibronectin and α-SMA was determined by Western blot. The activation of NF-κB/TGF-β1/Smad3 pathway was also evaluated by Western blot. RESULTS: CSE increased the levels of TNF-α, IL-1β, IL-6 and TGF-β1 and the expression of collagen IV, fibronectin and α-SMA, and decreased the expression of miR-181a in the HBECs (P<0.05). However, transfected with miR-181a mimic partially prevented the releases of TNF-α, IL-1β, IL-6 and TGF-β1, and inhibited the expression of collagen IV, fibronectin and α-SMA (P<0.05). Additionally, the activation of NF-κB/TGF-β1/Smad3 evoked by CSE was attenuated after transfected with miR-181a mimic. CONCLUSION: Up-regulation of miR-181a prevents the releases of CSE-induced pro-inflammatory factors and expression of collagen IV, fibronectin and α-SMA in the HBECs, and its mechanism may be related to the inhibition of NF-κB/TGF-β1/Smad3 pathway.     

模拟城市SO2污染对晚松生理特性的影响

以2年生晚松盆栽苗为供试材料,从抗大气环境中SO₂污染方面对晚松的抗逆能力进行了定量评价,为晚松的园林应用提供理论依据。研究表明,晚松叶绿素a、b对模拟SO₂污染反应敏感,叶绿素b所受伤害大于叶绿素a,类胡萝卜素较不敏感。过氧化物酶活性的提高有助于减轻膜脂过氧化造成的伤害,从而使其在SO₂污染胁迫较轻(10 mmol/L)时免受伤害。     

核桃主栽品种新新2和温185光合特性与其品质和产量的关系

【目的】 研究核桃光合特性与其品质和产量的关系。【方法】 以新疆优良核桃品种新新2和温185为材料,对照分析2个品种的光合作用及其叶绿素荧光特性,与其果实品质和产量的关系。【结果】 温185和新新2光合速率与量子效率没有显著差异,这与2个品种单株产量和单产一致。2个品种在出仁率上有显著差异,即温185出仁率显著高于新新2,温185呼吸速率也显著高于新新2,出仁率可能与其代谢特性有关。总叶绿素含量之间无显著差异,而温185的叶绿素a/b值和类胡萝卜素含量均极显著高于新新2。【结论】 温185和新新2的产量与其光合特性有关,而其品质与其它代谢有关。温185比新新2具有更强的光保护能力。     

不同矮化中间砧木对库尔勒香梨光合特性的影响

【目的】 研究中矮1号、中矮2号、中矮3号和杜梨对库尔勒香梨光合荧光特性的影响,为筛选适宜库尔勒香梨矮化密植栽培的砧木提供理论基础。【方法】 采用Li-6400光合仪及FMS-2脉冲调制式荧光仪对4种不同矮化中间砧木库尔勒香梨叶片进行光合及荧光参数的测定,分析不同砧木的库尔勒香梨的光合特性和叶绿素荧光特性差异。【结果】 不同矮化中间砧木库尔勒香梨的Pn日变化均呈“双峰”曲线,都有明显的光合“午休”现象,光合“午休”现象是由非气孔限制因素导致;中矮1号的净光合速率(Pn)日均值、水分利用率(WUE)日均值均高于其他3个砧木的Pn日均值和WUE日均值;不同矮化中间砧木对库尔勒香梨的气孔导度(Gs)没有显著影响;中矮1号的实际光化学效率(ΦPSII)、非光化学淬灭系数(CNPQ)、PSII电子传递速率(ETR)的值为最高;不同矮化中间砧木对库尔勒香梨的光化学淬灭系数(CqP)、PSⅡ潜在光化学活性(Fv/Fo)值没有显著影响。【结论】 中矮1号作为库尔勒香梨的砧木优于其他2个矮化砧木和杜梨,可以作为库尔勒香梨的优良砧木。