储粮害虫磷化氢抗性分子遗传学研究进展

由于长期单一使用磷化氢,储粮害虫对其产生的抗药性日益增强,已成为威胁粮食安全储藏的重大问题。自从发现两个磷化氢抗性基因rph1和rph2,以及完成赤拟谷盗全基因测序以来,储粮害虫磷化氢抗性基因的相关研究迅速发展。近年来,在抗性基因的鉴定、表达、进化、突变等研究方面取得了一定成果,为储粮害虫磷化氢抗性治理提供了新的科学指导。本文就储粮害虫磷化氢抗性分子遗传学的研究进展进行综述。     

规模化蛋鸡场病原菌溯源与生物安全防控研究

细菌病严重威胁蛋鸡健康,以抗生素为主导的细菌病传统防控方式会导致病原菌耐药性严重、鸡蛋药物残留,影响蛋品安全。要保证蛋鸡产蛋期不用抗菌药物而又能生产出"无菌、无抗"的鸡蛋,是国内外技术难题。本团队以规模化蛋鸡场全封闭鸡舍为研究对象,基于PFGE、MLST、全基因组测序等技术,探明了蛋鸡病原菌的种类及基因组特征;对病原菌的耐药性研究表明,蛋鸡病原菌多重耐药严重,依靠药物防控的方式亟待转变;对规模化蛋鸡场细菌溯源研究结果表明,鸡舍生物媒介(鼠、苍蝇等)和非生物媒介(空气、饮水、饲料、鸡粪等)是蛋鸡病原菌的重要传染来源和传播途径;对规模化蛋鸡场生物安全量化评价结果表明,外部和内部生物安全权重高达92.2%。根据溯源和生物安全权重分析结果,提出了在规模化全封闭蛋鸡舍将蛋鸡细菌病从药物防控转变为生物安全防控的新策略;创新了规模化蛋鸡场细菌病防控的系统技术,实现蛋鸡规模养殖整个产蛋期不用抗生素,为生产"无菌、无抗"安全鸡蛋提供技术支撑。     

不同温度下TMV侵染枯斑三生烟的LncRNA差异表达

【目的】 筛选不同温度下烟草花叶病毒（Tobacco mosaic virus,TMV）侵染后枯斑三生烟（Nicotiana tabacum var. Samsun NN）差异表达的长链非编码RNA（long non-coding RNA,lncRNA）,研究lncRNA在枯斑三生烟抗性反应中的作用。【方法】 N基因的温度敏感性使枯斑三生烟在25℃时具备对TMV的抗性、在31℃抗性丧失,在这两个温度条件下对枯斑三生烟接种TMV和磷酸盐缓冲盐水（phosphate buffered saline,PBS）,48 h后提取系统叶总RNA,构建链特异性文库后进行深度测序。对测序结果进行过滤后利用HTSeq将有效数据与近缘品种TN90（N. tabacum var. TN90）基因组比对,筛选得到lncRNA后利用FPKM法估计lncRNA的表达水平。通过edgeR筛选差异表达lncRNA（differentially expressed lncRNA,DElncRNA）,并利用qRT-PCR技术对这一结果进行验证。通过共定位及共表达分析预测DElncRNA的靶基因,通过参考基因组注释、GO和KEGG富集分析研究靶基因的功能。【结果】 4个处理共12个样本经lncRNA-seq各测得约8 000万条clean reads,共获得4 737条已知lncRNA、40 169条新lncRNA。其中64个lncRNA在不同温度条件下TMV侵染后存在差异表达,qRT-PCR测定结果显示这些lncRNA的测序正确率在80%左右,表明本研究所得测序数据具备较高的可信度。对DElncRNA进行靶基因预测,发现一些基因同时被25℃下调和31℃上调的DElncRNA靶向。靶基因注释功能丰富,主要参与植物抗病、激素和代谢等生理过程。部分可能与激素通路相关的lncRNA,在25℃下TMV侵染时呈现下调趋势,而在31℃下TMV侵染则呈现上调趋势。GO富集分析显示靶基因主要参与构成膜、囊泡等组分,具备钙、钾离子通道抑制剂活性等分子功能,使相应离子得以转运引发随后的反应,同时也参与发病、抗原加工和呈现、细胞分裂素代谢等生理过程。KEGG分析发现靶基因显著富集在植物激素信号转导通路,25℃下调和31℃上调的DElncRNA靶基因同时富集在激素信号传导、ABC运输蛋白、苯丙烷类生物合成等通路。【结论】 不同温度（25℃和31℃）条件下TMV侵染枯斑三生烟后,长链非编码RNA差异表达,DElncRNA通过作用于激素信号传导、物质转运等过程参与寄主系统获得性抗性反应。研究结果可为揭示植物系统获得性抗性中lncRNA的调控功能以及新型抗病毒技术开发提供依据。     

利用分子标记辅助选择改良春恢350稻瘟病抗性

水稻抗稻瘟病基因Pigm(t)抗谱广、抗性强。春恢350是超级早稻春光1号的恢复系,该恢复系配合力强,丰产性好,但不抗稻瘟病。本研究以携带抗稻瘟病基因Pigm的谷梅4号为抗源,以春恢350为轮回受体亲本,在回交选育过程中,通过表形筛选与分子标记辅助选择相结合,将Pigm(t)基因导入到春恢350中,获得3个带有目标基因的改良恢复系纯合株系。以江西近年来具有代表性的20个菌株对这3份材料进行抗性鉴定,其抗性频率为85%~100%,而原始对照春恢350的抗性频率仅为5%,表明抗性基因已成功导入春恢350中并表达;并用不育系江农早4号A与改良的春恢350测配,其杂种一代田间表现优势强,抗性强。     

拟南芥响应3-羰基己酰基高丝氨酸内酯的膜蛋白质组分析

为了探索植物响应细菌信号N-酰基高丝氨酸内酯(AHLs)的分子机制。本文利用二维差异凝胶电泳技术分析了3-羰基己酰基高丝氨酸内酯(3OC6-HSL)处理引起拟南芥膜蛋白质组的变化情况,结果表明3OC6-HSL共诱导53个蛋白点发生显著变化,涉及天然免疫、逆境胁迫响应、碳代谢、能量代谢、光合作用等多种生物学功能,暗示了3OC6-HSL在调控植物生长发育和抗逆性方面发挥着重要作用,可为阐明植物-细菌跨界通讯的分子机制提供更多的功能蛋白质信息。     

两种血清抗性检测方法的比较分析

采用流式细胞术法对大肠杆菌的血清抗性进行检测,并与传统涂板法相比较,探讨流式细胞术在血清抗性检测方法中的应用。结果表明,两者呈正相关关系(P=0.0340.05),流式细胞术法可以代替涂板法对大肠杆菌的血清抗性进行检测。该方法可以减少由于涂板不均匀、人工计数等造成的误差,且易操作、计算少、耗时少,数据更准确。     

益生菌大肠杆菌Nissle 1917抗逆性能、猪肠上皮细胞黏附率及抑菌效果研究

本试验旨在研究益生菌大肠杆菌Nissle 1917(Ec N)抗逆性能、猪肠上皮细胞黏附率及抑菌效果。采用体外法对Ec N进行生长曲线绘制和耐酸、耐胆盐、耐热性能的测定;以猪肠上皮细胞IPEC-J2细胞为体外细胞模型,考察了Ec N对该细胞的黏附率以及对致病菌大肠杆菌K88的黏附抑制率;同时通过蛋白质印迹法检测了Ec N对IPEC-J2细胞β-防御素-2和Toll样受体4的水平的影响。结果表明:1)Ec N对高酸、高胆盐和高温环境具有一定耐受能力。2)Ec N对IPEC-J2细胞的黏附作用以对数期最佳,黏附率达33.96%,显著高于迟缓期、稳定期和衰亡期(P0.05)。3)Ec N对致病菌大肠杆菌K88具有良好的抑制效果,黏附抑制率达87.84%。4)Ec N还能上调IPEC-J2细胞β-防御素-2和Toll样受体4水平。结果提示,益生菌Ec N具有较好的抗逆性能,能够良好地黏附猪肠上皮细胞,对致病菌大肠杆菌K88具有良好的抑制作用。     

Screening of Drought Resistance Identification Indexes for Rice (Oryza sativa L.) Cultivars in Ningxia

The morphological characters, physiological characters, yield traits and yield per plant of total 49 rice cultivars from Ningxia were investigated under conditions of water stress and non-water stress so as to determine the relationship between each trait and yield per plant under water stress and the relationship between each relative character and drought resistance coefficient under water stress and non-water stress. The correlation, grey correlation, stepwise regression and path analyses showed that the til er number per plant, plant height, grain density, effective panicle number per plant and grain number per pan-icle, total 5 traits, were significantly correlated with the drought resistance of rice, and they could be used to identify the drought resistance of rice in Ningxia. In addition, the drought resistance of rice was graded qualitatively according to the subordinate function value of corresponding drought resistance coefficient. The results showed that among the 49 rice cultivars from Ningxia, 6 rice cultivars were highly drought resistant, and 9 rice cultivars were moderately drought resistant, suggesting that the evalua-tion method was feasible and effective.     

Foliage‐applied sodium nitroprusside and hydrogen peroxide improves resistance against terminal drought in bread wheat

Terminal drought is threatening the wheat productivity worldwide, which is consumed as a staple food by millions across the globe. This study was conducted to examine the influence of foliage‐applied stress signalling molecules hydrogen peroxide (H₂O₂; 50, 100, 150 μm ) and nitric oxide donor sodium nitroprusside (SNP; 50, 100, 150 μm ) on resistance against terminal drought in two bread wheat cultivars Mairaj‐2008 and BARS‐2009. These stress signalling molecules were applied at anthesis stage (BBCH 61); drought was then imposed by maintaining pots at 35% water holding capacity. Terminal drought caused significant reduction in grain yield of both tested bread wheat cultivars; however, foliage application of both stress signalling molecules at either concentration improved the performance of both bread wheat cultivars. Maximum improvement in 100‐grain weight (12.2%), grains per spike (19.7%), water‐use efficiency (WUE; 19.8%), chlorophyll content index (10.7%), total soluble phenolics (21.6%) and free leaf proline (34.3%), and highest reduction in leaf malondialdehyde contents (20.4%) was recorded when H₂O₂ was foliage‐applied at 100 μm . Foliage application of SNP enhanced the grains per spike, 100‐grain weight and grain yield by 14.9%, 11.3% and 20.1%, respectively, than control. The foliage‐applied stress signalling molecules improved the accumulation of soluble phenolics, proline and glycine betaine with simultaneous reduction in malondialdehyde contents, which enabled wheat plants to sustain the biological membranes under stress resulting in better stay green (high chlorophyll contents) under drought. This helped improving the grain number, grain weight, grain yield, WUE and transpiration efficiency. In crux, foliage‐applied H₂O₂ and SNP, at pre‐optimized rate, may be opted to lessen the drought‐induced yield losses in bread wheat in climate change conditions.     

低温胁迫下草坪草抗寒性的研究进展

通过介绍低温对草坪草的影响和作用机理,系统地综述了在低温胁迫下草坪草的光合特性、水分生理代谢、细胞质膜相对透性、保护酶系变化、碳水化合物贮藏、可溶性蛋白质含量、以及脂肪酸的变化在低温胁迫方面的研究进展,提出了今后进行草坪草抗寒性研究的趋势和建议。