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。     

Biochar addition mitigates nitrogen loss induced by straw incorporation and nitrogen fertilizer application

Biochar has been shown to be potentially beneficial for enhancing yields and soil properties, and diminishing nitrogen (N) losses. However, it remains unclear how biochar regulates soil carbon (C) and N to mitigate N losses induced by straw mixing with N fertilizer in dryland soils. Therefore, we investigated the effects of straw mixing (S₁), S₁ with biochar (SB) and no straw inputs (S₀), and routine urea application rates (N₁) and 70% of routine rates (N_0.7) on yields and N losses, and identify the relationship between N losses and soil C and N compounds. Results showed that N_0.7 and N₁ were suitable for the maize and wheat seasons, respectively, contributing to mitigating N losses without reducing crop yields. Moreover, in the maize season, N_0.7-SB significantly mitigated the straw-induced NH₃-N and N₂O-N emissions by 106% and 81%, respectively. In the wheat season, N₁-SB reduced the straw-induced NH₃-N and N₂O-N emissions by 35% and 66%, respectively. In addition, N_0.7-SB sharply reduced soil inorganic N (SIN) storage in the maize season. Furthermore, the NH₃-N and N₂O-N emission rates were negatively correlated with dissolved organic carbon/SIN content (0–20 cm) (DOC/SIN_0-20). N losses (N₂O-N and NH₃-N emissions and SIN storage) were positively correlated with SIN_0-20, but negatively correlated with soil organic carbon / SIN_0-20 (SOC/ SIN_0-20). This study provides further evidence that biochar with an appropriate N application rate decreased SIN_0-20 and increased DOC/SIN_0-20, thus reducing SIN storage and the straw-induced gaseous N emissions without decreasing crop yields.     

‘红羽1号’油茶新品种营养成分动态及相关关系分析

为了解‘红羽1号’油茶新品种的营养成分动态及其相关关系，以‘红羽1号’油茶果实近熟期的叶片与果皮、种壳、种仁为材料，测定其还原糖含量、黄酮含量、总酚含量及果皮与叶片中原花青素含量。结果表明，‘红羽1号’油茶中各部位还原糖含量、黄酮含量、总酚含量、原花青素含量具有极显著性差异；在果实近熟期，果实相对成熟，叶片与果皮、种壳、种仁各营养成分动态变化规律明显，但变化幅度不大。叶片原花青素含量与总酚含量间存在显著的线性关系，叶片原花青素含量与黄酮含量和果皮原花青素与总酚含量之间存在显著曲线关系，种壳中总酚含量与黄酮含量极显著正相关。     

鲁西南地区10个小麦品种丰产性和稳产性分析

为更科学的分析评价鲁西南地区主栽的10个小麦品种丰产性和稳定性,本研究利用联合方差分析、线性回归分析和AMMI模型对 3个试验地点产量数据进行分析。结果表明,菏麦20、济麦22和周麦22丰产性和稳定性表现优异,周麦18、烟农999和山农20属于丰产性较好稳产性一般的品种,良星99和山农28属于稳产性好丰产性一般的品种,青农2号稳产性较好丰产性较差,但矮抗58丰产性和稳产性均较差但在E3具有特定区域适应性。     

三种水稻栽培模式对主要害虫种群发生动态的影响

轻简化和机械化是现代农业发展的趋势。本文于2 0 1 3 —2 0 1 5 年连续进行了常规人工栽插、机械直播、机械栽插等3种栽培模式对水稻主要害虫种群发生动态的影响研究,旨在为农业轻简、机械化模式的进一步推广提供依据。结果表明,3种栽培模式对稻飞虱、稻纵卷叶螟种群发生的影响差异不明显。比较3 种栽培模式下的水稻螟虫(二化螟为主, 大螟较少) 发生和为害可知,机械直播田的螟虫前期株为害率较高,且后期仍维持较高的为害率,明显高于人工栽插和机械插秧田(p<0.05)。分析原因,很可能是机械直播水稻暴露在大田的时间要稍长于栽插模式(机械插秧和人工栽插),有利于二化螟的提前入田为害。因此在推广机械直播时,应考虑调整播期,尽可能地避开或减少螟虫为害。     

磁化水灌溉对保护地土壤质量及尖椒品质的影响

[目的]探讨磁化水处理技术对长期保护地栽培土壤矿质养分、生物酶活性、物理结构等土壤理化性状及果实品质的影响,为减缓保护地栽培土壤次生盐渍化进程及土壤质量改善提供理论依据,为磁化水处理技术的应用提供理论基础。[方法]以山东省寿光市洛城镇长期保护地栽培土壤和尖椒为试材,采用田间小区试验设计,设置磁化水灌溉处理,以非磁化水灌溉处理为对照,测定0~15cm土层中土壤全量养分、有效性养分和交换性养分含量、土壤脲酶、磷酸酶、蔗糖酶和过氧化氢酶活性以及土壤容重、孔隙度、土壤颗粒以及pH等土壤理化性质和果实品质指标,分析其变化特征。[结果]结果表明：(1)磁化水灌溉提高了尖椒果实中维生素C、花青素、还原糖含量和可溶性糖含量,提高比例分别为1.76~46.85%；降低了果实中有机酸含量。(2)磁化水灌溉处理促进了Fe、Cu、N和P在叶片中的累积,其含量高于果实,而Mn和Zn含量则略低于果实中含量；Fe含量提高比例最高,在叶片和果实中平均值为57.20%,其次为全磷含量,平均值为53.49%；且磁化水灌溉处理尖椒果实和叶片矿质元素含量均高于非磁化水灌溉处理。(3)磁化水灌溉保护地栽培土壤中N、P、Mn和Zn等元素全量含量降低,为2.04%~20.61%；Fe和Cu等元素全量含量提高为4.64%-6.48%；土壤交换性K、Na和Ca等离子含量降低,为2.15%~4.95%,而交换性Mg离子含量则提高；铵态氮、硝态氮、速效磷和有机质等有效性养分含量提高。(4)磁化水灌溉保护地栽培土壤脲酶、蔗糖酶、磷酸酶和过氧化氢酶等生物酶活性均有不同幅度的提高,为0.27%~24.75%。(5)磁化水灌溉保护地栽培土壤容重降低、孔隙度增加,黏粒和粉粒等细小颗粒含量增加,而砂粒含量则有所下降。[结论]综上所述,磁化水处理技术有利于土壤物理结构的改善,对土壤有效性养分累积和供应强度以及土壤生物酶活性有明显的促进作用,从而提高尖椒果实的鲜食品质；并通过调整交换性离子含量和组成以及土壤pH的调节,减缓保护地土壤次生盐渍化的发生。     

Role of cytokeratin 8 on changes of intestinal epithelial permeability induced by corticotropin-releasing factor

AIM: To investigate the role of cytokeratin 8 (CK8) on the change of intercellular permeability of intestinal epithelial cells induced by corticotropin-releasing factor (CRF). METHODS: The expression levels of CRF receptor 1 (CRFR1) and CRFR2 on human colon adenocarcinoma HT29 cell surface were determined by immunofluorescence staining. After treatment with 100 nmol/L CRF for 72 h, the translocation of FITC-labelled dextran was measured in a Transwell chamber. The structural changes of tight junctions were observed under transmission electron microscope. The expression levels of CK8, and tight junction proteins ZO-1 and occludin were determined by Western blot. The activity of protein kinase C (PKC) was detected by ELISA. Furthermore, the effects of CRF on intestinal epithelial permeability were examined in CK8-silencing HT29 cells, which were constructed by infection with sh-CK8 lentivirus. RESULTS: CRF treatment increased the permeability of FITC-labelled dextran (P<0.05), caused the opening of tight junctions, and induced increased fluorescence intensity of CK8. The expression levels of occludin and ZO-1 were down-regulated (P<0.05). PKC activity was decreased at 1 h after CRF treatment (P<0.05). CRF-induced increase in the permeability and down-regulation of occludin were not blocked by CK8 silencing. Nevertheless,CK8 silencing blocked the effects of CRF regarding the decrease in the expression levels of ZO-1 and the increase in PKC activity (P<0.05). CONCLUSION: CK8 may be involved in CRF-induced increase in intestinal epithelial permeability by inhibiting the activity of PKC, and there may be other signaling pathways involved.     

宜宾市农业干旱危险性演变特征分析

基于四川省宜宾市1951—2010年夏季(6—8月)降水距平资料,以30年为窗口、1年为滑动步长,通过构造降水距平百分率和农业干旱危险性等级(轻旱、中旱、重旱、严重干旱)之间的模糊隶属函数,引入区间平均隶属度,将信息扩散理论与之结合,研究了宜宾近60年农业干旱危险性的演变特征。结果表明,轻度干旱危险性水平变化趋势总体为波动下降;中度干旱危险性水平在10.5%处保持振荡平衡;重度干旱危险性水平在前期有下降趋势,之后迅速上升并持续保持高位振荡;严重干旱危险性水平长期保持低位振荡后,在第24序列之后骤增;中度以下农业干旱危险性水平总体呈振荡下降趋势,重度以上农业干旱危险性水平总体呈上升趋势。本研究针对不同等级农业干旱危险性分别计算各自的模糊概率,对该地防灾减灾实践具有重要价值。     

水分胁迫对不同抗旱型玉米自交系苗期生长的影响

通过水分胁迫处理对不同抗旱型玉米自交系苗期的影响研究,为抗旱育种选择自交系提供选择参考。采用人工控水的方法,研究干旱胁迫对夏玉米抗旱型和对敏感型自交系苗期株高、地上部和地下部分干重、叶面积、比叶重、叶绿素和净光合速等的影响。结果表明：不同抗旱型自交系苗期对水分胁迫的响应不同。抗旱玉米自交系从5叶期开始在干旱胁迫期间的株高、地上部和地下部分干重、叶面积、叶绿素和净光合速率等指标均高于对水分敏感的玉米自交系,且比叶重相对稳定。这表明干旱条件下,抗旱型自交系较普通自交系在苗期能够表现出更好的适应性,本研究对加强抗旱自交系的筛选和选育,进一步提高耐旱节水玉米新品种选择效率具有一定参考价值。     

Down-regulation of miR-153-3p attenuates H2O2-induced H9C2 cell injury by promoting Nrf2 expression

AIM To study the effect of microRNA-153-3p (miR-153-3p) knock-down on oxidative injury of H9C2 cells induced by H₂O₂ and its specific mechanism. METHODS The oxidative stress injury of H9C2 cell model was induced by H₂O₂, and then the cell viability and the expression of miR-153-3p were detected by MTT assay and RT-qPCR, respectively. The effects of miR-153-3p knock-down on the H9C2 cell injury under oxidative stress were studied by RNA interference technology. The targets of miR-153-3p were identified by Western blot and dual-luciferase reporter assay. RESULTS MTT assay showed that the viability of H9C2 cells was decreased with the increase in H₂O₂ concentration (P<0.05). The results of RT-qPCR showed that the expression of miR-153-3p was increased with the increase in H₂O₂ concentration (P<0.05). Knock-down of miR-153-3p increased the viability of H9C2 cells under oxidative stress, decreased the cell apoptosis and the content of malondialdehyde (MDA), and increased the activity of superoxide dismutase (SOD). The expression of nuclear factor E2-related factor 2（Nrf2） and antioxidant response element（ARE） activity were increased with the increase in H₂O₂ concentration (P<0.01). TargetScan analysis and dual-luciferase reporter assay showed that Nrf2 was one of the potential target genes of miR-153-3p. The results of Western blot further showed that over-expression of miR-153-3p inhibited the expression of Nrf2 (P<0.01), while down-regulation of miR-153-3p increased the expression of Nrf2 (P<0.01). Dual interference with Nrf2 and miR-153-3p significantly reduced H9C2 cell viability， promoted the apoptosis, increased MDA content, and decreased SOD activity in the presence of H₂O₂ (P<0.01). CONCLUSION Inhibition of miR-153-3p expression attenuates the injury of H9C2 cells induced by H₂O₂ through up-regulating Nrf2/ARE signaling pathway.