分子标记技术在工业大麻性别分化研究中的应用进展

工业大麻性别表达影响着工业大麻种子、纤维和药用成分产量,其性别表达除了受到遗传物质控制外,还受到外界环境条件的影响,这种复杂性使得性别研究一直是工业大麻研究的难点和热点。分子标记技术能在DNA水平上鉴定工业大麻性别,开发与工业大麻性别紧密连锁的分子标记可为大麻性别早期鉴定和特定性别分子辅助选择育种提供理论依据。本文就分子标记在工业大麻性别研究中的应用进展进行了总结,并对应用前景进行了展望。     

β-carotene attenuates weaning-induced apoptosis via inhibition of PERK-CHOP and IRE1-JNK/p38 MAPK signalling pathways in piglet jejunum

Weaning may cause oxidative injury, immune response impairment, apoptosis and other injuries in piglets. Oxidative and endoplasmic reticulum stress (ERS) can elicit inflammatory responses, and persistent oxidative and ERS also may lead to apoptotic cascades, which is associated with the pathogenesis of multiple diseases. β-carotene, a natural carotenoid, has potential anti-inflammatory and antioxidant functions. However, the effect of β-carotene on apoptosis in weaned piglets and the detailed molecular mechanism remain unclear. In this study, we found that β-carotene decreased malondialdehyde (MDA) levels and increased the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in piglet serum. β-carotene could inhibit the mRNA levels of caspase-3 significantly, but had no significant inhibitory effect of the mRNA levels of caspase-9 and caspase-12 in the piglet jejunum. In addition, β-carotene decreased the activation of GRP78, CHOP, and JNK/p38 MAPK and the ratio of Bax/Bcl-2. Furthermore, β-carotene had a significant influence on the activation of ERS and apoptosis-related signals in TG-induced IPEC-J2. In the present study, β-carotene pre-treatment attenuated the ratio of Bax/Bcl-2 and prevented TG-induced increases in the level of PERK-CHOP and IRE1-JNK/p38 MAPK pathway activation in a dose-dependent manner. Overall, these findings indicate that β-carotene may protect weaning-induced apoptosis through inhibiting ERS.     

硅钙钾镁肥配施黄腐酸钾对土壤酶活性及桃幼树生长的影响

为探究硅钙钾镁肥配施黄腐酸钾对桃树生长的效应,以盆栽一年生瑞蟠21号毛桃(Amygdalus persica Linn.)为试验材料,设置3种不同施肥方式(T1:硅钙钾镁;T2:硅钙钾镁+黄腐酸钾;CK:空白对照),在桃幼树生长季进行追肥处理,研究硅钙钾镁肥及硅钙钾镁肥配施黄腐酸钾对土壤酶活性、土壤养分状况及桃幼树根系构型和植株生长的影响。结果表明,与CK相比,T1、T2均显著提高了桃幼树生长季各时期土壤酸性磷酸酶、脲酶和过氧化氢酶活性,以及土壤中碱解氮、有效磷、速效钾和有效硅等含量,且配施黄腐酸钾后作用更明显。与CK相比,T1、T2细根比重分别提高了11.2%和26.2%,根系活力分别提高了12.2%和18.1%,根系总长度分别增加了28.1%和61.9%,且有效延缓了根系衰老;T1、T2植株总干重分别提高了1.2倍和2.0倍,根冠比分别提高了9.4%和17.0%,且差异显著。与CK相比,T1、T2各器官中全氮、全磷、全钾含量均显著增加,地上部硅含量分别提高了61.3%和91.9%,地下部硅含量分别提高了62.3%和102.5%,且差异显著。同时,T1、T2桃幼树叶片叶绿素含量和净光合速率均显著提高。综上所述,硅钙钾镁肥可有效提高土壤酶活,促进桃幼树细根的生长,延缓根系衰老进程,增加氮、磷、钾、硅等营养元素积累量,提高叶片光合速率,促进植株生长;硅钙钾镁肥配施黄腐酸钾后,提高了土壤有效硅及植株硅含量,比单施硅钙钾镁肥效果更加显著。本研究结果为桃树合理施用硅钙钾镁肥提供了理论依据。     

贵州省城乡人口变化与建设用地扩张的脱钩关系

[目的]研究贵州省人口与建设用地变化的关系,以促进土地资源优化配置。[方法]利用贵州省2000—2017年城乡人口及建设用地数据,采用脱钩模型和GIS空间分析方法,研究城乡人地关系演化过程。[结果]贵州省城镇人口持续增加,乡村人口逐年减少;城乡建设用地面积不断扩大,城镇和乡村建设用地同向增长。2000—2008年,贵州省城乡人口变化与建设用地扩张关系为强负脱勾类型,乡村人口与乡村建设用地变化为强负脱钩类型,城镇人口与城镇建设用地变化为扩张负脱钩类型。农村人口减少伴随着建设用地增加,农村人地关系处于较不合理状态;城镇人口和建设用地二者均增长,但建设用地增幅快于人口增幅。[结论]贵州省应积极推动农村宅基地流转,提高城乡建设用地集约利用水平,促进城乡融合。     

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.     

Effect of soil acidification on the growth and nitrogen use efficiency of maize in Ultisols

Purpose

To examine the effect of soil acidification on growth and nitrogen (N) uptake by maize in Ultisols.

Materials and methods

A clay Ultisol derived from Quaternary red earth and a sandy Ultisol derived from tertiary red sandstone were used in this study. A pot experiment was conducted with maize growing in the two Ultisols acidified to different pH values. Urea with ¹⁵N abundance of 10.11% was used to investigate the distribution of N fertilizer between soil and plant. Total N content and ¹⁵N abundance in plant and soil samples were determined by elemental analysis-isotope mass spectrometry.

Results and discussion

Critical soil pHs of 4.8 and 5.0 were observed for maize growing in the clay and sandy Ultisols, respectively. Below the critical soil pH, increasing soil pH significantly increased maize height and the yield of maize shoots and roots (both P < 0.05), but changes in soil pH showed no significant effect on maize growth above the critical soil pH in both Ultisols. Maize growing in the sandy Ultisol was more sensitive to changes in soil pH than in the clay Ultisol. Increase in the pH in both Ultisols also increased N accumulation in maize, the N derived from fertilizer in maize, physiological N use efficiency, and N use efficiency (NUE) by maize. Changes in soil pH had a greater effect on these parameters below the critical soil pH, compared to above. The change in soil pH had a greater effect on N accumulation in maize, the N derived from fertilizer in maize, and NUE in the sandy Ultisol than in the clay Ultisol. The NUE increased by 24.4% at pH 6.0, compared with pH 4.0 in the clay Ultisol, while the NUE at pH 5.0 was 4.8 times that at pH 4.0 in the sandy Ultisol. The increase in soil pH increased the ratio of N accumulation in maize/soil residue N and decreased the potential loss of fertilizer N from both Ultisols.

Conclusions

Soil acidification inhibited maize growth, reduced N uptake by maize, and thus, decreased NUE. To maintain soil pH of acidic soils above the critical values for crops is of practical importance for sustainable food production in acidic soils.

     

响应面法优化绿色木霉产孢子固体发酵培养基

为探讨绿色木霉固体发酵产孢子的最佳发酵培养基。运用响应面法设计试验,筛选出主要影响因子碳酸钙、硫酸铵及氯化锰,以孢子量为响应值建立二次回归方程,并借助Design Expert 8.0.6软件进行分析数据并确定优化条件。结果表明,在温度28℃,接种量15%,料水比1:0.70的培养条件下发酵7天,孢子量为8.505×10⁹CFU/g。通过优化,最终确定最佳发酵配方为：麸皮（过40目筛）50%,稻壳粉（过20目筛）30%,玉米粉（过80目筛）20%,硫酸铵1.08%,黄豆饼粉2.00%,玉米浆干粉2.00%,碳酸钙0.55%,氯化锰0.37%,pH 6.0~7.0。该配方所用原料廉价易得,发酵水平高,为大规模工业化生产提供保障。     

羊IL-1Ra基因全长cDNA克隆及生物信息学分析

试验旨在对羊白介素1受体颉颃因子（interleukin-1 receptor antagonist，IL-1Ra）基因进行全长cDNA克隆及生物学分析。根据布鲁氏菌感染羊白细胞层SSH cDNA文库中的IL-1Ra基因序列信息及已知核苷酸序列（GenBank登录号：KC425613.1）设计引物，利用RT-PCR结合RACE方法扩增并克隆IL-1Ra基因，对其进行序列及相关分子特性分析。结果表明，羊IL-1Ra基因全长为1 228 bp，可编码174个氨基酸，含有1个完整的IL-1保守结构域和1个IL-1Ra结构域（PHA02651结构域）。IL-1Ra分子质量为19 765.8 u，等电点（pI）为5.72，其分子式为C₈₈₅H₁₃₈₅N₂₃₅O₂₅₆S₁₁，且含有信号肽。二级结构分析显示，IL-1Ra分子存在较多的β折叠及受体结合位点，与三级结构预测结果一致，且与人/鼠IL-1Ra分子的空间结构相似度极高，达到90%以上。羊IL-1Ra有IL-1特有的三叶草结构，其酶结合结构域位于TYR⁴⁷至GLU⁶⁶之间。将羊IL-1Ra与牛、虎鲸、宽吻海豚、猪、家犬、人、鼠等14个物种进行蛋白质同源性比对，发现在各物种间存在5个高度同源的半胱氨酸位点。系统进化树分析发现，羊IL-1Ra基因全长cDNA所编码的氨基酸序列同山羊、牛单独形成一个分支。本研究结果为今后深入研究IL-1Ra基因的生物学功能奠定了基础。     

草果AtNCED基因克隆、表达和植物表达载体构建

9-顺式环氧类胡萝卜素双加氧酶(9-cis-epoxycarotenoid dioxygenase, NCED)是ABA生物合成的限速酶。诸多研究表明NCED基因表达量的变化与ABA含量显著相关,在种子休眠过程中发挥重要作用。为探究草果AtNCED基因的序列特征和功能,本研究以草果(Amomum tsaoko Crevost et Lemarie)种子转录组为基础,采用RT-PCR技术克隆了一个AtNCED基因。该基因全长2 372 bp,包含一个1 830 bp的开放阅读框(open reading frame, ORF),编码610个氨基酸。生物信息学分析显示,AtNCED基因编码的蛋白在N-末端包含典型的叶绿体转运肽序列,在催化结构域含有4个高度保守的组氨酸残基。AtNCED蛋白与芭蕉科马来西亚蕉同源性较高,与其他单子叶植物处在同一进化分支。实时荧光定量PCR分析结果表明,AtNCED基因表达量随着层积时间的增加逐渐下降,该基因可能正向调控种子休眠,在草果种子休眠诱导与维持中发挥重要作用。进一步地,利用同源重组方法,成功构建了pBI121-AtNCED过表达载体,并转化至农杆菌EHA105。该研究结果为弄清ABA激素信号在种子休眠中的调控作用提供了帮助。