不同水稻种质中渗透胁迫抗性基因DREB2A的遗传多样性分析

本研究旨在分析转录因子DREB2A基因在不同水稻种质中的遗传多样性,以期为水稻耐渗透胁迫遗传改良提供分子工具。利用单倍型分析、系统进化树、遗传距离和密码子偏好性分析,对85份不同类型水稻种质中DREB2A基因的功能性核苷酸序列变异及遗传多样性进行了研究。共鉴定出55个核苷酸变异位点,其中12个位于编码区,43个位于非编码区;鉴定出12个DREB2A等位基因型,其中来自非洲栽培稻(Oryza glaberrima)的3个等位基因型表现大片段变异;根据DREB2A基因的序列变异鉴定出39个单倍型,其中33个为新单倍型;将所有单倍型分为三组（Group I、II和III）,其中非洲栽培稻(Oryza glaberrima)中的10个单倍型单独分为一组(Group III);系统进化树、遗传距离和密码子偏好性分析均表明Group III与其他两组具有较大差异。对85份不同类型水稻种质中DREB2A基因的序列分析表明,非洲栽培稻(Oryza glaberrima)中的DREB2A等位基因在序列变异、系统进化关系和密码子偏好性方面均明显不同于其他种质材料中的等位基因。     

甜樱桃花芽不同发育时期内参基因的筛选与验证

为了筛选甜樱桃花芽不同发育时期均稳定表达的内参基因,以甜樱桃桑提娜和黔樱一号不同发育时期花芽为材料,通过qRT-PCR技术检测28S rRNA、EF1-a1、EF1-a2、UBC、RPL13、18S rRNA、RSP3、CYP40、ACT2和α-TUB3等10个常用看家基因的表达水平,并利用GeNorm、NormFinder和BestKeeper综合评价其表达稳定性。结果表明,EF-1a2和RSP3在所有样品中稳定性最好。分别以EF-1a2、RSP3及EF-1a2+RSP3作为内参基因检测不同发育时期花芽生长素运输载体AUX1基因及生长素响应因子ARF基因的表达模式,该2个基因在不同内参基因标定下表达模式相同。表明EF-1a2、RSP3及EF-1a2+RSP3可作为甜樱桃花芽不同发育时期的内参基因。     

不同介质对真姬菇菌种保藏效果的对比

选择适宜的菌种保藏介质,是有效保障真姬菇菌种品质的重要一环,不仅关系到真姬菇栽培者的直接利益,也关系到种质资源的保存与延续。通过测定菌丝生长速率和菌丝脱氢酶活力,对22种真姬菇菌种保藏介质的保藏效果进行了对比分析。结果表明0.1%的PEG6000溶液对真姬菇菌种保藏6个月和9个月的整体保藏效果最好,其次是0.1%的PEG10000溶液与超纯水,再次是0.8%葡萄糖溶液和矿泉水。0.6%葡萄糖溶液仅适于真姬菇菌种9个月的保藏,纯净水仅适于真姬菇菌种6个月的保藏。其他介质的保藏效果略差。     

中雄4号杨选育研究

从中国林业科学研究院林业研究所引入39个美洲黑杨×甜杨杂交无性系,在黑龙江西部高纬度高寒地区对各无性系苗期生长、越冬及抗病虫性等性状进行多年测试,选择繁殖能力强、苗期长势好、无严重病虫害、抗逆性强、能安全越冬的优良品系进行大量繁殖,建立品系评比林进行复选。经20 a时间,开展苗期品系评比、区域试验与示范,筛选出杨树新品种中雄4号杨(Populus deltoides×P.suaveolens)。该品种为雄株,具有树干通直圆满、速生、材质好、耐寒、抗旱、抗病虫和不飞絮等优良特性,适合作纸浆材等工业用材林、防护林及绿化树种。     

Periplaneta americana peptide decreases apoptosis of pig-ovary granulosa cells induced by H2O2 through FoxO1

Oxidative stress can induce apoptosis of granulosa cells and lead to follicular atresia, thereby reducing the number of pigs giving birth. The aim of this study was to investigate the protective effect of Periplaneta americana peptide (PAP) on the apoptosis of the granulosa cells of pig ovaries (PGCs) induced by hydrogen peroxide (H₂O₂) via FoxO1. PGCs were treated with H₂O₂ to establish a cell apoptosis model. Cell viability was measured using the cell counting kit-8 (CCK-8) assay, and cell apoptosis was detected using flow cytometry. The malondialdehyde (MDA) level and nitric oxide (NO) content were detected to reflect the oxidative stress. Western blotting, qRT-PCR and overexpression were undertaken to determine the expression of FoxO1 and caspase-3, and immunofluorescence was used to detect FoxO1 in the nucleus and cytoplasm. PGCs were treated with 100 μM H₂O₂ for 6 hr, which resulted in oxidative damage and apoptosis and an apoptosis rate for PGCs of 32.95%. Next, PGCs were treated with 400 μg/ml PAP for 24 hr to repair the apoptosis induced by H₂O₂. PAP improved cell viability in H₂O₂-stimulated PGCs, the increased MDA level and NO content caused by H₂O₂ stimulation were reversed and the apoptotic rate of PGCs was reduced. The qRT-PCR and Western blotting results indicated that PAP decreased the H₂O₂-induced apoptosis and the expression of FoxO1 and caspase-3 in PGCs. The effect of PAP was the same following FoxO1 overexpression. FoxO1 was expressed in the nucleus when stimulated by H₂O₂ or overexpression; however, it migrated to the cytoplasm following PAP treatment. PAP decreased the apoptosis of PGCs induced by H₂O₂ by regulating FoxO1 expression and nuclear translocation.     

MCP-1/CCR2 signaling pathway mediates alcohol-induced angiogenesis in breast cancer

AIM To investigate the role of monocyte chemoattractant protein-1 (MCP-1) and its receptor CC chemokine receptor 2 (CCR2) in ethanol-promoted breast cancer angiogenesis and the underlying mechanism. METH?ODS: A mouse model of transplanted breast tumor with moderate alcohol consumption was established. The correlations between the expression of MCP-1/CCR2 and the expression of angiogenesis markers [platelet endothelial cell adhesion molecule-1 (PECAM-1) and vascular endothelial growth factor (VEGF)] in tumor tissues were examined by immunohistochemistry. In vitro, a 3D tumor-endothelial co-culture system was established to observe tumor angiogenesis and the role of MCP-1/CCR2 signaling pathway in alcohol-mediated angiogenesis. The cell migration ability was detected to clarify whether MCP-1/CCR2 enhanced cell mobility to form new vessels. RESULTS MCP-1 and CCR2 were both highly expressed in the breast tumor tissues of tumor-bearing mice consuming alcohol, and their expression levels were consistent with the angiogenic markers PECAM-1 and VEGF (P<0.05). The interaction between mouse breast cancer E0771 cells and endothelial cells was observed to promote angiogenesis in the 3D tumor-endothelial co-culture system with or without alcohol stimulation. MCP-1 promoted this kind of tumor angiogenesis, while CCR2 antagonist effectively inhibited the tumor angiogenesis and especially blocked alcohol-induced angiogenesis. Activation of MCP-1/CCR2 signaling pathway enhanced the migration ability of endothelial cells. CONCLUSION The MCP-1/CCR2 signaling pathway plays an important role in promoting the angiogenesis of breast cancer stimulated by alcohol. The mechanism might be that MCP-1 improves the migration of endothelial cells and then promotes angiogenesis.     

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.     

不同灌水下限设施番茄土壤CO2排放特征及其影响因素研究

【目的】探讨不同灌水下限设施土壤CO2排放特征及其影响因素,为调控设施土壤水分和碳排放提供理论依据。【方法】在番茄生育期内采用LI-8100A土壤碳通量自动测定仪观测不同灌水下限[20 kPa(D20)、30 kPa(D30)、40 kPa(D40)]下的土壤CO2排放速率,并分析其影响因素。【结果】在番茄生育期内,不同灌水下限设施土壤CO2排放速率变化趋势基本一致,D20处理最高,平均速率为2.759μmol/(m2·s),其次是D30处理,为2.601μmol/(m2·s),D40处理最低,为2.559μmol/(m2·s)。在土壤CO2累积排放量方面,D20处理显著高于其他2个处理,而D30和D40处理之间无显著差异。就单因素模型而言,不同灌水下限处理的土壤CO2排放速率与15 cm土壤温度呈指数回归关系,且均达显著水平(P<0.05);不同灌水下限处理的土壤CO2排放速率与15 cm土壤含水率均呈显著二次回归关系(P<0.05);与单因素模型相比,土壤温度和土壤含水率的双因素复合模型(68.5%~83.8%)可以更好地解释土壤CO2排放的变化。土壤温度敏感系数Q10值在1.442~1.498之间,其中D20处理最敏感,D40处理最不敏感。相关分析结果表明,土壤CO2累积排放量与0~20 cm土层土壤有机质量、pH值、全氮量、速效磷量、速效钾量、碱解氮量和微生物量碳呈显著相关关系。采用PCA分析提取出的2个主成分累积贡献率为85.79%。【结论】灌水下限影响设施土壤CO2的排放,其中D20处理促进了设施土壤CO2的排放。     

SiO2纳米颗粒对杉木幼苗生长发育的影响

【目的】近年来,硅对于的植物影响受到人们广泛关注,人们试图将硅确定为植物生长发育的的“必要元素”。因此如何提高植物对硅的吸收利用来增加植物的农艺性状和产量,是当前农业和林业研究的一个重要课题。【方法】以黄枝杉Cunninghamia lanceolata和罗田垂枝杉C.lanceolata var.luotian为材料,采用盆栽的方式,研究施加不同浓度的纳米SiO2颗粒(0、1、2、4 g/盆)对杉木幼苗光合参数、叶绿素荧光、叶片厚度、氮磷钾含量及其分配、生物量累积的影响。【结果】本研究结果显示,SiO2纳米颗粒对杉木根系的生长发育有很大的促进作用,使之能更好的吸收养分,加强了氮磷钾含量的累积。且叶片的厚度也有所增加,在较低的浓度下,气孔导度、光合作用机制、以及光合作用效率增加;地上的生物量也随着施入SiO2纳米颗粒浓度的增加而增加。【结论】我们的实验表明,SiO2纳米颗粒通过促进根系发育加强对营养的吸收和提升叶片厚度,以及提高净光合速率和气孔导度来提高植物的生长机理,进而增加植物的生物量来提升产量。鉴于其对植物生长发育及产量的多种积极影响,硅纳米材料将会为农林植物增产有重要的影响和广阔的前景。     

3种介导猪NR2F2基因转染PK15细胞方法的比较研究

试验旨在研究3种转染方法对猪肾上皮细胞（PK15）的转染效率，为以PK15细胞为模型研究外源基因的功能提供参考。本研究以PK15细胞为研究对象，用脂质体、电穿孔、慢病毒3种转染方法转染后，在荧光显微镜下观察绿色荧光蛋白的表达，采用实时荧光定量PCR检测EGFP和NR2F2基因的表达情况，采用CCK-8检测细胞存活率，进而比较3种方法的转染效果。结果显示，转染PK15细胞后，电穿孔和慢病毒方法转染效率极显著高于脂质体（P<0.01），但电穿孔方法和慢病毒方法之间差异不显著（P>0.05）；EGFP和NR2F2基因的实时荧光定量PCR结果显示慢病毒方法效果最好，脂质体方法较差，与细胞转染效率基本一致；CCK-8结果显示，电穿孔转染后细胞存活率最低，极显著低于对照组（P<0.01），脂质体方法显著低于对照组（P<0.05），慢病毒方法与对照组间差异不显著（P>0.05）。综合考虑转染效率及细胞活性，本研究认为慢病毒转染方法最适合转染PK15细胞，为今后高效转染PK15细胞提供了理想的方法。