外源5-氨基乙酰丙酸对低温胁迫下茶树叶片光合及生理特性的影响

通过对陕茶1号（耐低温型）和金牡丹（低温敏感型）2个茶树品种在冬季自然低温胁迫下喷施不同浓度（0、10、30、50 mg·L^-1）的外源5-氨基乙酰丙酸（5-aminolevulinic acid,ALA）,探究外源ALA对低温胁迫下茶树叶片光合荧光特性及生理特性的调控作用。结果表明,适宜浓度的外源ALA对低温胁迫下茶树叶片净光合速率、气孔导度、胞间CO₂浓度、蒸腾速率、PSⅡ最大光化学效率及PSⅡ潜在活性具有促进作用,能够提高水浸出物、咖啡碱、游离氨基酸、儿茶素类物质、茶氨酸、可溶性糖、可溶性蛋白等生理活性物质含量的累积;外源ALA能够提高低温胁迫下茶树光合作用的能力并改善茶叶品质,其中50 mg·L^-1的ALA处理能有效提高陕茶1号应对低温胁迫的能力,10 mg·L^-1和30 mg·L^-1的外源ALA对金牡丹应对低温胁迫具有较好的缓解效用。     

响应面法优化香水莲花甾醇的超声提取工艺

为了分析香水莲花的植物甾醇含量并优化其超声提取工艺,采用高效液相色谱（HPLC）法测定香水莲花中菜籽甾醇、菜油甾醇和β-谷甾醇的含量,再以上述3种甾醇提取量为评价指标,对液料比、超声提取温度和超声提取时间进行单因素试验,在此基础上,再采用Box-Behnken响应面法优化香水莲花甾醇提取工艺并进行验证。结果表明,影响香水莲花甾醇提取量的主次因素是液料比>提取时间>提取温度,其最优工艺条件为以95%乙醇为提取溶剂、超声功率100 W、液料比30∶1（mL/g）、超声提取温度60 ℃、超声提取时间30 min;在最优工艺条件下进行验证实验,得出香水莲花中甾醇含量为菜籽甾醇64.61 mg/100 g、菜油甾醇40.77 mg/100 g、β-谷甾醇99.04 mg/100 g,总甾醇204.42 mg/100 g,综合评分与预测值相比差异不显著（P<0.05）。研究结果为香水莲花资源的开发和利用提供参考。     

桔小实蝇在云南蒙自地区的发生规律研究

桔小实蝇寄主范围广、繁殖量大、危害重,是世界性的重要检疫害虫。本文通过调查分析,研究了云南蒙自市的海拔、防治措施、果树种类三因素对桔小实蝇种群动态的影响。结果表明,桔小实蝇在1412 m低海拔地区的种群数量大于1608 m和1700 m高海拔地区的,这说明海拔越高,桔小实蝇种群数量越小,危害也越小。根据在相似海拔地区相同果园桔小实蝇种群数量的比较,发现严格按照本课题组探索的防治措施做好桔小实蝇防治能收到良好的效果。蒙自市的水果成熟期不同,随着各种水果的成熟,桔小实蝇种群动态会出现多个峰值,3~5月桔小实蝇种群高峰期主要出现在枇杷园,7~10月主要出现在石榴园、小红枣园和水蜜桃园。对桔小实蝇在蒙自种群动态的调查,能够为下一步防治措施的制定做好准备。     

野老鹳草Geranium carolinianum种子的萌发条件

为明确野老鹳草Geranium carolinianum种子的休眠特性及萌发适宜的环境条件,采用培养皿法和盆钵法研究野老鹳草种子萌发和出苗适宜的条件,记录种子萌发数,计算种子萌发率、平均萌发时间及萌发指数。结果显示,野老鹳草具有较长的休眠期,新采集的野老鹳草种子在室温干储210 d后可以解除休眠;在10~25℃的恒温条件下种子萌发率均在93.8%以上;有无光照及光周期的长短对种子的萌发无影响;对酸碱度不敏感,在pH 4~10范围内种子萌发率均在91.3%以上;对水势具有一定耐受力,抑制50%种子萌发率所需的水势为-0.42 MPa;盐分对野老鹳草种子萌发具有一定抑制作用,当盐浓度达到160 mmol/L时基本不能萌发;此外,种子在土壤垂直深度5 cm以内时仍然可以出苗,对播种深度适应性较强。表明野老鹳草种子具有较长休眠期,土壤层积可解除种子休眠;而且野老鹳草种子对温度、光照、土壤酸碱度、播种深度适应性较强,水势及盐分对种子萌发有一定影响。     

云南省水稻稻瘟病菌无毒基因AVR-Pia变异及水稻抗性基因Pia的有效性

为进一步了解田间稻瘟病菌Magnaporthe oryzae群体中AVR-Pia基因的分布及变异,利用水稻单基因系IRBLa-C水稻品种对自云南省13个市(州)采集分离得到的471株稻瘟病菌菌株进行抗性基因Pia有效性测定;利用无毒基因AVR-Pia特异性标记对471株稻瘟病菌菌株进行PCR检测和测序,并分析稻瘟病菌群体中无毒基因AVR-Pia的分布及DNA结构变异;利用有效性结果和PCR检测结果对471株菌株进行反应型划分,筛选鉴定菌株;利用鉴定菌株对云南省112份地方稻种进行Pia基因鉴定。结果表明,在471株稻瘟病菌菌株中,对含有Pia基因的水稻单基因系IRBLa-C表现为抗病和感病的菌株数分别为139株和332株,所占比例分别为29.5%和70.5%;在471株稻瘟病菌菌株中,分别有244株和227株菌株含有无毒基因AVR-Pia和不含有无毒基因AVR-Pia,所占比例分别为51.8%和48.2%,无毒基因AVR-Pia主要为完全缺失变异;在471株稻瘟病菌菌株中,A-和V+反应型菌株数分别为56株和161株,共217株,占总菌株数的46.1%,在13个市(州)稻瘟病菌群体中,A-和V+反应型菌株所占比例差异较大,其中在普洱市、红河哈尼族彝族自治州、昭通市、玉溪市4个市(州)的比例较大,分别为77.8%、57.1%、52.1%和50.0%;在112份云南省地方稻种质资源中,有20份地方稻品种含有抗性基因Pia,主要分布在9个市(州)中。表明云南省13个市(州)绝大部分水稻产区水稻Pia基因已丧抗性,含Pia基因的水稻种质在云南省分布较广。     

东北日光温室葡萄园水热通量特征及其对气象因子的响应

运用温室葡萄水热平衡观测资料,分析了东北日光温室葡萄的能量平衡和能量分量日变化、生育期变化以及分配规律,同时也分析了潜热通量（λET）对环境因子的响应。结果表明：水热通量各分量在整个生育期日变化总体上呈现为单峰趋势,净辐射（R_n）的峰值最大为618.75 W·m^-2,λET峰值最大为242.73 W·m^-2,感热通量（H）峰值最大为327.93 W·m^-2;在新梢生长期,白天λET较小,为34.55 W·m^-2,随着生育期推进,λET逐渐增大,在果实着色成熟期达到最大值（78.49 W·m^-2）之后减小;H在各生育期能量中均占了绝大部分;白天潜热通量占净辐射的比例（λET/R_n）在新梢生长期最小,为25.28%,在果实着色成熟期最大,为44.17%;感热通量占净辐射比例（H/R_n）整个生育期几乎都达50%以上,土壤热通量占净辐射比例（G/R_n）相对较小,变化范围为4.46~12.32 W·m^-2;在整个生育期能量比率大小依次为H/R_n>λET/R_n>G/R_n。在不同生育阶段瞬时尺度上,R_n是影响潜热变化最主要的气象因子,R²高达0.88。在日尺度上,各气象因子对潜热通量的影响在逐渐变弱,相对湿度（RH）与λET相关系数仅为0.28。但无论从瞬时尺度还是日尺度,R_n都是影响潜热通量最主要的气象因子。各气象因子对潜热通量的影响大小依次为：R_n>VPD>T_a>RH。     

玉米叶片表皮蜡质相关性状的全基因组关联分析

为发掘玉米叶片表皮蜡质合成与调控相关基因,以218份由温带、亚热带和热带自交系组成的关联群体为材料,在原阳对其3个生物学重复叶片表皮挂水能力进行调查,利用1.25 M覆盖玉米全基因组的SNPs进行Q、K和Q+K这3种模型下的全基因组关联分析。结果表明,Q模型较K模型和Q+K模型能更好地评价叶片表皮的挂水能力。Q模型下,共检测到88个覆盖玉米9条染色体的显著SNPs（P ≤ 2.04E-6）,88个SNPs分布于47个QTLs内,单个QTL可解释13.6%~45.6%的叶片挂水能力表型变异,47个QTL内共有97个候选基因,其中,77个具有功能注释。位于第2染色体上的转录因子NAC77（GRMZM2G018436）和第3条染色体上的亚油酸酯氧合酶（GRMZM2G156861）编码基因,是叶片表皮蜡质性状的重要候选基因。     

Sulodexide protects against hyperglycemia-induced retinal vascular injury via suppressing NOX4/NLRP3 pathway

AIM To investigate the effect of sulodexide (SDX) on high glucose-induced damage in retinal microvascular endothelial cells. METHODS (1) High-fat diet combined with intraperitoneal injection of streptozocin were used to induce type 2 diabetes mellitus (DM) followed by injection of saline or SDX in C57BL/6J male mice. Retinal microvascular leakage and density, and the protein levels of NLRP3 inflammasome-related proteins, zonula occludens-1 (ZO-1) and NADPH oxidase 4 (NOX4) were measured. (2) Human retinal microvascular endothelial cells (HRMECs) were treated with normal glucose or high glucose with or without SDX, and were further transfected with siRNA to knock down NOX4, or infected by adenovirus to over-express NOX4. The protein levels of ZO-1, VE-cadherin (VE-Cad), NOX4 and NLRP3 inflammasome-related proteins as well as the level of reactive oxygen species (ROS) were detected. RESULTS Treatment with SDX increased the protein level of ZO-1, attenuated retinal leakage and NLRP3 inflammasome activation, and enhanced the density of microvasculature and the number of ganglion cells in diabetic retinas. The protein levels of ZO-1 and VE-Cad were decreased, while the levels of NOX4, NLRP3 inflammasome-related proteins and ROS generation were increased in high glucose-treated HRMECs. Silencing of NOX4 inhibited high glucose-induced increases in NLRP3 inflammasome and ROS generation, and decreases in the protein levels of ZO-1 and VE-Cad. Over-expression of NOX4 significantly increased the levels of NLRP3 inflammasome-related proteins and ROS generation in HRMECs, and reduced the protein levels of ZO-1 and VE-Cad. Treatment with SDX partly reversed NOX4 over-expression-induced changes. CONCLUSION SDX alleviates hyperglycemia-induced retinal microvascular endothelial injury via inhibiting NOX4/ROS/NLRP3 pathways.     

Pyroptosis mediates high glucose-induced inflammation and injury in MC3T3-E1 osteoblasts

AIM To investigate whether pyroptosis contributes to the inflammation and injury in mouse embryonic osteoblastic cell line MC3T3-E1 induced by high glucose (HG; 45 mmol/L glucose). METHODS The cell viability was measured by CCK-8 assay. The protein expression levels of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) and caspase-1 (CASP1) were determined by Western blot. The secretion levels of interleukin-18 (IL-18) and IL-1β were measured by ELISA. The intracellular level of reactive oxygen species (ROS) was detected by 2',7'-dichlorodihydrofluorescein diacetate staining followed by photofluorography. Mitochondrial membrane potential (MMP) was examined by rhodamine 123 staining followed by photofluorography. The alkaline phosphatase (ALP) activity was determined using the ALP kit, and the number of mineralized nodules was detected by alizarin red S staining. RESULTS After the MC3T3-E1 osteoblasts were treated with HG for 24 h, the protein expression levels of NLRP3 and CASP1, and the secretion levels of IL-18 and IL-1β were significantly increased. The decrease in cell viability, and the increases in ROS generation and MMP loss were also observed. Moreover, the differentiation and mineralization of MC3T3-E1 osteoblasts were inhibited, evidenced by decreases in both ALP activity and mineralized nodule number. Knockdown of CASP1 by siRNA attenuated the HG-induced osteoblast inflammation and injury mentioned above. CONCLUSION Pyroptosis mediates HG-induced inflammation and injury in MC3T3-E1 osteoblasts.     

Role of PERK pathway in morphine-induced myocardial protection of H9c2 cells

AIM: To explore whether morphine protects oxidative stress-damaged myocardial cells by inhibiting the PERK pathway to reduce endoplasmic reticulum stress and prevent mitochondrial permeability transition pore (mPTP) opening. METHODS: Rat myocardial H9c2 cells were cultured to establish an oxidative stress model, and then randomly divided into control group, H₂O₂ group, H₂O₂+morphine group, H₂O₂+morphine+PERK pathway inhibitor GSK2656157 group, morphine group and GSK2656157 group. Immunohistochemical method was used to detect the effects of morphine on expression of glucose-regulated protein (GRP) 78 and GRP94 induced by oxidative stress. The protein levels of PERK signaling pathway-related molecules were determined by Western blot. Confocal microscopy was used to observe the effects of morphine on mPTP opening and endoplasmic reticulum induced by oxidative stress. Cellular toxicity was detected by lactate dehydrogenase (LDH) kit and cell viability was measured by MTT assay. RESULTS: Compared with control group, GRP78 and GRP94 proteins in H₂O₂ group were strongly expressed, and the brown-yellow particles were significantly increased, but morphine significantly inhibited this process. Compared with control group, the phosphorylation of PERK was significantly reduced with GSK2656157 treatment at different concentrations, among which 2 μmol/L had the most significant effect (P < 0.05). Oxidative stress significantly increased the protein levels of GRP78, GRP94, p-PERK and CHOP, but significantly decreased p-GSK-3β level. These changes were inhibited by morphine, and the effects of morphine were further enhanced by GSK2656157 (P < 0.05). Compared with control group, oxidative stress significantly reduced the fluorescence intensity of mitochondrial TMRE and ER-Tracker Red. Morphine significantly inhibited this effect even when mitochondrial membrane potential was reduced, mPTP was open, and endoplasmic reticulum was damaged, while GSK2656157 further enhanced the effect of morphine (P < 0.05). Compared with control group, H₂O₂ significantly increased cellular toxicity and decreased the cell viability. Morphine inhibited this effect and GSK2656157 significantly enhanced the effect of morphine (P < 0.05). CONCLUSION: Morphine protects cardiac H9c2 cells under oxidative condition by inhibiting endoplasmic reticulum stress through PERK pathway and preventing the mPTP opening via GSK-3β inactivation.