白屈菜CmFAD2基因的克隆及植物转化载体构建

通过对白屈菜低温应答过程的转录组分析发现膜脂不饱和化相关基因的表达在一定过程中发生变化,脂肪酸去饱和酶基因FAD2在随温度的变化趋势为正"V"型,且表达量变化显著。利用NCBI等在线软件对序列进行相关生物学信息分析,并对白屈菜FAD家族成员FAD2基因的完整开放阅读框(ORF)进行克隆,并命名为CmFAD2。选用克隆载体pMD-19-T,转化大肠杆菌DH5α,测序验证序列正确性及完整性。将目的基因与植物表达载体pRI-201-AN连接构建重组DNA pRI-201-AN-Cm FAD2,电击法转化农杆菌LBA4404,利用菌液PCR法验证成功。该基因可作为药用植物抗寒品种创制的候选基因。     

Zerumbone attenuates MPP+-induced cytotoxicity in human neuroblastoma SH-SY5Y cells by inhibition of oxidative stress

AIM: To investigate the role of zerumbone (ZER) in 1-methyl-4-phenylpyridinium (MPP⁺)-induced cytotoxicity of human neuroblastoma SH-SY5Y cells. METHODS: Human neuroblastoma SH-SY5Y cells were cultured in vitro and the protective effect of ZER against MPP⁺-induced cytotoxicity was measured by CCK-8 assay. Flow cytometry was used to determine the apoptosis and reactive oxygen species (ROS). The expression of Parkinson disease protein 7 (PARK7) was knocked-down by using PARK7-specific short hairpin RNA (shRNA). The protein levels of PARK7, nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were determined by Western blot. RESULTS: MMP⁺ remarkably reduced the cell viability in a dose-dependent and time-dependent manner. The SH-SY5Y cell injury model was established by treatment with MPP⁺ at 600 μmol/L for 24 h. ZER up-regulated the protein levels of PARK7 and Nrf2 (P<0.05), alleviated apoptosis (P<0.05), and reduced ROS production (P<0.05) in the SH-SY5Y cell injury model. Meanwhile, N-acetyl-L-cysteine (NAC) had the similar functions. Moreover, significant reductions in the protein levels of Nrf2 and HO-1 (P<0.05), and obvious increases in apoptosis (P<0.05) and ROS level (P<0.05) were demonstrated in PARK7-knockdown cells. CONCLUSION: ZER protects SH-SY5Y cells against MPP⁺-induced cytotoxi-city, which may be related to activation of PARK7/Nrf2/HO-1 pathway, and subsequent attenuation of oxidative stress and apoptosis.     

贮运条件对草莓主要色素成分稳定性的影响

草莓在贮藏和运输期间会产生褪色现象,花青素含量逐渐减少。为研究光照时间、温度、pH、金属离子对草莓花青素的影响,探索合适的加工与贮运条件,在花青素含量保持一致的情况下设置4种反应条件,探讨光照时间、温度、pH、金属离子对草莓花青素的影响。草莓花青素在自然光下照射8 h,吸光度值下降25%,温度高于60℃后,吸光度值下降67%。当pH>7时,吸光度值下降85%,添加Na⁺、Mg²⁺后的色素提取液反应2 h后所测吸光度值分别保持在0.070和0.050左右,而添加Fe³⁺后测得的吸光度值下降87%。实验结果表明,草莓在贮运过程中应避光、低温、保持酸性条件,且避免与铁制品接触。     

正交设计优化青蒿SRAP-PCR反应体系及引物筛选

为了建立青蒿的SRAP最佳扩增体系,并筛选出SRAP多态性引物,本研究以青蒿叶片DNA为模板,采用正交试验设计,以Mg^2+、dNTP Mix、Taq DNA聚合酶、引物和DNA模板5种因素5个水平,对青蒿SRAP反应体系进行研究。结果表明,青蒿SRAP-PCR最佳反应体系为:引物0.6μmol/L、Mg^2+2.0 mmol/L、模板DNA 5.1 ng、Taq DNA聚合酶2.0 U、dNTPs 0.25 mmol/L,总体积为25μL。各因素对扩增反应均有不同影响,其中引物浓度的影响最大,dNTPs的影响最小。运用该体系对不同种质资源的青蒿进行验证,证明该体系稳定可靠,并在30个引物组合中筛选出了25对扩增条带清晰,多态性丰富的引物组合。这一结论为今后利用SRAP标记技术进行青蒿分子遗传学研究提供了科学依据。     

橡胶园酸性、中性土壤交换性钙、镁测定方法研究

为了获得简单、易操作、准确度高的橡胶园酸性、中性土壤交换性钙、镁测定方法,本试验通过响应面优化试验,建立称样量、提取液体积、提取方式与交换性钙测定结果之间的数学模型。结果表明：在25℃条件下,以1 mol/L的乙酸铵为土壤浸提液,称样量为2 g、提取液体积175 mL、振荡36 min,为橡胶园酸性、中性土壤的最佳提取条件。将上述条件下得到的结果与标准方法条件下得到的结果相对比,两结果的相对误差在1.25~3.04之间,均属合理范围。利用标准样品对优化方法进行试验验证,结果与推荐值相符。改进后的方法分析成本低、操作简便、测定结果准确、稳定性好,可用于橡胶园酸性、中性土壤交换性钙、镁的测定。     

Spatio-temporal pattern evolution and spatial spillover effect of green development efficiency: Evidence from Shandong Province,China

Improving green development efficiency is urgently required, yet challenging, since it comprehensively reflects the bidirectional evolutionary relationship between regional development and resource and environmental consumption. The issue of regional green development efficiency has become a key topic; however, where the increase in efficiency originates and its spatial spillover effect remains unclear to date. Therefore, the spatial spillover effect of the green development efficiency of Shandong Province was quantified through the slack variable (SBM)-Undesirable and Spatial Durbin model. The green development efficiency showed clear spatial differentiation in Shandong Province. This was more obvious than the polarized development trend of the provincial capital circle that has Jinan at its core and the eastern coastal region with Qingdao at its core. Green development efficiency has a significant spatial spillover effect. The regression coefficient of the direct effect of the economic development level is positive. The regression coefficient of the indirect effect of the marketization level and government financial support level is negative. Science and technology played a positive role in the promotion of the green development efficiency of local and neighboring cities. This study contributes empirical evidence to the green development efficiency associated with regional development and resource and environment consumption of Shandong Province.     

豚草粗提物对杨树水泡型溃疡病的抑菌作用研究

为解决豚草提取物在林业病虫害防治中的推广与应用问题,以豚草叶片为试验材料,采用温浸-超声波联合提取法提取抑菌活性物质,并开展其对杨树水泡型溃疡病病原菌的室内外抑菌试验。室内抑菌试验表明,粗提物浓度为16 mg/mL时,抑菌率达到73.62%;林间防治试验表明,最小抑菌浓度为 40 mg/mL,抑菌率达到95.14%。通过以上抑菌试验确定豚草粗提物对杨树水泡型溃疡病有较好的抑制效果。     

直播稻田无人机全程防治病虫草害初探

针对不同设备设计不同喷液量,飞行参数验证防治效果,从而验证无人机在稻田全程应用的可能性,旨在探索2种小型多旋翼植保无人机低空喷洒在整个水稻生长时期病虫草害防治过程中的防效。结果表明:当喷洒水田封闭式除草剂的喷洒量为10650、15000 mL/hm²,其对杂草的株防效与鲜重防效与人工电动喷雾相近。在病虫害的防治试验中,使用推荐用量且喷洒量为10650、15000 mL/hm²时,以及使用70%推荐用量（减量30%）且喷洒量为15000 mL/hm²时,防治效果均与人工电动喷雾器无显著差异。通过验证,选择合适的药剂,配合科学的喷洒参数,可在保证药效的前提下减少农药使用量。多旋翼植保无人机在水稻田有进行大面积全程飞防植保应用有广阔的前景。     

Effect of chlorogenic acid on intestinal inflammation,antioxidant status,and microbial community of young hens challenged with acute heat stress

Heat stress in poultry is deleterious to productive performance. Chlorogenic acid (CGA) exerts antibacterial, anti-inflammatory, and antioxidant properties. This study was conducted to evaluate the effects of dietary supplemental CGA on the intestinal health and cecal microbiota composition of young hens challenged with acute heat stress. 100-day-old Hy-line brown pullets were randomly divided into four groups. The control group (C) and heat stress group (HS) received a basal diet. HS + CGA₃₀₀ group and HS + CGA₆₀₀ group received a basal diet supplemented with 300- and 600-mg/kg CGA, respectively, for 2 weeks before heat stress exposure. Pullets of HS, HS + CGA₃₀₀, and HS + CGA₆₀₀ group were exposed to 38°C for 4 h while the control group was maintained at 25°C. In this study, dietary CGA supplementation had effect on mitigate the decreased T-AOC and T-SOD activities and the increasing of IL-1β and TNFα induced by acute heat stress. Dietary supplementation with 600 mg/kg CGA had better effect on increasing the relative abundance of beneficial bacterial genera, such as Rikenellaceae RC9_gut_group, Ruminococcaceae UCG-005, and Christensenellaceae R-7_group, and deceasing bacteria genera involved in inflammation, such as Sutterella species. Therefore, CGA can ameliorate acute heat stress damage through suppressing inflammation and improved antioxidant capacity and cecal microbiota composition.     

昆明鼠卵母细胞的孤雌发育及对其干扰核移植试验的预防措施

本文着重研究了在昆明鼠细胞核移植过程中，卵母细胞孤雌发育的激活原因、形成过程，并提出了一些防止其干扰细胞核移植的措施。