过表达ZmIBH1-1提高玉米苗期抗旱性

【目的】干旱是严重影响玉米生长发育进程的一个重要因素。挖掘玉米抗旱相关基因,通过转基因功能验证和转录组分析,解析关键基因在响应干旱胁迫过程中的分子调控机制,为抗旱分子育种和遗传改良提供理论依据。【方法】以玉米自交系B104（WT）为背景材料,利用农杆菌介导方法构建过表达ZmIBH1-1转基因株系（ZmIBH1-1-OE）;通过对转基因植株进行草铵膦抗性筛选、标记基因和目的基因PCR检测,以及运用实时荧光定量PCR检测目的基因的表达情况,鉴定阳性植株和株系;以WT和ZmIBH1-1-OE转基因株系为材料,通过干旱处理（20% PEG6000）,进行表型鉴定和耐旱生理生化指标测定,验证ZmIBH1-1的抗旱功能;通过对干旱胁迫下玉米4叶期转录组的比较分析,鉴定出差异表达的基因（differentially expressed genes,DEGs）;结合DAP-seq（DNA affinity purification sequencing）分析,初步确定ZmIBH1-1蛋白直接调控与抗旱相关的下游靶基因,利用基因组可视化软件IGV（integrative genomics viewer）分析ZmIBH1-1蛋白结合候选靶基因的位置,然后通过Dual-Luciferase试验验证ZmIBH1-1蛋白与靶基因的调控关系。【结果】通过玉米遗传转化获得12个转化事件;T₃代中,能同时检测到标记基因Bar和目的基因ZmIBH1-1的植株有458个,实时荧光定量PCR检测结果表明,ZmIBH1-1-OE中ZmIBH1-1的表达量显著高于WT,株系3和株系8表达量最高,将其自交获得T₄代转基因株系用于后续试验。在干旱胁迫条件下,ZmIBH1-1-OE株系存活率、叶片相对含水量、叶绿素含量、可溶性蛋白含量及其生理生化指标（超氧化物歧化酶、过氧化物酶、过氧化氢酶活性）均显著高于WT,说明玉米中过量表达ZmIBH1-1赋予玉米更高的耐旱性。转录组分析结果表明,WT与ZmIBH1-1-OE株系在干旱胁迫下有1 214个差异表达基因;Gene Ontology（GO）功能富集分析结果表明,差异表达基因主要涉及生物过程、细胞组分和分子功能,如在生物过程中主要涉及到光合作用、应激响应、脱水响应等;KEGG富集分析表明,差异表达基因主要参与植物激素信号传导、新陈代谢等过程。结合转录组显著差异表达基因和DAP-Seq分析所得到ZmIBH1-1蛋白的靶基因,初步确定ZmIBH1-1蛋白直接调控与抗旱相关的11个候选靶基因,包括2个钙信号相关基因、3个半胱氨酸代谢相关基因、1个bHLH转录因子、1个应激响应蛋白、1个谷胱甘肽转移酶、1个氧化还原过程蛋白和2个乙烯响应因子;基因组可视化结果显示ZmIBH1-1蛋白可以结合靶基因启动子区;随后通过Dual-Luciferase试验进一步表明,ZmIBH1-1蛋白可以直接作用于11个候选靶基因,其中,ZmIBH1-1蛋白可以促进ZmCa-M、ZmSYCO、ZmbHLH54、ZmGlu-r1、ZmCLPB3和ZmP450-99A2的表达,抑制ZmAGD12、ZmCYS、ZmCYSB、ZmERF-107和ZmEIN3的表达。此外,在干旱胁迫下NAC、WRKY、MYB等转录因子在ZmIBH1-1-OE和WT株系中也存在差异表达。【结论】ZmIBH1-1的过表达可以增强玉米苗期的耐旱性;ZmIBH1-1蛋白通过直接调控乙烯信号通路中的ZmERF-107和ZmEIN3的表达提高玉米的耐旱性;ZmIBH1-1蛋白通过直接调控钙信号相关基因ZmCa-M和ZmAGD12增强玉米的耐旱性;ZmIBH1-1蛋白可能通过间接调控NAC、WRKY、MYB等转录因子响应干旱胁迫。     

动物饲料国际贸易、养殖密度与环境氮磷平衡的关系

粮食和饲料的国际贸易促进了许多国家农业生产系统的专业化和集约化。专业化动物养殖越来越多地依赖进口大豆和玉米,虽然提高了动物生产力,但也促使作物和动物生产系统在空间上的分离。本文综述了几十年来全球范围内大豆和玉米的贸易变化,并将其与养殖密度和整个食品系统中营养平衡的变化联系起来。这一变化与不同的营养管理条例的差异和动物饲养密度的空间变化有关。深入解析这些变化有助于理解动物饲料国际贸易、养殖密度与氮磷平衡之间的复杂关系。     

Exogenous application of allelopathic water extracts helps improving tolerance against terminal heat and drought stresses in bread wheat (Triticum aestivum L. Em. Thell.)

The allelopathic water extracts (AWEs) may help improve the tolerance of crop plants against abiotic stresses owing to the presence of the secondary metabolites (i.e., allelochemicals). We conducted four independent experiments to evaluate the influence of exogenous application of AWEs (applied through seed priming or foliage spray) in improving the terminal heat and drought tolerance in bread wheat. In all the experiments, two wheat cultivars, viz. Mairaj‐2008 (drought and heat tolerant) and Faisalabad‐2008 (drought and heat sensitive), were raised in pots. Both wheat cultivars were raised under ambient conditions in the wire house till leaf boot stage (booting) by maintaining the pots at 75% water‐holding capacity (WHC). Then, managed drought and heat stresses were imposed by maintaining the pots at 35% WHC, or shifting the pots inside the glass canopies (at 75% WHC), at booting, anthesis and the grain filling stages. Drought stress reduced the grain yield of wheat by 39%–49%. Foliar application of AWEs improved the grain yield of wheat by 26%–31%, while seed priming with AWEs improved the grain yield by 18%–26%, respectively, than drought stress. Terminal heat stress reduced the grain yield of wheat by 38%. Seed priming with AWEs improved the grain yield by 21%–27%; while foliar application of AWEs improved the grain yield by 25%–29% than the heat stress treatment. In conclusion, the exogenous application of AWEs improved the stay green, accumulation of proline, soluble phenolics and glycine betaine, which helped to stabilize the biological membranes and improved the tolerance against terminal drought and heat stresses.     

Comparing the behavioural thermoregulation response to heat stress by Atlantic salmon parr (Salmo salar) in two rivers

Climate change is expected to increase the frequency and magnitude of extreme thermal events in rivers. The Little Southwest Miramichi River (LSWM) and the Ouelle River (OR) are two Atlantic salmon (Salmo salar) rivers located in eastern Canada, where in recent years, water temperatures have exceeded known thermal limits (~23°C). Once temperature surpasses this threshold, juvenile salmon exploit thermal heterogeneity to behaviourally thermoregulate, forming aggregations in coolwater refuges. This study aimed to determine whether the behavioural thermoregulation response is universal across rivers, arising from common thermal cues. We detailed the temperature and discharge patterns of two geographically distinct rivers from 2010 to 2012 and compared these with aggregation onset temperature. PIT telemetry and snorkelling were used to confirm the presence of aggregations. Mean daily maximum temperature in 2010 was significantly greater in the OR versus the LSWM (p = 0.005), but not in other years (p = 0.090–0.353). Aggregations occurred on 14 and 9 occasions in the OR and LSWM respectively. Temperature at onset of aggregation was significantly greater in the OR (T_onset = 28.3°C) than in the LSWM (T_onset = 27.3°C; p = 0.049). Logistic regression models varied by river and were able to predict the probability of aggregation based on the preceding number of hours >23°C (R² = 0.61 & 0.65; P₅₀ = 27.4°C & 28.9°C; in the OR and LSWM respectively). These results imply the preceding local thermal regime may influence behaviour and indicate a degree of phenotypic plasticity, illustrating a need for localised management strategies.     

矾根主要品种的叶色表型性状分析

通过目测进行分类的方式很难准确界定矾根品种叶色。为建立一套更为科学的基于叶色表型的矾根品种分类体系,笔者利用色差仪对72个矾根品种共计432个样品的叶色进行测定,对获得的L、a、b 3个色度值进行聚类分析,将72个矾根品种分为褐色系、深绿色系、黄色系、灰绿色系、浅绿色系5类色系。对矾根品种各色系的L、a、b值进行相关性分析,发现叶片正面的L值与a值、a值与b值呈负相关,L值与b值呈正相关,但相关性不明显;叶片背面的L值与a值、a值与b值呈负相关,L值与b值呈显著正相关。从多重比较的结果中可以看出,矾根品种各色系之间的L、a、b色度值差异显著,尤其是各色系间叶正面的颜色差异较大,在矾根品种选育工作中,叶片正面的颜色将作为主要参考。     

盐碱交互胁迫对生姜产量和品质的影响及其数学模型的建立

采用二次饱和D- 最优设计，研究营养液pH 与盐交互胁迫对生姜产量和品质的影响，建立了以pH、盐水平为变量因子，生姜产量和品质为目标函数的二元二次数学模型。模型解析表明，pH 与盐水平对生姜的产量和品质存在显著的交互效应，盐水平为产量的主效应因子，pH 为品质的主效应因子。较低水平的pH 变化对生姜产量无显著影响，当pH 升至7.70时，生姜产量随pH 升高呈快速下降趋势；随盐含量的升高，生姜产量持续降低。随pH 的升高，生姜品质呈缓慢降低的趋势；盐含量在低于77.91 mmol · L^-1 时，生姜品质随盐含量的增加而升高，之后则随盐含量的增加而降低。     

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.     

大白菜TPS基因家族鉴定及其在高温胁迫下的表达分析

为明确大白菜TPS(BrTPS)家族成员信息及对高温胁迫信号的响应,本研究利用生物信息学方法,在大白菜基因组数据库中鉴定了BrTPS基因家族成员,并对其理化性质、进化特征、蛋白结构及在高温胁迫下的表达模式进行分析。结果表明,大白菜全基因组含有15个TPS基因家族成员,分布于8条染色体上。除BrTPS14和BrTPS15外,其余成员各含有1个TPS和1个TPP结构域,并且所含motif的排列顺序也完全一致。理化性质分析发现,15个成员的氨基酸长度介于129~1459 aa之间,分子量大小在14.73~165.83 kD之间,大部分BrTPS蛋白为酸性蛋白和亲水蛋白,以无规则卷曲作为二级结构主要构成元件。进化分析表明,大白菜TPS基因家族成员可分为2类,其中ClassⅠ包含5个成员,ClassⅡ包含10个成员。本研究对高温胁迫前后不同组织和持续高温胁迫下叶片中的表达分析,发现大部分的BrTPS基因可对高温胁迫产生响应,但在表达规律上存在差异。这些研究结果为后续研究大白菜TPS基因提供一定参考。     

畜牧养殖污染现状及综合治理对策

畜牧养殖业污染问题是畜牧养殖生产过程中的常见问题,对于养殖业也有一定的影响,直接影响畜牧产品质量及动物疫病防治工作。该文针对畜牧养殖业污染现状进行分析研究,阐述畜牧养殖污染问题现状,并综合新疆地区的畜牧养殖业发展状态,提出畜牧养殖污染问题综合治理的有效对策。