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。     

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.     

Determination of critical nitrogen dilution curve based on leaf area index for winter wheat in the Guanzhong Plain,Northwest China

Excessive use of nitrogen(N) fertilizers in agricultural systems increases the cost of production and risk of environmental pollution. Therefore, determination of optimum N requirements for plant growth is necessary. Previous studies mostly established critical N dilution curves based on aboveground dry matter(DM) or leaf dry matter(LDM) and stem dry matter(SDM), to diagnose the N nutrition status of the whole plant. As these methods are time consuming, we investigated the more rapidly determined leaf area index(LAI) method to establish the critical nitrogen(N_c) dilution curve, and the curve was used to diagnose plant N status for winter wheat in Guanzhong Plain in Northwest China. Field experiments were conducted using four N fertilization levels(0, 105, 210 and 315 kg ha-1) applied to six wheat cultivars in the 2013–2014 and 2014–2015 growing seasons. LAI, DM, plant N concentration(PNC) and grain yield were determined. Data points from four cultivars were used for establishing the N_c curve and data points from the remaining two cultivars were used for validating the curve. The N_c dilution curve was validated for N-limiting and non-N-limiting growth conditions and there was good agreement between estimated and observed values. The N nutrition index(NNI) ranged from 0.41 to 1.25 and the accumulated plant N deficit(N_(and)) ranged from 60.38 to –17.92 kg ha~(-1) during the growing season. The relative grain yield was significantly affected by NNI and was adequately described with a parabolic function. The N_c curve based on LAI can be adopted as an alternative and more rapid approach to diagnose plant N status to support N fertilization decisions during the vegetative growth of winter wheat in Guanzhong Plain in Northwest China.     

Impact of soil water content on maize responses to the plant growth-promoting rhizobacterium Azospirillum lipoferum CRT1

Members of the bacterial genus Azospirillum are root-associated bacteria that increase yield in cereals by promoting growth and alleviating drought stress. How plants integrate the many bacterium-derived growth-promoting stimuli with other environmental factors to generate a coordinated response remains unresolved. Using a commercial Azospirillum strain, A. lipoferum CRT1 and two host maize cultivars, it was observed that bacterization reduced the drought-induced increase in lateral root growth and enhanced the flood-induced increase in lateral root growth in the more drought- and flood-sensitive cultivar. In the other one, A. lipoferum CRT1 only elicited a moderate root growth response under low soil water potential. The photosynthetic potential and activity were increased in the earlier cultivar and decreased in the later one, irrespective of the soil water content. No impact of the bacterium was seen on the growth of the leaves of both cultivars under both stresses until the third leaf stage, therefore suggesting that it is a consequence of multiple primary adaptations to biotic and abiotic stresses. It is suggested that host–bacteria recognition leads to a stress-specific modulation of the root response and a differential stress-independent effect on photosynthesis. This is the first report of the impact of Azospirillum under flood conditions.     

蔬菜幼苗叶面积快速测定方法筛选与优化

为得到一种快速且准确测定蔬菜幼苗叶面积的方法,以番茄幼苗为试材,以方格法为对照,研究扫描分析法、叶面积仪法、打孔称重法测定叶面积的准确性、精确性与测定速度,然后采用黄瓜、辣椒叶片及纸片的面积对测定结果进行验证,最后基于不同扫描分辨率对筛选出的方法进行优化。结果表明,相比叶面积仪法和打孔称重法,扫描分析法测定结果更接近真实值,且精度更高,用时更短。扫描分析法不同分辨率下测定结果无显著差异,且扫描分辨率越低所需测定时间越短。扫描分辨率50 dpi下的扫描分析法可作为一种快速准确测定蔬菜幼苗叶面积的方法。     

黑龙江省国有林区脆弱异质性家庭对停伐政策的态度及影响因素研究

基于2016年黑龙江省重点国有林区348户家庭的微观调查数据,实证分析黑龙江省国有林区脆弱异质性家庭对停伐政策的态度及影响因素。采用因子分析与综合评价法将348户职工家庭分为脆弱性家庭和非脆弱性家庭,描述性分析2组样本家庭对停伐政策的态度及认知情况,利用多元有序logit模型分析脆弱性与非脆弱性家庭对停伐政策态度的影响因素。结果表明:非脆弱性家庭对停伐政策的了解程度及执行情况的评价均高于脆弱性家庭,脆弱性家庭相对非脆弱性家庭对停伐政策的认可度较高,但是应对风险冲击的能力相对较低。脆弱性家庭和非脆弱性家庭对停伐政策的态度受户主特征、家庭特征以及政策认知特征变量的显著影响,但脆弱性家庭变量参数估计绝对值相对更大,因而停伐政策对脆弱性家庭有着更为敏感和强烈的冲击。     

NO对大鼠中脑腹侧被盖区DA神经元的调节

实验观察了微量注射 NOS抑制剂 L -NAME及微量联合注射 NO的前体 L-精氨酸( L-Arg) L-NAME于大鼠中脑腹侧被盖区( VTA)对该部位多巴胺神经元的调节。发现VTA注射 L -NAME( 1 mg/ 5μL )后 ,伏隔核( Acb)多巴胺 ( DA )代谢产物—双羟苯乙酸( DOPAC)水平升高到注射前的 1 2 2 .5% ( P <0 .0 0 1 ) ,小剂量注射 L-NAME( 0 .2 mg/ 5μL)对伏隔核 DOPAC水平无明显影响 ;同样方法联合注射 L-Arg( 3 0 0 μg/ 5μL) L-NAME( 1 mg/5μL)后 ,伏隔核 DOPAC水平无明显变化。结论 :VTA微量注射 L-NAME兴奋了该部位的DA神经元 ,而 L -Arg L -NAME联合注射 ,却不能影响 DA神经元的活动 ,说明 NO可以通过L-Arg-NOS-NO途径参与 VTA多巴胺神经元的调节     

我国农业区域开发简史及历史经验

本文讨论了中国农区开发之始、几个主要农区的深开发、黄淮江湖的连片开发、海河流域的开发、其它农区的开发和农区开发的历史经验与教训。完好的灌溉系统、导致深开发的人工运河和农业技术是开发成功的要素。     

氮吸附法分析生物质焦孔隙结构

采用氮吸附法对4种生物质焦(稻壳、树叶、玉米秆、棉花秆)的孔隙结构进行测量,结果表明,不同种类焦样的比表面积和孔径分布有明显差别,树叶的比表面积最大,为242.21 m2·g-1,玉米秆的比表面积最小,为0.81 m2·g-1.850℃时,稻壳、树叶、玉米秆焦样的孔径分布曲线在微孔和中孔范围各有一个分布峰,而棉花秆焦样的孔径分布曲线只在中孔范围内出现一个分布峰.热解温度是影响孔隙结构的一个重要因素,在高温条件下,同步热解得到的焦样的比表面积较大,微孔较多.在本研究中,600℃、850℃的稻壳焦样和850℃的树叶焦样具有较大的比表面积,比较适合做吸附剂.     

Acclimation abilities of three tropical rainforest seedlings to an increase in light intensity

Current practices in many nurseries involve the germination of tropical rainforest seedlings in shaded conditions and transferral, at a later date, to environments with greater light intensities. Determination of the ability of these seedlings to acclimate to increased light intensities will allow seedling stock to be processed with maximum efficiency within the nursery. The acclimation abilities of three species, Agyrodendron actinophyllum, Cardwellia sublimis, and Flindersia brayleyana, commonly found in north Queensland's rainforests were investigated in this study. These particular species are highly valued for their cabinet timber qualities and are being reared in nurseries for use in reforestation trials and programs in north Queensland. Seedlings were initially raised in greenhouse conditions under two layers of shade cloth (16% of full sunlight) and then transferred into full sunlight at three different ages (3 weeks, 10 weeks, 14 weeks). Upon transfer, organ ratios and the direction of dry matter distribution was determined for each species and age group. Approximately 3 months after the seedlings were transferred, the acclimation ability of each species and age group was then determined. Dry matter distribution was found to change with age, irrespective of light environment. Individuals within a species with larger root systems and thicker or more dense leaves had a greater acclimation ability than those with smaller root systems and thinner or less dense leaves. Furthermore, individuals within a species whose dry matter distribution upon transfer was directed towards developing a large root system, and a small photosynthetic area and mass, had a greater acclimation ability than those whose dry matter distribution was directed away from such morphologies. Awareness of these relationships allows a better understanding of seedling response to gap formation in natural forests, and also allows plant nursery operators to make a more informed decision about when to move seedlings to environments with a higher light intensity.