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.     

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.     

小麦雪霉叶枯病产量损失的初步研究

人工接种试验结果表明,雪腐格氏霉(Gerlachia nivalis)引起的小麦成株期叶枯、鞘枯和穗腐,鞘枯使小麦穗粒重减少2.1％～27.1％,千粒重降低9.5％～16.9％;穗腐使小麦穗粒重减少9.6～33.9％,千粒重降低7.8％～17.9％;鞘枯—穗腐复合症状使穗粒重减少10.2％～55.2％,千粒重下降9.5％～14.8％;叶斑—鞘枯—穗腐使穗粒重减少11.7％～47.1％,千粒重下降12.1％～18.3％。     

杉木林地套种绿肥筛选初报

收集了三十种绿肥,在杉木当年造林地上进行套种试验,根据其生长状况、生物量、覆盖度及对杉木生长的影响,把供试绿肥分成三类,初步筛选出羽扁豆、决明豆、日木草为适宜杉木林地种植的林用绿肥。同时对今后林用绿肥的选择提出了有益的建议。     

Stoffbildung, CO2-Gaswechsel, Kohlenhydrat- und Stickstoffgehalt von Winterweizen bei erhöhter CO2-Konzentration und Trocken-streß

Dry Matter Production, CO₂ Exchange, Carbohydrate and Nitrogen Content of Winter Wheat at Elevated CO₂ Concentration and Drought Stress
Methods of mathematical modelling and simulation are being used to an increasing degree in estimating the effects of rising atmospheric CO₂ concentration and changing climatic conditions on agricultural ecosystems. In this context, detailed knowledge is required about the possible effects on crop growth and physiological processes. To this aim, the influence of an elevated CO₂ concentration and of drought stress on dry matter production, CO₂ exchange, and on carbohydrate and nitrogen content was studied in two winter wheat varieties from shooting to milk ripeness. Elevated CO₂ concentration leads to a compensation of drought stress and at optimal water supply to an increase of vegetative dry matter and of yield to the fourfold value. This effects were caused by enhanced growth of secondary tillers which were reduced in plants cultivated at atmospheric CO₂ concentration. Analogous effects in the development of ear organs were influenced additionally by competitive interactions between the developing organs. The content and the mass of ethanol soluble carbohydrates in leaves and stems were increased after the CO₂ treatment and exhausted more completely during the grain filling period after drought stress. Plants cultivated from shooting to milk ripeness at elevated CO₂ concentration showed a reduced response of net photosynthesis rate to increasing CO₂ concentration by comparison with untreated plants. The rate of dark respiration was increased in this plants.     

Evaluation of Wheat (Triticum aestivum) and Legume Intercropping Under 1:1 and 2:1 Row-replacement Series System

A field experiment was conducted during the winter season of 1992–93 and 1993–94 at Agricultural Experimental Farm, Giridih, Bihar to evaluate the intercropping systems of legumes, gram ( Cicer arietinum L.), pea ( Pisum sativum ) and lentil ( Lens culinaris ) with wheat ( Triticum aestivum ) in 1:1 and 2:1 'row replacement series'. Intercropping systems were assessed on the basis of new indices termed as actual yield loss (AYL) and intercropping advantage (IA). This paper highlights the comparative effectiveness of evaluating the intercropping systems through the existing intercropping indices like LER, RCC, aggressivity. The indices AYL and IA seem to be more appropriate particularly when per plant yield is considered. Intercropping reduced the yield of component crops compared with respective pure stands. Wheat + pea in 1:1 row replacement series gave the highest wheat yield equivalent value (3.02 t ha⁻¹) followed by wheat + lentil (2.91 tha⁻¹). When the actual sown proportion was considered wheat + lentil (1:1) resulted in maximum AYL (+0.610) and IA (+0.279) values. This treatment also gave the maximum monetary advantage (Rs 5985.45 ha⁻¹).     

Long-term application of animal slurries to grassland alters soil cation balance

Abstract. Soil samples from a 32-year grassland field experiment were taken from 0–5, 5–10, and 10–15 cm soil depths in February 2002. Plots received annual treatments of unamended control, mineral fertilizer, three rates of pig slurry and three rates of cow slurry, each with six replicates. Samples were analysed for cation exchange capacity (CEC), exchangeable cations (Na⁺, K⁺, Ca²⁺, Mg²⁺), pH and Olsen P. Exchangeable sodium percentage (ESP) was calculated as a sodicity indicator. Mean ESP was generally greater for slurry treatments than the control, with a trend of increasing ESP with application rate. This was particularly marked for cow slurry. At 0–5 cm depth ESP increased from 1.18 in the control to 1.75 at the highest rate of pig slurry and 5.60 at the highest rate of cow slurry. Similar trends were shown for CEC, exchangeable Na⁺, K⁺ and Mg²⁺, Ca²⁺ and Olsen P. The build-up of soil P due to slurry applications, together with this combination of physical and chemical factors, may increase the risk of P loss to surface waters, particularly from soils receiving high rates of cow slurry.     

微波电路电测及其计算方法

从实际工程的角度出发，对电路电测方法进行了论述。在微通信工程中，需要获得障碍物的位置和高度，最合理的天线高度组合和障碍物绕射损耗等参数，为此论述了微波电路电测方法及技术要求。在大庆－秦皇岛输油管道通信工程中采用了Ａ，Ｂ测试法进行测试，得到高度－损耗曲线。这种曲线给出相对自由空间接收电平与收，发天线高度之间的关系。在测试所得的高度－损耗曲线与断面图相结合的基础上，由绕射损耗计算公式推出了几种常见障碍     

湖北省分地区水灾损失等级划分初探

建立了湖北省分地区水灾损失灾度等级的划分标准。基于模糊模式识别理论，按最大隶属原则和择近原则对灾度等级进行了模糊综合评判。