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1.
【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。 相似文献
2.
Exogenous application of allelopathic water extracts helps improving tolerance against terminal heat and drought stresses in bread wheat (Triticum aestivum L. Em. Thell.) 下载免费PDF全文
M. Farooq F. Nadeem M. Y. Arfat M. Nabeel S. Musadaq S. A. Cheema A. Nawaz 《Journal of Agronomy and Crop Science》2018,204(3):298-312
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. 相似文献
3.
Sonia Czarnes Pierre-Edouard Mercier Damien G. Lemoine Jihane Hamzaoui Laurent Legendre 《Journal of Agronomy and Crop Science》2020,206(5):505-516
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. 相似文献
4.
浙江省油菜花期降水量风险评估 总被引:1,自引:0,他引:1
对浙江省66个县市(区)1971—2014年油菜花期降水量及油菜产量资料进行统计分析,并基于信息扩散的风险评估模型,对浙江省油菜花期降水量在各级降水量下的超越概率进行计算分析,评估各地油菜花期面临的旱涝风险;同时分析了浙江省油菜花期各级降水量风险值与油菜产量变化之间的关系。结果表明:油菜花期降水量严重过剩的风险高值区主要位于浙西南地区,降水量短缺风险高值区主要位于浙江省沿海岛屿及宁波的部分县市;浙江省油菜花期降水量短缺基本可以通过灌溉得到改善,但油菜减产与花期降水量过多显著相关,油菜花期降水量300 mm是油菜是否减产的阈值。该结果可为油菜种植合理布局及农业保险政策提供科学依据。 相似文献
5.
本文对光箨篌竹笋期的出笋规律、成竹规律以及竹笋的生长发育规律进行了研究。初步认为,光箨篌竹林分的母竹立竹度一般应保持在2500株/亩左右、Ⅰ~Ⅱ级母竹株数保持在40%以上时,才能保证林分的优质高产与稳产;抽枝成竹历期长短受竹笋笋级大小、出土早晚或立地环境条件的影响可相应延长或推迟2~4d左右,一般历时25d,竹笋开始抽枝成竹时的高度一般为5m左右;竹笋在生长发育高峰期明显受到气候因子的影响,尤其是对气温更为敏感。 相似文献
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7.
J. Müller 《Journal of Agronomy and Crop Science》1993,171(4):217-235
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. 相似文献
Methods of mathematical modelling and simulation are being used to an increasing degree in estimating the effects of rising atmospheric CO
8.
奎屯地区农七师位于天山北麓,准噶尔盆地的西南缘,总面积5986km2,其中作物种植面积120万亩,随着农七师产业结构的调整,2003年实施了“万头澳牛引进工程”,养牛业的发展使农七师畜牧业对玉米的需求量增加,尤其是澳牛养殖业对青贮玉米需求量更大,因此,研究分析气象要素对玉米播种及苗期的影响是重要工作之一。 相似文献
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10.
采取盆栽遮雨人工定量灌水的方法,探讨了常规晚稻和杂交晚稻前后两大生育时期不同灌水量对生育和产量的影响.结果表明:两类型晚稻品种前后两个生育时期节水灌溉都能达到高产之目的,但不同类型和不同生育时期的最佳灌水量和节水效果不同. 相似文献