过表达ZmIBH1-1提高玉米苗期抗旱性     

朱芳芳  董亚辉  任真真  王志勇  苏慧慧  库丽霞  陈彦惠 《中国农业科学》2021,54(21):4500-4513

【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   

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   

欧洲葡萄‘粉红亚都蜜’NAC基因DRL1负向调节植物抗旱性     

朱自果  阴启忠  张庆田  韩真  张倩  李勃 《园艺学报》2020,47(12):2290-2300

以欧洲葡萄‘粉红亚都蜜’（Vitis vinifera‘Yatomo Rose’）为材料，利用荧光定量PCR技术和转基因技术研究葡萄NAC转录因子DRL1基因对逆境的响应。欧洲葡萄‘粉红亚都蜜’在激素和逆境胁迫下，DRL1表达呈下降趋势，其中以ABA和干旱胁迫处理最为显著。在ABA处理下DRL1转基因烟草株系种子萌发率和根长均高于野生型。干旱处理下，转基因植株对干旱的耐受性降低，同时胁迫相关基因NtLEA5、NtP5CR1、NtPSCS1、NtERD10C和NtDREB3的表达水平比野生型显著下降。此外，DRL1转基因烟草茎中柱发育受到抑制，尤其是导管横切面积仅为野生型的58%。以上结果表明，DRL1基因可能作为1个负向调节子参与植物的干旱胁迫。  相似文献   

Comparison of the microbial communities of alpacas and sheep fed diets with three different ratios of corn stalk to concentrate     

Ruimin Chao  Chengqiang Xia  Caixia Pei  Wenjie Huo  Qiang Liu  Chunxiang Zhang  Youshe Ren 《Journal of animal physiology and animal nutrition》2021,105(1):26-34

The objective of this study was to investigate the characteristics of ruminal microbial communities of alpacas (Lama pacos) and sheep (Ovis aries) fed three diets with varying ratios of roughage (corn stalk) to concentrate, 3:7 (LS), 5:5 (MS) and 7:3 (HS). Six alpacas (one-year-old and weighing 29.5 ± 7.1 kg) and six sheep (one-year-old and weighing 27.9 ± 2.7 kg) were used in this study, in a replicated 3 × 3 Latin square experiment. Total protozoa concentration was determined under the microscope; total fungi and methanogens were assessed using quantitative polymerase chain reaction and expressed as a percentage of total bacterial 16S rRNA gene copies; bacterial communities were investigated by targeted 16S rRNA gene (V3–V4 region) sequencing. The percentage of fungi was significantly higher in alpacas than in sheep under the LS diet, while the concentration of protozoa was significantly lower in alpacas under HS, MS and LS diets. The alpha diversity including Shannon, Chao l and ACE indices of bacterial communities was higher in alpacas than in sheep, under the LS diet. A total of 299 genera belonging to 22 phyla were observed in the forestomach of alpaca and sheep, with Bacteroidetes and Firmicutes dominating both animal species. Phyla Armatimonadetes and Fusobacteria, as well as 64 genera, were detected only in alpacas, whereas phyla Acidobacteria and Nitrospira, as well as 44 genera, were found only in sheep. The abundance of cellulolytic bacteria, including Butyrivibrio and Pseudobutyrivibrio, was higher in alpacas than in sheep under all three diets. These differences in the forestomach microbial communities partly explained why alpacas displayed a higher poor-quality roughage digestibility, and a lower methane production. Results also revealed that the adverse effects of high-concentrate diets (70%) were lesser in alpacas than in sheep.  相似文献   

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

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   

湟源县草地资源基本特点及开发利用     

殷咸芳  刘志林 《青海草业》2001,10(4):21-22,15

湟源县草地面积有 85 1 85 .1 3hm2 ,占全县土地总面积的 5 6.45 % ,草地资源丰富 ,牧草种类较多 ,农业生产秸秆来源广泛 ,但由于利用不当 ,造成了草地严重破坏和秸秆的浪费 ,如何合理利用草地和秸秆资源 ,扼制草地退化 ,是促进该县畜牧业发展的重要因素  相似文献   

中西药结合预防肉鸡腹水综合征的临床试及防治机理探讨     

杨慧萍  岳治权  王建华 《中国家禽》2004,8(Z1):69-71

将一日龄艾维因肉仔鸡(雌雄各半)180只在同一舍内饲养,14日龄时将其随机分成4组,其中Ⅰ、Ⅱ组饲料按1％分别添加我们研制的方剂一和方剂二,连续用药15d,为药物预防组;Ⅲ组在饲料中按0.05％添加肾肿腹水消,连用7d(药品要求的预防剂量),为药物对照组;Ⅳ组饲料中不添加任何药物,为空白对照组。Ⅰ-Ⅳ组发病率分别为0、10％、3％和20％,42d时Ⅰ、Ⅳ组体重高于其它组,Ⅰ平均体重为1972.92g,Ⅳ组平均体重为1989.91g。试验表明方剂一为该试验条件下预防肉鸡腹水综合征最有效的方剂,其保护率可达100％,且不影响出栏重。对方剂一预防肉鸡腹水综合征的机理进行探讨。  相似文献   

大豆室内耐冷筛选及其在田间的应用研究   总被引：4，自引：1，他引：3  

李育军  常汝镇 《大豆科学》1992,11(1):49-57

本试验把生长箱大豆耐低温萌发筛选与田间早春播研究相结合,验证耐低温发芽的可靠性。在实验室,萌发期表现抗冷的大豆,田间出昔期仍表现抗冷或较抗冷。实验表明,大豆出苗的生物学最低温度平均为7.1℃,所需有效积温136℃。地温的高低直接影响大豆早春出苗的多寡,两者呈密切的正相关。黄、青、黑豆中,以黑豆抵抗早春不良气候条件的能力最强。  相似文献   

四川省退耕还林模式初探     

赵安玖  胡庭兴  李贤伟 《四川林勘设计》2001,(3)

根据自然地理分异规律、社会经济技术原理、生态学原理,分析了四川省的水土流失概况,提出了川中丘陵、盆周山地区、川西南干热干旱河谷区、川西北高山高原区的退耕还林模式。  相似文献   

初冬早春南昌郊区灯下常见蛾类活动情况纪实     

章士美  赵凤霞 《江西农业大学学报》1994,16(1):47-51

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