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   

延迟收获对长江中游春玉米农艺性状及机收质量的影响     

刘志辉  展茗  梁如玉  王燕  刘永忠  黄益勤  杨仁能  尚春辉 《中国农业大学学报》2021,26(11):10-22

为探明长江中游玉米籽粒机械直收适宜品种与配套农艺措施，2018—2019年选用不同玉米品种，测定不同机收时间下玉米关键农艺性状、产量及机收质量指标。结果表明，收获时间对春玉米机收产量与机收质量均有显著影响。延迟1周收获后籽粒容重显著增加，机收产量显著提高，2年平均提高9.72%；而延迟2周收获则有降低机收产量的趋势。2年收获时杂质率总体≤3%，而机收籽粒破碎率与损失率均>5%，是该区域春玉米籽粒机收面临的主要问题。籽粒厚度、籽粒含水率和百粒重是影响机收籽粒破碎率的关键性状，三者与机收籽粒破碎率均呈显著的倒二次曲线关系；玉米的倒伏率、穗位高和重心高度是影响机收损失率的关键性状，倒伏率与机收损失率呈显著正相关，而穗位高和重心高度与机收损失率均呈显著的二次曲线关系。延迟收获能显著降低籽粒含水量，从而降低籽粒破碎率，但继续延迟收获有增加倒伏的风险。综上，长江中游春玉米成熟后适时延迟7~10 d收获，可有效降低籽粒含水量与机收籽粒破碎率，提高玉米籽粒机收产量。  相似文献   

Effects of Artificial Warming from Late-Winter to Early-Spring on Photosynthesis and Flag Leaf Senescence of Winter Wheat     

YAN Peng  SUN XiaoNuo  DU Xiong  GAO Zhen  BIAN DaHong 《中国农业科学》2019,52(15):2581-2592

  相似文献   

我国牧草种植成本收益变化与比较     

石自忠  王明利  胡向东  崔姹 《草业科学》2017,34(4)

本研究以牧草种植户牧草及粮食作物的调研数据为基础,计算出2010-2014年主要牧草和粮食作物的成本收益基本信息,对其变化进行统计分析,并对各作物的成本收益进行比较。分析发现,2014年苜蓿(Medicago sativa)的经济效益有所下降;黑麦草(Lolium perenne)和青贮玉米(Zea mays)的纯收益和收益率有所增长;苜蓿的纯收益和收益率要高于黑麦草和青贮玉米;马铃薯(Solanum tuberosum)和苜蓿的经济效益更具优势,种植小麦(Triticum aestivum)经济效益较差;黑麦草和青贮玉米人工和土地成本更高,苜蓿水电费最高。建议调整农业产业结构,推行"粮+经+饲+草"四元种植结构,全面扶持牧草产业发展,建立健全牧草生产资料补贴制度。  相似文献   

Detect unique molecular structure associated with physiochemical properties in CDC varieties of oat grain with unique nutrient traits [Feed Type vs. Milling Type] in comparison with barley grain using advanced molecular spectroscopy as a non-destructive biological tool     

《Journal of Cereal Science》2017

The objective of this study was to determinate grain unique protein inherent molecular structure that are related physiochemical and nutrient profiles in CDC developed oat varieties [CDC Nasser (Feed Type) and CDC Seabiscuit (Milling Type)] grown in cool climate condition in western Canada in comparison with conventional barley variety of CDC Meredith as a control using advanced molecular spectroscopy. Multivariate analyses, including an agglomerative hierarchical cluster analysis (CLA) and principal component analysis (PCA), were performed to identify protein molecular structural differences among the grains. The results revealed that CDC Seabiscuit contained greater (P < 0.05) protein structural Amide I and II than CDC Nasser and CDC Meredith, while the greater (P < 0.005) structural Amide I to II area and height ratios was detected in CDC Meredith. New oat grains had greater (P < 0.05) β-sheet height than barley grains, however, there was no difference in α-helix to β-sheet ratio values among the varieties. In conclusion, CDC Nasser and CDC Meredith had no difference in protein molecular structural features, while CDC Seabiscuit contains different protein structural characteristics as compared to CDC Meredith grain. The molecular structure features are highly associated with physiochemical and nutrient profiles in grains, which indicate that it also affect nutrient utilization and availability.  相似文献   

Identification of an H1N1 subtype of swine influenza virus and serological analysis     

Fa-chao SUN  Min TAN  Yuan-chao ZHANG  Yu-chao WANG  Sheng-liang CAO  Guo-fei DING  Fang-yuan CONG  Li-hong GUO  Si-dang LIU  Yi-hong XIAO 《农业科学学报》2019,18(7):1436-1442

To investigate the epizootic of swine influenza virus(SIV), 60 nasal swabs were collected from a clinical cases of pig farm in Tai'an City, Shandong Province of China in April 2017. SIV was isolated by inoculating into 10-day-old Special Pathogen Free embryonated eggs and the whole genome was sequenced. An H1N1 subtype SIV was isolated and designated as A/swine/Shandong/TA04/2017(H1N1). Phylogenetic analysis showed that apart from the polymerase A(PA) fragment belonging to the 2009 pandemic H1N1 branch, seven genome segments belonged to avian-like H1N1 influenza virus lineage. The cleavage site sequence of the hemagglutinin(HA) protein was PSIQSR↓G, which is a typical molecular biological characteristic. Five potential N-glycosylation sites(N14, N26, N277, N484 and N543) were found in the HA gene. To further investigate the epidemiology of SIV in this farm, the 995 serum samples were assessed with EAH1N1 2009 pandemic H1N1 and H3 N2 antigens. The results showed that the total positive rate was 65.43%. The positive rates of single virus infection detected by EAH1N1, 2009 pdmH1N1 and H3 N2 for serum HI(Hemagglutination inhibition) were 48.35, 30.85 and 7.47%, respectively. The results showed that SIV in Shandong Province has been reassorted in some segments and the SIV-positive rate was high on the SIV outbreak farm. These data provide evidence of an epizootic of SIV.  相似文献   

Effect of Heat Stress on Grain Growth, Starch Synthesis and Protein Synthesis in Grains of Wheat (Triticum aestivum L.) Varieties Differing in Grain Weight Stability     

C. Viswanathan  & Renu Khanna-Chopra 《Journal of Agronomy and Crop Science》2001,186(1):1-7

The effect of heat stress on the components of grain weight was analysed in wheat ( Triticum aestivum L.) varieties differing in grain weight stability. The wheat varieties PBW154, Sonalika and Hindi62 were raised in the field (New Delhi, India; 77°12' E, 28°40' N, 228.6 m a.s.l.) at three dates of sowing: 19 November 1992 (DOS I), 14 December 1992 (DOS II) and 18 January 1993 (DOS III). The late-sown crop (DOS III) experienced 6–8 °C warmer temperatures during grain development than the crop sown at the normal time (DOS I). The heat susceptibility index (S) revealed that grain weight was less susceptible to heat in Sonalika and PBW154 than in Hindi62. Heat stress reduced both the grain growth duration (GGD) and the grain growth rate (GGR). The grain weight reduction in PBW154 and Sonalika was mainly due to a reduction in GGR, while that of Hindi62 was due to a reduction in GGD. In vivo studies on starch and protein synthesis in excised endosperm at 15, 25 and 35 °C revealed that both processes were more thermotolerant in Hindi62 than in PBW154. The grain starch content was stable in Hindi62 while that in PBW154 was significantly reduced under heat stress. The grain nitrogen content at maturity increased in both varieties under heat stress. It was concluded that the heat susceptibility of grain weight in Hindi62 was mainly due to a reduction in GGD, although GGR, starch and protein synthesis were more thermotolerant in developing grains of Hindi62 than in those of PBW154.  相似文献   

导入AGPase基因的转基因可育水稻及其经济性状的研究   总被引：8，自引：0，他引：8  

宋敏  李援亚  张云孙 《华北农学报》2001,16(4):11-14

以农杆菌介导法将淀粉合成关键酶AGPase(腺嘌呤二磷酸葡萄糖焦磷酸化酶）基因导入云南地方优质水稻合系35，38，39，41，42中，获得了大量的可育转基因植株。PCR扩增检测，外源基因已稳定整合进水稻基因组中。大量的农艺性状及淀粉含量研究表明，转基因水稻种子的千粒重比对照明显提高，但总淀粉含量没有显著变化。  相似文献   

Effects of Benzyl Adenine and Abscisic Acid on Grain Yield and Yield Components in Triticale and Wheat     

T. Sivakumar  Virendranath  & G. C. Srivastava 《Journal of Agronomy and Crop Science》2001,186(1):43-46

Grain yield and yield components (grains per ear, grain weight, 1000-grain weight, ear weight, ear seed ratio and dry matter partitioning between ear and seed) were examined in a wheat genotype (PBW-343) with well-filled grains and a Triticale genotype (DT-46) with poorly filled grains (showing grain shrivelling) grown in pots. Six days after anthesis (DAA), benzyladenine (BA) @2 µg ear⁻¹ and abscisic acid (ABA) @4 µg ear⁻¹ were injected at the base of the mother shoot ear in both species. It was observed that, in both wheat and Triticale , BA increased the grain weight, grain number and partitioning of dry matter between ear and seed, whereas ABA decreased the grain weight, grain number and dry matter partitioning between ear and seed. However, these decreases were slower in Triticale than in wheat. BA treatment increased the grain dry matter accumulation, which in turn resulted in better filling of grains and increased the grain weight in both wheat and Triticale . The average grain weight of Triticale was lower than that of wheat. Thus, it appears that variation in grain weight between wheat and Triticale might be due to different availabilities of growth-promoting phytohormones such as cytokinins and assimilates.  相似文献   

辛亥革命前后张謇与袁世凯的交往     

王志军  王瑞芳 《信阳农业高等专科学校学报》2005,15(4):51-54

张謇与袁世凯的交往长达三十余年,他从最初的相识荐袁、拥袁合作、相互借重到后来的分离,张謇曾幻想依靠袁世凯实现自己“实业救国”的抱负,却以失败而告终。二人分分合合,充满波折的交往过程反映了中国近代资本主义发展的艰难与坎坷。  相似文献