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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-MZmSYCOZmbHLH54ZmGlu-r1ZmCLPB3ZmP450-99A2的表达,抑制ZmAGD12ZmCYSZmCYSBZmERF-107ZmEIN3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-107ZmEIN3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-MZmAGD12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.
Journal of Plant Diseases and Protection - Recent observations of increasing incidences of potato plants showing blackleg symptoms have caused serious concern for potato growers and traders in...  相似文献   
3.
羔羊全营养代乳粉对羔羊增重影响的试验   总被引:1,自引:0,他引:1  
为了研究羔羊出生后采取人工饲喂代乳粉溶液的效果,对10日龄羔羊称重,以平均日增重与母乳组比较,用t检验方法检验差异显著性。研究结果显示羔羊平均日增重试验组为0.097-0.126kg,母乳组为0.071-0.113kg,二者没有显著差异。在生产中,代乳粉能替代绵羊母乳饲喂羔羊,对多胎品系羊的推广应用以及解决绵羊缺乳、少乳或无乳问题具有重要意义。  相似文献   
4.
Thirty-two crossbred cattle (steers = 17; heifers = 15) exhibiting an ultrasound fat thickness at the 12 to 13th rib region of at least 10 mm were selected from a slaughter shift at a commercial packing plant. After splitting, alternating sides of each carcass were trimmed of 1) subcutaneous fat in excess of 6.4 mm; 2) all kidney, pelvic, and heart fat; and 3) all cod or udder fat and fat in the flank region. Both sides of each carcass were fabricated into subprimals (final trim level of 6.4 mm) according to normal industry procedures. Effect of hot-fat trimming, yield grade (3, 4, and 5), and gender on hot-fat trim, fabrication fat trim, major subprimal, and total subprimal yield of untrimmed and trimmed carcasses were determined. Higher numerical yield grade (YG) corresponded with higher (P less than .05) percentages of hot-fat trim. Hot-fat trimming increased (P less than .05) the difference in fabrication fat trim between steers and heifers and between YG 3 and YG 5. Steers and heifers differed (P less than .05) in percentage of major subprimals and total subprimals when processed conventionally, whereas hot-fat trimming eliminated this difference (P less than .05). Untrimmed YG 3 carcasses had 3.1 and 5.0% higher major subprimal yield (P less than .05) than untrimmed YG 4 and YG 5 carcasses, respectively, whereas hot-fat trimming reduced this difference to 2.5% for YG 4 and to 3.7% for YG 5.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   
5.
中国果树类病毒的发生及其研究进展(英文)   总被引:4,自引:0,他引:4  
王国平  洪霓  Ahmed Hadidi 《果树学报》2005,22(1):51-54,F003
概述了在中国发生且己鉴定明确的5种果树类病毒,即苹果绣果类病毒(Apple scar skin viroid,ASSVd)、梨泡状溃疡类病毒(Pear blister canker viroid,PBCVd)、葡萄黄斑类病毒-1(Grapevine yellow speckle viroid-1,GYSVd-1)、柑橘裂皮类病毒(Citrus exocortis viroid,CEVd)和桃潜隐花叶类病毒(Peachlatent mosaic viroid,PLMVd)的研究进展,包括病害的首次发现、症状特征、发病规律、检测方法与防治对策以及这些类病毒的生物学与分子生物学特性。  相似文献   
6.
Two bacterial isolates and one strain of Trichoderma harzianum were tested alone and in combination with chitin for efficacy in control of root rot disease caused by Phytophthora capsici and Rhizoctonia solani in pepper plants under greenhouse conditions. These bacteria (Bacillus subtilis HS93 and B. licheniformis LS674) were isolated from repeatedly washed roots of pepper plants. In in vitro assays, HS93, LS674 and T. harzianum were antagonistic against P. capsici and R. solani and produced high levels of chitinase. Seed treatment and root drenching with bacterial suspensions of HS93 with 0.5% chitin was more effective against Phytophthora and Rhizoctonia root rot than addition of the organisms without chitin. LS674 and T. harzianum reduced Rhizoctonia but not Phytophthora root rot. In two greenhouse tests, seed treatment and root drenching with HS93 amended with chitin enhanced its biocontrol activity against P. capsici but not on R. solani. The effects of LS674 and T. harzianum against R. solani were significantly enhanced when they were used as suspensions with 0.5% chitin for root drenching, but this had no effect on P. capsici. In both greenhouse experiments, the use of 0.5% chitin alone for root drenching reduced Rhizoctonia root rot. Reduction of root rot disease was accompanied by increased yield. These results show that the antagonistic activity of HS93, LS674 and T. harzianum may be stimulated by chitin resulting in significant improvements in their effectiveness against pathogens.  相似文献   
7.
Leaf explants of Caladium ‘Pink Cloud’ were cultured in vitro on MS medium containing various auxins (NAA, IBA, IAA, 2,4,5-T and 2,4-D) in combination with cytokinin (BA). NAA gave the most vigorous in vitro propagation of this plant, and only 15% of the plants were leaf-colour variants on the medium containing 0.5 μmol NAA. Leaf colour variation was observed in all plants regenerated on the medium containing 2,4-D at 0.5–4.5 μmol. In hormone-free medium, only a few leaf-colour variants (6%) occurred, but the rate of plant regeneration was very low. Application of 0.5 μmol NAA together with 4.5 μmol BA seemed to be the most appropriate for in vitro propagation of Caladium ‘Pink Cloud’ with only a few leaf-colour variants.  相似文献   
8.
The objective of this study was to determine the effects of dietary supplementation of Nigella sativa L. seeds, Rosmarinus officinalis L. leaves and their combination on rumen metabolism, nutrient intake and digestibility, growth performance, immune response and blood metabolites in Dorper lambs. Twenty-four entire male Dorper lambs (18.68?±?0.6 kg, 4–5 months old) were randomly assigned to a concentrate mixture containing on a dry matter basis either, no supplement (control, T1), 1% R. officinalis leaves (T2), 1% N. sativa seeds (T3) or 1% R. officinalis leaves +1% N. sativa seeds (T4). The lambs had ad libitum access to urea-treated rice straw (UTRS) and were raised for 90 days. Supplemented lambs had greater (P?< 0.05) intake of DM and UTRS than the control lambs. The T4 lambs had lower (P?< 0.05) nutrient digestibility than those fed other treatments. Total and daily weight gain was greater (P?< 0.05) in T2 lambs than those fed other diets. The T3 and T4 lambs had greater (P?< 0.05) ruminal pH than the T1 and T2 lambs. Supplemented lambs had lower (P?<?0.05) ruminal total volatile fatty acids, acetate, propionate, NH3-N and C18:0 than the control lambs. The T4 lambs had lower (P?< 0.05) population of Fibrobacter succinogenes, Ruminococcus albus, methanogens and total protozoa compared with those fed other diets. Supplemented lambs had lower (P?< 0.05) neutrophils, basophils and serum urea and greater (P?<?0.05) serum IgA and IgG compared with the control lambs. The current results emphasised the variation in the efficacy of medicinal plants in ruminant nutrition.  相似文献   
9.
The aims of this study were to establish optimal doses of doxycycline (dox) against Haemophilus parasuis on the basis of pharmacokinetic–pharmacodynamic (PK‐PD) integration modeling. The infected model was established by intranasal inoculation of organism in pigs and confirmed by clinical signs, blood biochemistry, and microscopic examinations. The recommended dose (20 mg/kg b.w.) was administered in pigs through intramuscular routes for PK studies. The area under the concentration 0‐ to 24‐hr curve (AUC0–24), elimination half‐life (T½ke), and mean residence time (MRT) of dox in healthy and H. parasuis‐infected pigs were 55.51 ± 5.72 versus 57.10 ± 4.89 μg·hr/ml, 8.28 ± 0.91 versus 9.80 ± 2.38 hr, and 8.43 ± 0.27 versus 8.79 ± 0.18 hr, respectively. The minimal inhibitory concentration (MIC) of dox against 40 H. parasuis isolates was conducted through broth microdilution method, the corresponding MIC50 and MIC90 were 0.25 and 1 μg/ml, respectively. The Ex vivo growth inhibition data suggested that dox exhibited a concentration‐dependent killing mechanism. Based on the observed AUC24 hr/MIC values by modeling PK‐PD data in H. parasuis‐infected pigs, the doses predicted to obtain bacteriostatic, bactericidal, and elimination effects for H. parasuis over 24 hr were 5.25, 8.55, and 10.37 mg/kg for the 50% target attainment rate (TAR), and 7.26, 13.82, and 18.17 mg/kg for 90% TAR, respectively. This study provided a more optimized alternative for clinical use and demonstrated that the dosage 20 mg/kg of dox by intramuscular administration could have an effective bactericidal activity against H. parasuis.  相似文献   
10.
为确定临沂市机播条件下红玫瑰彩色马铃薯的最佳播种密度,开展了机播马铃薯相同行距(90 cm)、不同株距(14、18、22、26 cm)的密度试验,分析了不同密度处理对红玫瑰马铃薯出苗期、现蕾期、开花期、成熟期、株高、茎粗、主茎数、马铃薯晚疫病发病情况、产量、商品薯率的影响。结果表明:随着播种密度的增加,红玫瑰物候期无明显变化,株高、马铃薯晚疫病株数呈升高趋势,茎粗和主茎数呈降低趋势,商品薯率和商品薯产量呈下降趋势。以行距90 cm、株距22 cm密度下商品薯667 m~2产量最高,为2 129.25 kg,并且马铃薯晚疫病未见发生。  相似文献   
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