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1.
苯丙氨酸解氨酶(phenylalanine ammonia-lyase,PAL)是植物中广泛存在的苯丙烷代谢途径的第一个关键酶,在生物化工领域可利用其逆向催化性质生产L-苯丙氨酸(L-Phe),而筛选活力高和稳定性好的PAL酶源是提高L-Phe产量的重要途径。本研究以苦荞(Fagopyrum tataricum)苯丙氨酸解氨酶基因(FtPAL)为材料,构建其原核表达融合载体pET-30b(+)-FtPAL,并转入大肠杆菌(Escherichia coli)BL21(DE3)中诱导表达,表达产物纯化后分析其逆向催化的酶学性质。结果FtPAL基因获得了有活性的表达,正向催化比活力在第5小时最高达24.17 U/mg,纯化后正向和逆向催化比活力分别为158.74 U/mg和194.11U/mg,薄层层析也证明,FtPAL可逆向催化肉桂酸氨化为L-Phe。酶学性质分析表明,FtPAL逆向催化的最适温度为30℃,热不稳定,最适反应pH为10,此时有较好的稳定性,除Mg2+对其具有激活作用外(P0.01),其它金属离子(Na+、Mn2+、Fe2+、Fe3+、Cu2+和Ca2+)均具有抑制作用(P0.05或P0.01),Hg2+则可使该酶完全失活。本研究明确了该酶具有合成L-Phe的性质,也为对进一步采用基因工程手段提高其催化能力和稳定性提供了基础资料。  相似文献   

2.
水稻化感物质作用特性的研究   总被引:8,自引:0,他引:8  
在对水稻化感作用生理生化特性研究的基础上,进行了水稻叶片浸提液的GC-MS分析和相关酶的活性测定。结果表明,水稻叶片中苯丙氨酸裂解酶(PAL)和肉桂酸-4-羟化酶(CA4H)活性的大小与酚的含量密切相关,同时水稻化感作用强度与其肉桂酸含量呈正相关关系。而与其水杨酸含量呈负相关关系,因此水稻化感作用是由许多化感物质共同作用的结果。  相似文献   

3.
稻瘟菌细胞壁热解物诱导水稻抗稻瘟病的研究   总被引:1,自引:0,他引:1  
以水稻近等基因系和广东省稻瘟病菌优势小种ZC13的细胞壁热解物(HDS)为材料进行诱导抗病性的研究。HDS通过细胞壁121°C 20 m in热解获得。HDS中葡萄糖质量浓度为176.83 m g/mL。HDS在体外对稻瘟病菌的孢子萌发和菌丝生长均无抑制作用,但处理水稻后可以显著提高水稻对稻瘟病菌的抗病性。在亲和性品系中,诱导抗病效果最高可达82.47%,且浓度与活性间存在正相关趋势;在非亲和品系中,热解物处理引起HR反应。HDS处理后,水稻过氧化物酶(POD)和苯丙氨酸解氨酶(PAL)活力明显升高。  相似文献   

4.
L-半胱氨酸(L-cysteine)作为生物体中的常见氨基酸,已有研究发现其能有效延缓青脆李果实的衰老和品质的下降。然而,L-半胱氨酸处理对李果实中苯丙烷代谢途径合成酚类物质的影响尚不清楚。因此,该研究以青脆李果实为试材,采用1 g/L L-半胱氨酸浸泡处理后于(20±1)℃贮藏,研究贮藏期间苯丙烷代谢途径中关键酶活性以及总酚、总黄酮等相关代谢产物的变化规律,同时测定果实中糖酸含量和抗氧化活性的变化。结果表明,李果实在贮藏过程中可溶性固形物(Total soluble solids,TSS)含量和可滴定酸(Titratable acid,TA)呈逐渐降低的趋势,L-半胱氨酸处理显著延缓了李果实TSS和TA的下降(P0.05)。在贮藏期间,苯丙烷代谢途径中关键酶活性均呈逐渐上升的趋势,与对照组相比,处理组中苯丙氨酸解氨酶、4-香豆酸辅酶A连接酶活性更高。果实中总酚、总黄酮含量在贮藏期间先降低后上升,在贮藏前三天,处理组中总酚、总黄酮含量显著高于对照组(P0.05)。对酚类物质单体含量的测定发现,处理后的果实中绿原酸、咖啡酸、丁香酸、芦丁等酚类物质单体含量显著高于对照组(P0.05)。抗氧化活性与总酚、总黄酮的变化趋势一致,经L-半胱氨酸处理后的果实保持了较高的抗氧化活性。相关性分析结果表明,果实中苯丙氨酸解氨酶、肉桂酸-4-羟基化酶和4-香豆酸辅酶A连接酶与果实酚类物质含量及抗氧化活性显著相关(P0.05)。总体来说,1 g/L L-半胱氨酸浸泡处理能够延缓青脆李果实贮藏品质下降,同时能够激活苯丙烷代谢途径关键酶,促进果实中酚类物质的积累。  相似文献   

5.
罗自生 《农业工程学报》2004,20(Z1):178-181
研究了MA贮藏对采后竹笋内源激素含量及木质化相关酶活性的影响.结果表明竹笋贮藏期间,GA3和IAA含量逐渐减少,ABA含量先增加后减少,苯丙氨酸解氨酶(PAL)、肉桂酸-4-羟化酶(C4H)和过氧化物酶(POD)活性先增加后减少,木质素含量和组织硬度逐渐增加,而可食用率则逐渐下降.MA贮藏能延缓内源GA3和IAA含量的降低,并抑制PAL、C4H和POD活性的上升,降低竹笋的木质素含量和硬度.MA贮藏抑制竹笋木质化可能与提高内源激素(GA3、IAA)含量和降低PAL、C4H及POD活性有关.  相似文献   

6.
为探讨钙氮营养对番茄抗叶霉病效果的影响,采用沙培的方法,研究了钙氮比为1∶1、1∶2、1∶4和1∶8条件下,番茄接种叶霉病病原菌后叶片中苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)、过氧化物酶(POD)和过氧化氢酶(CAT)的活性变化、番茄发病率和番茄体内钙的积累量。结果表明,接种后各处理的番茄PAL、PPO、POD和CAT酶活性均显著提高,其中钙氮比为1∶4条件下酶活性增加幅度最大,峰值最高,其次是钙氮比为1∶2处理,钙氮比为1∶8处理番茄POD、PPO酶活性峰值较其他处理酶活性峰值出现晚。钙氮比为1∶2和1∶4下番茄的病情指数显著低于其它处理。在钙氮比为1∶2~1∶4条件下,番茄体内钙的积累量较高,钙素在番茄根、茎、叶中的分配平衡。研究结果表明,钙氮比为1∶2~1∶4有利于番茄体内钙含量的积累,钙在根、茎、叶中均衡分配,提高番茄抗叶霉病的能力。过量的氮会减少番茄植株内钙的含量,降低番茄抗病性。  相似文献   

7.
茉莉酸甲酯对蓝莓贮藏品质及抗病相关酶活性的影响   总被引:4,自引:0,他引:4  
研究不同浓度(0、10、20、30μmol·L-1)茉莉酸甲酯(MeJA)熏蒸处理对蓝莓贮藏品质及抗病相关酶活性的影响,为蓝莓采后保鲜技术研究和应用提供参考。结果表明:蓝莓在5℃贮藏条件下,MeJA处理能保持较高的好果率,且能抑制果实硬度下降和糖酸含量的降低,其中10、20μmol·L-1MeJA处理贮藏品质较好;MeJA处理对蓝莓果实中苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、多酚氧化酶(PPO)有激活作用,其中20μmol·L-1MeJA浓度处理更有利于诱导抗病酶的活性,提高果实抗病性,为蓝莓采后较适宜的处理浓度。  相似文献   

8.
氨基酸和壳聚糖对烤烟生理特性及香味品质的影响   总被引:4,自引:0,他引:4  
本文研究了叶面喷施氨基酸和壳聚糖对烤烟烟叶质体色素含量、酶活性的变异以及对致香物质含量的影响.结果表明:打顶后氨基酸处理烟叶叶绿素、类胡萝卜素、苯丙氨酸解氨酶(PAL)活性和脂氧合酶(LOX)活性较高,壳聚糖处理影响不显著;氨基酸处理的烟叶致香物质均比壳聚糖处理和CK高,尤其是对苯丙氨酸类、美拉德反应产物类影响显著,增幅分别达到20.9%、23.1%.壳聚糖对苯丙氨酸类致香物质有较大提高,增幅为17.6%.从整体上看,氨基酸处理调整了发育后期质体色素组分比例和相关酶类的活性,对烟叶香味品质的影响效果要好于壳聚糖.  相似文献   

9.
山药PAL基因全长cDNA序列的克隆、表达与分析   总被引:2,自引:0,他引:2  
为了阐明苯丙氨酸解氨酶(PAL,EC4.3.1.5)基因在山药地下块茎酚类物质积累过程中的分子机理,本研究运用普通PCR、RACE和TAIL-PCR技术从大薯(Dioscorea alataL.)’Zixiao’地下块茎中克隆到同一个PAL基因的3个片段,即长度分别为1 145bp、1 151bp和424bp的中间片段、3’端和5’端的cDNA序列;将3段cDNA序列拼接后获得大小为2 376bp的PAL基因全长cDNA序列,该序列含有一个1986bp的最大开放阅读框(ORF),一个28bp的5’端非翻译区,一个362bp含25 nt的Poly(A+)尾的3’端非翻译区,最大ORF可推测编码一个包括PAL酶活性中心的特征序列(GTITASGDLVPLSYIA)在内的661个氨基酸的多肽,分子量为71.974kDa,等电点6.310;大薯地下块茎的PAL基因分别在核苷酸与氨基酸水平上和GenBank中所选已知其他物种的同源性为73%~77%和76%~82%,说明PAL基因在系统进化上相对保守;RT-PCR分析表明该基因仅在地下块茎中表达,而且表达丰度在收获前逐渐降低。  相似文献   

10.
我们曾分离到一株高抗多种重金属盐的微紫青霉菌(Penicillium janthinellum)菌株GXCR。本研究在该菌株中建立了基于携带苯菌灵抗性基因(Benr)的质粒pCPXBN-1、聚乙二醇(PEG)介导的原生质体转化体系。原生质体的产量和再生的最佳条件是酶解液中的酶浓度在0.6%~0.8%(纤维素酶∶蜗牛酶=1∶1)、孢子萌发后培养24 h的菌体和0.05 mol/L二硫苏半糖醇(DTT)预处理。最佳转化组合条件为3×108原生质体/mL,50μg质粒DNA组合条件下,40%PEG4000,山梨糖醇(sorbitol)-三(羟甲基)氨基甲烷盐酸盐(Tris-HCl)-氯化钙(CaCl2)缓冲液(STC)[1 mol/L sorbitol,50 mmol/L Tris-HCl(pH 7.0),50 mmol/L CaCl2],室温转化20 min。在此条件下的转化效率为0.3个转化子/μg DNA。  相似文献   

11.
Aluminum (Al) toxicity represents one of the main yield-limiting factors for crops in acid soils. Silicon (Si) is known to increase tolerance in higher plants. This study was conducted to determine whether treatment with Si could improve nutrient uptake by peanut under Al stress. Peanut (Arachis hypogaea L. cv Zhonghua 4) was raised with or without Si (1.5 mM) in the growth chamber under 0 and toxic Al (0.3 mM) levels. Aluminum stress significantly decreased the root- and total-dry weight by 52.4% and 32.0%, respectively. The content of nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), and magnesium (Mg) was significantly decreased, but that of Al increased markedly in shoots and roots after Al exposure at seedling, flower-needle, and pod-setting stage. Silicon alleviates Al toxicity in peanut plants in relation to Al distribution and allocation of tissue P, K, Ca, and Mg by favoring the partitioning of dry mass to roots.  相似文献   

12.
The objective of this study was to evaluate the influence of phosphate deficiency on enzymatic activities and on compounds of the secondary metabolism linked with the production of root exudates in bean (Phaseolus vulgaris L.) plants cv. ‘Bianco di Bagnasco.’ Phosphate deficiency induced a decrease in root total soluble phenolic content, but also caused an increase in total soluble phenolic content of exudates. Alteration of phenolic production was paralleled by increased activity of L-phenylalanine ammonia-lyase (PAL) and decreased activity of chalcone isomerase (CHI) in roots. Two isoflavonoids daidzein and naringenin, have been detected in roots; they are considered signal molecules in the chemical trafficking with soil microorganisms. Naringenin content was reduced by phosphorus (P) deficiency, due to lower activity of CHI, the enzyme responsible for its biosynthesis. On the other hand, daidzein content was higher in –P plants than in the control. The results are discussed, considering the possible roles of phenolics and flavonoids in plant-soil microbe interactions and rizosphere modification.  相似文献   

13.
Inductively coupled plasma-mass spectroscopic (ICP-MS) analysis of leaves from 22 cabbage crops in the Sa P? and B?c Hà districts of Láo Cai Province, North-Western Vi?t Nam, revealed unexpectedly high concentrations of chromium (Cr) and nickel (Ni). The concentrations were strongly linearly related (r2 = 0.94), indicating sample contamination during grinding through a stainless-steel hammer mill. We tested this hypothesis in two ways. First, brown rice ground through the same mill was contaminated not only by Cr and Ni, but also cobalt (Co), iron (Fe) and molybdenum (Mo). Second, scanning electron microscopy/energy dispersive analysis of x-rays (SEM/EDS) of the ground samples revealed small fragments with co-located Fe, Cr and Ni, consistent with stainless steel wear fragments. Other grinders may perform differently and we suggest that quality assurance protocols for trace metal analysis of plants should include testing for grinder wear metals. Lastly, brown rice appears to be convenient for investigating contamination of plant tissues during grinding.  相似文献   

14.
模拟降雨条件下污染土壤中重金属元素径流迁移特征   总被引:6,自引:0,他引:6  
探明污染土壤中重金属元素随地表径流迁移特征,对于防控重金属污染物的迁移扩散具有重要意义。通过室内模拟降雨试验,研究不同降雨强度下(55,100,120mm/h)污染土壤中Cd、Pb和Cr随地表径流迁移的特征。结果表明:径流中3种重金属元素总量在产流的前20min内逐渐降低,随后趋于稳定;产流初期(产流10min内)颗粒态重金属占径流重金属总量的比例均在80%以上;随产流时间延长,颗粒态Cd和Cr对径流重金属的贡献率快速降低,其颗粒态/溶解态的比值范围分别为47.0~0.4和12.9~0.4,而Pb则主要以颗粒态的形式存在,溶解态Pb对其随径流迁移的贡献率可忽略不计;随降雨强度增大,径流中总Cd和Pb含量显著降低,而径流中总Cr含量以及溶解态重金属含量无明显变化规律。  相似文献   

15.
Information is scant on the effect of humic acid (HA) on physiological, antioxidant and photosynthesis attributes of gerbera plants undergoing nutrient deficiency in culture solution. Gerbera plants cv. Malibu were grown in a factorial experiment based on a completely randomized design with 3 replications, using 3 different nutrient solutions [complete nutrient solution (NSc), 25% NSc (NS1), and 50% NSc (NS2)] treated with 2 levels of humic acid [0 (HA0) and 500 mg/l (HA1)].The interaction effect of HA and NS showed that HA improved the flower number in NSc, the transpiration in NS1+HA1, photosynthesis rate in NSc+HA1, stomatal conductance (gs) in NS2, mesophyll conductance of leaves in all NS levels and photosynthetic water use efficiency in NSc+HA1. The interaction effect of nutrient solution and HA on antioxidant activity was inconclusive, malondialdehyde content was the highest in NS2 and the lowest in NS1+HA1. The peroxidase activity increased in complete nutrient solution with and without HA and there were no differences among other treatments. Superoxide dismutase activity increased in NS1 and complete nutrient solution with HA and reached the highest in NSc. Humic acid was more effective in nutrient uptake, i.e., nitrogen, phosphorus, potassium, calcium, zinc, and iron (N, P, K, Ca, Zn, and Fe) in complete nutrient solution compared to NS1 and NS2. Conclusively, humic acid can compensate the nutrient deficiency stress of the culture solution in regards to protein synthesis, photosynthesis attributes regardless of the nutrient uptake of gerbera.  相似文献   

16.
ABSTRACT

Delayed sowing and imprecise application of irrigation water to cotton has been the major hurdle in sustaining cotton yield in north-western India. Therefore, studies were initiated to ascertain the impact of heavy or normal level of presowing irrigation (PSI), scheduling time of first postsowing irrigation (POSI) under two sowing dates (SDs) on cotton arranged in a split block design replicated thrice. PSIh (100 mm) recorded 23.2% higher seed cotton yield over PSIn (70 mm) owing to better yield attributes and higher stand. First POSI at 4 weeks after sowing (WAS) resulted the highest yield (3072 kg ha?1), while one at 6 WAS (POSI6) recorded least due to poor population and reduced yield attributes. Water productivity under POSI6 was lesser by 29.1%, 25.2%, and 16.2% as compared to POSI3, POSI4, and POSI5, respectively. April sown crop out yielded the May sown cotton crop by 939 kg ha?1 due to better yield attributes. Nitrogen factor productivity (NFP) among SD remained higher by 23.3% for April as compared to May sowing. PSIh exhibited better NFP over PSIn. POSI6 recorded least NFP by 24.4%, 28.5%, and 16.3% as compared to POSI3, POSI4, and POSI5, respectively and was indicative of relatively poor utilization of N under delayed schedules. Therefore, planting in April after heavy PSI and scheduling first POSI at 4 WAS is best strategy for sustaining cotton yield and maximizing farmer profitability.  相似文献   

17.
采用田间小区试验研究“推荐配比”(N:P2O5:K2O比率为190:90:100 kg hm-2)、“高氮量配比”(N:P2O5:K2O比率为210:90:100 kg hm-2)、“低氮量配比”(N:P2O5:K2O比率为170:90:100 kg hm-2)以及“常规配比”(N:P2O5:K2O比率为216:112.5:202.5 kg hm-2)4种氮、磷、钾配比施肥对饲用稻威优198蔗糖合成酶(SUS)、腺苷二磷酸焦磷酸化酶(AGPase)、硝酸还原酶(NR)、谷氨酰胺合成酶(GS)以及产量和糙米蛋白质的影响。结果表明:“推荐配比”能提高不同生育时期水稻功能叶(旗叶)和粒籽中碳、氮代谢关键酶的活性,这些关键酶活性的变化显著影响水稻产量和糙米全氮以及蛋白氮的含量。统计(P0.05)结果证实“推荐配比”能提高水稻产量达到8200 kg hm-2,与“常规配比”相比产量提高了24.81%;“推荐配比”糙米全氮和蛋白氮含量分别达到22.70 g kg-1和21.98 g kg-1,与“常规配比”相比差异显著,并且其全氮和蛋白氮含量分别提高17.01%,18.38%。  相似文献   

18.
ABSTRACT

High bicarbonate (HCO3 ?) of irrigation water can be detrimental to plant growth in sustainable horticultural production systems. The ability of arbuscular mycorrhizal fungi (AMF), ZAC-19, (composed of Glomus albidum, Glomus claroideum, and Glomus diaphanum) to enhance tolerance to HCO3 ? was tested on Rosa multiflora cv. Burr. Arbuscular mycorrhizal colonized and non-inoculated (non-AMF) plants were treated with 0, 2.5, 5, and 10 mM HCO3 ?. Increasing HCO3 ? concentration and associated high pH and electrical conductivity (EC)—reduced plant growth, nutrient uptake, and acid phosphatase activity, while increasing alkaline phosphatase activity (ALP). Inoculation with AMF enhanced plant tolerance to HCO3 ?, as indicated by greater growth (leaf, stem, and total plant dry weight, leaf area and leaf area ratio), leaf elemental concentration [nitrogen (N), phosphorus (P), potassium (K), calcium (Ca), iron (Fe), zinc (Zn), aluminum (Al), boron (B)], leaf chlorophyll concentration, higher mycorrhizal inoculation effect, lower root Fe reductase activity, and generally lower soluble ALP activity. While AMF colonization was reduced by increasing HCO3 ? concentration, colonization still occurred at high HCO3 ? concentration. At 2.5 mM HCO3 ?, AMF plant growth was comparable to plants at 0 mM HCO3 ?, further indicating the beneficial effect of AMF for alleviation of HCO3 ? plant stress.  相似文献   

19.
四种常规方法提取伊利石有效钾的机制比较   总被引:1,自引:0,他引:1  
采用化学分析、X射线衍射、中红外光声光谱以及原子力显微镜的方法,比较了0.2 mol L~(-1)四苯硼钠法、1 mol L~(-1)沸硝酸法、2 mol L~(-1)冷硝酸法和2 mol L~(-1)热盐酸法浸提伊利石中有效钾的机制。结果表明,四苯硼钠法浸提时,伊利石中钾素释放量达到全钾量的59.5%,且基本均通过层间交换反应予以释放,结构离子铁、铝和硅释放量极低;采用三种酸溶液浸提时,其钾素释放量仅占全钾量的1.53%~2.46%,通过层间交换反应释放的钾量占释放量的比例为88.4%~94.0%。四苯硼钠浸提时伊利石层间距扩大,产生次生过渡矿物,并形成富硅表层,但在伊利石表面无溶蚀特征;三种酸溶液浸提时伊利石结构无改变,但其结晶度降低,且表面有明显的溶蚀特征。因此,土壤矿物层间钾是作物可利用有效钾的主要来源,三种酸溶液浸提方法一方面低估了有效钾容量,另一方面提取了一部分不能为植物所利用的结构态钾,不适宜于用来评价伊利石及土壤有效钾库容量。  相似文献   

20.
ABSTRACT

Understanding the potential of clinoptilolite (CLI) for adsorption of NH4+ and K+, providing appropriate fertilizer formula, and evaluation of the produced zeolitic nutrient sources (ZNSs) to meet the plant need are the main objectives of this study. Three ZNSs (NH4+-saturated, K+-saturated and dual-purpose NH4+-K+ saturated CLI) were produced, assessed, and compared with commercial N and K fertilizers (CFs) on corn growth in a greenhouse. The results indicated that CLI can potentially adsorb both NH4+ and K+ to the maximum values of 25.00 mg-NH4+ g?1 and 47.61 mg-K g?1, respectively, and chemisorptions mainly followed the process of adsorption. Saturation of zeolite by NH4+ and K+ occurred after 10 and 15 d which lead to ZNSs with 2% and 5% of N and K, respectively. NH4+-K+ saturated CLI contained 1% N and 1% K. The greenhouse experiment showed no significant difference between ZNSs and CFs on plant growth. However, the application of both N and K in the form of zeolitic sources significantly increased the uptake of N by the plant. The highest uptake of K (2.05 g pot?1) occurred in plants supplied with both natural zeolite and CFs. The present results may benefit the future utilization of ZNSs in environmental friendly farming practices.  相似文献   

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