不同氮效率茄子氮代谢相关酶活性的差异

以3个不同氮效率基因型茄子为供试材料,采用大田培养,研究了正常供氮和低氮胁迫下,茄子幼苗期到结果期的氮代谢相关酶活性,探讨氮代谢相关酶活性的差异.结果表明,与正常供氮相比,低氮胁迫下,不同氮效率基因型茄子的硝酸还原酶活性、谷氨酰胺合成酶活性均降低,且大多数指标达到显著水平.与低氮高效基因型07-860及氮双低效基因型07-857相比,氮双高效基因型07-862具有较强的氮代谢相关酶活性.在供氮水平相同条件下,通过对幼苗期至结果期不同氮效率基因型进行GS及NR活性测定,选择具有相对高活性GS及NR的氮效率基因型是对氮高效基因型的有效早期选择.     

灌水对不同追氮水平下夏玉米氮代谢及产量的影响

为研究玉米生育后期补灌和追氮对氮代谢及产量的耦合效应,采用大田试验,测定了拔节后灌水对不同追氮水平下硝酸还原酶（NR）、谷氨酰胺合成酶（GS）等氮代谢酶活性、产量及产量构成因素的影响。结果表明,追氮和灌水使花后穗位叶片硝酸还原酶（NR）活性增加;随追氮量增加,谷氨酰胺合成酶（GS）活性反而降低;灌水处理和自然降水处理对GS活性变化影响差异较小。相同灌水下,追氮可明显提高玉米产量;相同追氮下,补灌处理产量要高于自然降水处理。追肥对产量的增加幅度要大于灌水。     

玉米穗位叶碳氮代谢的关键指标测定

碳、氮代谢是植物体内最主要的两大代谢过程,玉米一生中碳、氮代谢的协调在很大程度上决定着产量结果。实验用可见分光光度法对不同施氮水平下玉米穗位叶的碳氮代谢指标进行了检测。结果表明,施氮量在120～180 kg?hm-2间明显促进玉米穗位叶蔗糖的积累,在120～240 kg?hm-2间明显促进碳代谢的关键酶蔗糖磷酸合成酶（SPS）、氮代谢的关键酶硝酸还原酶（NR）和谷氨酰胺合成酶（GS）活性的增强。     

低氮条件下木薯谷氨酰胺合成酶(GS)酶活及GS家族基因表达分析

谷氨酰胺合成酶(GS)是植物氮代谢中的关键酶,在氨同化和谷氨酰胺生物合成中起着重要的作用。木薯中谷氨酰胺合成酶基因家族有四个基因,分别是细胞质型cassava4.1_008086m(Me GS1.1),cassava4.1_010581m(Me GS1.2),cassava4.1_010597m(Me GS1.3)和叶绿体型cassava4.1_008019m(Me GS2)。实验采用q RT-PCR技术分析铵盐和硝酸盐两种不同的氮源低氮胁迫下耐低氮品种(双高CH16)和低氮敏感型品种(双低G16)GS基因的表达差异。结果表明,在低铵盐氮源处理下木薯两类品种的GS酶活在叶中要高于正常氮源处理,根中低于正常氮源处理,而低硝酸盐氮源处理下木薯两类品种的GS酶活在叶和根中要低于正常氮源。在不同低氮源处理下,两类材料叶绿体型GS表达量在叶中始终与GS酶活存在相反现象。因此,推测木薯叶片中叶绿体型GS表达量可能与GS酶活呈负相关关系。而细胞质型GS表达量差异较大,与叶绿体型GS共同影响GS酶活,影响着木薯的氮利用效率。     

大豆谷氨酰胺合成酶基因的分类及根瘤特异表达GmGS1β2基因功能的初步分析

从Phytozome数据库中获得包括大豆在内的12种植物的谷氨酰胺合成酶(glutamine synthetase, GS)氨基酸序列,利用MEGA5.10软件进行多序列比对、构建进化树。进化分析表明,植物GS可以分成胞质型(GS1)和质体型(GS2)两大类,GS1可进一步分成分5个亚类,包括双子叶植物为主的I、II和III亚类、低等植物类(IV)和单子叶植物类(V)。这5亚类中,第II类是豆科植物特有的一类,大豆的4个GS1 (GmGS1β1/2和GmGS1γ1/2)属于该亚类;利用qPCR在大豆盛花期分析GS1基因的组织表达特异性,结果表明不同类型GmGS1基因在表达部位和表达丰度上存在较大差异,而同一类基因之间具有相似的表达规律;4个豆科植物特有的GS1基因在大豆根瘤中都有较高的表达量,其中位于大豆第18染色体上的GmGS1β2基因表达丰度最高;利用原核表达系统体外表达GmGS1β2蛋白,诱导出分子量大小与理论预测值一致的目标蛋白,酶活性分析表明GmGS1β2可以与底物发生催化反应,具有谷氨酰胺合成酶活性,推测该基因在大豆根瘤氮素同化代谢中具有重要作用。     

不同烤烟品种成熟期氮代谢及次生代谢差异研究

为研究不同烤烟品种成熟期氮代谢及次生代谢的差异性，探究不同烤烟品种合适的施氮量。以‘NC71’、‘云烟105’、‘豫烟13号’为试验材料，设置低氮(30 kg/hm² )、中氮(45 kg/hm²)、高氮(60 kg/hm²)3个处理，对成熟期氮代谢关键酶活性及其产物、次生代谢产物的含量进行测定。结果表明，‘NC71’高氮处理、‘豫烟13号’低氮处理、‘云烟105’中氮处理的硝酸还原酶(NR)活性分别为1.474、1.395、1.533 U/g；谷氨酸合成酶(GOGAT)活性分别为218.447、192.276、203.052 nmol/(mg·min)；总酚含量分别为6.652%、6.539%、6.076%，差异均未达到显著水平。相关性分析表明，NR活性与类胡萝卜素的相关系数达到0.855，达到极显著水平，与游离氨基酸、烟碱和总酚的相关系数均在0.6以上，呈显著正相关；谷氨酰胺合成酶(GS)活性与其次生代谢产物的相关系数均在0.5以上，且均达到显著水平。改变NR、GS活性会影响氮代谢次生代谢产物含量，有利于烟叶品质改善。表明不同烤烟品种适宜氮用量存在显著差异，在成熟期利用NR、GS活性去衡量施氮量具有指导意义。     

镉对植物光合作用及氮代谢影响研究进展

镉是毒性最强的环境污染物之一。植物中积累的镉可通过食物链进入人体,给人类健康带来潜在危害。镉能抑制植物生长,能使植物形态、生理生化及结构发生改变。镉可减少净光合速率,损伤光合器官;使叶绿素含量下降;抑制气孔开放,从而影响植物的光合作用;镉通过改变植物硝酸还原酶(NR)、谷胺酰氨合成酶(GS)、谷氨酸合成酶(GOGAT)以及谷氨酸脱氢酶(GDH)的活性,进一步影响植物氮代谢过程。光合作用及氮代谢在植物生长发育过程中起着十分重要的作用,综述了重金属镉对植物光合作用及氮代谢的影响,并对其机理进行了探讨。     

水氮互作下水稻氮代谢关键酶活性与氮素利用的关系

以杂交稻冈优527为材料,设“淹水灌溉”(W₁)、“前期湿润灌溉+孕穗期浅水灌溉+抽穗至成熟期干湿交替灌溉”(W₂)和“旱种”(W₃)3种灌水及不同的施氮量处理,研究对水稻氮代谢酶活性及氮素吸收利用的影响,并探讨各生育期水稻氮代谢酶活性与氮素吸收利用及产量间的关系。结果表明,水与氮对水稻各生育期氮代谢酶活性及氮素吸收利用有显著互作作用,W₂相对于其他灌水处理有助于拔节至抽穗期水稻吸氮量的增加,提高氮素干物质生产效率及稻谷生产效率,而且与施氮量为180 kg hm^-2 耦合能达到提高氮代谢酶活性、增产、提高氮肥利用效率的目的,为本试验最佳的水氮耦合运筹模式;施氮量达270 kg hm^-2 时水氮互作优势减弱,不利于3种灌水方式下硝酸还原酶(NR)、谷氨酰胺合成酶(GS)、谷氨酸合酶(GOGAT)活性的提高,还会导致产量及氮效率的下降。相关分析表明,水氮互作下各氮代谢酶活性与氮素利用特征及产量间存在显著或极显著的相关性,据此可将各生育期功能叶GS活性作为准确判断水稻各生育期氮素积累量的指标;并可将抽穗期剑叶中NR、GS、GOGAT及内肽酶(EP)活性作为综合评价水稻产量及氮效率的指标。     

烤烟根系激素水平、GS、PMT对氮素形态的响应

通过水培试验,探讨了不同氮素形态对烤烟苗期根系激素水平与氮代谢酶的影响。研究结果表明,外源铵态氮对烤烟根系及叶片的谷氨酰胺合成酶(GS)具有一定的诱导作用,饼肥浸提液对烤烟叶片GS有激活作用、对根系GS有抑制作用;外源铵态氮对根系烟碱合成关键酶N-甲基腐胺转移酶(PMT)活性有激活作用;不同氮素形态对根系激素的影响不同,纯铵态氮处理的IAA含量最高,纯硝态氮处理的ABA含量最高,铵态氮与硝态氮1∶1处理的ZR含量最高,浸提液对IAA,ABA,ZR,IAA/ABA,ZR/ABA有明显的激活作用。     

长期施肥对夏玉米叶片氮代谢关键酶活性的影响

以“国家褐潮土土壤肥力与肥料效益长期监测基地”的大田肥料长期试验为平台,研究了长期施肥对夏玉米叶片中硝酸还原酶（NR）、谷氨酰胺合成酶(GS)、谷氨酸合成酶(GOGAT)和谷氨酸脱氢酶(GDH）活性的影响。结果表明,长期N、P、K化肥配合（NPK处理）以及N、P、K化肥与有机肥或秸秆配合施用（NPKM与NPKS处理）,叶片氮代谢酶（NR、GS、GOGAT和GDH）在籽粒灌浆时期均具有较高的活性,有利于玉米籽粒产量的形成;长期N、P、K化肥非平衡施肥的处理（NP、N、NK、PK处理）以及不施肥的CK,叶片氮代谢酶活性变化差异较大,且活性高峰期和籽粒灌浆关键期时间不一致,植株整体生产能力较弱,玉米籽粒产量较低。不同的酶对籽粒产量的贡献不同,因而系统研究长期定位配比施肥处理下玉米叶片的氮代谢酶活性具有重要的意义。     

蛋白和核酸合抑制剂对氮素诱导甜菜谷氨酰胺合成酶基因表达的影响

谷氨酰胺合成酶(GS)家族是甜菜等高等植物体内氨态氮同化酶, 也是氮利用与循环的核心构件。为了揭示在氮素诱导下, 放线菌素D(AMD)和放线菌酮(CHM)对甜菜GS基因调控表达的影响。采用半定量RT-PCR技术, 对甜菜的胞液型谷氨酰胺合成酶基因(GS1)和质体型谷氨酰胺合成酶基因(GS2)进行mRNA的表达检测, 同时进行GS活性的测定。结果表明, 甜菜幼苗经过低浓度AMD处理2~6 h, GS活性略有增加, 9 h后, 高和低浓度AMD处理下的GS活性都下降, 且随着浓度的增加下降幅度加大, 同时GS1mRNA和GS2mRNA的相对量随浓度的增加而下降。CHM处理甜菜幼苗9 h后, 随着浓度的增加和处理时间的延长, GS活性下降幅度增加, 但GS1mRNA和GS2mRNA的相对量在不同CHM浓度处理间变化不显著。     

Effect of Different Crop Densities of Winter Wheat on Recovery of Nitrogen in Crop and Soil within the Growth Period

Previous experiments have shown that, at harvest of winter wheat, recovery of fertilizer N applied in early spring [tillering, Zadok’s growth stage (GS) 25] is lower than that of N applied later in the growth period. This can be explained by losses and immobilization of N, which might be higher between GS 25 and stem elongation (GS 31). It was hypothesized that a higher crop density (i.e. more plants per unit area) results in an increased uptake of fertilizer N applied at GS 25, so that less fertilizer N is subject to losses and immobilization. Different crop densities of winter wheat at GS 25 were established by sowing densities of 100 seeds m^–2 (S_low), 375 seeds m^–2 (S_cfp= common farming practice) and 650 seeds m^–2 (S_high) in autumn. The effect of sowing density on crop N uptake and apparent fertilizer N recovery (aFNrec = N in fertilized treatments ? N in unfertilized treatments) in crops and soil mineral N (N_min), as well as on lost and immobilized N (i.e. non‐recovered N = N rate ? aFNrec), was investigated for two periods after N application at GS 25 [i.e. from GS 25 to 15 days later (GS 25 + 15d), and from GS 25 + 15d to GS 31] and in a third period between GS 31 and harvest (i.e. after second and third N applications). Fertilizer N rates varied at GS 25 (0, 43 and 103 kg N ha^–1), GS 31 (0 and 30 kg N ha^–1) and ear emergence (0, 30 and 60 kg ha^–1). At GS 25 + 15d, non‐recovered N was highest (up to 33 kg N ha^–1 and up to 74 kg N ha^–1 at N rates of 43 and 103 kg N ha^–1, respectively) due to low crop N uptake after the first N dressing. Non‐recovered N was not affected by sowing density. Re‐mineralization during later growth stages indicated that non‐recovered N had been immobilized. N uptake rates from the second and third N applications were lowest for S_low, so non‐recovered N at harvest was highest for S_low. Although non‐recovered N was similar for S_cfp and S_high, the highest grain yields were found at S_cfp and N dressings of 43 + 30 + 60 kg N ha^–1. This combination of sowing density and N rates was the closest to common farming practice. Grain yields were lower for S_high than for S_cfp, presumably due to high competition between plants for nutrients and water. In conclusion, reducing or increasing sowing density compared to S_cfp did not reduce immobilization (and losses) of fertilizer N and did not result in increased fertilizer N use efficiency or grain yields.     

玉米不同组织器官谷氨酰胺合成酶同工酶表达差异及聚合方式

谷氨酰胺合成酶(GS)是作物氮同化及转移利用的关键酶,本试验研究了玉米灌浆期不同组织器官的GS同工酶表达特性,鉴定了玉米GS同工酶的聚合方式。Western blot结果表明,玉米不同组织器官的GS同工酶亚基表达存在明显差异,分子量约40 kD的GS1亚基在所有组织中均大量表达,39 kD的GS1亚基仅在穗位节及穗柄中大量表达,分子量约44 kD的GS2亚基在叶片等光合组织中微量表达。通过改进BNE技术,结合胶内转移酶活性的测定,分析了玉米GS同工酶全酶的大小;利用2-D胶结合Western blot鉴定了GS同工酶相应的亚基组成。结果表明,在玉米组织鉴定出3种分子量不同的GS同工酶,GS2全酶分子量约460 kD,为十聚体;GS1全酶有2种聚合状态,一种是分子量约410 kD的十聚体,另一种是分子量约240 kD的五聚体形式,可见玉米GS同工酶表达存在多种方式。     

Prospects for genomic selection in forage plant species

Genomic selection (GS) is a powerful method for exploitation of DNA sequence polymorphisms in breeding improvement, through the prediction of breeding values based on all markers distributed genome‐wide. Forage grasses and legumes provide important targets for GS implementation, as many key traits are difficult or expensive to assess, and are measured late in the breeding cycle. Generic attributes of forage breeding programmes are described, along with status of genomic resources for a representative species group (ryegrasses). Two schemes for implementing GS in ryegrass breeding are described. The first requires relatively little modification of current schemes, but could lead to significant reductions in operating cost. The second scheme would allow two rounds of selection for key agronomic traits within a time period previously required for a single round, potentially leading to doubling of genetic gain rate, but requires a purpose‐designed reference population. In both schemes, the limited extent of linkage disequilibrium (LD), which is the major challenge for GS implementation in ryegrass breeding, is addressed. The strategies also incorporate recent advances in DNA sequencing technology to minimize costs.     

Effects of High Temperature on Key Enzymes Involved in Starch and Protein Formation in Grains of Two Wheat Cultivars

High temperature is a major determinant of grain growth and yield formation in wheat. The present study was undertaken to investigate the effects of high temperature regimes on the activities of key regulatory enzymes involved in starch and protein accumulation in grains of two winter wheat (Triticum aestivum L.) cultivars Yangmai 9 and Xuzhou 26 with different protein contents. Four day/night temperature regimes of 34 °C/22 °C, 32 °C/24 °C, 26 °C/14 °C and 24 °C/16 °C were established after anthesis, resulting in two daily temperature levels of 28 °C and 20 °C and two diurnal temperature differences of 12 °C and 8 °C. The activities of glutamine synthase (GS) in flag leaves and glutamate pyruvic aminotransferase (GPT), sucrose synthase (SS), soluble starch synthase (SSS) and granule‐bound starch synthase (GBSS) in grains were measured during the periods of grain filling. High temperature reduced both content and yield of starch in grains, while enhanced protein content and reduced protein yield in grains. High temperature significantly enhanced the activities of SS and GBSS on 14 days after anthesis (DAA). High temperature affected SSS slightly in Yangmai 9, but reduced SSS activity markedly in Xuzhou 26 on 14 DAA. However, at the middle and late stages of grain filling, high temperature reduced the activities of SS, GBSS and SSS significantly in the two wheat cultivars. High temperature reduced GPT activity in grains in the two wheat cultivars, but reduced GS activity in flag leaves of Yangmai 9 and enhanced GS activity of Xuzhou 26 on 14 DAA. In addition, under the same high temperature level, SS activity was higher at 34 °C/22 °C, whereas the activities of SSS and GBSS were higher at 32 °C/24 °C. Also, diurnal temperature differences affected GPT and GS activities differently between the two cultivars. Under optimum temperature level, the activities of key enzymes for starch and protein synthesis were higher at 26 °C/14 °C. The activities of SS, SSS and GBSS significantly correlated with starch accumulation in grains, except for GBSS activity to starch content on 14 DAA. GPT activity was positively correlated with protein yield, and GS activity was negatively correlated with protein yield on 14 DAA, while the activities of both GPT and GS were negatively related to protein content in grains.     

Genomic prediction for yields,processing and nutritional quality traits in cultivated potato (Solanum tuberosum L.)

Current potato breeding approaches are hampered by several factors including costly seed tubers, tetrasomic inheritance and inbreeding depression. Genomic selection (GS) demonstrated interesting results regardless of the ploidy level, and can be harnessed to circumvent these problems. In this work, three GS models were evaluated using 50,107 informative SilicoDArT markers and 11 traits in two values for cultivation and use (VCU) potato trials. Two key breeding problems modelled included predicting the performance of (i) new and unphenotyped clones (cross‐validation) and (ii) a VCU using another as training set (TS). GS models performed comparably. Cross‐validation accuracy was high for D35, D45, DMW and BVAL, in ascending order. Prediction accuracies of the VCUs were highly correlated, but the best prediction was obtained for the smaller VCU using the bigger as TS. Cross‐validation and VCU prediction accuracies were higher when bigger TSs were used. The findings herein indicate that GS can be attractively integrated in potato breeding, particularly in early clonal generations to predict and select for traits with low heritability which would otherwise require more testing years, environments and resources.     

镁营养对高温强光下烟草碳氮代谢的影响研究

为探究镁营养对高温强光胁迫下烟草碳氮代谢的影响,以烟草‘翠碧一号’为材料,设置4个镁浓度(0、12.0、48.0、120.0 mg/L)与33℃高温下2个光强[600、1200 µmol/(m²·s)]处理,分析不同镁供应浓度和光强处理对烟草植株糖类物质积累、碳氮代谢生理以及钾钙含量的影响。结果表明：（1）镁供应和光强均显著影响烟株地上和地下部的蔗糖和可溶性糖含量,且光强对烟株淀粉含量也有显著影响。强光能使烟株地上和地下部可溶性糖累积,在镁营养为48.0 mg/L时地上部可溶性糖达到最大值。（2）随着镁离子浓度的增加,烟株的蔗糖合成酶(SS)、蔗糖磷酸合成酶(SPS)、酸性转化酶(AI)、谷氨酰胺合成酶(GS)和硝酸还原酶(NR)等碳氮代谢关键酶的活性呈现先升高后降低的趋势,在48.0 mg/L时活性达到最大,且强光处理下活性均高于正常光。镁浓度、光强及镁浓度×光强对烟株地上部NR、AI活性的影响均达1%极显著水平。（3）高温强光促进烟株对钾、钙离子的吸收,但随着镁离子浓度的增加,吸收受到抑制。镁浓度、光强及镁浓度×光强对地下部钾和钙含量的影响均达5%显著水平。适宜的镁营养(48.0 mg/L)增强了烟株碳氮关键酶的活性,促进了碳氮代谢,提高了可溶性糖的积累,从而缓解高温强光对烟株的伤害。     

乙矮合剂对不同密度夏玉米花粒期叶片氮素同化与早衰的影响

为探讨乙矮合剂调控夏玉米氮素同化和防止后期早衰的生理机制,为建立华北夏玉米区密植高产稳产化学调控技术提供理论依据,2013—2014年在中国农业科学院新乡试验站,以中单909和浚单20为材料,设置乙矮合剂(ECK)和密度梯度处理,研究密度梯度对花后玉米穗位叶氮同化特征和早衰的影响,以及ECK的化学调控效应。结果表明,灌浆期(花后0~40 d),穗位叶硝酸还原酶(NR)活性和谷氨酰胺合成酶(GS)活性随密度增加而显著下降;灌浆后期(花后30~40 d),谷草转氨酶(GOT)和谷丙转氨酶(GPT)活性随种植密度增加而显著降低。穗位叶叶绿素相对含量、可溶性蛋白含量和游离氨基酸含量在灌浆中后期(花后20~40 d)随种植密度增加而显著降低;两品种产量在7.5万株hm–2密度达最大值,7.5~10.5万株hm–2密度群体产量下降,高密群体易发生早衰。ECK处理显著提高了各密度群体灌浆中后期(花后20~40 d)穗位叶NR活性、GS活性、游离氨基酸含量、可溶性蛋白含量和叶绿素含量;显著提高了高密群体(7.5~10.5万株hm–2)GOT活性和GPT活性;较高密群体下(7.5~10.5万株hm–2),中单909和浚单20较各自对照的增产幅度分别为5.59%~6.63%和6.73%~8.10%。ECK处理提高了高密群体夏玉米穗位叶片氮代谢关键酶活性及其产物含量,保证密植群体氮代谢正常进行,有效防止早衰及提高产量。综上所述,采用合理的种植密度并结合喷施乙矮合剂可作为华北夏玉米区高产栽培的重要技术措施。     

水稻生理生化特性对氮肥的反应及与氮利用效率的关系

选用水稻氮高效基因型IR72和9311及氮低效基因型Lemont和PECOS,采用土培方法,在5个施氮量（0、0.51、1.02、1.53、2.04 g N 钵^-1,分别相当于0、75、150、225、300 kg N hm^-2）处理下,研究了生理生化特性对氮肥的反应及与氮效率的关系。结果表明,在幼穗分化期,氮高效基因型水稻的可溶性蛋白含量相对低,而谷氨酰胺合成酶（GS）活性高;不同氮效率基因型间1,5-二磷酸核酮糖羧化酶/加氧酶(Rubisco)含量的差异不大;两种氮效率基因型间的净光合速率（P_n）在幼穗分化期差异不明显。而在齐穗期,氮低效基因型的P_n比高效基因型的低28.66%左右;氮低效基因型在两个时期的单位叶绿素光合速率（P_n/Chl）比氮高效基因型分别低18.51%和29.67%左右。在成熟期,氮高效基因型干物质积累能力强,籽粒产量高。这些结果说明氮效率不同的基因型对氮肥的生理反应差异大。相关性分析表明,低氮水平时(0~1.53 g N 钵^-1), GS酶活性与收获时生物量呈显著或极显著正相关;氮肥偏生产力（PFP）、氮肥农学利用率（AE）及氮素生理利用率（NUEb）分别与GS活性、P_n/Chl和齐穗期的Pn呈显著正相关,而与可溶性蛋白含量、Rubisco含量显著负相关;氮肥吸收效率（RE）与这些生理指标没有显著相关。结果表明水稻光合特征及氮代谢与水稻氮效率间存在紧密的关系,GS活性和可溶性蛋白含量对评价水稻氮肥利用率具有重要的参考价值。