植物谷氨酰胺合成酶研究进展及其应用前景

氮素是制约作物产量的主要营养元素之一,谷氨酰胺合成酶(Glutamine synthase,GS;EC 6.3.1.2)是氮素代谢途径中的关键酶。目前,拟南芥、水稻、小麦和玉米等植物中的GS成员均已被分离鉴定。研究表明,超表达GS能够提高植物对氮素的利用效率,从而在植株的生长发育特别是产量形成过程中发挥重要作用,但是其功能在不同植物上并不完全一致,可能与GS基因受到转录和翻译后等水平的调控有关。以下综述了植物GS基因分类、QTL定位、对氮素代谢响应、组织表达特异性、生物学功能及其分子调控机制等方面的研究进展,并展望了植物GS基因的应用前景,以期为利用GS基因来提高植物氮素利用效率提供具有参考价值的信息。     

Can less yield more? Is reducing nutrient input into the environment compatible with maintaining crop production?

Plant scientists have long recognized the need to develop crops that absorb and use nutrients more efficiently. Two approaches have been used to increase nutrient use efficiency (NUE) in crop plants. The first involves both traditional breeding and marker-assisted selection in an attempt to identify the genes involved. The second uses novel gene constructs designed to improve specific aspects of NUE. Here, we discuss some recent developments in the genetic manipulation of NUE in crop plants and argue that an improved understanding of the transition between nitrogen assimilation and nitrogen recycling will be important in applying this technology to increasing crop yields. Moreover, we emphasize the need to combine genetic and transgenic approaches to make significant improvements in NUE.     

Differential responses of system of rice intensification (SRI) and conventional flooded-rice management methods to applications of nitrogen fertilizer

Background

Rising food demand, slowing productivity growth, poor N-use efficiency in rice, and environmental degradation necessitate the development of more productive, environmentally-sound crop and soil management practices. The system of rice intensification (SRI) has been proposed as a methodology to address these trends. However, it is not known how its modified crop-soil-water management practices affect efficiency of inorganic nitrogen applications.

Methods

Field experiments investigated the impacts of SRI management practices with different N-application rates on grain yield, root growth and activity, uptake of N and its use-efficiency, leaf chlorophyll content, leaf N-concentration, and photosynthetic rate in comparison with standard management practices for transplanted flooded rice (TFR).

Results

Overall, grain yield with SRI was 49 % higher than with TFR, with yield enhanced at every N application dose. N-uptake, use-efficiency, and partial factor productivity from applied N were significantly higher in SRI than TFR. Higher leaf nitrogen and chlorophyll contents during the ripening-stage in SRI plants reflected delayed leaf-senescence, extension of photosynthetic processes, and improved root-shoot activities contributing to increased grain yield.

Conclusions

Rice grown under SRI management used N fertilizer more efficiently due to profuse root development and improved physiological performance resulting in enhanced grain yield compared to traditional flooded rice.     

Inadequate root uptake may represent a major component limiting rice to use urea as sole nitrogen source for growth

Background and aims

Urea represents over 50 % of nitrogen fertilizers applied worldwide to crop production, however one-third of nitrogen fertilized could be recovered by crops. Previous studies have provided useful knowledge of urea-related plant nitrogen-nutrition, whereas information about crop growth-response to urea without its external degradation seems limiting. We thus assess the ability of rice seedlings to use urea at a physiological level.

Methods

Rice growth on urea versus other nitrogen regimes was tested under aseptic conditions. Activity of urease and GS was analyzed; urea, ammonium, total nitrogen and expression of a nitrogen limitation-responsive gene OsDUR3 were examined.

Results

Growth phenotyping revealed urea-dose-dependent growth improvement but significant growth reduction associated with nitrogen-deficiency of plants compared to those on other nitrogen-sources, indicating a physiological impediment of effective urea utilization by rice. Enzymatic assay showed that activities of urease and GS were well expressed in plants supplied with urea. Low concentrations of urea and ammonium were detected in rice (particularly in roots) on 1 mM urea or other nitrogen-forms, and a less nitrogen-content was determined in urea-fed plants. Additionally, the strongest OsDUR3-expression occurred in seedlings on no-nitrogen or 1 mM urea.

Conclusions

We suggest that insufficient urea-absorption but not assimilation represents likely a factor contraining rice to use urea as sole nitrogen-source.     

The challenge of remobilisation in plant nitrogen economy. A survey of physio-agronomic and molecular approaches

In this article, we discuss the ways in which our understanding of the controls of nitrogen remobilisation in model species and crop plants have been increased through classical physiological studies and the use of transgenic plants or mutants with modified capacities for nitrogen or carbon assimilation and recycling. An improved understanding of the transition between nitrogen assimilation and nitrogen recycling will be vital, if improvements in crop nitrogen use efficiency are to reduce the need for excessive input of fertilisers and improve or stabilise yield. In this review, we present an overall view of past work and more recent studies on this topic, using different plants systems and models depicting the biochemical and molecular events occurring during the transition between sink leaves and source leaves. These models may provide a way to identify the nature of the metabolic or developmental signals triggering in a coordinate manner nitrogen and carbon recycling during leaf senescence. Another way of developing crop varieties with improved nitrogen use efficiency, and identifying key elements controlling the process of nitrogen remobilisation, is the use of quantitative genetics. We present and discuss recent findings on the genetic variability and basis of nitrogen use efficiency in crops in general and in maize in particular. A genetic approach using maize recombinant inbred lines was undertaken allowing the detection of Quantitative Trait Loci (QTLs) for morphological traits, grain yield and its components under high nitrogen or low nitrogen input. Co‐mapping was observed between genes encoding enzymes involved in nitrogen assimilation (nitrate reductase, glutamine synthetase) and these Quantitative Trait Loci. All coincidences were consistent with the expected physiological function of the corresponding enzyme activities. This work strongly suggests that in maize, nitrogen use efficiency can be improved both by marker‐assisted selection and genetic engineering.     

开放式空气CO2浓度增高对水稻N素吸收利用的影响

在大田栽培条件下 ,研究空气中CO2 浓度增高 (FACE) 2 0 0 μmol·mol-1对水稻N素吸收及其利用效率的影响 .结果表明 ,FACE处理使水稻不同生育时期的植株含N率显著下降 ;由于干物质生产量显著增大 ,FACE处理使水稻不同生育时期的N素累积量有所提高 ,但无显著影响 ;FACE处理能够显著提高移栽后 2 8d、抽穗期以及成熟期单位N素的干物质生产效率、单位N素的籽粒生产效率和显著提高水稻的N素收获指数 .高N处理的植株含N率、N素累积量均有所增加 ,但使N素生产效率呈现下降趋势 .     

Nitrogen fixation in rice systems: state of knowledge and future prospects

Rice is the most important cereal crop. In the next three decades, the world will need to produce about 60% more rice than today's global production to feed the extra billion people. Nitrogen is the major nutrient limiting rice production. Development of fertilizer-responsive varieties in the Green Revolution, coupled with the realization by farmers of the importance of nitrogen, has led to high rates of N fertilizer use on rice. Increased future demand for rice will entail increased application of fertilizer N. Awareness is growing, however, that such an increase in agricultural production needs to be achieved without endangering the environment. To achieve food security through sustainable agriculture, the requirement for fixed nitrogen must increasingly met by biological nitrogen fixation (BNF) rather than by using nitrogen fixed industrially. It is thus imperative to improve existing BNF systems and develop N₂-fixing non-leguminous crops such as rice. Here we review the potentials and constraints of conventional BNF systems in rice agriculture, as well as the prospects of achieving in planta nitrogen fixation in rice.     

农业N肥投入与生态经济效益的协调增长

精细养分管理多年的试验结果证明水稻养分投入至少可以比目前减少25％,而产量可上升8％,同时每公顷可以降低淋溶量和气化量近30％。利用TechnoGIH模型计算养分利用效率,其结果表明：目前水稻N的表观吸收利用率仅达到0．2,而采用精细养分管理可提高到0．3,如果采用更先进的技术可以将其提高到0．4,甚至0．5～0．6。而目前对于非稻类作物蔬菜,N的表观吸收利用率仅为0．14。如果改进技术将其提高到0．20,N肥的投入可以减少32％,N的丧失可以减少29％。如果能进一步提高到0．25,至少还可以减少14％的N投入和16％的N丧失。这样淋溶和气化对环境产生的影响可大大降低,从而避免过多的养分通过淋溶渗入土壤和直接挥发到大气中,影响生态环境,影响农业投入产出效益。因此,需要从经济收益、环境效益、社会效益等多方面综合考虑未来农业生态效益协调增长方式。     

Much Improved Water Use Efficiency of Rice under Non-Flooded Mulching Cultivation

Water shortage is increasingly limiting the luxury use of water in rice cultivation. In this study, non-flooded mulching cultivation of rice only consumed a fraction of the water that was needed for traditional flooded cultivation and largely maintained the grain yield. We also investigated the growth and development of rice plants and examined grain yield formation when rice was subjected to non-flooded mulching cultivation. One indica hybrid rice combination was grown in a field experiment and three cultivation methods, traditional flooding （TF）, non-flooded straw mulching cultivation （SM） and non-flooded plastic mulching cultivation （PM）, were conducted during the whole season. Grain yield showed that there was no significant difference between SM and TF rice, but the grain yield of SM cultivation was significantly higher than that of PM. The tiller numbers were inhibited in the early stage under non-flooded mulching cultivation, but the situation was reversed at the later period. Both SM and PM rice reduced dry matter accumulation of shoot, but increased root dry weight, enhanced the remobilization of assimilates from stems to grains and increased the harvest index. During the middle and later grain filling period, mulched plants showed a faster decrease in chlorophyll concentrations, photosynthetic rates of flag leaves and root activity than TF rice, indicating that non-flooded mulching cultivation enhanced plant senescence. In comparison, SM treatment produced higher grain yield and, more dry matter accumulation and panicle numbers than the PM treatment. The overall results suggest that high yield of non-flooded mulching cultivation of rice can be achieved with much improved irrigaUonal water use efficiency.     

不同水分和氮素处理对寒地水稻生育及产量的影响

为了探讨不同水分和氮素处理对寒地水稻生长发育及产量的影响,以水稻品种空育131、龙粳21为试验材料,于2010—2011年度在黑龙江建三江进行水分、氮素处理大田试验,水分为雨养、间歇灌溉、水层灌溉3个水平,氮素为不施氮、常规施氮(112—135 kg/hm2)、高氮(142—173 kg/hm2)3个水平。结果表明:与水层灌溉相比,雨养水稻生育期缩短1—5 d,生长指标明显降低,产量显著降低,间歇灌溉水稻生育期、生长指标与其相似,产量差异不显著。与常规施氮相比,不施氮生育期缩短2—5 d,高氮条件下延长2—4 d;施氮量增加,生长指标增大,产量显著增加;低氮条件下,水分不足的限制作用明显,高氮能一定程度弥补水分的限制,促进水稻生长。增加施氮量及灌溉水平可以显著地提高有效穗数、每穗粒数。在试验条件下,水氮互作效应不显著。间歇灌溉及高氮管理具有较好的增产效应及资源利用率,研究可为寒地水稻生产进行水氮科学管理、实现高产高效提供理论依据。     

长期施肥对黄土高原黄绵土氮肥利用率的影响

氮肥利用率是确定推荐施氮量和施氮效果评价的关键参数.本文通过黄土高原黄绵土区持续34年(1981—2015年)的长期定位试验,研究了长期不同施肥处理对氮肥当季利用率和累积利用率的影响及氮肥当季利用率与累积利用率的关系.结果表明: 除试验起始年(1982)外,不同施肥处理对小麦、油菜和胡麻3种作物氮肥的当季利用率和累积利用率都有显著影响,其中小麦以氮磷钾配施(NPK)处理平均氮肥当季利用率最高,其次为氮磷配施(NP)处理,分别较单施氮肥(N)处理提高了77.7%和62.0%;油菜也以氮磷钾配施(NPK)处理平均氮肥当季利用率最高,其次为有机肥与氮磷钾配施(MNPK)处理,分别较单施氮肥处理提高了93.7%和65.6%.有机肥与氮肥配施(MN)处理氮肥当季利用率较单施氮肥(N)处理显著增加,而有机肥与氮磷配施(MNP)和氮磷钾配施(MNPK)处理氮肥当季利用率较相应氮磷配施(NP)和氮磷钾配施(NPK)处理明显降低.氮肥当季利用率与作物籽粒产量存在显著的线性正相关关系,而累积利用率与作物籽粒产量的相关关系不明显.表明与氮肥累积利用率相比,当季利用率能较及时地反映特定土壤肥力、作物种类、品种和环境条件下的肥料利用率特征.     

有机无机肥配施提高麦-稻轮作系统中水稻氮肥利用率的机制

采用盆栽试验,研究了有机无机肥配施对麦-稻轮作系统中水稻氮素累积动态和土壤氮素供应动态的影响,并从微生物学角度探讨了有机无机肥协同提高水稻氮肥利用率的机制.结果表明：有机无机肥配施处理的土壤微生物生物量碳、氮和矿质态氮在水稻分蘖期前低于化肥处理,而在抽穗期至灌浆期显著高于其他处理.土壤氮素供应动态与水稻吸收利用氮素规律吻合程度最高,促进了水稻产量、生物量和氮素累积量的增加,显著提高了水稻的氮肥利用率.其主要机制是有机无机肥配施促进了土壤微生物繁殖,使其在水稻生育前期固持了较多的矿质氮,在水稻生育中、后期这些氮素逐渐被释放以供水稻吸收利用,较好地满足了水稻各阶段生长发育对氮素养分的需求.     

Physiological nitrogen efficiency in rice: Nitrogen utilization,photosynthesis, and yield potential

Characteristics of rice (Oryza sativa) as a crop plant are briefly introduced, and the relationship between formation of yield potential and nitrogen (N) nutrition is described on the basis of studies using 15N as a tracer. In addition, the relationship between the leaf photosynthetic capacity and leaf N, and the factors limiting leaf photosynthesis under different growth conditions are reviewed. Finally, targets for improving rice yield potential are discussed with a focus on the role of increased photosynthesis efficiency in relation to leaf N status and the photosynthetic components in the leaves.     

Iron homeostasis and fortification in rice

Iron (Fe) is an important micronutrient used by all living organisms for proper development. A deficiency in that element causes a metabolic imbalance that is deleterious to plant growth, making it a significant worldwide health concern. In response to limited Fe supplies, rice plants use a combination of strategies to take up iron from the soil. As a major staple crop, rice varieties that contain high levels of Fe would be vital for improving public nutritional status. Therefore, understanding the molecular components for Fe homeostasis and fortification in rice is crucial to the production of high-iron grains.     

开放式空气CO2浓度增高对水稻N素吸收利用的影响

在大田栽培条件下,研究空气中CO₂浓度增高(FACE)200μmol·mol^-1对水稻N素吸收及其利用效率的影响.结果表明,FACE处理使水稻不同生育时期的植株含N率显著下降;由于干物质生产量显著增大,FACE处理使水稻不同生育时期的N素累积量有所提高,但无显著影响;FACE处理能够显著提高移栽后28d、抽穗期以及成熟期单位N素的干物质生产效率、单位N素的籽粒生产效率和显著提高水稻的N素收获指数.高N处理的植株含N率、N素累积量均有所增加,但使N素生产效率呈现下降趋势.     

Expression of a Late Embryogenesis Abundant Protein Gene,HVA1, from Barley Confers Tolerance to Water Deficit and Salt Stress in Transgenic Rice

A late embryogenesis abundant (LEA) protein gene, HVA1, from barley (Hordeum vulgare L.) was introduced into rice suspension cells using the Biolistic-mediated transformation method, and a large number of independent transgenic rice (Oryza sativa L.) plants were generated. Expression of the barley HVA1 gene regulated by the rice actin 1 gene promoter led to high-level, constitutive accumulation of the HVA1 protein in both leaves and roots of transgenic rice plants. Second-generation transgenic rice plants showed significantly increased tolerance to water deficit and salinity. Transgenic rice plants maintained higher growth rates than nontransformed control plants under stress conditions. The increased tolerance was also reflected by delayed development of damage symptoms caused by stress and by improved recovery upon the removal of stress conditions. We also found that the extent of increased stress tolerance correlated with the level of the HVA1 protein accumulated in the transgenic rice plants. Using a transgenic approach, this study provides direct evidence supporting the hypothesis that LEA proteins play an important role in the protection of plants under water-or salt-stress conditions. Thus, LEA genes hold considerable potential for use as molecular tools for genetic crop improvement toward stress tolerance.     

根际微生物促进水稻氮利用的机制

根际微生物影响植物的生长及环境适应性。不同种属、不同种群的植物影响其环境微生物群落;反之,根际微生物也影响宿主植物生长发育与生态适应性。植物与根际微生物的互作现象及其机制,是生命科学研究关注的热点,也是农业微生物利用的关键问题。近期,中国科学家在该领域取得了突破性进展。通过对不同籼稻(Oryza sativa subsp. indica)和粳稻(O. sativa subsp. japonica)品种的根际微生物组进行研究,发现籼稻根际比粳稻根际富集更多参与氮代谢的微生物群落,且该现象与硝酸盐转运蛋白基因NRT1.1B在籼粳之间的自然变异相关联。通过对籼稻接种籼稻根际特异富集的微生物群体可以提高前者对有机氮的利用,促进其生长。该研究揭示了籼稻和粳稻根际微生物分化的分子基础,展示了利用根际微生物提高水稻营养高效吸收的应用前景。     

Molecular mechanisms underpinning phosphorus‐use efficiency in rice

Orthophosphate (H₂PO₄^?, Pi) is an essential macronutrient integral to energy metabolism as well as a component of membrane lipids, nucleic acids, including ribosomal RNA, and therefore essential for protein synthesis. The Pi concentration in the solution of most soils worldwide is usually far too low for maximum growth of crops, including rice. This has prompted the massive use of inefficient, polluting, and nonrenewable phosphorus (P) fertilizers in agriculture. We urgently need alternative and more sustainable approaches to decrease agriculture's dependence on Pi fertilizers. These include manipulating crops by (a) enhancing the ability of their roots to acquire limiting Pi from the soil (i.e. increased P‐acquisition efficiency) and/or (b) increasing the total biomass/yield produced per molecule of Pi acquired from the soil (i.e. increased P‐use efficiency). Improved P‐use efficiency may be achieved by producing high‐yielding plants with lower P concentrations or by improving the remobilization of acquired P within the plant so as to maximize growth and biomass allocation to developing organs. Membrane lipid remodelling coupled with hydrolysis of RNA and smaller P‐esters in senescing organs fuels P remobilization in rice, the world's most important cereal crop.     

根际微生物促进水稻氮利用的机制

根际微生物影响植物的生长及环境适应性。不同种属、不同种群的植物影响其环境微生物群落; 反之, 根际微生物也影响宿主植物生长发育与生态适应性。植物与根际微生物的互作现象及其机制, 是生命科学研究关注的热点, 也是农业微生物利用的关键问题。近期, 中国科学家在该领域取得了突破性进展。通过对不同籼稻(Oryza sativa subsp. indica)和粳稻(O. sativa subsp. japonica)品种的根际微生物组进行研究, 发现籼稻根际比粳稻根际富集更多参与氮代谢的微生物群落, 且该现象与硝酸盐转运蛋白基因NRT1.1B在籼粳之间的自然变异相关联。通过对籼稻接种籼稻根际特异富集的微生物群体可以提高前者对有机氮的利用, 促进其生长。该研究揭示了籼稻和粳稻根际微生物分化的分子基础, 展示了利用根际微生物提高水稻营养高效吸收的应用前景。