氮肥水平对不同基因型水稻氮素利用率、产量和品质的影响

以不同基因型的水稻品种日本晴、N70、N178和OM052为供试品种,氮肥采用尿素,按基肥(70%)和蘖肥(30%)两次施用,设置3个施氮水平(N用量设0、120、270 kg·hm^-2)的田间小区试验,研究氮素水平对水稻产量、氮素利用效率和稻米品质的影响,以期为氮肥合理施用和氮高效水稻品种创制提供科学依据.结果表明:施氮能增加水稻品种产量的原因是提高了有效穗数和每穗实粒数;与对照(0 kg·hm^-2)相比,当施氮量为120和270 kg·hm^-2时,OM052籽粒产量在4个品种中增幅最大,分别为41.1%和76.8%;品种产量增幅不同是由于氮素利用效率的差异,在120、270 kg·hm^-2氮处理下,4个供试品种中,日本晴籽粒产量和氮素农学利用率(40.90 g·g^-1、18.56 g·g^-1)都最低,为氮低效品种,OM052籽粒产量和氮素农学利用率(145.9 g·g^-1、81.24 g·g^-1)都最高,为氮高效品种.施N能够增加各品种的直链淀粉和蛋白质含量,使胶稠度变长,降低垩白率、垩白度和碱消值;随施氮量增加,热浆黏度、峰值黏度、回复值和崩解值递减,而消碱值递增.相关性分析表明,低N水平下,供试品种产量及产量构成因子与外观品质、蒸煮食味的相关性更显著.综上,OM052是一个籽粒产量和氮素利用效率“双高”基因型品种,合理施用氮肥可以显著增加水稻的有效穗数和每穗粒数,改善稻米籽粒品质,实现高产和优质的协同.     

Blocking amino acid transporter OsAAP3 improves grain yield by promoting outgrowth buds and increasing tiller number in rice

Amino acid transporters (AATs) play indispensable roles in nutrient allocation during plant development. In this study, we demonstrated that inhibiting expression of the rice amino acid transporter OsAAP3 increased grain yield due to a formation of larger numbers of tillers as a result of increased bud outgrowth. Elevated expression of OsAAP3 in transgenic plants resulted in significantly higher amino acid concentrations of Lys, Arg, His, Asp, Ala, Gln, Gly, Thr and Tyr, and inhibited bud outgrowth and rice tillering. However, RNAi of OsAAP3 decreased significantly Arg, Lys, Asp and Thr concentrations to a small extent, and thus promoted bud outgrowth, increased significantly tiller numbers and effective panicle numbers per plant, and further enhanced significantly grain yield and nitrogen use efficiency (NUE). The promoter sequences of OsAAP3 showed some divergence between Japonica and Indica rice, and expression of the gene was higher in Japonica, which produced fewer tillers than Indica. We generated knockout lines of OsAAP3 on Japonica ZH11 and KY131 using CRISPR technology and found that grain yield could be increased significantly. These results suggest that manipulation of OsAAP3 expression could be used to increase grain yield in rice.     

Genetic engineering of improved nitrogen use efficiency in rice by the tissue-specific expression of alanine aminotransferase

Nitrogen is quantitatively the most essential nutrient for plants and a major factor limiting crop productivity. One of the critical steps limiting the efficient use of nitrogen is the ability of plants to acquire it from applied fertilizer. Therefore, the development of crop plants that absorb and use nitrogen more efficiently has been a long-term goal of agricultural research. In an attempt to develop nitrogen-efficient plants, rice ( Oryza sativa L.) was genetically engineered by introducing a barley AlaAT ( alanine aminotransferase ) cDNA driven by a rice tissue-specific promoter ( OsAnt1 ). This modification increased the biomass and grain yield significantly in comparison with control plants when plants were well supplied with nitrogen. Compared with controls, transgenic rice plants also demonstrated significant changes in key metabolites and total nitrogen content, indicating increased nitrogen uptake efficiency. The development of crop plants that take up and assimilate nitrogen more efficiently would not only improve the use of nitrogen fertilizers, resulting in lower production costs, but would also have significant environmental benefits. These results are discussed in terms of their relevance to the development of strategies to engineer enhanced nitrogen use efficiency in crop plants.     

氮肥处理对氮素高效吸收水稻根系性状及氮肥利用率的影响

2011—2012年在土培条件下,以氮素吸收效率差异较大的15个常规籼稻为供试材料,研究氮肥运筹对不同氮效率品种根系性状、成熟期吸氮量及氮肥利用率的影响,分析影响氮高效水稻氮素吸收的主要根系性状。结果表明:(1)各氮肥处理下,成熟期吸氮量均表现为氮高效品种氮中效品种氮低效品种。适量增施氮肥及基肥+促花肥处理有利于氮高效品种吸氮量的增加,氮素吸收受品种、氮肥处理的显著影响。(2)在施氮量处理下,氮高效品种单株不定根数、单株根干重、单株不定根总长大或较大,单株根活力在常氮(N2)、高氮(N3)处理下有一定的优势;在施氮时期处理下,氮高效品种单株不定根数、单株不定根总长、单株根干重、单株根系总吸收面积、单株根系活跃吸收面积、抽穗期冠根比多数处理有优势;增施氮肥有利于促进氮高效品种单株不定根总长和单株根活力的提高,适量施氮有利于单株不定根数、单株根干重增加,前期施氮可促进不定根的发生和伸长,后期施氮有利于不定根的充实和根系生理性状的提高。此外,增施氮肥可提高各类品种冠根比;(3)在常氮、高氮处理下,氮高效品种氮肥利用率大于氮中效、氮低效品种。(4)提高单株不定根数、单株不定根总长、单株根活力及抽穗期冠根比有利于各类品种吸氮量的提高,增加根干重对氮高效品种吸氮量的提高也有显著的促进作用。结合相关分析与通径分析结果,抽穗期冠根比及单株不定根数、单株根活力、单株不定根总长、单株根干重是影响氮高效品种吸氮能力的主要根系性状。     

Seasonal and within‐canopy variation in shoot‐scale resource‐use efficiency trade‐offs in a Norway spruce stand

Previous leaf‐scale studies of carbon assimilation describe short‐term resource‐use efficiency (RUE) trade‐offs where high use efficiency of one resource requires low RUE of another. However, varying resource availabilities may cause long‐term RUE trade‐offs to differ from the short‐term patterns. This may have important implications for understanding canopy‐scale resource use and allocation. We used continuous gas exchange measurements collected at five levels within a Norway spruce, Picea abies (L.) karst., canopy over 3 years to assess seasonal differences in the interactions between shoot‐scale resource availability (light, water and nitrogen), net photosynthesis (A_n) and the use efficiencies of light (LUE), water (WUE) and nitrogen (NUE) for carbon assimilation. The continuous data set was used to develop and evaluate multiple regression models for predicting monthly shoot‐scale A_n. These models showed that shoot‐scale A_n was strongly dependent on light availability and was generally well described with simple one‐ or two‐parameter models. WUE peaked in spring, NUE in summer and LUE in autumn. However, the relative importance of LUE for carbon assimilation increased with canopy depth at all times. Our results suggest that accounting for seasonal and within‐canopy trade‐offs may be important for RUE‐based modelling of canopy carbon uptake.     

Development and field performance of nitrogen use efficient rice lines for Africa

Nitrogen (N) fertilizers are a major input cost in rice production, and its excess application leads to major environmental pollution. Development of rice varieties with improved nitrogen use efficiency (NUE) is essential for sustainable agriculture. Here, we report the results of field evaluations of marker‐free transgenic NERICA4 (New Rice for Africa 4) rice lines overexpressing barley alanine amino transferase (HvAlaAT) under the control of a rice stress‐inducible promoter (pOsAnt1). Field evaluations over three growing seasons and two rice growing ecologies (lowland and upland) revealed that grain yield of pOsAnt1:HvAlaAT transgenic events was significantly higher than sibling nulls and wild‐type controls under different N application rates. Our field results clearly demonstrated that this genetic modification can significantly increase the dry biomass and grain yield compared to controls under limited N supply. Increased yield in transgenic events was correlated with increased tiller and panicle number in the field, and evidence of early establishment of a vigorous root system in hydroponic growth. Our results suggest that expression of the HvAlaAT gene can improve NUE in rice without causing undesirable growth phenotypes. The NUE technology described in this article has the potential to significantly reduce the need for N fertilizer and simultaneously improve food security, augment farm economics and mitigate greenhouse gas emissions from the rice ecosystem.     

氮肥运筹对小麦产量、氮素利用效率和光能利用率的影响

连续2年在西南冬麦区的重庆、仁寿、广汉、西昌4个地点,开展3种施氮水平(每公顷纯氮0、120、180 kg,简写为N₀、N₁₂₀、N₁₈₀)和3种氮肥分配模式(N_A:底肥100%;N_B:底肥70%+苗肥30%;N_C:底肥60%+拔节肥40%)的田间试验,监测小麦花后冠层叶片SPAD值、群体光合速率(CAP)、光能利用等生理参数和籽粒产量,计算氮素利用效率、光能利用率等.结果表明: 随施氮水平增加,小麦上三叶SPAD值、CAP、光合有效辐射(PAR)截获率和产量均呈增加趋势,而氮肥农学利用效率、生产效率、吸收效率和利用效率呈降低趋势.氮肥后移的增效作用因施氮水平而异,SPAD于N₁₈₀增效明显,而CAP于N₁₂₀增效明显,不同氮肥管理模式的光能利用率因地点而异.氮肥后移能明显提高小麦氮肥农学效率、生产效率、吸收效率和氮素表观回收率,但氮肥利用效率则略有减少.氮肥后移效果N_C总体优于N_B处理.不同地点比较,广汉的SPAD值、CAP、PAR截获率、氮肥利用参数较高,其产量也相应最高;西昌的产量、SPAD值及氮素利用效率较高,但其光能利用率和CAP较低;重庆和仁寿的SPAD值、光能利用率及氮素利用效率均较低,其产量也最低.小麦生物产量与各地点的籽粒产量、CAP、SPAD值和PAR累积截获量均呈显著或极显著的正相关关系.表明不同生态区域增施氮肥都能促进小麦增产,氮肥后移可进一步优化产量结构、改善氮肥和光能利用效率,但存在年份和地点差异,各地需要制定有针对性的氮肥管理模式.     

Overexpression of the leucine‐rich receptor‐like kinase gene LRK2 increases drought tolerance and tiller number in rice

Drought represents a key limiting factor of global crop distribution. Receptor‐like kinases play major roles in plant development and defence responses against stresses such as drought. In this study, LRK2, which encodes a leucine‐rich receptor‐like kinase, was cloned and characterized and found to be localized on the plasma membrane in rice. Promoter–GUS analysis revealed strong expression in tiller buds, roots, nodes and anthers. Transgenic plants overexpressing LRK2 exhibited enhanced tolerance to drought stress due to an increased number of lateral roots compared with the wild type at the vegetative stage. Moreover, ectopic expression of LRK2 seedlings resulted in increased tiller development. Yeast two‐hybrid screening and bimolecular fluorescence complementation (BiFC) indicated a possible interaction between LRK2 and elongation factor 1 alpha (OsEF1A) in vitro. These results suggest that LRK2 functions as a positive regulator of the drought stress response and tiller development via increased branch development in rice. These findings will aid our understanding of branch regulation in other grasses and support improvements in rice genetics.     

有机肥替代部分化肥对水稻光合速率、氮素利用率和产量的影响

农业生产中合理施用有机肥对实现化肥零增长、提高土壤肥力和保证粮食稳产高产至关重要.本试验在辽宁省沈阳市稻田以'沈农9816'为供试材料,设置7种不同处理,分别为不施氮肥(CK)、低氮150 kg ? hm-2(LN)、中氮240 kg ? hm-2(MN)、高氮330 kg·hm-2(HN)、中氮有机肥替代10％(OM...     

TAC4 controls tiller angle by regulating the endogenous auxin content and distribution in rice

Tiller angle, an important component of plant architecture, greatly influences the grain yield of rice (Oryza sativa L.). Here, we identified Tiller Angle Control 4 (TAC4) as a novel regulator of rice tiller angle. TAC4 encodes a plant‐specific, highly conserved nuclear protein. The loss of TAC4 function leads to a significant increase in the tiller angle. TAC4 can regulate rice shoot gravitropism by increasing the indole acetic acid content and affecting the auxin distribution. A sequence analysis revealed that TAC4 has undergone a bottleneck and become fixed in indica cultivars during domestication and improvement. Our findings facilitate an increased understanding of the regulatory mechanisms of tiller angle and also provide a potential gene resource for the improvement of rice plant architecture.     

长期施肥对水稻光合特性及水分利用效率的影响

在实施了27年的长期田间定位试验区,研究了长期不同施肥对红壤区水稻光合特性及水分利用效率的影响.结果表明:在不施肥(CK)、无机肥(N、NP、NPK)、有机肥(猪粪+紫云英绿肥,M)和无机肥与有机肥配施(NPKM)处理中,长期施用肥料,特别是有机肥与无机肥配施提高水稻各生育期剑叶叶绿素含量、净光合作用速率、气孔导度、蒸腾速率、水分利用效率和水稻产量,降低水稻剑叶胞间CO_2浓度;水稻剑叶叶绿素含量、净光合作用速率、气孔导度、蒸腾速率随发育阶段演进而减小,孕穗期>齐穗期>乳熟期,而胞间CO_2浓度相反;水分利用效率以齐穗期为最大;水稻发育阶段叶绿素含量、净光合速率和水稻产量之间均呈显著正相关;长期施用肥料,特别是有机肥与无机肥配施更有利于红壤区水稻的生长发育、产量和水分利用效率的提高.

Abstract：

A field experiment has being conducted for 27 years in Jinxian County, Institute of Red Soil in Jiangxi Province (116°20'24" E, 28°15'30" N) to study the effects of fertilization on the rice photosynthetic traits and water use efficiency. Four treatments were installed,i. e., no fertilization (CK), chemical fertilization (N, NP, NPK), organic fertilization (M), and chemi-cal and organic fertilization (NPKM). Long-term fertilization, especially treatment NPKM, in- creased the flag leaf chlorophyll content, net photosynthetic rate, stomatal conductance, transpi-ration rate, and water use efficiency of rice at its all growth stages and the rice yield, and de-creased the flag leaf intercellular CO_2 concentration. With the growth of rice, the chlorophyll content, net photosynthetic rate, stomatal conductance, and transpiration rate decreased, but the intercellular CO_2 concentration increased. The water use efficiency was the greatest at full-head-ing stage. There were significant positive correlations between the chlorophyll content and net photosynthetic rate at various growth stages and the rice yield, Long-term fertilization, especially the combined chemical and organic fertilization, was favorable to the rice growth and develop-ment, water use efficiency, and yield production in red soil region.     

Precipitation‐use efficiency along a 4500‐km grassland transect

Aims Clarifying the spatiotemporal variations in precipitation‐use efficiency (PUE), the ratio of vegetation above‐ground productivity to annual precipitation, will advance our understanding of how ecosystems' carbon and water cycles respond to climate change. Our goal is to investigate the variations in PUE at both regional and site scales along a 4500‐km climate‐related grassland transect. Location The Inner Mongolian Plateau in northern China and the Qinghai‐Tibetan Plateau. Methods We collected data on 580 sites from four data sources. The data were acquired through field surveys and long‐term in situ observations. We investigated the relationships between precipitation and PUE at both regional and site scales, and we evaluated the effects of the main biotic and climatic factors on PUE at both spatial scales. Results PUE decreased with decreasing mean annual precipitation (MAP), except for a slight rise toward the dry end of the gradient. The maximum PUE showed large site‐to‐site variation along the transect. Vegetation cover significantly affected the spatial variations in PUE, and this probably accounts for the positive relationship between PUE and MAP. However, there was no significant relationship between inter‐annual variations in precipitation or vegetation cover and PUE within given ecosystems along the transect. Conclusions The findings of this research contradict the prevailing view that a convergent maximum PUE exists among diverse ecosystems, as presented in previous reports. Our findings also suggest the action of distinct mechanisms in controlling PUE at different spatial scales. We propose the use of a conceptual model for predicting vegetation productivity at continental and global scales with a sigmoid function, which illustrates an increasing PUE with MAP in arid regions. Our approach may represent an improvement over use of the popular Miami model.     

施氮和肥料添加剂对水稻产量、氮素吸收转运及利用的影响

在苏南太湖地区开展田间试验,研究了施氮和肥料添加剂对水稻产量、氮素吸收转运及利用的影响.结果表明:施氮对水稻产量、各生育时期植株累积吸氮量、阶段氮累积量和花后氮素转运量具有显著的促进作用(P＜0.01),当施氮量高于200 kg·hm-2时,增施氮肥的增产效应不显著(P＞0.05);花后氮素转运率和氮肥利用率均随施氮量的增加而降低.施用肥料添加剂可进一步提高水稻产量、累积吸氮量、花后氮素转运量和氮肥利用率,且该效应在高施氮量( ≥200 kg·hm-2)条件下表现更明显.本试验条件下不施用肥料添加剂时,施氮150kg·hm-2可同时获得较高的产量和氮肥利用率.     

减量施氮下基肥后移对南方冬小麦产量和氮素利用效率的影响

过量施用氮肥导致氮肥利用率降低,环境风险加大.合理降低施氮量、优化氮肥运筹对于小麦高产高效栽培具有重要意义.本研究采用大田试验,以常规施氮方式(240 kg N·hm^-2, 基肥∶拔节肥∶孕穗肥=5∶3∶2)为对照,研究了不同施氮量(240、180、150 kg N·hm^-2,分别用N₂₄₀、N₁₈₀、N₁₅₀表示)及基苗肥施用时期(基施、4叶期施、6叶期施,分别用L₀、L₄、L₆表示)对小麦产量和氮素利用效率的影响.结果表明: 小麦籽粒产量随施氮量的降低而降低,但N₁₈₀与N₂₄₀处理相比无显著差异,而N₁₅₀处理显著降低;氮肥农学效率和吸收效率均以N₁₈₀处理最高.不同施肥时期间,L₄处理的籽粒产量和氮肥利用率最高.N₁₈₀四叶施肥(N₁₈₀L₄)处理的产量与对照无显著差异,但氮肥利用率显著提高.N₁₈₀L₄处理叶面积指数、旗叶光合速率、叶片氮含量、旗叶硝酸还原酶和谷氨酰胺合成酶活性、拔节后干物质和氮素积累量较对照未显著降低.适量降低氮肥用量配合基肥后移能够提高生育后期光合生产能力和氮素吸收同化能力,在保持高产的条件下实现氮素利用效率的同步提高.     

花期渍水逆境下氮素对油菜产量及氮肥利用效率的影响

温室盆栽试验条件下,设置渍水和对照2个水分处理,每个水分处理下设置3个施氮水平(0.05、0.2、0.3 g N·kg^-1土),研究了花后渍水逆境下氮素营养对两个氮高效基因型‘Monty’、‘湘油15’和两个氮低效基因型‘R210’、‘Bin270’油菜产量、产量性能及氮肥利用效率的影响.结果表明:与对照相比,花后渍水处理显著降低了油菜的单株角果数、千粒重、每角粒数和籽粒产量.在适宜水分条件下,增施氮肥显著增加了油菜籽粒产量,而在渍水逆境处理下,增施氮肥对油菜籽粒产量的形成贡献不大.氮高效基因型较氮低效基因型对花后渍水逆境下的籽粒灌浆充实具有一定的促进作用.在同一水分处理下,花后渍水明显降低了油菜氮肥利用率、氮肥偏生产力、氮肥农学利用率、氮素吸收效率和氮收获指数,渍水显著影响了不同基因型油菜的氮素吸收利用能力,而氮高效基因型在渍水逆境下较氮低效基因型更有利于将氮素转运、再分配到角果中,提高籽粒生产效率.油菜产量性能参数存在显著的水氮互作效应,水分、氮肥及水氮互作对油菜籽粒产量和产量性能参数的影响因基因型的不同而异.     

Increasing the efficiency of CRISPR‐Cas9‐VQR precise genome editing in rice

Clustered regularly interspaced short palindromic repeats‐associated protein 9 (CRISPR‐Cas9) is a revolutionary technology that enables efficient genomic modification in many organisms. Currently, the wide use of Streptococcus pyogenes Cas9 (SpCas9) primarily recognizes sites harbouring a canonical NGG protospacer adjacent motif (PAM). The newly developed VQR (D1135V/R1335Q/T1337R) variant of Cas9 has been shown to cleave sites containing NGA PAM in rice, which greatly expanded the range of genome editing. However, the low editing efficiency of the VQR variant remains, which limits its wide application in genome editing. In this study, by modifying the single guide RNA (sgRNA) structure and strong endogenous promoters, we significantly increased the editing efficiency of the VQR variant. The modified CRISPR‐Cas9‐VQR system provides a robust toolbox for multiplex genome editing at sites containing noncanonical NGA PAM.