The role of OsBRI1 and its homologous genes, OsBRL1 and OsBRL3, in rice

Since first identifying two alleles of a rice (Oryza sativa) brassinosteroid (BR)-insensitive mutant, d61, that were also defective in an orthologous gene in Arabidopsis (Arabidopsis thaliana) BRASSINOSTEROID INSENSITIVE1 (BRI1), we have isolated eight additional alleles, including null mutations, of the rice BRI1 gene OsBRI1. The most severe mutant, d61-4, exhibited severe dwarfism and twisted leaves, although pattern formation and differentiation were normal. This severe shoot phenotype was caused mainly by a defect in cell elongation and the disturbance of cell division after the determination of cell fate. In contrast to its severe shoot phenotype, the d61-4 mutant had a mild root phenotype. Concomitantly, the accumulation of castasterone, the active BR in rice, was up to 30-fold greater in the shoots, while only 1.5-fold greater in the roots. The homologous genes for OsBRI1, OsBRL1 and OsBRL3, were highly expressed in roots but weakly expressed in shoots, and their expression was higher in d61-4 than in the wild type. Based on these observations, we conclude that OsBRI1 is not essential for pattern formation or organ initiation, but is involved in organ development through controlling cell division and elongation. In addition, OsBRL1 and OsBRL3 are at least partly involved in BR perception in the roots.     

Engineering OsBAK1 gene as a molecular tool to improve rice architecture for high yield

Generating a new variety of plant with erect-leaf is a critical strategy to improve rice grain yield, as plants with this trait can be dense-planted. The erect-leaf is a significant morphological trait partially regulated by Brassinosteroids (BRs) in rice plants. So far, only a few genes can be used for molecular breeding in rice. Here, we identified OsBAK1 as a potential gene to alter rice architecture. Based on rice genome sequences, four closely related homologs of Arabidopsis BAK1 ( AtBAK1 ) gene were amplified. Phylogenetic analysis and suppression of a weak Arabidopsis mutant bri1-5 indicated that OsBAK1 (Os08g0174700) is the closest relative of AtBAK1. Genetic, physiological, and biochemical analyses all suggest that the function of OsBAK1 is conserved with AtBAK1 . Overexpression of a truncated intracellular domain of OsBAK1 , but not the extracellular domain of OsBAK1 , resulted in a dwarfed phenotype, similar to the rice BR-insensitive mutant plants. The expression of OsBAK1 changed important agricultural traits of rice such as plant height, leaf erectness, grain morphologic features, and disease resistance responses. Our results suggested that a new rice variety with erect-leaf and normal reproduction can be generated simply by suppressing the expression level of OsBAK1 . Therefore, OsBAK1 is a potential molecular breeding tool for improving rice grain yield by modifying rice architecture.     

种植密度对夏玉米产量和源库特性的影响

以高产玉米品种郑单958(ZD958)和登海661(DH661)为试验材料,在4个不同区域(山东农业大学、汶口、兖州和莱州)设置22500、45000、67500、90000和112500株.hm-25个种植密度,研究了种植密度对夏玉米产量及源库特性的影响.结果表明:两品种在112500株.hm-2密度条件下玉米籽粒产量和生物产量最高,分别为19132和36965kg.hm-2,与22500和67500株.hm-2密度相比,籽粒产量分别增加了72%和48%,生物产量分别增加了152%和112%.两品种单株叶面积、最大花丝数、穗粒数和千粒重随密度增大而减小,但叶面积指数随密度增大而显著提高.收获指数与粒叶比随密度增大而显著减小,当密度超过67500株.hm-2时差异不显著,表明高密度条件下玉米通过增加群体库来提高产量.     

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.     

水稻冠层光截获、光能利用与产量的关系

以两优培九和武香粳14号水稻品种为材料,在不同栽插密度和施氮水平下进行2年田间试验,研究水稻冠层光合有效辐射(PAR)截获率、光能利用率与水稻产量的关系.结果表明:分蘖期至成熟期,各处理水稻冠层平均PAR反射率为3.45％,其中,分蘖期至抽穗期的冠层反射PAR占冠层总PAR损失的10.90％,显著小于抽穗期至成熟期的22.06％.分蘖期至成熟期的冠层PAR转化率随栽插密度的增加而减少,随施氮量的增加而增大;分蘖期至抽穗期的冠层PAR转化率高于抽穗期至成熟期.在分蘖期至成熟期,冠层PAR利用率随栽插密度和施氮量的增加而增大,各处理中两优培九的平均PAR利用率(1.83 g· MJ-1)显著高于武香粳14(1.42 g·MJ-1);武香粳14因生育期较长,分蘖期至成熟期的入射PAR及中、高栽插密度处理的PAR截获量均高于两优培九.水稻不同生长阶段冠层PAR截获率和利用率与产量呈显著正相关,PAR转化率与产量也呈正相关,但相关性不显著.因此,在保持较高PAR截获率的基础上提高冠层PAR转化率,进而提高冠层PAR利用率,有利于水稻高产.     

Over-expression of rice OsAGO7 gene induces upward curling of the leaf blade that enhanced erect-leaf habit

High-yield cultivars are characterized by erect leaf canopies that optimize photosynthesis and thus favor increased biomass. Upward curling of the leaf blade (called rolled leaf) can result in enhanced erect-leaf habit, increase erect duration and promote an overall erect leaf canopy. The rice mutant R05, induced through transferred DNA (T-DNA) insertion, had the rolled-leaf trait. The leaves in the wild type demonstrated natural drooping tendencies, resulting in decreasing leaf erection indices (LEIs) during senescence at the 20th day after flowering. Conversely, LEIs of the leaves in R05 remained high, even 20-day post-flowering. We applied T-DNA tagging and isolated a rolled-leaf gene from rice which, when over-expressed, could induce upward curling of the leaf blade. This gene encodes for a protein of 1,048 amino acids including the PAZ and PIWI conserved domains, belonging to the Argonaute (AGO) family. There are at least 18 members of the AGO family in rice. According to high-sequence conservation, the rolled-leaf gene in rice could be orthologous to the Arabidopsis ZIP/Ago7 gene, so we called it OsAGO7. These results provide a possible opportunity for implementing OsAGO7 gene in crop improvement. ZhenYing Shi and Jiang Wang contributed equally to this work.     

RNAi-mediated disruption of squalene synthase improves drought tolerance and yield in rice

About one-third of the world's rice area is in rain-fed lowlands and most are prone to water shortage. The identification of genes imparting tolerance to drought in the model cereal plant, rice, is an attractive strategy to engineer improved drought tolerance not only rice but other cereals as well. It is demonstrated that RNAi-mediated disruption of a rice farnesyltransferase/squalene synthase (SQS) by maize squalene synthase improves drought tolerance at both the vegetative and reproductive stages. Twenty-day-old seedlings of wild type (Nipponbare) and seven independent events of transgenic RNAi lines showed no difference in morphology. When subjected to water stress for a period of 32 d under growth chamber conditions, transgenic positives showed delayed wilting, conserved more soil water, and improved recovery. When five independent events along with wild-type plants were subjected to drought at the reproductive stage under greenhouse conditions, the transgenic plants lost water more slowly compared with the wild type, through reduced stomatal conductance and the retention of high leaf relative water content (RWC). After 28 d of slow progressive soil drying, transgenic plants recovered better and flowered earlier than wild-type plants. The yield of water-stressed transgenic positive plants ranged from 14-39% higher than wild-type plants. When grown in plates with Yoshida's nutrient solution with 1.2% agar, transgenic positives from three independent events showed increased root length and an enhanced number of lateral roots. The RNAi-mediated inactivation produced reduced stomatal conductance and subsequent drought tolerance.     

普通野生稻中增产相关QTL的发掘

普通野生稻(Oryza Rufipogon)是重要的遗传资源,发掘其优良等位基因将对水稻遗传改良产生重要影响。文章从以珍汕97为轮回亲本,普通野生稻为供体的BC2F1群体中选择一个与珍汕97表型明显不同的单株BC2F1-15,经过连续自交获得回交重组自交系BC2F5群体。均匀分布于12条染色体的126个多态性SSR(Simplesequence repeats)标记基因型分析,发现BC2F1-15单株在30%的标记位点为杂合基因型;利用该群体共检测到4个抽穗期、3个株高、4个每穗颖花数、2个千粒重和1个单株产量QTL。在第7染色体RM481-RM2区间,检测到抽穗期、每穗颖花数和产量QTL,野生稻等位基因表现增效作用;其他3个每穗颖花数QTL位点,野生稻等位基因也均具有增效作用。结果表明野生稻携带有增产相关的等位基因,这些有利等位基因无疑是水稻遗传改良可资利用的新资源。     

水稻冠层光截获、光能利用与产量的关系

以两优培九和武香粳14号水稻品种为材料,在不同栽插密度和施氮水平下进行2年田间试验,研究水稻冠层光合有效辐射(PAR)截获率、光能利用率与水稻产量的关系.结果表明： 分蘖期至成熟期,各处理水稻冠层平均PAR反射率为3.45%,其中,分蘖期至抽穗期的冠层反射PAR占冠层总PAR损失的10.90%,显著小于抽穗期至成熟期的22.06%.分蘖期至成熟期的冠层PAR转化率随栽插密度的增加而减少,随施氮量的增加而增大;分蘖期至抽穗期的冠层PAR转化率高于抽穗期至成熟期.在分蘖期至成熟期,冠层PAR利用率随栽插密度和施氮量的增加而增大,各处理中两优培九的平均PAR利用率(1.83 g·MJ^-1)显著高于武香粳14(1.42 g·MJ^-1);武香粳14因生育期较长,分蘖期至成熟期的入射PAR及中、高栽插密度处理的PAR截获量均高于两优培九.水稻不同生长阶段冠层PAR截获率和利用率与产量呈显著正相关,PAR转化率与产量也呈正相关,但相关性不显著.因此,在保持较高PAR截获率的基础上提高冠层PAR转化率,进而提高冠层PAR利用率,有利于水稻高产.     

Responses of Super Rice (Oryza sativa L.) to Different Planting Methods for Grain Yield and Nitrogen-Use Efficiency in the Single Cropping Season

To break the yield ceiling of rice production, a super rice project was developed in 1996 to breed rice varieties with super high yield. A two-year experiment was conducted to evaluate yield and nitrogen (N)-use response of super rice to different planting methods in the single cropping season. A total of 17 rice varieties, including 13 super rice and four non-super checks (CK), were grown under three N levels [0 (N0), 150 (N150), and 225 (N225) kg ha⁻¹] and two planting methods [transplanting (TP) and direct-seeding in wet conditions (WDS)]. Grain yield under WDS (7.69 t ha⁻¹) was generally lower than TP (8.58 t ha⁻¹). However, grain yield under different planting methods was affected by N rates as well as variety groups. In both years, there was no difference in grain yield between super and CK varieties at N150, irrespective of planting methods. However, grain yield difference was dramatic in japonica groups at N225, that is, there was an 11.3% and 14.1% average increase in super rice than in CK varieties in WDS and TP, respectively. This suggests that high N input contributes to narrowing the yield gap in super rice varieties, which also indicates that super rice was bred for high fertility conditions. In the japonica group, more N was accumulated in super rice than in CK at N225, but no difference was found between super and CK varieties at N0 and N150. Similar results were also found for N agronomic efficiency. The results suggest that super rice varieties have an advantage for N-use efficiency when high N is applied. The response of super rice was greater under TP than under WDS. The results suggest that the need to further improve agronomic and other management practices to achieve high yield and N-use efficiency for super rice varieties in WDS.     

一份水稻叶片反卷突变体的遗传分析及电镜显微观察

叶片是植物进行光合作用的重要器官。叶片适度卷曲能够提高水稻(Oryza sativa)生长中后期群体基部的光能利用率, 因而有利于水稻产量的提高。该研究首先在水稻T-DNA插入突变体库中发现一份叶片反卷的突变体。遗传分析表明, 该性状受到1对隐性核基因控制。扫描电镜观察结果显示, 突变体成熟叶片上下表皮的气孔发生了畸变; 且叶片上表皮气孔数目增多, 而下表皮气孔数目与野生型基本相同。叶片横切面电镜观察结果表明, 与野生型相比, 突变体叶片的泡状细胞数目和面积在早期(二叶期)就开始增加, 在成熟期更加明显, 这可能是导致叶片反卷的主要原因。     

紧凑大穗型玉米品种陕单902群体源库性状与产量形成的研究

通过对紧凑大穗型玉米品种陕单902的研究发现,保证适宜种植密度,增加群体总粒数是陕单902高产的基础;建立合理的群全结构和干物质生产体系,提高吐丝至成熟期的干物质生产能力是陕单902高产的关键;协调群体库源关系,提高成粒率是陕单902高产的根本。采用合理密植（不超过60000株／hm^2),宽窄行和双株栽培,保证足量氮肥和适宜氮,磷,钾配比等主要配套技术可改善大穗型玉米品种陕单902的群体库源性状,提高产量潜和。     

Optimizing Hill Seeding Density for High-Yielding Hybrid Rice in a Single Rice Cropping System in South China

Mechanical hill direct seeding of hybrid rice could be the way to solve the problems of high seeding rates and uneven plant establishment now faced in direct seeded rice; however, it is not clear what the optimum hill seeding density should be for high-yielding hybrid rice in the single-season rice production system. Experiments were conducted in 2010 and 2011 to determine the effects of hill seeding density (25 cm×15 cm, 25 cm×17 cm, 25 cm×19 cm, 25 cm×21 cm, and 25 cm×23 cm; three to five seeds per hill) on plant growth and grain yield of a hybrid variety, Nei2you6, in two fields with different fertility (soil fertility 1 and 2). In addition, in 2012 and 2013, comparisons among mechanical hill seeding, broadcasting, and transplanting were conducted with three hybrid varieties to evaluate the optimum seeding density. With increases in seeding spacing from 25 cm×15 cm to 25 cm×23 cm, productive tillers per hill increased by 34.2% and 50.0% in soil fertility 1 and 2. Panicles per m² declined with increases in seeding spacing in soil fertility 1. In soil fertility 2, no difference in panicles per m² was found at spacing ranging from 25 cm×17 cm to 25 cm×23 cm, while decreases in the area of the top three leaves and aboveground dry weight per shoot at flowering were observed. Grain yield was the maximum at 25 cm×17 cm spacing in both soil fertility fields. Our results suggest that a seeding density of 25 cm×17 cm was suitable for high-yielding hybrid rice. These results were verified through on-farm demonstration experiments, in which mechanical hill-seeded rice at this density had equal or higher grain yield than transplanted rice.     

Transformation and Functional Expression of the rFCA-RRM2 Gene in Rice

The primary aim of the present study was to investigate the overexpression of the rice (Oryza sativa L. subsp. japonica var. Zhonghua 11) flowering control gene (rFCA-RRM2) in monocotyledonous model rice. Constitutive expression of rFCA-RRM2 from the Actl-5 rice promoter caused late flowering in transgenic rice and increased grain weight that was more than 50% higher than that of control plants, which is the first demonstration of rFCA-RRM2 being able to increase rice production. Late flowering was accompanied by strong phenotype and some morphological modifications. These observations suggest that rFCA-RRM2 is a useful tool for phenotype improvement and yield enhancement in cereal crops.     

Identification and characterization of OsEBS,a gene involved in enhanced plant biomass and spikelet number in rice

Common wild rice (Oryza rufipogon Griff.) is an important genetic reservoir for rice improvement. We investigated a quantitative trait locus (QTL), qGP5‐1, which is related to plant height, leaf size and panicle architecture, using a set of introgression lines of O. rufipogon in the background of the Indica cultivar Guichao2 (Oryza sativa L.). We cloned and characterized qGP5‐1 and confirmed that the newly identified gene OsEBS (enhancing biomass and spikelet number) increased plant height, leaf size and spikelet number per panicle, leading to an increase in total grain yield per plant. Our results showed that the increased size of vegetative organs in OsEBS‐expressed plants was enormously caused by increasing cell number. Sequence alignment showed that OsEBS protein contains a region with high similarity to the N‐terminal conserved ATPase domain of Hsp70, but it lacks the C‐terminal regions of the peptide‐binding domain and the C‐terminal lid. More results indicated that OsEBS gene did not have typical characteristics of Hsp70 in this study. Furthermore, Arabidopsis (Arabidopsis thaliana) transformed with OsEBS showed a similar phenotype to OsEBS‐transgenic rice, indicating a conserved function of OsEBS among plant species. Together, we report the cloning and characterization of OsEBS, a new QTL that controls rice biomass and spikelet number, through map‐based cloning, and it may have utility in improving grain yield in rice.