Gibberellin-responsive and -insensitive dwarfing alleles on wheat performance in contrasting tillage systems

Near-isogenic lines (NILs) of winter wheat varying for alleles for reduced height (Rht), gibberellin (GA) response and photoperiod insensitivity (Ppd-D1a) in cv. Mercia background (rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht8c+Ppd-D1a, Rht-D1c, Rht12) and cv. Maris Widgeon (rht (tall), Rht-D1b, Rht-B1c) backgrounds were compared to investigate main effects and interactions with tillage (plough-based, minimum-, and zero-tillage) over two years. Both minimum- and zero- tillage were associated with reduced grain yields allied to reduced harvest index, biomass accumulation, interception of photosynthetically active radiation (PAR), and plant populations. Grain yields were optimized at mature crop heights of around 740mm because this provided the best compromise between harvest index which declined with height, and above ground biomass which increased with height. Improving biomass with height was due to improvements in both PAR interception and radiation-use efficiency. Optimum height for grain yield was unaffected by tillage system or GA-sensitivity. After accounting for effects of height, GA insensitivity was associated with increased grain yields due to increased grains per spike, which was more than enough to compensate for poorer plant establishment and lower mean grain weights compared to the GA-sensitive lines. Although better establishment was possible with GA-sensitive lines, there was no evidence that this effect interacted with tillage method. We find, therefore, little evidence to question the current adoption of wheats with reduced sensitivity to GA in the UK, even as tillage intensity lessens.

Endophytic bacterial community composition in wheat (Triticum aestivum) is determined by plant tissue type, developmental stage and soil nutrient availability

Aims: To understand effects of tissue type, growth stage and soil fertilisers on bacterial endophyte communities of winter wheat (Triticum aestivum cv. Hereward). Methods: Endophytes were isolated from wheat grown under six fertiliser conditions in the long term Broadbalk Experiment at Rothamsted Research, UK. Samples were taken in May and July from root and leaf tissues. Results: Root and leaf communities differed in abundance and composition of endophytes. Endophytes were most abundant in roots and the Proteobacteria were most prevalent. In contrast, Firmicutes and Actinobacteria, the Gram positive phyla, were most prevalent in the leaves. Both fertiliser treatment and sample time influenced abundance and relative proportions of each phylum and genus in the endosphere. A higher density of endophytes was found in the Nil input treatment plants. Conclusions: Robust isolation techniques and stringent controls are critical for accurate recovery of endophytes. The plant tissue type, plant growth stage, and soil fertiliser treatment all contribute to the composition of the endophytic bacterial community in wheat. These results should help facilitate targeted development of endophytes for beneficial applications in agriculture.

The identification of QTL controlling ergot sclerotia size in hexaploid wheat implicates a role for the Rht dwarfing alleles

The fungal pathogen Claviceps purpurea infects ovaries of a broad range of temperate grasses and cereals, including hexaploid wheat, causing a disease commonly known as ergot. Sclerotia produced in place of seed carry a cocktail of harmful alkaloid compounds that result in a range of symptoms in humans and animals, causing ergotism. Following a field assessment of C. purpurea infection in winter wheat, two varieties ‘Robigus’ and ‘Solstice’ were selected which consistently produced the largest differential effect on ergot sclerotia weights. They were crossed to produce a doubled haploid mapping population, and a marker map, consisting of 714 genetic loci and a total length of 2895 cM was produced. Four ergot reducing QTL were identified using both sclerotia weight and size as phenotypic parameters; QCp.niab.2A and QCp.niab.4B being detected in the wheat variety ‘Robigus’, and QCp.niab.6A and QCp.niab.4D in the variety ‘Solstice’. The ergot resistance QTL QCp.niab.4B and QCp.niab.4D peaks mapped to the same markers as the known reduced height (Rht) loci on chromosomes 4B and 4D, Rht-B1 and Rht-D1, respectively. In both cases, the reduction in sclerotia weight and size was associated with the semi-dwarfing alleles, Rht-B1b from ‘Robigus’ and Rht-D1b from ‘Solstice’. Two-dimensional, two-QTL scans identified significant additive interactions between QTL QCp.niab.4B and QCp.niab.4D, and between QCp.niab.2A and QCp.niab.4B when looking at sclerotia size, but not between QCp.niab.2A and QCp.niab.4D. The two plant height QTL, QPh.niab.4B and QPh.niab.4D, which mapped to the same locations as QCp.niab.4B and QCp.niab.4D, also displayed significant genetic interactions.

Redistribution of Nickel, Cobalt, Manganese, Zinc, and Cadmium via the Phloem in Young and Maturing Wheat

The phloem mobility of heavy metals is relevant to the redistribution of micronutrients and pollutants and, ultimately, to the quality of harvested plant parts. The relative mobility in wheat may vary considerably between different cations. In the study reported here, radio-labeled nickel (Ni), cobalt (Co), manganese (Mn), zinc (Zn) and cadmium (Cd) were introduced into either intact young winter wheat (Triticum aestivum L. cv. Arina) via a leaf flap, or detached maturing shoots via the cut stem. Elements fed into the lamina of the second leaf of 21-day-old plants were translocated to the younger (expanding) leaves and to the roots but not or only in trace amounts to the first (already fully expanded) leaf. The 63Ni and 65Zn were exported more rapidly compared with the other heavy metals. Most of 54Mn was retained in the originally labeled leaf. The peduncle of some maturing shoots was steam-girdled below the ear to distinguish between xylem and phloem transport. This phloem interruption reduced the content of 63Ni in the ear to about 25%. Intermediate effects were observed for 65Zn, 57Co, and 109Cd. Total 54Mn accumulation in the ear was hardly affected by steam-girdling, indicating a transport of this element within the xylem to the ear. These results suggest that the relative phloem mobility of Ni and Zn in young wheat plants and in maturing wheat shoots is higher than the mobility of Co and Cd, whereas the mobility of Mn is very low.

Long-distance transport of alkali metals in maturing wheat

The alkali metals cesium, rubidium, lithium and sodium were introduced together with strontium via flaps into leaf laminas or into the stem of maturing, intact winter wheat (Triticum aestivum L. cv. Arina) grown in a field. Long-distance transport of these elements and the influence of the application date and of different application positions were investigated. The phloem-immobile Sr served as a marker for the distribution of the xylem sap in the plants. Dry matter accumulation in the grains and the transpiration per shoot were not markedly affected by the treatments as compared to control plants. The phloem mobility was rather high for Cs and Rb. Li was almost immobile in the phloem (similarly to Sr). An application into the cut stem xylem below the second leaf node contributed more to the contents in the grains than an application into the flag leaf. An earlier feeding date led to a higher accumulation in the grains. The marked losses of the elements applied during maturation (most pronounced for Li) can be explained by leakage in the rain.

Long-distance transport of cobalt and nickel in maturing wheat

Cobalt, nickel and strontium were introduced via flaps into leaf laminas or into the stem of maturing, intact winter wheat (Triticum aestivum L., cv. `Arina') grown under natural conditions in a field. Long-distance transport of these elements and the influence of the application date and of different application positions were investigated. The dry-matter accumulation in the grains was not markedly affected by the treatments as compared to untreated control plants. The phloem-immobile strontium served as a marker for the distribution of the xylem sap in the plants. After foliar application, nickel accumulated more rapidly and in higher quantities in the grains than cobalt. Therefore, nickel has a slightly better phloem mobility than cobalt. Regardless of the application date, a higher percentage of the two elements was transported from the flag leaf lamina than from the second or third lamina from the top to the grains. These results indicate that the leaf position is highly relevant for the transfer of the heavy metals investigated to the ear. Introduction into the stem led to a higher accumulation of nickel and cobalt in the grains than introduction into one of the leaves. An earlier feeding date caused a higher accumulation of nickel and cobalt in the grains when introduced into the stem. In contrast, no major differences between earlier and later feeding dates were detected when the elements were introduced into the leaves. Losses of the applied elements were detected during maturation and can be explained by leakage in the rain.

Examining wheat yield sensitivity to temperature and precipitation changes for a large ensemble of crop models using impact response surfaces

Impact response surfaces (IRSs) depict the response of an impact variable to changes in two explanatory variables as a plotted surface. Here, IRSs of spring and winter wheat yields were constructed from a 25-member ensemble of process-based crop simulation models. Twenty-one models were calibrated by different groups using a common set of calibration data, with calibrations applied independently to the same models in three cases. The sensitivity of modelled yield to changes in temperature and precipitation was tested by systematically modifying values of 1981-2010 baseline weather data to span the range of 19 changes projected for the late 21st century at three locations in Europe.

Selecting crop models for decision making in wheat insurance

In crop insurance, the accuracy with which the insurer quantifies the actual risk is highly dependent on the availability on actual yield data. Crop models might be valuable tools to generate data on expected yields for risk assessment when no historical records are available. However, selecting a crop model for a specific objective, location and implementation scale is a difficult task. A look inside the different crop and soil modules to understand how outputs are obtained might facilitate model choice. The objectives of this paper were (i) to assess the usefulness of crop models to be used within a crop insurance analysis and design and (ii) to select the most suitable crop model for drought risk assessment in semi-arid regions in Spain. For that purpose first, a pre-selection of crop models simulating wheat yield under rainfed growing conditions at the field scale was made, and second, four selected models (Aquacrop, CERES- Wheat, CropSyst and WOFOST) were compared in terms of modelling approaches, process descriptions and model outputs. Outputs of the four models for the simulation of winter wheat growth are comparable when water is not limiting, but differences are larger when simulating yields under rainfed conditions. These differences in rainfed yields are mainly related to the dissimilar simulated soil water availability and the assumed linkages with dry matter formation. We concluded that for the simulation of winter wheat growth at field scale in such semi-arid conditions, CERES-Wheat and CropSyst are preferred. WOFOST is a satisfactory compromise between data availability and complexity when detail data on soil is limited. Aquacrop integrates physiological processes in some representative parameters, thus diminishing the number of input parameters, what is seen as an advantage when observed data is scarce. However, the high sensitivity of this model to low water availability limits its use in the region considered. Contrary to the use of ensembles of crop models, we endorse that efforts be concentrated on selecting or rebuilding a model that includes approaches that better describe the agronomic conditions of the regions in which they will be applied. The use of such complex methodologies as crop models is associated with numerous sources of uncertainty, although these models are the best tools available to get insight in these complex agronomic systems.

Temperature and precipitation effects on wheat yield across a European transect: a crop model ensemble analysis using impact response surfaces

This study explored the utility of the impact response surface (IRS) approach for investigating model ensemble crop yield responses under a large range of changes in climate. IRSs of spring and winter wheat Triticum aestivum yields were constructed from a 26-member ensemble of process-based crop simulation models for sites in Finland, Germany and Spain across a latitudinal transect. The sensitivity of modelled yield to systematic increments of changes in temperature (-2 to +9°C) and precipitation (-50 to +50%) was tested by modifying values of baseline (1981 to 2010) daily weather, with CO2 concentration fixed at 360 ppm. The IRS approach offers an effective method of portraying model behaviour under changing climate as well as advantages for analysing, comparing and presenting results from multi-model ensemble simulations. Though individual model behaviour occasionally departed markedly from the average, ensemble median responses across sites and crop varieties indicated that yields decline with higher temperatures and decreased precipitation and increase with higher precipitation. Across the uncertainty ranges defined for the IRSs, yields were more sensitive to temperature than precipitation changes at the Finnish site while sensitivities were mixed at the German and Spanish sites. Precipitation effects diminished under higher temperature changes. While the bivariate and multi-model characteristics of the analysis impose some limits to interpretation, the IRS approach nonetheless provides additional insights into sensitivities to inter-model and inter-annual variability. Taken together, these sensitivities may help to pinpoint processes such as heat stress, vernalisation or drought effects requiring refinement in future model development.

Effect of environment and genotype on bread-making quality of spring-sown spring wheat cultivars in China

Improvement of end-use quality in bread wheat depends on a thorough understanding of current wheat quality and the influences of genotype (G), environment (E), and genotype by environment interaction (G x E) on quality traits. Thirty-nine spring-sown spring wheat (SSSW) cultivars and advanced lines from China were grown in four agro-ecological zones comprising seven locations during the 1998 and 1999 cropping seasons. Data on 12 major bread-making quality traits were used to investigate the effect of G, E, and G x E on these traits. Wide range variability for protein quantity and quality, starch quality parameters and milling quality in Chinese SSSW was observed. Genotype and environment were found to significantly influence all quality parameters as major effects. Kernel hardness, flour yield, Zeleny sedimentation value and mixograph properties were mainly influenced by the genetic variance components, while thousand kernel weight, test weight, and falling number were mostly influenced by the environmental variance components. Genotype, environment, and their interaction had important effects on test weight, mixing development time and RVA parameters. Cultivars originating from Zone VI (northeast) generally expressed high kernel hardness, good starch quality, but poor milling and medium to weak mixograph performance; those from Zone VII (north) medium to good gluten and starch quality, but low milling quality; those from Zone VIII (central northwest) medium milling and starch quality, and medium to strong mixograph performance; those from Zone IX (western/southwestern Qinghai-Tibetan Plateau) medium milling quality, but poor gluten strength and starch parameters; and those from Zone X (northwest) high milling quality, strong mixograph properties, but low protein content. Samples from Harbin are characterized by good gluten and starch quality, but medium to poor milling quality; those from Hongxinglong by strong mixograph properties, medium to high milling quality, but medium to poor starch quality and medium to low protein content; those from Hohhot by good gluten but poor milling quality; those from Linhe by weak gluten quality, medium to poor milling quality; those from Lanzhou by poor bread-making and starch quality; those from Yongning by acceptable bread-making and starch quality and good milling quality; and those from Urumqi by good milling quality, medium gluten quality and good starch pasting parameters. Our findings suggest that Chinese SSSW quality could be greatly enhanced through genetic improvement for targeted well-characterized production environments.

A study of environmental and management drivers of non-CO2 greenhouse gas emissions in Australian agro-ecosystems

Australian climate, soils and agricultural management practices are significantly different from those of the northern hemisphere nations. Consequently, experimental data on greenhouse gas production from European and North American agricultural soils and its interpretation are unlikely to be directly applicable to Australian systems. A programme of studies of non-CO2 greenhouse gas emissions from agriculture has been established that is designed to reduce uncertainty of non-CO2 greenhouse gas emissions in the Australian National Greenhouse Gas Inventory and provide outputs that will enable better on-farm management practices for reducing non-CO2 greenhouse gas emissions, particularly nitrous oxide. The systems being examined and their locations are irrigated pasture (Kyabram Victoria), irrigated cotton (Narrabri, NSW), irrigated maize (Griffith, NSW), rain-fed wheat (Rutherglen, Victoria) and rain-fed wheat (Cunderdin, WA). The field studies include treatments with and without fertilizer addition, stubble burning versus stubble retention, conventional cultivation versus direct drilling and crop rotation to determine emission factors and treatment possibilities for best management options. The data to date suggest that nitrous oxide emissions from nitrogen fertilizer, applied to irrigated dairy pastures and rain-fed winter wheat, appear much lower than the average of northern hemisphere grain and pasture studies. More variable emissions have been found in studies of irrigated cotton/vetch/wheat rotation and substantially higher emissions from irrigated maize.

Methane and nitrous oxide fluxes in annual and perennial land-use systems of the irrigated areas in the Aral Sea Basin

Land use and agricultural practices can result in important contributions to the global source strength of atmospheric nitrous oxide (N2O) and methane (CH4). However, knowledge of gas flux from irrigated agriculture is very limited. From April 2005 to October 2006, a study was conducted in the Aral Sea Basin, Uzbekistan, to quantify and compare emissions of N2O and CH4 in various annual and perennial land-use systems: irrigated cotton, winter wheat and rice crops, a poplar plantation and a natural Tugai (floodplain) forest. In the annual systems, average N2O emissions ranged from 10 to 150 μg N2O-N m−2 h−1 with highest N2O emissions in the cotton fields, covering a similar range of previous studies from irrigated cropping systems. Emission factors (uncorrected for background emission), used to determine the fertilizer-induced N2O emission as a percentage of N fertilizer applied, ranged from 0.2% to 2.6%. Seasonal variations in N2O emissions were principally controlled by fertilization and irrigation management. Pulses of N2O emissions occurred after concomitant N-fertilizer application and irrigation. The unfertilized poplar plantation showed high N2O emissions over the entire study period (30 μg N2O-N m−2 h−1), whereas only negligible fluxes of N2O (<2 μg N2O-N m−2 h−1) occurred in the Tugai. Significant CH4 fluxes only were determined from the flooded rice field: Fluxes were low with mean flux rates of 32 mg CH4 m−2 day−1 and a low seasonal total of 35.2 kg CH4 ha−1. The global warming potential (GWP) of the N2O and CH4 fluxes was highest under rice and cotton, with seasonal changes between 500 and 3000 kg CO2 eq. ha−1. The biennial cotton–wheat–rice crop rotation commonly practiced in the region would average a GWP of 2500 kg CO2 eq. ha−1 yr−1. The analyses point out opportunities for reducing the GWP of these irrigated agricultural systems by (i) optimization of fertilization and irrigation practices and (ii) conversion of annual cropping systems into perennial forest plantations, especially on less profitable, marginal lands.

运用减法杂交、差异筛选与信使RNA差别显示聚合酶链式反应克隆冬小麦春化相关基因

对于某些一年生或二年生高等植物，春化作用是诱导其成花的一个重要的环境因子。冬小麦春化进程中存在着一个核酸代谢的关键期，利用分子生物学技术分离特异表达的基因是研究春化诱导成花机理的一个突破口。 利用TRIzol试剂快速提取冬小麦燕大1817(Triticum aestivum L. cv Yanda 1817)未春化、春化4d、春化20d、5d脱春化的胚芽中的总RNA，去除污染的DNA后，将引物P_1(5'TTTTTTTTTTTCA3')、P_2(5'TTTTTTTTTTCC3')与10个碱基的随机引物OPF_1-OPF_(20)、OPG_1-OPG_(20)组成80个引物对，对不同来源的RNA进行差别显示，共显示了大约10,000种mRNA，结果发现了两个仅在春化20d这一关键期表达而在未春化、春化4d、5d脱春化时不表达的春化相关基因(VRG)VRG49与VRG54。Northern分析进一步表明这两个基因仅与春化20d的冬小麦RNA有杂交信号。将VRG49与VRG54亚克隆于pGEM-4Z载体上，利用T_7测序系统获得了VRG49和VRG54的DNA序列，它们的长度分别为307bp与169bp。 春化21d的冬小麦京冬1号(T. aestivum L. cv Jingdong No. 1)胚芽的mRNA在逆转酶作用下反转录成sscDNA杂交，将过量的未春化、脱春化的mRNA与sscDNA杂交，运用磁珠法分离出未杂交上的sscDNA，以特异的sscDNA为模板，用DNA聚合酷I合成了dscDNA。通过对dscDNA内部EcoRI位点的甲基化、末端补平、EcoRI接头的安装、连接进入λgt10载体的EcoRI位置，以及运用包装系统进行体外包装，建立了库容为4 * 10~6pfu的富集低温诱导的冬小麦cDNA噬菌体文库。用来源于未春化、春化21d、脱春化的冬小麦mRNA合成3种cDNA探针，对噬菌斑进行原位杂交，结果筛选出了3个春化相关基因(VRG)VRG79、VRG111和VRG231。Dot blotting与Northern分析表明VRG79仅在冬小麦春化关键期21d表达。运用PCR方法从λgt10DNA中扩增出VRG79片断并亚克隆于PUC18载体上，通过T_7测序系统获得了VGR79的序列，其包括349个碱基。 通过Internet将VRG49、VRG54、VRG79与GenBank、EMBL、DDBJ、PBD中的序列进行同源性分析，结果发现这些基因至少是在植物中新发现的基因，对这些基因推测的一些功能也进行了讨论。

冬小麦春化作用相关基因Verc203的功能鉴定

本文应用反义基因技术对减法杂交和差异筛选所获得的春化相关基因CDNA克隆( Verc203)的功能进行鉴定。将Vcrc203插入PB1221质粒的CaMV35S启动子与GLIS基因之间构成表达载体，采用花粉管途径转基因法导入冬小麦胚，得到300粒种子。将上述处理的冬小麦种子萌发后在2-4℃处理28天，转入自然条件下栽培。以幼叶为材料提取基因组DNA，以Gus基因片断为探针，用点杂交法筛选获得转基因植株．转基因植株在自然条件下生长三个月未见抽穗， 而同样栽培条件下的对照（未经转基因处理的冬小麦，经相同条件春化处理）则全部抽穗。以Vcrc203和Gus基因的部分序列为引物，在转基因材料的基因组DNA中扩增出Vcrc203反义基因序列，Gus活性检测表明Gus基因在转基因小麦的茎尖和叶片中得到表达。上述结果表明Vcrc203的反义基因已整合到冬小麦基因组内，并得到高效表达。转基因植株的发育受到严重影响，显示Vcrc203基因可能参与了冬小麦春化作用对植物发育进程的调控．

植物克隆化基因功能分析及蛋白编码基因分离的研究

直到九十年代，以利用拟南芥菜和金鱼草突变体克隆到花器官发育基因为重要标志的植物发育研究取得了重大突破。但由于开花决定过程分子控制机理研究进展却较为缓慢。以mRNA人手利用差异筛选技术获得春化相关基因克隆(verc203，verc17)，Northem blot初步证明这些克隆对春化处理的特异性。根据反义RNA理论和技术对克隆化verc203基因功能进行研究。另外，从纯化蛋白人手，在金针菇中克隆到新的溶血性毒蛋白编码基因，并在细菌中得到表达，这一实验为利用此途径来克隆发育调控蛋白的编码基因进行了有益的技术路线探索。本研究的具体结果如下： 1． 春化相关基因(verc203)对冬小麦花发育调控 根据反义RNA的理论和技术进一步证明verc203在冬小麦开花启动和花发育过程中的可能控制作用。将verc203 DNA序列插入带有CaMV35S启动子和GUS基因的pBI221表达载体， 使之能在植物中高效表达反义RNA，并以转正义基因植株作为对照。通过花粉管途径转基因方法，得到326粒反义质粒冬小麦转化种子和1 98粒正义质粒冬小麦转化种子．所有转化处理种子和正常小麦种子萌发后，经春化处理，与未春化冬小麦植株一起培养直到对照植株达到成熟期。实验观察发现，转反义基因植株的抽穗受到明显的阻抑，开花过程大大推迟。其中表达较强的6株转基因植株甚至晚花五十多天。还发现一些表达较弱的植株尽管晚花20到30天，但花器官发育却受到影响，如，雄蕊缺失，穗发育异常等。Southemblot和PCR实验结果表明外源基因整合到转基因植株基因组。 Northrenblot以及报告基因(GUS)活性等实验证明反义基因在转反义基因植株RNA水平上得到高效表达。同时，GUS基因在蛋白质水平上得到表达。基于上述实验，有理由认为ver203基因可能是控制冬小麦春化诱导的成花启动和花发育过程中的主要基因之一。 2． 金针菇毒蛋白flammutoxin编码基因的克隆及其在细菌中的表达 利用色谱分离技术及SDS．PAGE电泳技术从金针菇(Flammuleina velutips)分离得到一种分子量为31 kDa、对人血红细胞具有溶血活性的蛋白质(flammutoxin)，通过蛋白质序列微量分析技术测得其N-端3 1个氨基酸顺序。利用氨基酸序列推测其编码基因序列信息，通过RT - PCR克隆到一个由973个核苷酸组成的编码基因，5’端93个核苷酸编码肽链序列与测得flammutoxin N-端氨基酸序列完全相同。基因序列同源性分析表明在GenBank (USA)，EMBL Database (Europe)和DDBJ (Japan)基因序列数据库中未找到与之同源的基因序列。这个基因与载体p-半乳糖苷酶部分基因编码的融合蛋白在E coli细胞中有效地得到表达。表达产物的SDS-PAGE，Western blot和溶血活性实验表明它是flammutoxin蛋白原编码基因。并对融合蛋白的溶血活性降低原因作了讨论。