Lipid composition and deposition during grain filling in intact barley (Hordeum vulgare) mutant grains as studied by H HR MAS NMR

Conventional barley has an excellent nutritional value, but a lower content of lipids than oat. Nevertheless, barley lipids deserve a focused interest due to their fatty acid composition and their vitamin E composition. In this study, ¹H HR MAS NMR was used to monitor the deposition of lipids during grain filling of conventional barley, and of two barley endosperm mutants with increased lipid content. The lipids in the mutants are primarily synthesized between days 9 and 13, whereas the lipid biosynthesis in the conventional barley took place ten days later. GC analysis on barley flour lipids showed that the mutants and the control exhibit comparable relative levels of the major fatty acids: C16:0, C18:1 and C18:2 despite the higher content of lipids in the mutants. Differences in the minor fatty acid composition between control and mutants were also observed. The degree of unsaturation was found to be increasing during grain filling for both conventional barley and the two mutants.     

Predicting pre-harvest sprouting susceptibility in barley: Looking for “sensitivity windows” to temperature throughout grain filling in various commercial cultivars

Pre-harvest sprouting (PHS) is common in cereals that lack grain dormancy if maturing grain is exposed to rain. This phenomenon leads to immediate loss of seed viability, and since the malting process requires germination, its occurrence is highly undesirable in malting barley crops. Dormancy release rate is genetically and environmentally controlled. We evaluated the effect of temperature during grain filling on the dormancy release pattern (and then on the PHS susceptibility) of grains from five malting barley (Hordeum vulgare L.) cultivars widely sown in Argentina, with the aim of predicting PHS susceptibility of a barley crop from easy-to-gather data. Barley cultivars (Quilmes Ayelén, Q. Palomar, Q. Painé, B1215 and Scarlett) were sown on different dates over a 3-year period for generating variability in the thermal environment during grain filling. The period from pollination to physiological maturity (PM) was adjusted to a thermal time (TT) scale, which was then arbitrarily divided into 50 °C d intervals. Mean air temperature within each interval and for the whole filling period was calculated for the different sowing dates. Dormancy release pattern was followed by determining a weighed germination index (GI) throughout grain filling and maturation. We sought a linear relationship between temperature during grain filling and GI at some moment after PM. For all barley cultivars, except B1215, a significant (p < 0.001) and positive correlation was found between the GI of grains with 10–20% moisture content (fresh basis) and mean temperature within TT intervals located at the last stages of seed development. Then, simply temperature-based models for predicting crop PHS susceptibility were generated for each barley cultivar. Moreover, we intended a single, universal prediction model constructed with data from all cultivars. Two general forms were proposed, but the relationships were slightly less tight when each barley cultivar model was used. A preliminary validation for each cultivar model was done for three genotypes with independent data from four sites of the major barley production area in Argentina. When comparing experimental and field data regressions we did not find significant differences in slope for any cultivar (p > 0.25). However, most of the observed GIs were higher than predicted. This upwards displacement of GI–temperature relationship suggests the role of other environmental factors (i.e. water and soil N availability, day length), differing among tested locations. We are currently evaluating and quantifying the effect of these factors with the aim of improving PHS susceptibility prediction in malting barley crops.     

Phloem sugars and amino acids as potential regulators of hordein expression in field grown malting barley (Hordeum vulgare L.)

The commercial quality of malting barley is dependent on the content and composition of grain proteins which are subjected to nitrogen (N) and sulfur (S) control. In nutrient deficient soils, grain protein content is mainly a consequence of the remobilization efficiency. In order to evaluate the effect of N and S soil availability on phloem amino acid and sugar export rates and B- and C-hordein gene expression during grain filling, a factorial combination of N and S fertilization assay was carried out under field conditions. Besides, several carbon (C) and N metabolites were analyzed in leaves and spikes. We observed that, even under soil N and S availabilities that do not limit yield, N and S fertilization induced significant changes in N and C metabolism. N phloem remobilization was promoted by S fertilization independently of N fertilization and C remobilization was also promoted by S fertilization but only in N fertilized plants. The B- and C-hordein gene expression correlated positively with sugar and amino acid exudation rate, respectively. Our results suggest an important role of the export rate of sugars and amino acids in the regulation of grain prolamine expression.     

Effects of free air carbon dioxide enrichment and nitrogen supply on growth and yield of winter barley cultivated in a crop rotation

The increase in atmospheric CO₂ concentration [CO₂] has been demonstrated to stimulate growth of C₃ crops. Although barley is one of the important cereals of the world, little information exists about the effect of elevated [CO₂] on grain yield of this crop, and realistic data from field experiments are lacking. Therefore, winter barley was grown within a crop rotation over two rotation cycles (2000 and 2003) at present and elevated [CO₂](375 ppm and 550 ppm) and at two levels of nitrogen supply (adequate (N2): 262 kg ha⁻¹ in 1st year and 179 kg ha⁻¹ in 2nd year) and 50% of adequate (N1)). The experiments were carried out in a free air CO₂ enrichment (FACE) system in Braunschweig, Germany. The reduction in nitrogen supply decreased seasonal radiation absorption of the green canopy under ambient [CO₂] by 23%, while CO₂ enrichment had a positive effect under low nitrogen (+8%). Radiation use efficiency was increased by CO₂ elevation under both N levels (+12%). The CO₂ effect on final above ground biomass was similar for both nitrogen treatments (N1: +16%; N2: +13%). CO₂ enrichment did not affect leaf biomass, but increased ear and stem biomass. In addition, final stem dry weight was higher under low (+27%) than under high nitrogen (+13%). Similar findings were obtained for the amount of stem reserves available during grain filling. Relative CO₂ response of grain yield was independent of nitrogen supply (N1: +13%; N2: +12%). The positive CO₂ effect on grain yield was primarily due to a higher grain number, while changes of individual grain weight were small. This corresponds to the findings that under low nitrogen grain growth was unaffected by CO₂ and that under adequate nitrogen the positive effect on grain filling rate was counterbalanced by shortening of grain filling duration.     

Comparing the interactive effects of water and nitrogen on durum wheat and barley grown in a Mediterranean environment

The understanding of the interactive effect of water and N availability, associated with the ability of crops to efficiently use these resources, is a crucial issue for stabilizing cereal production in Mediterranean areas. A 3-year side by side experiment on durum wheat and barley, under different water regimes and nitrogen levels, was carried out in a typical Mediterranean environment of Southern Italy, to identify the outstanding features of these species that contribute to enhanced grain yield and improved water and nitrogen use efficiency.     

大麦气孔发育相关基因HvSPCH的克隆及初步功能分析

气孔是植物与外界进行气体交换的通道,对植物的生长发育至关重要。SPCH转录因子与气孔的发育关系密切,为揭示其在大麦中的功能,以大麦G1614为材料,利用PCR方法克隆大麦SPCH（HvSPCH）基因后,进行生物信息学分析,并对该基因在干旱胁迫条件下的表达模式进行分析。结果表明,HvSPCH基因编码区长为1 098 bp,可编码365个氨基酸,相对分子量为90 504.73 Da,理论等电点为4.96,为疏水的、不稳定的碱性蛋白质。氨基酸序列分析表明,HvSPCH蛋白的氨基酸序列不含有跨膜结构域和信号肽,含有40个磷酸化位点和27个糖基化位点。二级结构分析表明,HvSPCH蛋白包含35.62%的α-螺旋、8.77%的延长链、2.47%的β-转角和53.13%的无规则卷曲。系统进化分析表明,大麦HvSPCH蛋白与水稻、野生二粒小麦、二穗短柄草的SPCH蛋白亲缘关系较近。qRT-PCR分析结果表明,干旱处理4 h和12 h时,HvSPCH基因的相对表达量显著升高,说明其有可能参与大麦的干旱逆境响应过程。     

青稞法尼基转移酶β亚基编码基因HbERA1的克隆及表达分析

ERA1(Enhanced response to ABA)基因编码法尼基转移酶(Farnesyl transferase)β亚基,该酶在干旱胁迫下对ABA信号负向调控因子的修饰起着关键作用。本研究以青稞(Hordeum vulgare subsp.vulgare)抗旱品种喜马拉雅10号为材料,利用RT-PCR技术克隆获得了ERA1基因全长cDNA序列,命名为HbERA1(登录号:KJ699392)。生物信息学分析表明,该基因全长1 401bp,可编码466个氨基酸序列,蛋白分子量为51.14kD,等电点为5.00。Prosite Scan分析结果表明,HbERA1含有多个干旱胁迫响应蛋白的作用位点,如酪蛋白激酶Ⅱ磷酸化位点、N-糖基化位点、蛋白激酶C磷酸化位点及N-豆蔻酰化位点。利用实时定量PCR方法研究了HbERA1在干旱胁迫条件下及复水后不同时间点的表达情况,发现在水分过剩处理下(土壤绝对含水量15.5%),HbERA1在土壤绝对含水量为33.4%时表达量最高,并随着土壤绝对含水量的下降而下调表达;进行干旱胁迫后(15.5%)基因表达量也明显下调表达;复水后表达逐渐恢复,复水8h时超过正常表达水平,表明HbERA1基因可能参与调控水涝和干旱胁迫双重信号传导。     

Antiviral Sulfoquinovosyldiacylglycerols (SQDGs) from the Brazilian Brown Seaweed Sargassum vulgare

Total lipids from the Brazilian brown seaweed Sargassum vulgare were extracted with chloroform/methanol 2:1 and 1:2 (v/v) at room temperature. After performing Folch partition of the crude lipid extract, the lipids recovered from the Folch lower layer were fractionated on a silica gel column eluted with chloroform, acetone and methanol. The fraction eluted with methanol, presented a strong orcinol-positive band characteristic of the presence of sulfatides when examined by TLC. This fraction was then purified by two successive silica gel column chromatography giving rise to fractions F4I86 and F4II90 that exhibited strong activity against herpes simplex virus type 1 and 2. The chemical structures present in both fractions were elucidated by ESI-MS and ¹H/¹³C NMR analysis HSQC fingerprints based on their tandem–MS behavior as sulfoquinovosildiacylglycerols (SQDGs). The main SQDG present in both fractions and responsible for the anti-herpes activity observed was identified as 1,2-di-O-palmitoyl-3-O-(6-sulfo-α-d-quinovopyranosyl)-glycerol.     

Persistence and efficacy of spinosad on wheat,maize and barley grains against four major stored product pests

The insecticidal and residual effect of spinosad on wheat, maize and barley grain was evaluated in the laboratory against adults of Sitophilus oryzae (F.) (Coleoptera: Curculionidae), Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae), Tribolium confusum (DuVal) (Coleoptera: Tenebrionidae), Cryptolestes ferrugineus (Stephens) (Coleoptera: Laemophloeidae) as well as against larvae of T. confusum. Spinosad was applied as a solution to 2 kg lots of each commodity at three concentrations, 0.1, 0.5 and 1 ppm, and the treated grain quantities were kept at 25 °C and 65% RH. Samples were taken from each concentration-commodity combination at the day of storage and every 30 d for 6 consecutive months (6 bioassays). The test species were exposed for 14 d to the samples and mortality and reproduction were assessed over this exposure interval. With the exception of T. confusum, 1 ppm of spinosad was highly effective against the remainder of the tested species and provided protection for a period of storage at least 4 months. Although in general, spinosad performance was not very much affected by the grain type, efficacy on maize was less stable over the 6-month period of storage and declined sooner compared to the other commodities. Spinosad almost suppressed progeny production of R. dominica during the storage period, but did not suppress progeny of the other species, since progeny were recorded even 30 d post application especially with the lowest of the tested concentrations. The results of this study indicated that spinosad may provide suitable protection for 6 months against S. oryzae or R. dominica, but is not suitable for long-term protection against T. confusum or C. ferrugineus.     

Efficacy of entomopathogenic nematodes (Nematoda: Rhabditida) and insecticide mixtures to control Spodoptera frugiperda (Smith, 1797) (Lepidoptera: Noctuidae) in corn crops

The main insect pest in Brazilian corn is fall armyworm, Spodoptera frugiperda (Smith, 1797) (Lepidoptera: Noctuidae). Entomopathogenic nematodes (EPNs) can be used to control this pest, and can be applied together with various insecticides. Thus, the objective of this work was to evaluate the efficacy of mixtures of EPNs and insecticides to control S. frugiperda in corn crops. In laboratory bioassays three species of EPNs were tested (Heterorhabditis indica, Steinernema carpocapsae and Steinernema glaseri) together with 18 registered insecticides to control S. frugiperda in corn. Efficacy of association between insecticides and EPNs on S. frugiperda larvae was evaluated against the insect's third instar, 2 and 4 days after applications in laboratory. Experiments in the field were performed in two consecutive years, with located application of H. indica and S. carpocapsae (250 IJs/cm²) mixed with chlorpyrifos (0.3 L/ha) and lufenuron (0.15 L/ha) on the corn husk. In laboratory, after two days exposure the interaction between chlorpyrifos and H. indica was synergistic, while interaction with cypermethrin, spinosad, methoxyfenozide and deltamethrin + triazofos was additive, as was interaction between lufenuron, chlorpyrifos and cypermethrin with S. carpocapsae. In contrast, the interaction between chlorpyrifos (Vexter™ and Lorsban™) and lufenuron with S. glaseri was synergistic. In the field, the best treatment was the mixture of H. indica with lufenuron (0.15 L/ha), with 62.5% and 57.5% larval mortality in the two evaluation years in the field, respectively.     

Compatibility of entomopathogenic nematodes (Nematoda: Rhabditida) with registered insecticides for Spodoptera frugiperda (Smith, 1797) (Lepidoptera: Noctuidae) under laboratory conditions

The fall armyworm, Spodoptera frugiperda (Smith, 1797) (Lepidoptera: Noctuidae) is considered the main key pest of corn crops in Brazil. Entomopathogenic nematodes (EPNs) may be used to control this pest, applied together with other different entomopathogen agents or phytosanity products in the spraying mixture. Thus, the objective of work was to evaluate the compatibility of EPNs with different insecticides used of S. frugiperda control in laboratory conditions. Three species of EPNs (Heterorhabditis indica, Steinernema carpocapsae and Steinernema glaseri) and 18 insecticides registered to control of S. frugiperda in corn crops were tested. Compatibility of the insecticides with EPNs was evaluated by observing mortality and infectivity of infecting juveniles (IJs) 48 h after immersion in solution of the insecticide formulations. Among all insecticides tested, Lorsban™ (chlorpyrifos), Decis™ (deltamethrin), Match™ (lufenuron), Deltaphos™ (deltramethrin + triazophos), Dimilin™ (diflubenzuron), Stallion™ (gamacyhalothrin), Karate Zeon™ (lambdacyhalothrin) Tracer™ (spinosad), Vexter™ (chlorpyrifos), Galgotrin™ (cypermethrin), Certero™ (triflumuron), and Talcord™ (permethrin) were compatible (class 1) with the three nematode species tested under laboratory conditions.     

Waxy endosperm accompanies increased fat and saccharide contents in bread wheat (Triticum aestivum L.) grain

The contents of fat, starch, pentosan, fructan, β-glucan and several mono- and oligosaccharides in grain were evaluated to find out the possible effects of the Wx-D1 gene of bread wheat using two sets of near-isogenic waxy and non-waxy lines and two low-amylose mutant lines with a common genetic background of Kanto 107. These materials have two non-functional Wx-A1b and Wx-B1b alleles in common. Waxy near-isogenic lines with a non-functional Wx-D1d allele showed consistently increased contents of fat, total fructan, β-glucan, glucose, fructose, sucrose, 1-kestose, 6-kestose, neokestose, nystose and bifurcose compared with non-waxy lines with a functional Wx-D1a allele throughout three growing/harvest seasons. Starch and total pentosan contents were inconsistently influenced by the allelic status of the Wx-D1 locus, while water-soluble pentosan and raffinose contents were not affected. The compositional changes of a low-amylose mutant line with an almost non-functional Wx-D1f allele were closely similar to those of waxy near-isogenic lines, while significantly different changes were barely observed in another low-amylose mutant line with a partly functional Wx-D1g allele in two seasons. These results showed that the Wx-D1 gene has pleiotropic effects on the fat and saccharide contents of bread wheat grain.     

大麦醇溶蛋白启动子HorD的克隆及其种子特异性表达分析

种子特异性启动子的克隆和功能分析不仅有助于阐明作物种子发育和胚乳特异性基因的表达调控机制,也是进行作物品质改良和种子生物反应器在内的植物基因工程改造的基础。本研究从大麦品种Golden pormise中克隆到大麦胚乳特异性启动子HorD,生物信息学分析表明,其具备启动子的基本元件,如A-box、TATA-box、CAAT-box等,此外还含有胚乳特异性表达所需要的元件Prolamin-box。为验证HorD启动子的表达特性,以pU1300载体为骨架,将HorD启动子和新型冠状病毒（SARA-CoV-2）刺突蛋白基因LPS通过双酶切连接的方式构建表达载体phorD-LPS,并利用农杆菌介导的遗传转化试验验证其种子表达特性。结果表明,HorD启动子能驱动LPS基因在转基因大麦各组织中表达,但在根、茎、叶及颖壳中表达量较低,在种子中表达量较高。这说明HorD启动子可以驱动外源目的基因在大麦种子中特异高效表达。     

Enzyme inhibitory and antioxidant activity of Melia azedarach L. naturalized in Anatolia and its phenolic acid and fatty acid composition

In the present study, acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and tyrosinase inhibitory and antioxidant activities of the fruit and leaf extracts of Melia azedarach L. (Meliaceae) of Turkish origin were evaluated. Enzyme inhibitory activity of the extracts was tested in vitro using ELISA microplate reader. Antioxidant activity of the extracts was tested using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferrous ion-chelation, and ferric-reducing antioxidant power (FRAP) assays. Phenolic composition of the extracts was elucidated by high performance liquid chromatography (HPLC) and fatty acid compositions of the fatty oils of the fruits and leaves were elucidated by gas chromatography-mass spectrometry (GC-MS). The ethyl acetate extract from the leaves showed the highest inhibition against AChE (33.63 ± 1.40%) and BChE (92.89 ± 3.05%). The methanol extract from the leaves exerted the best antioxidant activity in DPPH radical scavenging and FRAP assays, while the ethyl acetate extracts of the fruits and leaves had the most notable effect in metal-chelation assay.     

Characterization of Solanum chomatophilum resistance to 2 aphid potato pests, Macrosiphum euphorbiae (Thomas) and Myzus persicae (Sulzer)

The aphids Macrosiphum euphorbiae (Thomas) and Myzus persicae (Sulzer) (Hemiptera: Aphididae) are responsible for yield reduction in potato (Solanum tuberosum) production by direct phloem feeding and by spreading viruses. Breeding resistant traits from Solanum chomatophilum into the potato germplasm provides alternative means to control aphid infestations. Integrated pest management strategy, using plant resistance, benefits from the characterization of the resistance and of its impact on aphid biology. Our objective was to characterize the resistance of S. chomatophilum by assessing the effects of accessions, plant parts on aphid performance, and by assessing the impact of the resistance factors on different aphid developmental stages and on alate morph production. Detailed aphid performance was obtained by measuring fecundity, survival, percentage of nymphs that reached adult moult, and population growth using whole plant and clip cage experimental designs. Accession and plant physiological age, but not aphid developmental stage, influenced all life-history parameters, except for alate morph production which was not induced on the resistant accessions. Plant part influence was independent of plant species and accession. Both experimental designs resulted in congruent resistance levels at the accession level for each of the two aphid species, supporting the use of any of them in S. chomatophilum resistance screening. PI243340 was resistant to both aphid species, while PI365324 and PI310990 were also resistant to M. euphorbiae and M. persicae, respectively.     

Relationship of physicochemical characteristics and hydrolyzed hydroxycinnamic acid profile of barley varieties and nutrient availability in ruminants

The objectives of this study were to (1) investigate the relationship between physicochemical characteristics (mean/median particle sizes, physical hull content) and hydrolyzed hydroxycinnamic acid profile (ferulic acid (FA), para-coumaric acid (pCA), and their ratio) of barley varieties and in situ rumen degradability in dairy cattle; and (2) investigate rumen degradation kinetics of FA and pCA of CDC barley varieties grown in western Canada. Barley variety had a significant effect (P < 0.05) on rumen undegradable fraction of DM, FA, pCA, neutral (NDF) and acid detergent fiber (ADF) at 12 and/or 24 h of rumen incubation. FA in barley grain was more degradable than the pCA (P < 0.05). There were no differences (P > 0.05) in effective degradability of DM (EDDM) and EDFA, but significant differences in EDpCA (P < 0.05). Barley hull was strongly correlated to NDF, ADF, ADL, hemicellulose and cellulose (R > 0.78, P < 0.001) and correlated to FA (R = 0.57, P < 0.05) but not to pCA (R = 0.42, P > 0.05) in original samples. FA and pCA were highly and positively correlated to NDF, ADF, ADL, hemicellulose, cellulose, mean/median particle sizes, and rumen indigestible DM, NDF and ADF at either 12 or 24 h (P < 0.05). Mean/median particle size of barley grain positively influenced the rumen indigestible DM, but not others (FA, pCA). The results implied that reduction of barley hull, FA and pCA contents could increase the degradability of barley grain in rumen. Multi-regression with best model variable selection analysis revealed that FA was the factor most inhibiting to DM degradability of barley in rumen, and was the most effective factor to predict DM degradability, while hull was the most effective factor to determine NDF degradability in rumen. Both hull and FA affected ADF degradability in the rumen. The results indicate that breeding or identifying barley varieties with lower hull and FA contents would result in higher degradability, higher energy density and higher quality barley and improve nutrient availability of barley.     

Comparative overwintering host range of three Adelphocoris species (Hemiptera: Miridae) in northern China

In northern China, Adelphocoris suturalis, Adelphocoris lineolatus and Adelphocoris fasciaticollis (Hemiptera: Miridae) are common pests of cotton and several other crops. These species have vastly diverse geographic distribution, seasonal dynamics and abundance, the underlying causal factors of which are poorly understood. In this study, the importance of a broad range of plant species as overwintering hosts for each Adelphocoris sp. was compared. Nymphal emergence from a total of 126 plant species was monitored at two distinct locations. The eggs of A. suturalis successfully eclosed from un-plowed cotton field soil and 115 plant species, primarily pastures, weeds and agricultural crops. The eggs of A. lineolatus successfully eclosed from 40 plant species, mainly pastures and weeds. Finally, A. fasciaticollis overwintered on 35 plant species, primarily tree species, weeds and agricultural crops. In conclusion, the most common and widely distributed mirid species, A. suturalis, overwintered on a comparatively broader range of plants compared to the other two species. These observations help to understand the differences in geographical distribution and abundance of the three Adelphocoris species, and constitute the basis for forecasting and pest management protocols for Adelphocoris spp. in China.