绿色荧光蛋白基因在小麦禾谷镰刀菌中的表达与鉴定

为了获得稳定表达绿色荧光蛋白（GFP）的我国代表性小麦禾谷镰刀菌菌株,用于小麦抗赤霉病接种鉴定与分析,以禾谷镰刀菌武昌菌株Fg175原生质体为受体,经PEG介导将序列优化的水母GFP基因转入该菌株中,分析鉴定了转基因菌株GFP基因的表达。PCR分析表明,Fg175基因组中整合了GFP基因,Western blot检测到高效表达的GFP特异蛋白;激光共聚焦显微镜及体视镜观察进一步证实,转基因菌株Fg175-GFP的分生孢子、菌丝及接种小麦麦穗后,均可发出清晰的绿色荧光。     

Aggressiveness variation of Fusarium graminearum isolates from Argentina following point inoculation of field grown wheat spikes

Aggressiveness variation among isolates of Fusarium graminearum from Argentina was analyzed by following disease development on point inoculated spikes of field grown wheat. Two aspects of the capacity of the isolates to induce disease were observed: (i) the infection efficiency reflected by the ability to produce symptoms and evaluated as the number of symptomatic spikes over the inoculated ones; and (ii) the size of the lesion provoked, which is reflected by the number of symptomatic spikelets over its total number. One hundred and twelve isolates were found to induce significantly different levels of disease severity, reflecting variation in aggressiveness. Differences were also found between the isolates for the thousand kernel weight (TKW) and the area under the disease progress curve (AUDPC) and the correlations between disease severity, AUDPC, and TKW were highly significant. Based on disease severity, isolates were clustered as low, medium and highly aggressive and both lesion size and infection efficiency were significantly different between these groups. The movement of the pathogen in the spike occurred primarily downwards the point of inoculation. Movement toward the upper portion was related with the aggressiveness of the isolates and the premature ripening of the spike appeared to be a function of isolate aggressiveness. In this work point inoculation allowed us to study not only the aggressiveness of a large collection of F. graminearum isolates but also to analyze the development of Fusarium head blight symptoms in large numbers of spikes under conditions similar to those in which this disease naturally occurs.     

Gramineous and non-gramineous weed species as alternative hosts of Fusarium graminearum,causal agent of Fusarium head blight of wheat,in Argentina

Weeds and wild plants around and within crops could serve as alternative hosts of fungal pathogens. In this work we describe the isolation of Fusarium graminearum, the main causal agent of Fusarium head blight (FHB) in Argentina from the inflorescences of healthy weed plants belonging to sixty seven gramineous and non-gramineous species, which showed no symptoms of Fusarium infection, sampled throughout a year. Fifty four of the weed species considered, belonging to 19 botanical families, were first identified as alternative hosts of F. graminearum in the present work. Furthermore, the trichothecene chemotype of a group of isolates was analysed and strains belonging to 15-acetyldeoxynivalenol, 3-acetyldeoxynivalenol and nivalenol chemotypes were found. The information provided could prove valuable to study further the epidemiological role of weeds in FHB epidemics, which might help to improve management of the disease in wheat growing areas.     

Assessment of biochemical markers identified in wheat for monitoring barley grain tissue

The possible use of specific biochemical compounds identified in wheat grains was evaluated for monitoring barley grain tissues during fractionation. First barley grain anatomy was studied through microscopic observation and quantification of the relative proportion of each anatomical part in four distinct barley samples from both hulled and hulless genotypes. As expected from cereal phylogeny and irrespective of the possible presence of hull, common features were observed between barley and wheat grains, but the aleurone layer predominated in the outer layers. The specific location of the compounds identified in wheat was established. Phytic acid was specifically localized in the aleurone layer and alkylresorcinols in the composite layer containing the testa, even if their concentration differed from that observed in wheat grain tissues. Thus, these two markers identified in wheat can be used to monitor the corresponding barley tissues, independent of the presence of hulls. Conversely, phenolic compounds, either ferulic acid trimer or p-coumaric acid, cannot be used to monitor respectively the outer pericarp or the aleurone cell walls in barley grains. p-coumaric acid was identified as an efficient marker of the hull and could be used to distinguish hulled or hulless barley grains and to help monitor the dehulling process.     

Recent findings on certain bioactive components in whole grain wheat and rye

Whole grain wheat and rye are important sources of many bioactive compounds and contribute significantly to the total intake of cereals in many countries. Alkylresorcinols, benzoxazinoids, lignans, phenolic acids, phytosterols and tocols are common bioactive compounds present in these cereals. In this review, we report recent findings (mainly from 2010 onwards) regarding their content, composition, effects of food processing and their uptake, elimination and bioactivities with implications for health.     

Molecular and biochemical impacts of environmental factors on wheat grain development and protein synthesis

Wheat grain yield and flour quality are strongly influenced by the effects of environment during grain fill. Environmental variables such as temperature, water and fertilizer influence the rate and duration of wheat grain development, protein accumulation and starch deposition in unique ways, and by different mechanisms. The effects of environment are superimposed on the intrinsic temporal patterns of gene expression during grain development. Integration of genomic and proteomic studies with developmental studies under controlled environmental conditions should make it possible to resolve complex patterns of gene expression during grain development, pinpoint key regulatory processes that are influenced by the environment, and reveal the molecular basis for environmental impacts on flour composition and quality.     

Effect of tolerance to Septoria tritici blotch on grain yield,yield components and grain quality in Argentinean wheat cultivars

Septoria tritici blotch (STB) caused by Zymoseptoria tritici (Mycosphaerella graminicola) is a major disease of wheat worldwide due to significant losses in grain yield and quality. Disease tolerance is the ability to maintain yield performance in the presence of disease symptoms. Therefore, it could be a useful tool in the management of the disease. Although it is known, that there is disease tolerance to STB in some wheat cultivars, this aspect has not been studied among Argentinean cultivars. The aims of this study were to evaluate genotypic differences in tolerance to STB among Argentinean cultivars, considering the relationship between the area under disease progress curve or the green leaf area or the non-green leaf area duration with the grain yield. In addition the effect of the disease on yield, yield components, test weight, grain protein concentration, wet and dry gluten concentration and the influence of tolerance on these traits was investigated. Field experiments were carried out with ten cultivars in a split-split-plot design during 2010 and 2011. Inoculation treatments were the main plots and cultivars, the subplots. STB significantly reduced grain yield, their components, test weight and increase grain protein and gluten concentration. Cultivar Baguette 10 showed major tolerance to STB, indicated by a consistent low regression slope between the green area duration and yield, while Klein Chaja was non-tolerant due to a high regression slope. However, many cultivars such as Buck Brasil, Buck 75 Aniversario, Klein Escorpion and Klein Flecha had considerably similar regression slopes to Baguette 10, provided good levels of tolerance. Other cultivars presented no significant differences. The correlation coefficient between tolerance and grain yield potential was not significant, suggesting that tolerant high-yielding cultivars can be obtained. No relationship was found between quality group or tolerance with the increase in protein and gluten concentration due to STB either.     

玉米穗腐病原菌禾谷镰孢的分离鉴定与致病力测试

利用形态学和分子鉴定技术,从玉米穗腐病病穗上分离并鉴定到17个禾谷镰孢菌株。以玉米自交系X178、593和B73为材料,对其中5个产孢能力强的菌株通过玉米发芽抑制试验进行致病力测试。结果表明,菌株FG14对测试自交系的平均发芽抑制率达54.10%;FG7最低,仅有23.22%,两者相差2.3倍。选用5个菌株在田间分别接种B73和LH59果穗,两个自交系对菌株FG14和FG21的抗性评价均为感,FG7为中抗。种子发芽和田间接种鉴定结果均表明,禾谷镰孢菌株间致病力存在显著差异。试验明确不同菌株对穗腐病抗性鉴定的影响,获得的菌株为玉米种质的穗腐病抗性筛选提供理论参考。     

禾谷镰刀菌Homeobox基因结构及其演化关系分析

对致病菌禾谷镰刀菌(Fusarium graminearum)全基因组序列中可能的Homeobox基因进行生物信息学分析.结果表明:该菌含有12个可能的Homeobox基因;它们所编码的蛋白都具有同源异型盒结构域,且该结构域中的L15、P28、W51、R56氨基酸残基最保守;所有的同源异型盒结构域蛋白都定位于细胞核中.分子进化分析表明,禾谷镰刀菌Homeobox基因在进化过程中出现分支,并且其分支早于真核生物的分化.研究结果将为深入分析该基因家族的生物学功能奠定基础.     

渥堆过程中黑茶生化成分变化及其影响因子研究进展

渥堆过程中,黑茶中的生化成分在湿热作用下发生剧烈变化,如茶多酚因多酚氧化酶引起的氧化聚合反应而含量锐减;儿茶素因微生物的酶促氧化与非酶促作用转化,其组分及比例发生明显改变;氨基酸和可溶性糖含量亦有所变化等。这些变化为形成黑茶外形棕褐油润,汤色橙黄或红浓,滋味醇和等独特品质奠定了重要的物质基础。本文对渥堆过程中,黑茶主要品质成分变化以及渥堆工艺关键影响因子等进行了综述,并对渥堆工艺研究趋势作了展望。     

Changes in carbohydrate and nitrogenous components and amylase activities during germination of grain amaranth

Grain amaranth (Amaranthus hypochondriacus), Yercaud local variety, was soaked overnight and germinated for 192 h taking the soaked grains as the zero time (0 h) sample. The changes in the activities of - and -amylases, starch, sugar, protein and lysine contents during germination are reported. Activity of -amylase was high in the 0 h soaked grains, while -amylase activity was high in 72 h germinated grains. The joint action of the amylases resulted in a decrease of starch content from 0 to 192 h in germinated grains and an increase in total sugars during the initial period of germination.Protein nitrogen was found to decrease from 48 h to 192 h accompanied by an increase in free amino acid and non-protein nitrogen contents. Total lysine content was found to be increased by 31% in 24 h germinated grain amaranth. Protein fractionation of raw, soaked and 24 h germinated grain amaranth showed that the distribution of different types of proteins varied during germination of the grains. An increase of water soluble protein content was noticed in 24 h germinated grains.     

Changes in common wheat grain milling behavior and tissue mechanical properties following ozone treatment

Ozone treatment (10 g/kg) of common wheat grains with a new patented process, Oxygreen^®, used before milling was found to significantly reduce (by 10–20%) the required energy at breaking stage whatever the grain hardness and without changes in the flour yield. Detailed study of each of the milling steps undertaken on a hard type cultivar showed that both the breaking and the reduction energy were decreased. Reduction of the coarse bran yield was also observed concomitantly with an increase in the yield of white shorts. Biochemical characterization of the milling fractions pointed out changes in technological flour properties as starch damage reduction, aleurone content enrichment and increase of insoluble glutenin polymers. Measurement of wheat grain tissue mechanical properties showed that ozone treatment leads to reduction of the aleurone layer extensibility and affects the local endosperm resistance to rupture. These data as well as the direct effect of ozone oxidation on biochemical compounds could explain the observed changes in milling energy, bran and shorts yield and flour composition.     

Wheat grain tissue proportions in milling fractions using biochemical marker measurements: Application to different wheat cultivars

Measurement of biochemical markers allows the quantification of wheat (Triticum spp.) grain tissue proportions in milling fractions. In order to evaluate the ability of extending this methodology to an unknown wheat grain batch, the variability of the markers in the different tissues was assessed on various wheat cultivars. Ferulic acid trimer amounts in the outer pericarp ranged from 0.97 to 1.67 μg mg⁻¹ (dm) with an average value equal to 1.31 μg mg⁻¹ (dm). Alkylresorcinols amounts in a composite layer, including the testa, the inner pericarp and the nucellar epidermis, ranged from 10.5 to 16.7 mg g⁻¹ (dm), with an average value equal to 14.0 mg g⁻¹ (dm). In the aleurone layer, phytic acid amounts ranged from 94.9 to 187.2 mg g⁻¹ (dm) with an average value equal to 152 mg g⁻¹ (dm) whereas, para-coumaric acid ranged from 0.08 to 0.29 μg mg⁻¹ with an average level of 0.18 μg mg⁻¹. In the embryonic axis, wheat germ agglutinin ranged from 879 μg g⁻¹ to 2086 μg g⁻¹ with an average value of 1487 μg g⁻¹. The impact of this variability on tissue proportion determination was evaluated and a strategy to decrease the prediction error was suggested. Percentages of the outer pericarp, intermediate layer (including the testa), aleurone layer and embryonic axis within grains were calculated and their variability discussed.     

Introgression of a major gene for high grain protein content in some Indian bread wheat cultivars

In bread wheat, high grain protein content (GPC) determines nutritional value, processing properties and quality of the end-product. In view of this, marker-assisted selection (MAS) was performed for introgression of a major gene for high GPC (Gpc-B1) into 10 wheat genotypes. As a result, 124 BC₃F₅/F₆ progenies with Gpc-B1 were developed and evaluated in multi-location field trials. Significant interaction of Gpc-B1 with the recipient parent genotypes and the environment was noticed. However, a total of seven MAS-derived progenies with significantly higher GPC (14.83-17.85%) than their recipient parental genotypes and having no yield penalty were obtained. In these selected progenies, no significant negative correlation of grain yield with GPC (%) or protein yield was observed suggesting that GPC could be improved without yield penalty. This study thus suggested that MAS in combination with phenotypic selection is a useful strategy for development of wheat genotypes with high GPC associated with no loss in yield.     

小麦品种望水白的抗赤霉病性遗传分析

为了研究小麦品种抗赤霉病性的遗传规律,以病小穗率为评价指标,运用单花滴注对安农8455/望水白重组自交系(R IL)群体的2001年F6代、2003年F8代、2004年F9代进行了赤霉病抗性评价,采用植物数量性状主基因 多基因混合遗传模型分离分析法,研究了该群体抗赤霉病的遗传规律。结果表明,安农8455/望水白重组自交系群体3年的抗赤霉病性分别符合E-2-6模型(两对主基因 多基因的加性-加性模型)、E-1-8模型(两对主基因 多基因,主基因间为重叠作用)、E-1-8模型(两对主基因 多基因,主基因间为重叠作用)。主基因的遗传率较高,分别为63.8%、69.02%、73.66%,多基因的遗传率分别为21.80%、21.38%、16.80%。说明望水白的抗赤霉病性由2对主基因控制,且符合两对主基因 多基因模型。还对望水白与另一感病品种A londra构成的重组自交系的抗性进行了比较。     

Performance of wheat and triticale varieties in a variable soil water environment IV. Yield components and their association with grain yield

Yield components (Y_c) and their association with grain yield (GY) were studied in seventeen varieties of Triticum aestivum, T. durum and triticale grown for five years in irrigated and unirrigated environments. The varieties were categorized into four contrasting groups based on their drought susceptibility indices (S) and yield potentials (Y_p). The slope (b) and intercept (a) of the linear regression between Y_c and the environmental index, and S values of Y_c were estimated. Their relationships with b, a and S of GY were also studied irrespective of the parameter used. Total dry matter (DM) and the number of grains per m² (GNO) were always positively correlated with GY. GWT (1000-grain weight) was not correlated with GY.There was considerable variation among the groups in their Y_c at different water-availability levels. The group of varieties having low S and moderate Y_p produced more spikes per m² (SPIKES) relative to other groups irrespective of the water availability. Simultaneously, their increased number of grains per spike (GNOSP) in drought environments relative to irrigated environments partially offset the accompanying reduction in SPIKES. The group of varieties characterized by high Y_p and low S had highest yield in irrigated and moderate drought environments because they had both more SPIKES and more GNOSP. By comparison, the varietal group having high Y_p and high S had higher GNOSP but lower SPIKES. Varieties which had low Y_p and were susceptible to drought had the lowest SPIKES, GNOSP and DM.It was concluded that maintenance of SPIKES would provide an advantage in environments varying in soil water availability. Performance in irrigated environments would depend on the maintenance of high GNOSP also. Higher DM production would be advantageous in all environments.     

Winter wheat genotypes under different levels of nitrogen and water stress: Changes in grain protein composition

Hard white winter wheat with superior and consistent quality is preferable for Asian markets. This study investigated the combined influences of moisture deficit during grain-fill and N management on protein quality, dough rheological properties, and protein molecular weight distributions in soft and hard winter wheats. Genotypes were grown under an irrigation gradient and two N-fertilization levels. Grain polyphenol oxidase (PPO) activity, SDS sedimentation, and Mixograph analyses were evaluated. Flour protein composition was characterized using SE-HPLC. Moisture stress during grain-fill increased flour protein content. N fertilization increased flour protein content. No significant correlation was found between flour protein and PPO. Changes in protein composition were related to general increases in protein content, regardless if the result of reduced irrigation or increased fertilization rate. The percentage of monomeric proteins increased more than the polymeric proteins as flour protein increased. Similarly, SDS sedimentation volume increased as a function of protein content. As expected, subunit GluD1 5+10 was associated with larger sedimentation volume and higher dough strength in genotypes as compared to those with subunit GluD1 2+12. Biplot analyses showed that genotypes of similar protein quality and composition responded similarly to N and irrigation treatments.