Geographical variation of β-amylase thermostability among varieties of barley (Hordeum vulgare) and β-amylase deficiency

Investigations onto the thermostability of β-amylase in 274 varieties of barley (Hordeum vulgare L.) indicated that all varieties except one were distributed into three types of high (type A), intermediate (type B), and low (type C) thermostability, respectively. One variety (TB29) from China showed no β-amylase activity. Geographical variation was observed in the thermostability of β-amylase. Type C varieties were not observed in East Asia (Japan, the Korean Peninsula, China and Nepal), although 36 out of 37 varieties in Ethiopia were type C. Most of the varieties were Type A in Japan, the Korean Peninsula and China, whereas the frequency of type A and type B were nearly equal in Nepal. Varieties in the other five areas (North America, North Africa, Southwest Asia, Turkey and Europe) consisted of types A, B and C. These results support the fact that East Asian cultivars are genetically different from those of the western regions, as previously reported.     

Worldwide distribution of β-amylase thermostability in barley

The thermostability of β-amylase in 6752 lines of worldwide barley genetic resources were investigated. Most of the lines were classified into high (type A), medium (type B) , and low (type C) thermostability. Subsequently the geographical distribution of these types was clarified. About 90% of the East Asian (Japan, the Korean Peninsula, China) lines were type A. More than 95% of Ethiopian barley was type C. The thermostability types of varieties in the western areas (north Africa, southwest Asia, Turkey, Europe) consisted of types A, B and C. These results suggest that there is a clear geographical differentiation in β-amylase thermostability, especially in East Asia and Ethiopia. The phenotype characteristics of each thermostability type line reflected the geographical differentiation. Besides types A, B and C, we found new thermostability types, including such useful mutants as a β-amylase-less mutant and highly-thermostable mutants than type A in both China and Nepal. This revised version was published online in August 2006 with corrections to the Cover Date.     

大麦β-淀粉酶活性对其种子在干旱胁迫下萌发影响的研究

为了鉴选出抗旱性较强的大麦品种,并以期找出β-淀粉酶活性对大麦抗旱性的影响,对β-淀粉酶活性不同的9个啤酒大麦品（系）种进行聚乙二醇(PEG-6000)模拟干旱胁迫,通过对种子发芽率及发芽势的测定来评价参试品种的抗旱性,并对β-淀粉酶活性与不同PEG浓度下的发芽率进行相关分析。结果表明,大麦β-淀粉酶活性与大麦抗旱性在PEG浓度为15%时,在0.01水平上达显著正相关。大麦种子发芽率随着干旱胁迫程度的增加而降低。5%浓度的PEG对大麦种子的萌发影响较小,10%、15%浓度显著抑制大麦种子萌发,20%浓度下,大麦个别品种发芽停止。处理样品间,‘沪麦16’综合表现出较好的抗旱性,其次A7-30,而‘邯98-58’、‘盐95137’为最差。由此得出,10%的PEG干旱胁迫剂浓度可作为大麦发芽期判断大麦抗旱性的适宜胁迫浓度,β-淀粉酶活性可作为大麦抗旱性早期筛选的指标之一。     

Identification of novel alleles from wild barley for the improvement of alpha‐amylase and related malt quality traits

Germplasms consisting of 64 wild barley accessions (Hordeum vulgare ssp. spontaneum) were examined for polymorphisms in α‐amylase using both isoelectric focusing (IEF) and thermostability assays. Wide variation was found for the high pI α‐amylase with 20 IEF band patterns identified. Enzyme activity and thermostability assays showed large differences among α‐amylase isoenzymes. Two wild accessions Tel‐Shoket CPI 77146‐32 and Afiq CPI 77128‐41 showed superior enzyme activity and thermostability compared with commercial varieties such as ‘Baudin’, ‘Flagship’ or ‘Navigator’. The functionality of the Tel‐Shoket allele was validated in backcross lines with ‘Flagship’ as the recurrent parent. The Tel‐Shoket allele at the amy1 locus increased α‐amylase thermostability at 75°C by 8.4% and α‐amylase activity in kilned malt by 18.7%. The introgression of the wild allele also led to significant improvements in fermentability, hot water extract and viscosity. Gene sequencing showed that there are three single nucleotide polymorphisms in the Tel‐Shoket amy1 sequence, which can be used as diagnostic markers for the selection of this allele in breeding programmes.     

Amy6-4基因遗传多样性及其与α-淀粉酶活性的关联分析

α-淀粉酶活性是影响大麦种子发芽和麦芽制作及啤酒酿造的重要性状,Amy6-4为编码高等电点的α-淀粉酶基因,挖掘其高活性的等位变异对啤酒大麦的品种改良具有指导意义。通过对58份大麦品种中Amy6-4基因的重测序,研究了该基因的核苷酸序列以及在品种间的遗传多样性,并在群体结构分析的基础上,进行了核苷酸多态性与α-淀粉酶活性的关联分析。结果表明,Amy6-4基因共存在7个单核苷酸变异位点(SNP),构成5种单倍型。其中,H_3单倍型最普遍,发生频率为51.7%(30/58);其次为H_1单倍型,发生频率为39.7%(23/58);其他3种单倍型发生的频率约为10%。SNP位点及其构成的单倍型均与酶活性无关联性。     

Use of barley seed vigour to discriminate drought and cold tolerance in crop years with high seed vigour and low trait variation

Seed vigour is a precondition for early and homogenous field emergence of barley, in addition to effective malting. This study aimed to assess the selection of barley varieties by using seed vigour as the indicator. Seed vigour of barley (quantified as the germination percentage) was evaluated under drought (?0.2 MPa) and temperature stress (10°C). At two locations over a 3‐year period, 1 population of 133 Derkado × B83‐12/21/5 doubled haploid (DH) lines (and parents) was evaluated for seed vigour, of which 108 DH lines were assessed for three malting parameters. The relatively high values of vigour during the 3‐year period (overall average 94–95%) probably impeded high variations in genetic potential. A total of 27 DH lines of the 133 evaluated showed transgression for vigour (up to 98%) in comparison with the parents (Derkado: 96%; B83: 92%). In conclusion, caution should be applied when selecting for seed vigour, even in good crop years with high levels of seed vigour and low trait variations. Such selection might improve vigour, particularly in crop years with unsuitable weather conditions.     

Geographical differentiation and diallel analysis of seed dormancy in barley

Seed dormancy is one of the most important parameters affecting the malting process and pre-harvest sprouting in barley (Hordeum vulgare L.). Variation of seed dormancy in 4365 cultivated and 177 wild barley (ssp. spontaneum) accessions derived from different regions of the world was investigated in Okayama University, Kurashiki, Japan. Seed dormancy of each accession was estimated from their germination percentages at 0, 5, 10 and 15 weeks post-harvest after-ripening periods. All of the wild barley accessions showed less than 10% germination at 0 week after-ripening period. Level of seed dormancy in 4365 cultivated barley accessions showed a clear geographical differentiation. Seventy seven percent of Ethiopian accessions showed high germination percentages, while 86% of Japanese, Turkish and North African accessions showed low germination percentages at 0 week after-ripening period. A half diallel cross using eleven barley accessions with different level of dormancy revealed that seed dormancy was predominately controlled by additive gene effects. These results suggest that large genetic diversity for seed dormancy in barley is explained as different levels of additive accumulation of genetic factors. Barley varieties showing appropriate dormancy could be developed by crossing among barley germplasm accessions used in the present study.     

Diversity and geographical distribution of seed lipoxygenase‐1 thermostability types in barley

To understand the diversity in the thermostability of the seed lipoxygenase‐1 (LOX‐1), 1040 cultivars of worldwide barley (Hordeum vulgare ssp. vulgare) genetic resources were investigated. The relative thermostability of LOX‐1 (LOX‐RTS) in these lines showed a bimodal frequency distribution and these lines were categorized into the high and low thermostability types (H‐type and L‐type, respectively). The H‐type lines predominated in the wild progenitor, ssp. spontaneum. The geographical distribution of these types in the cultivars was surveyed. The frequencies of the H‐ and L‐types were almost equal to one another in southwestern Asia. The occurrence of the H‐type predominated in eastern Asia and Africa, whereas in Europe and Turkey, the L‐type did. The predominance of the L‐type in Europe and Turkey can be understood through the hypothesis that the Fertile Crescent domestication contributed the majority of diversity in Europe. The uneven geographical distribution of the LOX‐1 thermostability types in the cultivars may reflect a polyphyletic origin of barley.     

The EEC Barley and Malt Committee Index for the Evaluation of Malting Quality in Barley and its Use in Breeding

A new statistical quality index, Q, is defined in order to facilitate the interpretation of the overall quality of malting barley varieties. Q ranges from O (feeding barley) to 9 (top quality malting barley), and is related to the Normal distribution. The varieties tested within the network of the European Brewery Convention are assessed for malting quality as an example of tire reliability of Q. In addition, the new index is used to study the genotype X environmental interaction for malting quality.     

大麦籽粒灌浆期β-淀粉酶积累动态与气象因子关系研究

为了明确大麦籽粒灌浆期β-淀粉酶动态与气象因子的内在联系,对两年间5个大麦品种籽粒灌浆期β-淀粉酶动态与气象因子进行了相关分析。结果表明,大麦籽粒灌浆期的β-淀粉酶活性呈升–降–升的动态过程,在成熟期达到最大值。偏相关分析和多元回归分析表明,大麦籽粒灌浆期β-淀粉酶活性与总积温呈极显著正相关,与总日照时数和日均日照时数的互作呈显著相关,与日均温、日均日照时数呈显著负相关,与总积温和日均温的互作、总雨量和日均日照时数互作呈极显著负相关,偏相关系数从大到小依次为总积温、总雨量和日均日照时数交互作用、总积温和日均温交互作用、总日照时数和日均日照时数交互作用、日均日照时数、日均温。     

Evaluation of Czech spring malting barleys with respect to the β-amylase allele incidence

Strong enzymatic activities in the germinating barley grain, together with protein and starch content, are crucial for high extraction values in the resulting malt and, therefore, barley malting quality. The efficient characterization of registered barley cultivars and genetic resources with respect to one of the relevant thermostability enzymes (β‐amylase) is an essential requirement. The template‐directed dye‐terminator incorporation (TDI) assay: based on flourescence resonance energy transfer (TDI‐FRET) ( Chen et al. 1995 ) was used to detect single nucleotide polymorphisms (SNPs) in the β‐amylase coding sequence resulting in low (Sd2L), intermediate (Sd1) and high (Sd2H and Sd3) thermostability enzyme across 84 Czech barley cultivars and genetic resources used over a period of time in the Czech Republic. The incidence of different alleles has changed during the last 100 years. Also the new resources with high thermostabile β‐amylase were identified. They can be used effectively to breed for malting quality improvement.     

Mapping a factor controlling the thermostability of seed lipoxygenase-1 in barley

Barley lipoxygenase (LOX)‐1 is believed to affect the stability of flavour and the foam of beer. The purpose of this study was to investigate the genetic variation of the LOX‐1 thermostability, and to analyse the mode of inheritance of this trait. A simple method was established to evaluate the LOX‐1 relative thermostability (LOX‐RTS). With this method, 153 barley cultivars were screened for LOX‐RTS. The frequency of the LOX‐RTS values was distributed in a bimodal manner. Based on these values, the barley lines were categorized into two groups: an H‐type with relatively thermostable LOX‐1 and an L‐type with relatively thermolabile LOX‐1. Using a ‘Steptoe’/‘Morex’ doubled haploid population, a major quantitative trait locus (QTL) associated with LOX‐RTS was identified on chromosome 4H of barley, explaining 82% of the variance. Mapping of a CAPS marker specific for the LoxA locus revealed co‐segregation with this QTL. In this study, the existence of the thermostability types of barley seed LOX‐1 and the locus controlling the thermostability were made clearer.     

大麦籽粒淀粉和β-葡聚糖积累特性研究

为研究大麦籽粒支链淀粉、直链淀粉和β-葡聚糖积累特性,及淀粉各组分与β-葡聚糖的关系,以2个皮大麦、1个裸大麦和1个糯裸大麦为试验材料,测定花后7、14、21、28d淀粉各组分及β-葡聚糖含量。结果表明,甘啤6号淀粉各组分含量随灌浆推进均逐渐升高,在成熟期达到最大值;甘垦啤7号和甘垦6号均呈先升高后降低趋势,在花后21d有1个峰值。糯大麦C 2-1直链淀粉含量显著低于非糯大麦;支链淀粉含量显著高于非糯大麦,整个灌浆期呈先升高后降低趋势。β-葡聚糖含量均随灌浆时间延长逐渐升高,成熟期含量最高;整个灌浆期糯大麦C 2-1含量显著高于非糯大麦。相关分析表明,直链淀粉/支链淀粉比值与β-葡聚糖含量呈极显著负相关,可以将直链淀粉/支链淀粉比值作为高β-葡聚糖品种选育的一个指标。Logistic方程拟合发现,直链淀粉、β-葡聚糖最终积累量与积累起势与有效积累时间有关,支链淀粉最终积累量取决于最高积累速率和平均积累速率。     

Mapping of Malt Endopeptidase, NADH Diaphorase and Esterase Loci on Barley Chromosome 3L

Isoelectric focusing (IEF) and immunoblotting were used to detect genetic variants of malt endopeptidase (MEP1), an enzyme related to malting quality in barley and coded by the CepB locus on barley chromosome 3 (= 3H). A variant was found in a Turkish accession of wild barley (Hordeum vulgare ssp. spontaneum). The self progeny of a hybrid between this accession and the barley cultivar ‘Clipper’ were analyzed for recombination between the CepB locus and other isozyme loci. The estimates of recombination linked the CepB locus to an NADH diaphorase locus (Ndh2), which in turn was linked to the seedling esterase complex (Est1, Est2, and Est4) situated near the distal end of the long arm of chromosome 3. The Ndh2 locus was independent of two other NADH dehydrogenase loci (Ndh3, Ndh5) which were mapped on barley chromosome 5 (= 1H) in relation to the three hordein loci (Hor1, Hor2, and Hor3).     

德国啤酒大麦品种性状评价及综合利用研究

本文对德国啤酒大麦品种的农艺性状、品质优势及产量潜力进行了详细的论述与评价,提出了直接应用与间接利用的指导性建议。根据多年的引种实践经验,我们有针对性地从德国引进啤酒大麦品种32份,经过连续近10年的试验研究,筛选出一批优异种质作为育种材料间接利用,选育出垦啤3号新品种已大面积推广应用,另外选育出1个新品系拟参加甘肃啤麦产区旱地试验示范。     

Diversity analysis for resistance of rice (Oryza sativa L.) to blast disease [Magnaporthe grisea (Hebert) Barr.] using differential isolates from the Philippines

A wide variation in resistance to blast disease caused by Magnaporthe grisea (Hebert) Barr. ( Rossman et al. 1992 ) was found using 922 rice ( Oryza sativa L.) varieties collected mainly from Asia. These were classified into six varietal groups, termed clusters A–F, according to Ward's hierarchical cluster analysis, based on the reaction pattern to 20 standard differential blast isolates from the Philippines. The most susceptible two clusters, B and C, dominated in varieties from the Far East (Japan). Varieties from East Asia and Southeast Asia occurred less frequently in B and C clusters than those of Japan, and more frequently in E and F clusters, which were the most resistant of the cluster groups. Varieties from South Asia showed the widest variation, occurring in all clusters but less frequently in cluster B. The cluster B varieties dominated in Japan and showed a high frequency of isozyme type VI, corresponding to Japonica type. In contrast, the frequency of cluster B was low in the groups with isozyme types I, II, III and V, which dominated in South Asia. Isozyme type I corresponds to Indica type varieties. The distribution of resistance corresponded to the geographical distribution of rice varieties and might be related to differentiation into Indica and Japonica types. These findings will provide useful information for understanding the variation in blast resistance at the global level.     

不同类型啤酒大麦品种遗传多样性及遗传差异的SSR分析

利用28对SSR引物对包括22份野生材料,8份耐盐碱材料,21份优质高产甘啤系列品种(系)材料和9份日本早熟材料在内的60份啤酒大麦品种资源的遗传多样性及其亲缘关系进行分析。结果表明,所用28对引物,有25对有多态性;聚类结果表明这些材料遗传相似系数分布在0.4792~0.9948之间。在遗传相似系数0.63的水平上,这些材料聚类为2大类,下分7个亚类。从聚类结果看出,不同类型材料基本各自聚为一类,表明不同类型品种(系)内遗传差异较小;不同类型品种(系)间遗传差异较大,杂交可能会产生较强的杂种优势和较高的产量潜力。     

Tolerance of barley seed germination to cold- and drought-stress expressed as seed vigour

In barley, high seed vigour is a precondition for rapid and homogenous field emergence and good malting quality. Seed vigour was defined as germination percentage under stress conditions (10°C, drought stress ‐ 2 Bars) in 7‐8 barley varieties grown in 7‐8 locations in the Czech Republic over 7 years. Three of the 7 years were not suitable for high seed quality, probably because of unsuitable weather, as average seed vigour reached only 61, 77 and 86%, respectively. In the remaining 4 years, the average vigour exceeded 94%. The impact of variety on seed vigour was higher in the ‘bad years’ and the impact of location was higher in the other years. Varieties with higher vigour from all locations in the ‘bad years’ were identified. Lower vigour was related to the high occurrence of fungi (indicated by ergosterol assays) and to lower field emergence rates of seed samples. The results support the possibility of selecting for improvement of barley seed vigour, which is related to tolerance to various conditions during emergence and to homogenous malting.     

The effect of water and nitrogen availability during grain filling on the timing of dormancy release in malting barley crops

Pre-harvest sprouting (PHS) causes immediate loss of seed viability, making barley (Hordeum vulgare L.) grains worthless for malting purposes. Grain dormancy release rate in barley crops is genetically and environmentally controlled. A 2 year experiment was conducted to evaluate the effect of soil nitrogen and water availability during grain filling on the dormancy release pattern (and then on the PHS susceptibility) for five malting barley commercial cultivars. Drought and well-irrigated control treatments were imposed from anthesis onwards, and contrast nitrogen fertilization treatments were applied at tillering. Nitrogen availability showed no effects on dormancy release. Drought during grain filling accelerated dormancy release with respect to well-irrigated control in 2004, but not in 2005 year. Mean temperatures during the last stages of grain filling were much higher (ca. 6°C) in 2005 than in 2004, indicating that high-dormancy loss promoting temperatures had masked drought effects on dormancy release.     

不同功能叶对啤酒大麦籽粒灌浆特性的影响

为明确不同功能叶对啤酒大麦籽粒灌浆特性的影响,采用单因素随机区组设计试验方法,对高产、优质啤酒大麦‘甘啤7号’的籽粒灌浆特性进行了研究,应用Logistic方程对灌浆进程进行了拟合。结果表明：不同光合器官处理下啤酒大麦灌浆进程符合Logistic生长曲线,呈“慢－快－慢”“S”型变化。不同处理对品种12项灌浆参数及麦芽品质均产生影响,但影响的具体参数和影响程度因处理而异。不同处理下产量性状指标筛选率和千粒重变化最大,且呈正相关关系。其中T4处理下,饱满粒(72.17%)和千粒重(42.37 g)与其他处理及CK产生显著差异。为选育高产、优质啤酒大麦,在育种选择实践中应综合考虑形态指标。