用一个SSR标记快速检测小麦杂种后代Waxy基因型

为了提高Wx蛋白的检测手段，通过对几个已知Waxy基因型的品种(系)的分子标记检测和蛋白电泳谱带分析，发现一个特异的SSR标记能同时对三个Wx基因位点进行检测；同时用这个特异的SSR标记对一个糯小麦Et-05及普通栽培小麦扬麦158的杂交后代F2进行分析，发现了4株糯小麦植株。     

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.     

Functionality of native tetraploid wheat starches: Effects of waxy loci alleles and amylose concentration in blends

Partial waxy (reduced amylose) and fully waxy (amylose-free) tetraploid durum wheats (Triticum turgidum L. var. durum) were used to investigate the relationships between both intra- and inter-granular variation in amylose concentration and starch functionality. Starches isolated from each genotype (intra-granular amylose variants) were compared to those of commercially available hexaploid wild-type and waxy starches, and functionalities compared to blends (inter-granular variants) of durum waxy and wild-type starches of 0, 6, 12, 18, 24, & 30% amylose content. Starch particle size distributions were not significantly different amongst partial waxy and wild-type genotypes; waxy samples had significantly smaller mean starch granule size. Few significant differences for crystallite melting and related intrinsic heat as determined by differential scanning calorimetry (DSC) were observed. The detected differences in starch gel color or gel strength generally were observed for the waxy samples relative to wild-type. Numerous significant differences were observed via Rapid-Visco Analysis (RVA). Pasting peak viscosity and breakdown were inversely proportional to % amylose. Wx-B1 null final viscosity differed significantly from that of all other blends and genotypes, demonstrating that genotypic differences exist amongst the partial waxy types, independent of amylose concentration per se.     

不同糯小麦品种(系)品质及其烘烤制品研究

为推动糯小麦的品质改良及开发利用,以国内不同育种单位育成的多个糯小麦为试验材料,以普通强筋、中筋及弱筋小麦作为对照,分析其淀粉组分、糊化特性、面团流变学特性,并对其制作的酥条进行比较.结果表明,糯小麦的淀粉含量及直链淀粉含量显著低于普通小麦;在淀粉糊化特性上,糯小麦的低谷黏度、最终黏度、回生值显著低于普通小麦,峰值温度...     

紫糯小麦籽粒石蜡切片制作技术优化及淀粉粒累积分析

为了探明糯性小麦授粉后籽粒淀粉粒的累积情况,以山农紫糯2号授粉后不同天数的籽粒为试验材料,通过对传统石蜡切片技术的脱水、透明和浸蜡等环节进行改进,制作切片后对小麦籽粒结构发育进行观察研究。结果显示,与传统石蜡切片方法相比较,技术改良70%乙醇和无水乙醇脱水时间与逐级透明和渐进式浸蜡环节后,小麦籽粒（授粉后0~14 d）不皱缩变形,石蜡切片完整,且染色分明,结构显示清晰。授粉后0~2 d,果皮中已经开始积累少量淀粉粒,随着授粉后天数推移而逐渐增多;授粉后第6天,胚乳内部逐渐由游离核期向细胞化过渡;授粉后第8天,胚乳细胞进一步增大,淀粉粒开始累积,但体积较小;授粉后第10~12天,胚乳细胞向无核化发展,不同部位胚乳细胞和淀粉粒的排列分布呈现明显差异;授粉后第14天,胚乳细胞中几乎完全充满淀粉粒。表明改进的石蜡切片制作方法适合于早期小麦籽粒的结构研究。     

Reduced height alleles (Rht) and Hagberg falling number of wheat

Near-isogenic lines varying for alleles for reduced height (Rht) and photoperiod insensitivity (Ppd-D1) in cv. Mercia (2005/6–2010/11; rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht8c+Ppd-D1a, Rht-D1c, Rht12) and cvs Maris Huntsman and Maris Widgeon (2007/8–2010/11; rht (tall), Rht-B1b, Rht-D1b, Rht-B1c, Rht-B1b+Rht-D1b, Rht-D1b+Rht-B1c) were compared at one field site, but within different systems (‘organic’, O, 2005/6–2007/8 v. ‘intensive’, I, 2005/6–2010/11). Further experiments at the site (2006/7–2008/9) compared 64 lines of a doubled-haploid (DH) population [Savannah (Rht-D1b) × Renesansa (Rht-8c+Ppd-D1a)]. Gibberellin (GA) insensitive dwarfing alleles (Rht-B1b; Rht-B1c; Rht-D1b; Rht-D1c) could reduce α-amylase activity and/or increase Hagberg falling number (HFN) but effects depended greatly on system, background and season. Only Rht-B1c increased grain dormancy despite producing plants taller than Rht-D1c. The GA-sensitive Rht8c+Ppd-D1a in Mercia was associated with reduced HFN but analysis of the DH population suggested this was more closely linked with Ppd-D1a, rather than Rht8c. The GA-sensitive severe-dwarfing allele Rht12 was associated with reduced HFN. Instability in HFN over season tended to increase with degree of dwarfing. There was a negative association between mean grain weight and HFN that was in addition to effects of Rht and Ppd-D1 allele.     

Waxy durum acts as a unique form of crumb softener in bread

Flour milled from waxy durum grain was incorporated into bread formulations and its effects on crumb softness and loaf volume compared to those of added fats and emulsifiers. Waxy durum had a small effect on increasing loaf volume, and little of this effect could be explained by dough height or oven spring; effects of most other additives were closely related to dough height or oven spring. For all additives, the relationship between crumb softness and loaf volume fitted a power law relationship. Waxy durum and some of the additives had a greater softening effect than predicted from their effect on volume while others produced a much firmer crumb. With only one exception, when combined with other additives waxy durum reduced compression of the crumb more than predicted on the basis of the effects on loaf volume of the additives alone. In common with some of the added lipids, during compression analysis, crumb prepared with waxy durum flour had similar levels of structural damage as crumb baked from the standard formulation; other lipids and emulsifiers produced more fragile crumb. Waxy durum flour appears to have unique effects on crumb softness.     

Detection of mutations in the 7A allele of wheat (Triticum aestivum) granule-bound starch synthase (Wx-7A) with a monoclonal antibody produced by targeted peptide immunisation

A peptide sequence unique for the granule-bound starch synthase (GBSS1) of hexaploid wheat (Triticum aestivum) was identified by sequence alignment of the three isoforms. Multiple antigenic peptides (MAP) of different length (16 and 20 AA) were synthesised against the selected sequence. The assessment of the immune response in mice against differently presented forms (resin-bound, free) and two types of adjuvants indicated that the free MAP with the longer peptide is more immunogenic. A 20AA MAP that elicited a strong immune response was used to produce a monoclonal antibody for the 7A isoform of GBSS 1. Epitope mapping of the selected Mab (F5-1F2) revealed that a 12mer partial sequence of the immunising peptide was specifically detected. Based on this Mab, a simple high throughput ELISA was developed that allows the quick identification of wheat lines carrying the 7A allele of GBSS 1 with minute amounts of sample.     

Partial purification and characterization of polyphenoloxidase from durum wheat (Triticum durum L.)

A bright yellow color of pasta is an important qualitative trait for the durum wheat industry. Final color is the result of the balance between yellow and brown components in semolina. Polyphenoloxidase (PPO) is implicated as playing a significant role in darkening. This study aimed to characterize PPO activity of durum wheats. PPO was extracted and partially purified by ion-exchange chromatography on a column packed with diethyaminoethyl cellulose (DEAE). This procedure led to 26.33-fold purification with 24.7% recovery. The optimum temperature and pH of PPO were found to be 40 °C and 6.5, respectively. Heat stability of durum wheat PPO decreased as the temperatures increased from 30 to 80 °C. The z-value was calculated as 23.4 °C. It increased to 26.3 and 48.4 °C in the presence of 40% sucrose and 1 M NaCl, respectively. Durum wheat PPO was shown to use several phenolic compounds as substrate. Among the substrates used, the greatest substrate specificity was observed with catechol. Durum wheat PPO was sensitive to inhibitors such as ascorbic acid, cysteine, oxalic acid and citric acid. Ascorbic acid was the most effective inhibitor.     

A novel Puroindoline b-2 variant present in Chinese winter wheat cultivar Yunong 202

The recently identified Puroindoline b-2 (Pinb-2) variants on the homoeologous group 7 chromosomes in bread wheat are Puroindoline-like genes that account for minor grain texture modulation and display activity against bacteria and phytopathogenic fungi. In this study, the coding sequence of the Pinb-2 gene in the Chinese winter wheat cultivar Yunong 202 (Triticum aestivum L.) was amplified with the universal primer, and the obtained 452-bp fragment was cloned and 36 plasmids encompassing the targeted fragment were sequenced from two strands. Results indicated that Yunong 202 possessed Pinb-2v1, Pinb-2v2, Pinb-2v3b and Pinb-2v4 alleles. A new Puroindoline-2 variant was discovered in Yunong 202 as well and was designated as Pinb-2v6. Full alignment showed that Pinb-2v6 possessed 74.0%, 95.4%, 94.7%, 92.3%, 98.7% and 98.0% identity at the DNA level with Pinb-D1a, Pinb-2v1, Pinb-2v2, Pinb-2v3a, Pinb-2v4 and Pinb-2v5 alleles, respectively. This study may provide useful information for further understanding the molecular and genetics basis of grain texture and for illustrating gene duplication events in wheat.     

opaque2对糯玉米子粒食味和营养品质的影响分析

玉米opaque2(o2)是胚乳发育的核心调控基因,对子粒发育至关重要。基于9组糯玉米o2近等基因系(o2-NILs)及对应轮回亲本,对鲜食期和成熟期子粒的种皮厚度、总淀粉含量、可溶性糖含量等性状进行分析。结果表明,黄糯2/o2o2和中63-2/o2o2鲜食期子粒的食味品质性状均有提高,其他o2-NILs子粒品质变化不一。鲜食期o2突变对种皮厚度影响较大,但与成熟期无直接相关性。o2突变使玉米子粒赖氨酸含量增加,营养价值提高,但多数近等基因系成熟期淀粉含量和百粒重降低,仅糯2/o2o2、淀粉含量和百粒重均不受影响。糯2/o2o2、SY1-2/o2o2、赵OP-6/o2o2中27 KD γ-zein表达量显著增加、子粒饱满度与轮回亲本相近,但均表现为粉质胚乳,因此认为,优质蛋白糯玉米硬质胚乳的恢复需多个o2胚乳修饰基因协同作用。     

Effects of Fusarium culmorum and water stress on durum wheat in Tunisia

The effects of water stress on Fusarium foot and root rot in durum wheat were investigated in growth chamber, greenhouse and field tests in Tunisia. In the seedling stage, emergence of six durum wheat cultivars in the growth chamber was significantly reduced by inoculation with Fusarium culmorum and water stress (P<0.0001), with more disease under drier conditions. Additionally, the tiller number per mature plant, the 1000 grain weight and disease severity in mature stage were reduced by inoculation in greenhouse studies. In a field test, inoculation with F. culmorum significantly reduced the yield (P<0.001), by more than 17% for Om Rabiaa and 38% for Karim, the two cultivars tested. Yield was also significantly affected by precipitation and irrigation levels. The severity of the disease, estimated by the percentage of white heads, was separately affected by the cultivar (P<0.001) and inoculation (P = 0.0004). Percentage of white heads was 1.5 and 2 × higher in inoculated plants than non-inoculated for Om Rabiaa and Karim cultivars, respectively. Disease severity was highest in treatments with the greatest water stress. This is the first detailed study of water stress and F. culmorum on durum wheat in Tunisia, and indicates that cultivar resistance and irrigation management may be important in the management of Fusarium foot rot.     

Development of STS markers and establishment of multiplex PCR for Glu-A3 alleles in common wheat (Triticum aestivum L.)

Low-molecular-weight glutenin subunits (LMW-GS) play a key role in determining the processing quality of the end-use products of common wheat. The objectives of this study were to identify genes at Glu-A3 locus, develop the STS markers, and establish multiplex PCR with the STS markers for Glu-A3 alleles. Gene-specific PCR primers were designed to amplify six near-isogenic lines (NILs) and Glenlea with different Glu-A3 alleles (a, b, c, d, e, f and g) defined by the protein electrophoretic mobility. Three Glu-A3 genes with complete coding sequence were cloned, designated as GluA3-1, GluA3-2 and GluA3-3, respectively. Seven dominant allele-specific STS (sequence tagged sites) markers were designed based on the SNPs (single nucleotide polymorphisms) among different allelic variants for the discrimination of the Glu-A3 protein alleles a, b, c, d, e, f and g. Four multiplex PCRs were established including Glu-A3b + Glu-A3f, Glu-A3d + Glu-A3f, Glu-A3d + Glu-A3g, and Glu-A3b + Glu-A3e. These markers and multiplex-PCR systems were validated on 141 CIMMYT wheat varieties and advanced lines with different Glu-A3 alleles, confirming that they can be efficiently used in marker-assisted breeding.     

Genetic characterization of kernel polyphenol oxidases in wheat and related species

Polyphenol oxidase (PPO) activity causes undesirable darkening of raw Asian noodles and other wheat products. In this study we investigate the genetic origins and diversity of wheat kernel PPO. PPO was characterized via activity assays, antigenic staining, and Southern blots in Triticum aestivum, Triticum dicoccoides, Triticum durum, Triticum dicoccum, Triticum monococcum, Triticum urartu, Aegilops speltoides, and Aegilops tauschii. Among these species, PPO activity was well-correlated with antigenic staining intensity toward a wheat kernel-type PPO antibody. High PPO activity was observed in all three T. monococcum accessions (A^m genome), one Ae. speltoides accession, one T. durum accession, and two hexaploid wheat cultivars. Southern blots suggested the presence of two or more kernel-type PPO genes in diploid progenitors of the hexaploid A, B, and D genomes. Whole-kernel PPO activity was evaluated in disomic substitution lines derived from three T. dicoccoides accessions in the background of T. durum ‘Langdon’. PPO activity was primarily associated with chromosome 2A and to a much lower degree with chromosome 2B. DNA sequence comparisons showed that the intron associated with the high PPO allele on chromosome 2AL of hexaploid wheat had 94% nucleotide identity with the homeologous intron found in T. monococcum, a species with high kernel PPO activity. This implies that the ancestral PPO allele on the A genome is one of the high activity, and the low PPO allele found in hexaploid wheat represents a relatively recent genetic alteration. Results confirm the presence of multiple kernel-type PPO genes in the diploid and tetraploid progenitors and relatives of hexaploid wheat. However, it is likely that relatively few of the many kernel-type PPO genes present in wheat contribute substantially to kernel PPO activity. A single genetic locus on homeologous group 2 chromosomes may be the primary cause of high PPO activity in wheat kernels.     

Amylolysis of wheat starches. II. Degradation patterns of native starch granules with varying functional properties

Scanning electron microscopy (SEM) and X-ray diffraction (XRD) were employed to investigate degradation patterns of native starch granules from wheat (Triticum aestivum L.) by different starch-degrading enzymes. The starches examined were from a waxy wheat and four varieties with slightly elevated amylose content, but with different functional properties. Differences in the digestion patterns after partial α-amylolysis of starch granules were noted between the starches. The waxy starch seemed to be degraded by endocorrosion, whereas the amylose-rich starches followed a slower mode of hydrolysis starting from the granular surface. X-ray diffractograms of the amylose-rich starches were not significantly altered by 2 h of α-amylolysis, whereas partial hydrolysis of the waxy starch decreased scattering intensity at higher 2θ angles, consistent with a different mode of attack by α-amylase in the initial digestion stages of granules of waxy and amylose-rich starches. We propose these differences are due to the combined effects of the change in packing density and partial preference for hydrolysis of amorphous material. The native starch granules were also attacked by beta-amylase, isoamylase and amyloglucosidase, which indicates that α-amylase is not the only starch-degrading enzyme that is able to initiate starch hydrolysis of native granules.     

Simulating multiple pest damage in varying winter wheat production situations

The production situation–injury profile paradigm can be used as a framework to assess the harmfulness of multiple-pest complexes in a changing agriculture. A mechanistic simulation model for wheat, WHEATPEST, was developed to incorporate drivers of (i) variable production situations and (ii) their related injury profiles. The model simulates the harmful effects of pathogens, pests, and weeds in a simple, open, generic manner. Simulation drivers were derived from published reports, in particular through a meta-analysis of highly detailed farmers’ field surveys in the United Kingdom and the Netherlands. Preliminary analysis of the model's performances indicates that WHEATPEST conforms with available published reports in a range of production situations and injury profiles. While improvement on some components of the model are discussed, this work points at the need for the collection of cross-disciplinary, reasonably accurate, and standardised data at a system's level, and at the usefulness of modelling tools for basic research and policy.     

Correspondence between two minor Glu-A3 genes of durum wheat and their encoded polypeptides by using a proteomic approach

The low molecular weight glutenin subunits (LMW-GS) are wheat storage proteins participating to the formation of glutenin polymers that, along with the other gluten proteins, allow the accumulation of a large quantity of protein in the endosperm tissue. The size and composition of the glutenin polymers are directly related to gluten visco-elastic properties. In particular, LMW-GS composition is the factor most influencing durum wheat quality.     

Are seeding densities an opportunity to increase grain yield of winter wheat in a living mulch of white clover?

Optimum plant densities are a key to maximise yields in most crops. However, such information is often lacking for more environmentally sound cropping systems, such as living mulches (LM) for small grains. In 2004 and 2005, three trials were conducted in the Swiss Midlands on fields managed in accordance with the Swiss organic farming guidelines. The objective of the study was to determine whether seeding density of winter wheat (Triticum aestivum L.) is a relevant factor for determining grain yield in a white clover (Trifolium repens L.) living mulch. The winter wheat cv. Titlis was directly sown in wide spaced rows (0.375 m) at densities of 300 (LM300), 450 (LM450) or 600 (LM600) viable grains m⁻² in a white clover living mulch established at a seeding rate of 15 kg ha⁻¹. A bare soil control treatment with a wheat density of 450 viable grains m⁻² (BS450) was also included in the trials. Mean grain yields of LM300, LM450, and LM600 never reached the values observed in BS450. This was mainly due to a lower ear density, which, nevertheless, increased linearly with the seeding density within the living mulch in all trials, but the rate of increase depended on the environment. The decrease of the grain weight brought about by the increasing seeding density had only a marginal impact on the grain yield, which was increased from 1.31, 1.98, and 4.09 Mg ha⁻¹ (LM300) to 1.97, 2.64, and 4.75 Mg ha⁻¹ (LM600) for each of the three trials in the study. Significantly higher protein contents were observed for LM300 compared to the higher densities in the living mulch and to BS450. Our research showed that an increase of the seeding density is an effective mean to increase the grain yield in living mulch systems with white clover. However, it is likely that the control of the living mulch to reduce competition with the main crop is a more relevant factor.     

Opaque2基因对糯玉米子粒品质的影响分析

玉米Opaque2(O2)基因突变显著影响子粒胚乳中蛋白体形成、淀粉含量、氨基酸组成等,其中,赖氨酸含量提高使营养品质大幅度提升。通过回交转育的方法构建17份糯玉米o2近等基因系,对子粒表型、种皮厚度、百粒重和粗淀粉含量等性状分析。结果表明,与对照相比,16个近等基因系子粒明显皱缩、胚乳变为粉质,6个近等基因系种皮厚度增加,11个近等基因系百粒重显著降低,12个近等基因系子粒粗淀粉含量显著降低。由此表明,o2基因的导入对糯玉米子粒表型、产量、淀粉含量等主要起负调控作用,影响程度与导入背景密切相关。