Effect of the grain protein content locus Gpc-B1 on bread and pasta quality

Grain protein concentration (GPC) affects wheat nutritional value and several critical parameters for bread and pasta quality. A gene designated Gpc-B1, which is not functional in common and durum wheat cultivars, was recently identified in Triticum turgidum ssp. dicoccoides. The functional allele of Gpc-B1 improves nitrogen remobilization from the straw increasing GPC, but also shortens the grain filling period resulting in reduced grain weight in some genetic backgrounds. We developed isogenic lines for the Gpc-B1 introgression in six hexaploid and two tetraploid wheat genotypes to evaluate its effects on bread-making and pasta quality. In common wheat, the functional Gpc-B1 introgression was associated with significantly higher GPC, water absorption, mixing time and loaf volume, whereas in durum wheat, the introgression resulted in significant increases in GPC, wet gluten, mixing time, and spaghetti firmness, as well as a decrease in cooking loss. On the negative side, the functional Gpc-B1 introgression was associated in some varieties with a significant reduction in grain weight, test weight, and flour yield and significant increases in ash concentration. Significant gene × environment and gene × genotype interactions for most traits stress the need for evaluating the effect of this introgression in particular genotypes and environments.     

Identification of novel glutelin subunits and a comparison of glutelin composition between japonica and indica rice (Oryza sativa L.)

Glutelin, a major protein in rice grains, is encoded by a multigene family. However, its protein composition is not well characterised. Here, we identified and characterised two novel glutelin subunits, GluBX and GluC. The individual glutelin subunits of japonica cv. Nipponbare and indica cv. 93-11 rice were analysed using 2-dimensional gel electrophoresis, LC–MS/MS, and Western blotting. Comparison of the glutelin profiles between three japonica and three indica cultivars indicated two distinct subunits (GluA-1 and GluA-3 isomers) and a distinction in the subunit composition (notably GluA-3 and Lys-rich GluB-1 components) of these two subspecies. Sequence alignment revealed different nutritional (Lys residues) and functional (Cys residues) characteristics between the type-A and type-B glutelin subfamilies. We also analysed amino acid and total protein contents of the grains in thirty-five cultivars, and we demonstrated that the Lys-rich glutelin composition of indica cultivars is superior to that of japonica cultivars. The Lys-rich and Cys-poor GluBX subunit is a native protein and is a high nutritional protein in grains. Our combined approaches for the identification of glutelin subunits have revealed the nutritional characteristics of individual subunits in rice, and this knowledge will provide new insights for improving grain quality during rice breeding.     

QTL mapping for crude protein and protein fraction contents in rice (Oryza sativa L.)

Improvement of rice storage proteins is important in rice breeding for high nutritional quality. Seventy-one recombinant inbred lines (RIL) derived from a cross between japonica variety Asominori and indica variety IR24 were used to study the inheritance of crude protein and protein fraction contents in rice. A total of 16 QTL were identified and mapped on eight chromosomes. Several QTL affecting contents of different protein fractions were mapped in the same chromosomal region. In particular, two QTL with a significant contribution were identified to simultaneously affect prolamin and glutelin contents. One QTL denoted as qCP-12 affecting crude protein content (CP) was located in the same region as QTL qGLT-12 affecting glutelin content, in agreement with the positive correlation between glutelin level and protein content. QTL with larger genetic effects were further confirmed using two sets of chromosome segment substitution lines (CSSL), where Asominori and IR24 were used as the recurrent parents. By QTL comparative analysis, two QTL for CP, three for globulin content and one for prolamin content were located in the vicinity of CP QTL previously identified in polished rice. Application of these results in rice breeding is discussed.     

High level of resistance to Sclerotinia sclerotiorum in introgression lines derived from hybridization between wild crucifers and the crop Brassica species B. napus and B. juncea

Sclerotinia rot caused by the fungus Sclerotinia sclerotiorum is one of the most serious and damaging diseases of oilseed rape and there is keen worldwide interest to identify Brassica genotypes with resistance to this pathogen. Complete resistance against this pathogen has not been reported in the field, with only partial resistance being observed in some Brassica genotypes. Introgression lines were developed following hybridization of three wild crucifers (viz. Erucastrum cardaminoides, Diplotaxis tenuisiliqua and E. abyssinicum) with B. napus or B. juncea. Their resistance responses were characterized by using a stem inoculation test. Seed of 54 lines of B. napus and B. juncea obtained from Australia, India and China through an Australian Centre for International Agricultural Research (ACIAR) collaboration programme were used as susceptible check comparisons. Introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum had much higher levels (P < 0.001) of resistance compared with the ACIAR germplasm. Median values of stem lesion length of introgression lines derived from the wild species were 1.2, 1.7 and 2.0 cm, respectively, as compared with the ACIAR germplasm where the median value for stem lesion length was 8.7 cm. This is the first report of high levels of resistance against S. sclerotiorum in introgression lines derived from E. cardaminoides, D. tenuisiliqua and E. abyssinicum. The novel sources of resistance identified in this study are a highly valuable resource that can be used in oilseed Brassica breeding programmes to enhance resistance in future B. napus and B. juncea cultivars against Sclerotinia stem rot.     

Association between line per se and hybrid performance under excessive soil moisture stress in tropical maize (Zea mays L.)

Excessive moisture (EM) stress during the summer–rainy season is one of the major production constraints for maize (Zea mays L.) in large areas of South and South-East Asia. A key question in breeding for tolerance to excessive moisture is the extent to which the performance of maize hybrids can be predicted on the basis of per se performance of inbred lines under excessive moisture. We attempted to identify the relationship between morpho-physiological traits and grain yield measured on inbred parents and their single cross progenies under EM stress. Responses of various morpho-physiological traits, except days to 50% anthesis, differ significantly under normal versus EM stress. Superiority of hybrid progenies over parental inbred lines increased under EM stress, suggesting that hybrids were comparatively more tolerant to EM stress than inbred progenies. Across moisture regimes, all morpho-physiological traits of hybrids, except lodging and root porosity under normal moisture, were found to be positively and significantly correlated with mid-parent traits. Our data suggest that per se performance of lines was a relatively more important factor in determining hybrid performance under EM stress, while under optimum soil moisture conditions mid-parent heterosis was relatively more important than per se performance of mid-parent. Phenotypic correlation between hybrid and mid-parent yields showed a strong relationship under EM stress (r = 0.66^**). The relationship was statistically significant under normal moisture as well, though it was comparatively weak (r = 0.41^*). Our findings suggest that performance of hybrid progenies under excessive moisture can be predicted and improved to some extent on the basis of their inbred parents that have been systematically selected and improved for EM stress.     

基于黄早四与掖478衍生的双向BC3回交群体的品质分析

以玉米骨干自交系黄早四与掖478分别作为轮回亲本构建的双向BC3回交群体为试验材料,系统分析在不同遗传背景下的玉米子粒淀粉、蛋白质、油分以及赖氨酸含量的变化及其相关性。结果表明,在不同遗传背景下,玉米子粒的淀粉含量与蛋白质、赖氨酸含量均呈极显著负相关,蛋白质与赖氨酸含量呈极显著正相关,蛋白质含量的改良在一定程度上可以促进赖氨酸含量的提高。     

Characterization of free amino acid QTLs in maize opaque2 recombinant inbred lines

The opaque2 (o2) mutation in maize (Zea mays L.) increases the content of free amino acids (FAA) in the endosperm. We investigated the basis of this trait by using recombinant inbred lines from a cross of Oh545o2 (high FAA) and Oh51Ao2 (low FAA) to identify quantitative trait loci (QTL) for FAA content and to determine their effect on FAA composition and protein accumulation. Mapping identified six QTLs that accounted for 71% of the phenotypic variation. Two QTLs in bins 4.01 and 7.02 are close to α-zein genes; high FAA individuals with these QTLs had reduced accumulation of α-zein 19 kDa isoforms and increased FAA abundant in α-zeins. A QTL in bin 3.03 is close to a gene encoding triose phosphate isomerase (tpi4) and a higher expression of this enzyme was found in high FAA individuals. Other differentially expressed proteins included vicilin-like globulins and the enzymes glyceraldehyde-3-phosphate dehydrogenase, enolase-2, sorbitol dehydrogenase and granule-bound starch synthase. The results suggest that the increased levels of FAA in o2 endosperm are mainly due to the reduction of storage proteins and the failure to incorporate their amino acids into other proteins, as well as the alteration of carbohydrate metabolism that may favor amino acid biosynthesis.     

Ear rot,aflatoxin accumulation,and fungal biomass in maize after inoculation with Aspergillus flavus

Aflatoxin, a toxin produced by the fungus Aspergillus flavus Link: Fries, occurs naturally in maize (Zea mays L.). Aflatoxin is a potent human carcinogen and is also toxic to livestock, pets, and wildlife. When contaminated with aflatoxin, the value of maize grain is markedly reduced. This investigation was conducted to compare ear rot, aflatoxin accumulation, and fungal biomass in maize single crosses with varying degrees of resistance to aflatoxin accumulation and to determine the relative importance of general combining ability (GCA) and specific combining ability (SCA) in the inheritance of resistance to ear rot, aflatoxin accumulation, and fungal biomass. Eight germplasm lines with different levels of resistance to aflatoxin accumulation were used as parents of a diallel cross. The cross was evaluated for visible ear rot, aflatoxin accumulation, and A. flavus infection in the grain. A. flavus infection was determined by quantitative real-time polymerase chain reaction (qRT-PCR) assays. Both GCA and SCA were significant sources of variation in the inheritance of the three traits although GCA accounted for a greater portion of the variation among single crosses. The interactions of GCA and SCA with years were highly significant for aflatoxin accumulation, but not significant for A. flavus infection. Estimates of GCA effects were highly significant for both reduced A. flavus infection and reduced aflatoxin accumulation for Mp313E, Mp715, and Mp717. Conversely, GCA effects associated with GA209 were significant for reduced levels of A. flavus infection and ear rot, but high levels of aflatoxin accumulation. Mp313E, Mp715, and Mp717 should be useful in breeding programs targeting both reduced levels of fungal infection and aflatoxin accumulation.     

Integrated management of Striga hermonthica, stemborers,and declining soil fertility in western Kenya

Striga hermonthica (Delile) Benth., stemborers, and declining soil fertility are serious threats to sustainable food production in the Lake Victoria zone of Kenya. To address these constraints, promising integrated crop management technologies were evaluated, using a multi-locational design in four sub-locations in Siaya and Vihiga district (western Kenya) for six cropping seasons. Technologies evaluated consisted of the traditional maize (Zea mays L.) – bean (Phaseolus vulgaris L.) intercrop, maize – Desmodium (Desmodium uncinatum (Jacq.) DC.) push–pull intercrop, Crotalaria (Crotalaria ochroleuca G. Don) – maize rotation, and soybean (Glycine max (L.) Merr) – maize rotation. Within each of these systems, imazapyr-coated herbicide-resistant maize (IR-maize) and fertilizer were super-imposed as sub-plot factors. The push–pull system was observed to significantly reduce Striga emergence and stemborer damage from the second season onwards. IR-maize reduced and delayed Striga emergence from the first cropping season. Differences in Striga emergence and stemborer damage between the other systems were not significantly different. After five cropping seasons, the Striga seedbank was significantly higher in the maize-bean intercrop system than in the push–pull system under both maize varieties while the rotational systems had intermediate values not different from the day zero values. Under IR-maize, the Striga seedbank was significantly lower than under local maize for all cropping systems. Maize yields varied between seasons, districts, and cropping systems. Yields in the push–pull system were higher than in the maize-bean intercrop after two seasons and in the absence of mid-season drought stress. Both maize and soybean responded significantly to fertilizer application for both districts and for most seasons. The various interventions did not substantially affect various soil fertility-related parameters after five seasons. In the short term, IR-maize integrated in a push–pull system is the most promising option to reduce Striga while the rotational systems may need a longer timeframe to reduce the Striga seedbank. Finally, farmer-led evaluation of the various technologies will determine which of those is really most acceptable under the prevailing farming conditions.     

玉米耐旱相关基因dbf1的序列变异分析

玉米dbf1 基因在干旱胁迫下的编码产物与DIP₁蛋白相结合,调控植物激素脱落酸（ABA）响应基因rab17 的表达,从而提高耐旱性。以B73的dbf1 基因组序列为模板,对104个玉米自交系的dbf1 基因进行测序,研究玉米dbf1 基因的序列多态性。多态性分析表明,在全长为2 118 bp的序列中共发现48个SNP和12个InDel。尽管大部分变异位点集中在非编码区,但编码区的1个InDel和3个非同义突变位点的变异可以产生氨基酸序列改变。玉米dbf1 基因的全长序列和编码区序列的变异位点可以分别将该基因划分成33和11种单倍型,并且编码7种蛋白质。在供试材料中,dbf1 基因至少经历了9次重组,中性检测表明,该基因的进化没有偏离中性选择。     

过量表达拟南芥GPX3基因提高玉米苗期抗旱性研究

通过农杆菌介导法将拟南芥抗旱基因AtGPX3导入玉米自交系郑58中,用PCR和RT-PCR法对转化玉米进行检测,在水分胁迫下对T₁代转基因玉米和非转基因玉米进行抗旱性分析。结果表明,共得到56株转化苗,检测获得9个株系的30株T₀代转基因阳性植株,抗性植株阳性率为53.6%。RT-PCR检测表明,T₁代有6个株系为稳定遗传阳性株系,并且AtGPX3基因在转基因玉米中表达量大幅度提高。耐旱性分析表明,非胁迫条件下,非转基因和转基因株系中游离脯氨酸（Pro）和丙二醛（MDA）的含量基本无显著差异。在干旱胁迫条件下,转基因玉米叶片的Pro含量高于非转基因玉米,比非转基因株系提高了46.2%;MDA含量低于非转基因玉米,比非转基因玉米下降了34%。通过导入AtGPX3基因,可以提高玉米苗期的耐旱性。     

Puroindoline genes and proteins in tetraploid and hexaploid species of Triticum

Six tetraploid (5 Triticum turgidum and 1 Triticum timopheevii) and four hexaploid (three Triticum aestivum and one Triticum kiharae) taxa of Triticum were studied in order to identify novel variation in Pin genes and proteins which can be exploited in the improvement of cultivated wheat. Western blotting with a highly specific antibody showed that puroindoline proteins were present in all of the hexaploid lines but were absent from the tetraploids. The immunoreactive bands differed slightly in their relative mobilities and their relative amounts, which could have resulted from variation in the allelic forms of Pin a and Pin b. This was supported by HPLC analyses which showed differences in the retention times and peaks heights of the putative puroindoline components in T. kiharae and T. timopheevii. Sequence analyses of cDNAs also showed variation in the sequences of expressed puroindoline genes. In particular, a sequence encoding a new form of Pin b was present in T. aestivum ssp. macha.     

玉米自交系铁7922苗期耐低氮胁迫原因探讨

以低氮胁迫(NO3--N0.2mmol/L)与正常供氮(NO3--N2mmol/L)条件下的生物量比值作为耐低氮能力指标,从12份玉米自交系中筛选出在低氮胁迫条件下苗期能够正常生长的自交系铁7922。通过不同玉米自交系对低氮胁迫反应差异的比较研究,从根系与氮利用效率对低氮环境的响应以及硝酸盐吸收动力学等3个方面对铁7922耐低氮原因进行了探讨。结果表明,铁7922对低氮环境的适应是多因素共同作用的结果,表现在氮胁迫浓度下根干重的增加、氮利用效率的显著提高以及较小的Km和低的Cmin。单纯的根干重的增加(掖515)或氮利用效率的提高(H21)或较小的Km(4F1、豫8701)并不能保证在低氮环境的正常生长。     

Swiss Flint maize landraces—A rich pool of variability for early vigour in cool environments

Today's modern Dent × Flint maize (Zea mays L. ssp. mays) hybrids have a high yield potential but often lack satisfying early vigour under typical low spring temperatures of temperate latitudes. Maize was introduced into Europe already in the 16th century and until the 1950s, landraces evolved and adapted well to various geographically restricted and climatically marginal regions in Europe. Therefore, the objective of this study was to assess representatives of the large pool of Swiss Flint maize landraces for their early vigour under cool conditions in the field. A set of 17 landrace accessions were tested for 2 years at sites on the Swiss plateau (450 and 550 m a.s.l.) and in the foothills of the Swiss Alps (830 and 870 m a.s.l.). Plant emergence (PE), emergence index (EI), the efficiency of the photosystem II (F_v/F_m), leaf greenness (SPAD) and plant dry weight at the three- and six-leaf stages (DW3_P, DW6_P) were measured.     

玉米骨干亲本对主要病害的抗性及其传递规律

选用5个玉米骨干亲本Mo17、黄早4、掖478、自330和丹340及其68个衍生品系,对大斑病、小斑病、丝黑穗病、弯孢叶斑病、禾谷镰孢茎腐病、矮花叶病和黑粉病的抗性进行鉴定.结果 表明,黄早四具有较好的多抗和兼抗性,60.7％衍生品系对大斑病达中抗以上水平,82.0％高感或感小斑病,对小斑病的抗性较易丢失,感丝黑穗病易...     

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.     

Stalk strength and reaction to infection by Macrophomina phaseolina of brown midrib maize (Zea mays) and sorghum (Sorghum bicolor)

Reduced lignin concentration in brown mid-rib mutants in both maize and sorghum have resulted in improved dry matter digestibility, increased milk yield and higher energy in lactating cows. However, the mutations were not widely deployed due to concern that reduced lignin concentration might increase vulnerability to lodging and stalk rot incidence. The objective of this study was to determine the effects of the mutations on stalk strength and stalk rot resistance in both sorghum and maize. Six brown midrib (bmr) sorghum, four brown midrib (bm) maize, and their normal isolines were evaluated for stalk strength and stalk rot disease reaction at two locations in four replications. Three randomly selected plants in each plot were inoculated with Macrophomina phaseolina at 14 d after flowering by using the toothpick inoculation technique. On 28 d after inoculation, the plants were rated for disease severity by measuring the length of necrotic lesions in the stalks. Stalk strength was determined from another three random plants in each plot by using a rind penetrometer. The mutations had no effect on stalk rot disease severity in either sorghum or maize though stalks strength was markedly affected by the mutations in both species. While maize bm entries broke easily in response to mild mechanical stress, bmr sorghums did not exhibit sign of stalk collapse in all backgrounds. The result suggests that the bmr genes in both maize and sorghum can be deployed without incurring losses to stalk rot disease.     

QTL identification of kernel composition traits with popcorn using both F2:3 and BC2F2 populations developed from the same cross

Quantitative trait loci (QTL) influencing three main kernel composition traits, starch, protein and oil concentrations, in unselected F_2:3 and selected BC₂F₂ maize populations derived from the same cross of a dent corn inbred Dan232 × an elite popcorn inbred N04 under the same conditions were detected. Four and two QTL for starch, four and three QTL for protein, and four and one QTL for oil were detected in the two populations, respectively, with three QTL jointly detected. The proportion of phenotypic variation explained by a single QTL was 5.2–10.6%, 5.0–14.3%, and 6.2–8.5% for the three traits. Ten QTL had favorable alleles contributed by Dan232. Several QTL detected in this study had identical or similar chromosome regions to those previously identified with other maize germplasms. No QTL with opposite effects for kernel composition traits and popping characteristics were detected in the same or near marker intervals. This reflected that some QTL detected in this study seemed to contribute to trait variation in a diverse array of maize populations and environments, and the opportunity existed for improving popcorn's nutritional quality while maintaining acceptable popping characteristics. Inconsistent broad sense heritability and trait correlation estimates were also observed in the two populations.     

玉米高赖氨酸含量基因型离体筛选方法与应用

设不同浓度赖氨酸(L)加苏氨酸(T)的培养基, 将opaqure-2等32个玉米自交系种子的胚接种在培养基上发芽, 同时测定种子中游离氨基酸和总氨基酸含量, 检测胚生长对L+T的抗性与种子中赖氨酸含量关系, 筛选鉴定玉米种子中高赖氨酸含量的有效方法。结果表明, opaqure-2对L+T有较强抗性, 子粒中有较高含量游离赖氨酸;普通玉米自交系抗L+T能力较弱, 种子中游离氨基酸含量较低;在旅大红骨类群中检测到对L+T有较强抗性的基因型, 子粒中有较高含量的赖氨酸和其他氨基酸。玉米中存在典型的天冬族氨基酸生物合成的反馈抑制代谢途径, 试管L+T胁迫方法可用于筛选和鉴定玉米高赖氨酸基因型。     

Stand establishment and early field vigour variation in a tropicalised shrunken-2 maize population

Low field emergence and early field vigour are major problems in shrunken-2 maize (sh-2). The genetic variability for stand-ability and early field vigour in a shrunken-2 maize population previously improved for adaptability to a tropical environment was investigated using a Design I mating system in which 40 randomly sampled males were crossed to four randomly sampled plants that served as females. Plants of the 160 crosses, representing full- and half-sib relationships, were evaluated in four sets, each composed of 40 crosses derived from 10 males during two growing seasons. Experimental design was randomized complete block with three replications. Difference in Emergence Index (EI) was not significant between the two seasons. Emergence Percentage (E%), Emergence Rate Index (ERI), and early field vigour traits viz. vigour score and seedling height determined at 28 days after planting (DAP) were, however, significantly (P < 0.05–P < 0.01) better in the first growing season. For all traits, the ‘female/male’ item was significant (P < 0.01) while ‘male’ was mostly non-significant. ‘Season × female/male’ interaction was significant for stand establishment traits but not for early field vigour traits. Averaged over all traits, dominance variance was 8.2 times higher than additive variance. Genetic variation for stand establishment and early field vigour in the tropicalised shrunken-2 maize population would be best exploited through the development of inbred lines, hybrids and synthetics. Broad-sense heritability was 53.0% for E%, 36.0% for EI, 33.0% for ERI, 71.5% for vigour score and 90.0% for seedling height. One hundred-seed weight ranged between 82.0 and 182.0 mg but correlation coefficients between seed weight on one hand, and E%, EI, ERI, vigour score and plant height on the other hand, were low (mostly <0.30) and, in general, non-significant. Emergence percentage had the highest average genotypic correlation value with the other traits studied (Mean Absolute Value = 0.71 in the first season and 0.45 in the second season). Emergence percentage, determined 10 days after planting, thus has potential for use as a selection index for stand establishment and early field vigour.