大豆属多年生野生种及栽培种种子蛋白质电泳分析

用SDS聚丙烯酰胺凝胶电泳(SDS—PAGE)法分析了5个多年生野生种及栽培种的种子水溶性蛋白。结果表明:各个种基本都有其特征电泳图谱。多年生野生种G.latifolia.G.microphylla及栽培种G.max的种内蛋白质图谱较为一致,而多年生野生种G.falcata,G.tabacinaG.tomentella种内蛋白质图谱则差异较大。种间蛋白质图谱比较表明:多年生野生种G.latifolia,G.micropylla、G.tabacina之间亲缘关系较近,而其他种间关系较远。野生种G.tomentella与栽培种G.max有较近的亲缘关系。     

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.     

Wild safflower species (Carthamus oxyacanthus): A possible source of resistance to the safflower fly (Acanthiophilus helianthi)

Safflower (Carthamus tinctorius L.) is an important oil seed crop with growing importance in many countries around the world. However, safflower fly (Acanthiophilus helianthi Rossi) is one of the main limiting factors to expand the production area of the crop in several countries. Since host plant-resistance is the most efficient method for pest management, we evaluated a germplasm of wild accessions (Carthamus oxyacanthus Bieb.) with mostly brown–black seed coat color along with some white-seeded cultivated safflower genotypes for resistance to safflower fly. The results showed that there was a significant difference between two species for safflower fly damage with all cultivated genotypes being susceptible. However, for the accessions of C. oxyacanthus, a wide range of resistance was observed. In a choice experiment and natural infestation, 35.3–78.9 percent of heads per plant were infested in genotypes of C. tinctorius while for the accessions of C. oxyacanthus this was much smaller and ranged from 1.6 to 13.6%. In addition, percentage of seed yield loss per infested plant was more drastic in cultivated safflower (29.0–72.8%) than the wild accessions (0.0–21.4%). In the no-choice experiment, fly population was dramatically decreased in the wild accessions due to larval mortality and possibility of antibiosis. There was a strong relationship between brown–black seed coat color and resistance to safflower fly indicating the possibility of using this trait in breeding programs of safflower to develop fly-resistant cultivars.     

Evaluation of the Ugandan sorghum accessions for grain mold and anthracnose resistance

Sorghum accessions from Uganda were evaluated for grain mold and anthracnose resistance during the 2005 and 2006 growing seasons at the Texas A&M University Research Farm, near College Station, TX. Accession PI534117 and SC719-11E exhibited the lowest grain mold severities of 2.4, whereas, accessions PI534117, PI534144, PI576337, PI297199, PI533833, and PI297210, with SC748-5 were highly resistant to anthracnose in both years. Accessions PI534117, PI297134, PI297156 exhibited low grain mold severities in 2006. Significant negative correlation was recorded between grain mold and percent germination and high temperature in both years. In 2006, daily precipitation recorded significant positive correlation with grain mold. The seed mycoflora was analyzed across sorghum lines and treatments. In 2005, Curvularia lunata and Fusarium thapsinum were the most frequently recovered fungal species with 31 and 21% incidence, respectively, followed by Alternaria spp. (19%) and F. semitectum (13%). In 2006, predominant colonizers were F. thapsinum (58%), followed by Alternaria and F. semitectum with 15 and 10, percent respectively, while C. lunata had a 6% incidence. In this study, PI534117 holds promise for multiple disease resistance, as it had the lowest disease severity of grain mold and was highly resistant to anthracnose in both years. It also has a high germination rate, a high seed weight, and its short stature is more advantageous for the new A-line conversion program.     

Hard seed breakdown patterns of unprocessed forage legume seed sown into dry soil in summer in southern Australia

The patterns and extent of hard seed breakdown of dormant seeds and pods following burial in the soil in February (summer) varied for up to 19 accessions of 12 species of annual forage legumes. The experiments, at six sites across southern Australia, were designed to identify legumes whose patterns of hard seed breakdown would suit them to agronomic evaluation in a summer sowing research program. For a subset of accessions, sufficient hard seed breakdown was demonstrated between February and April to allow a high proportion of seed to germinate in late autumn—a desirable pattern for summer sowing. Although there was a large G x E interaction, some cultivars of Ornithopus sativus Brot., O. compressus L. and Trifolium spumosum L. had a pattern that was reproducible at all sites. The experiments also identified accessions of Biserrula pelecinus L. and T. glanduliferum Boiss., which had the desirable pattern in regions of south‐eastern Australia, but not Western Australia. A pragmatic approach for selection of forage legume accessions in situ for evaluation within a summer sowing program was demonstrated.     

Application of a PCR-Based Cytoplasm Genotyping Method for Phylogenetic Analysis in Potato

We previously developed five polymerase chain reaction-based markers (T, S, SAC, D, and A) to distinguish potato cytoplasms into six types (M, P, A, W, T, and D). As the applicability of this genotyping method for phylogenetic studies had been questioned, we applied this method to species distantly related to cultivars (four accessions of two tomato species, and 176 accessions of 29 Solanum species). The T marker uncovered two insertions, which, along with a unique S marker band, supported independency of series Piurana. The A and D markers generated unique bands to A- and D-type cytoplasms, respectively, but the SAC marker generated a similar banding pattern to the cytoplasms of both cultivated and their distantly related species. Consequently, while the cytoplasm type definition is validated only among cultivated potatoes and their closely related wild species, the developed markers, with the exception of the SAC marker, could provide useful phylogenetic information.     

部分栽培稻和野生稻种子谷蛋白的电泳分析

 供试的不同属、不同种及不同生态型的水稻材料的种子各蛋白经SDS-PAGE电泳后可分辨为3类多肽，分子量分别为l8～23 kDa．30～36 kDa和49～100 kDa。不同属的材料各蛋白带型明显不同； 同属内基因组相同或相近种的各蛋白有特征多肽，AA基因组的有约Mt.50 kDa的多肽2～3条，CCDD基因组的有约Mt.23 kDa的多肽一条；籼型和粳型稻间带型存在差异，而爪哇稻、粳稻、籼稻间，前二者略近，并以粳型陆稻与爪哇稻最近似。同一生态型的各品种，除细弱带偶尔有差异外，带型基本一致。由此可见，水稻各蛋白电泳分析作为方法之一， 对于属间和稻属内各种及种内分类是有意义的。     

Characterization of Amaranthus cruentus L. seed proteins by 2-DE and LC/MS–MS: Identification and cloning of a novel late embryogenesis-abundant protein

Amaranth is taking great attention as an important cereal crop that could fulfill food requirements for the growing population, especially in developing countries. However, the protein composition of these seeds is not well known yet. We have used the proteomics tools to characterize amaranth seed proteome. About 400 proteins spots were resolved on two-dimensional gel electrophoresis (2-DE) and protein spots were analyzed by LC-MS/MS (liquid chromatography tandem mass spectrometry). Identified proteins were related to stress and defense responses, metabolic, respiratory, and oxide-reduction processes. One abundant spot was identified as a Late Embryogenesis Abundant (LEA) protein and the gene was cloned and characterized. The AcLEA cDNA contains a 418 bp open reading frame (ORF) encoding a polypeptide of 139 amino acids. Bioinformatics analysis showed that AcLEA belongs to LEAs Group 5. Proteomics is a powerful technique that could be used even in non-sequenced organisms such as amaranth. The obtained information reveals that amaranth seed, beyond the classical seed storage proteins, contains proteins related to protection against stress. The identification of these proteins opens the door to the application of new strategies to improve the quality of amaranth production.     

Measurement ofBeta-amylase in Malting Barley (Hordeum vulgareL.). I. Development of a Quantitative ELISA forBeta-amylase

A double antibody, sandwich enzyme-linked immunosorbent assay (ELISA) was developed using polyclonal antibodies specific tobeta-amylase to estimate the amount of ‘free’ (soluble in aqueous saline solution) or ‘combined’ (extracted with saline solution including reducing agent)beta-amylase protein in barley grain and malt. This ELISA was used to quantify the amount ofbeta-amylase in barley grain and malt from four varieties grown at nine sites in South Australia in 1993. The antibody used to develop the ELISA reacted differently withbeta-amylase depending on whether the source was barley grain or malt, and on thebeta-amylase band pattern in isoelectric focussing (IEF) of the barley variety. On the basis of their IEF band patterns barley varieties were divided into two types, designatedBmy1-Sd1 andBmy1-Sd2. Malting resulted in proteolytic cleavage of thebeta-amylase peptide with a reduction in the apparent molecular weight of up toM_r4000 and the appearance of new maltbeta-amylase IEF bands that were more basic. The new maltbeta-amylase IEF band patterns still allowed the identification of theBmy1-Sd1 andBmy1-Sd2 IEF types despite the change in molecular weight and pI. The data obtained using thebeta-amylase ELISA were highly correlated withbeta-amylase activity for both the free and combined fractions when the IEF band pattern and its source, barley grain or malt, were taken into account.     

Cultivar discrimination of Portuguese Lupinus albus by seed protein electrophoresis: the importance of considering “glutelins” and glycoproteins

The Osborne protein fractions of seeds of 17 Lupinus albus cultivars from different regions in Portugal (obtained from plants grown at the same location) were analysed. The objective of this work was to examine the variability in the seed protein fractions of such a L. albus germplasm sample in order to disclose existing genetic relationships and to obtain information useful for a breeding program. Globulins and, interestingly, the fraction extracted with NaOH (“glutelins”) were the major seed protein constituents. The several protein fractions were separated by electrophoresis (reducing SDS polyacrylamide gel electrophoresis) and the patterns examined by multivariate analysis. Bands of “glutelins” together with those of glycosylated polypeptides were found to have the highest discriminating capacity. Lupin populations were grouped according to their clinal geographical distribution from north (small-seeded, higher plant architecture and later flowering type) to south (large-seeded, shorter and early flowering type). It is also possible that polypeptide patterns could reflect microclimatic specificities related with altitude and temperature. In fact, one cultivar from the south but from high altitude was grouped with the north plant types adapted to colder and wetter weather, whereas two cultivars from a warmer north region were grouped with the southern types. The results show, in addition, that a general correlation exists between cultivar early flowering characteristics and higher amounts of both total seed protein and glutelins, and lower glutelin glycosylation. Since high seed protein content and early flowering are important aims of lupin breeding programs, these findings on the seed protein fractions could stimulate and direct future research in lupin improvement.     

Species relationships between a wild tetraploid potato species,Solanum acaule bitter, and its related species as revealed by rflps of chloroplast and nuclear DNA

Solanum acaule Bitter is a wild, disomic tetraploid potato species widely distributed in the South America. In this study, species relationships between eight accessions ofS. acaule and 108 accessions of 27 morphologically closely related species were investigated using chloroplast and nuclear DNA RFLPs. Chloroplast DNA types were determined for most of the accessions based on restriction digestion patterns of entire chloroplast DNA. C type chloroplast DNA was common inS. acaule andS. albicans (a hexaploid derivative fromS. acaule), which was also found inS. megistacrolobum, S. sanctae-rosae,S. raphanifolium, and several other Peruvian species in seriesTuberosa. Nuclear DNA was analyzed by Southern hybridization with 31 single-copy RFLP probes. One hundred and thirty-four bands were scored, from which a UPGMA dendrogram was constructed to show overall similarities among accessions.Solanum acaule andS. albicans (andS. demissum) formed a cluster with a high similarity, and then, this group was clustered withS. megistacrolobum andS. sanctae-rosae. The resulting cluster was well separated from other clusters. Thus, we suggest that an initial tetrasomic tetraploid arose from eitherS. megistacrolobum orS. sanctae-rosae. Interestingly, a Mexican hexaploid speciesS. demissum showed the closest similarity toS. acaule andS. albicans, suggesting that they shared a common ancestry.     

Manihot flabellifolia Pohl,wild source of resistance to the whitefly Aleurotrachelus socialis Bondar (Hemiptera: Aleyrodidae)

Aleurotrachelus socialis is one of the most important pests of cassava (Manihot esculenta Crantz) in the Neotropics. In Colombia, high whitefly populations can reduce crop yields by 79%; and although the farmers intensify the use of insecticides, this practice is highly contaminating, costly and leads to the development of resistance in the insect. An alternative for managing whitefly populations is to develop genetically resistant varieties. Wild parents of Manihot are a useful source of genes against pests for the cultivated species of cassava. Based on prior research that showed the existence of moderate-to-high levels of resistance to A. socialis in Manihot flabellifolia, a wild species of cassava, this study was proposed to characterize this new source of resistance, evaluating the biology and demographics of A. socialis on eight accessions of M. flabellifolia, a susceptible check (CMC-40) and a resistant (MEcu72) check. The averages of A. socialis longevity and fecundity on the accessions were not significantly different to MEcu72, but different from CMC-40 (P < 0.05). Development time was not significantly different, ranging from 35–40 days on accessions and MEcu72 and 33.5 days on CMC-40 (P < 0.05). In contrast, the population growth rate (r_m) was significantly lower on the M. flabellifolia accessions, with Fla 61 standing out with a growth rate 98 and 99% less than that obtained on MEcu72 and CMC-40, respectively. Once the resistant levels have been identified to A. socialis on the M. flabellifolia accessions, interspecific crosses of M. esculenta subsp. M. flabellifolia and backcross programs could be developed to incorporate the desirable characteristics from the wild relatives into elite progenitors of M. esculenta.     

Distribution of the 241 bp deletion of chloroplast DNA in wild potato species

The common potato (Solanum tuberosum L. ssp.tuberosum) has T-type chloroplast DNA characterized by a 241 bp deletion. To explore the maternal ancestry of the common potato, a total of 566 accessions of 35 wild species, collected mostly from central Bolivia to northern Argentina, were determined for presence or absence of the deletion by a simple PCR assay using primers flanking the deleted region of chloroplast DNA. Sixteen out of 80 accessions ofS. tarijense, S. berthaultii, andS. neorossii showed a shorter PCR amplified fragment. Sequencing of these fragments revealed that the same 241 bp was deleted at the same position in these accessions. This strongly suggests that the deletion event had occurred in wild species.     

Australian Oryza: Utility and Conservation

Australian Oryza are an understudied and underexploited genetic resource for rice improvement. Four species are indigenous: Oryza rufipogon, Oryza meridionalis, Oryza australiensis are widespread across northern Australia, whereas Oryza officinalis is known from two localities only. Molecular analysis of these wild populations is required to better define the distinctness of the taxa and the extent of any gene flow between them and rice. Limited collections of these wild populations are held in seed and DNA banks. These species have potential for domestication in some cases but also have many traits of potential value in the improvement of domesticated rice. Stress tolerance (biotic and abiotic) and grain quality characteristics in these populations may be useful.     

Tocols and carotenoids of einkorn,emmer and spring wheat varieties: Selection for breeding and production

Einkorn (Triticum monococcum L., subsp. monococcum), emmer (Triticum dicoccum Schuebl [Schrank], subsp. dicoccum) and spring bread wheat (Triticum aestivum L.) may be rich in lipophilic antioxidants (tocols and carotenoids), and therefore potential food sources with good nutritional properties. The aim of the present study was to determine the contents of major lipophilic antioxidants beneficial for human health in wheat varieties and landraces for breeding and production. In field experiments over two years, fifteen einkorn, emmer and spring wheat varieties were analysed for the contents of tocols and carotenoids in grain. A high carotenoid content (lutein, zeaxanthin, β-carotene) was typical for the selected einkorn genotypes. Among the analysed wheat species, the highest content was of β-tocotrienol, especially in the einkorn accessions. α-Tocotrienol was abundant in einkorn and emmer wheat species. Higher contents of α- and β-tocopherols were characteristic of spring and emmer wheats. δ-Tocotrienol has been detected for the first time in einkorn and some emmer accessions, although in low concentrations. Significant effects of genotype on the contents of carotenoids and tocols were found with einkorn differing from emmer and spring wheats. The year of cultivation had less effect on the contents of carotenoids and tocols. Selected accessions of einkorn with high contents of carotenoids and tocotrienols as well as spring and emmer wheats with higher contents of tocopherols are good sources of antioxidants with potential health promoting benefits for the production of nutritionally enhanced foods.     

Protein quality of developed home made weaning foods

Home made weaning foods developed from locally available foods like bajra, barley, green gram (Vigna radiata L.), amaranth grain (Amaranthus sp.) and jaggery using household technologies like roasting and malting had a PER ranging from 2.04 to 2.13, BV 79.56 to 80.68, NPU 66.75 to 67.86, NPR 2.13 to 2.76 and PRE 34.18 to 44.18. The values were comparable to that of cerelac—a commercial weaning food.     

Indirect changes associated with a selection program for increased seed-yield in wild species of Lesquerella (Brassicaceae): Are we developing a phenotype opposite to the expected ideotype?

Seed-yield stability, frequently associated with drought-tolerance strategies, is one of the main breeding objectives for the development of crops for semi-arid mediterranean-type environments. Since breeding of new industrial crops targeted for arid lands is not appreciably different from that of traditional crops, higher yield is achieved by increased harvest-index, at the cost of losing traits associated with drought-tolerance and reduced seed-yield stability. Using Lesquerella as a model we compared selected and unselected accessions of annual (L. gracilis and L. angustifolia) and perennial (L. pinetorum and L. mendocina) genotypes grown in field experiments in Patagonia, Argentina. Our objective was to assess the effects of breeding for increased seed-yield on traits related to the main characteristics that define the most common ideotype for mediterranean-type environments: early vigor, conservative growth strategy post-anthesis and reserves storage. Our specific question was: Have any of the attributes associated with seed-yield stability been indirectly selected during the domestication process? Our results show that these characteristics were reduced or lost, in selected lines compared to their wild relatives. Early vigor was lower in selected accessions and was associated mainly with reduced relative growth rate and CO₂ assimilation. During the reproductive period the growth strategy was changed by selection towards a non-conservative and more acquisitive resource use strategy. Traits associated with this strategy were linked to higher water use efficiency and growth capacity (higher CO₂ assimilation rate, specific leaf area, and leaf allocation), but also with loss of structural adaptation to low resource environments (i.e. low specific leaf area), an increase in nutrient and water demands, and reduced nutrient use efficiency. Carbohydrates accumulation pre-anthesis was lower in selected accessions of all four species, and also, just in perennials we found lower reserves storage post-anthesis. These changes in the pattern of carbohydrates accumulation could be associated to lower seed-yield stability due to the loss of buffer capacity linked with the use of pre-anthesis reserves for seed filling. On the other hand, in perennial species lower reserves storage after seed harvest could reduce plant longevity and survival. We conclude that indirect changes occurred during the preliminary domestication of both annual and perennial species of Lesquerella used in our experiment. These changes were against those required if these species were to be developed as crops for semi-arid, mediterranean environments and should result in low seed-yield stability.     

Characterization of seed proteome in Brachypodium distachyon

Brachypodium distachyon, an emerging model plant system for some economically important temperate grasses such as wheat, barley and switchgrass, has recently caught wide attention in modern biological research. In the current study, the glutenin, albumin and globulin components of 13 B. distachyon accessions were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) followed by peptide mass finger printing (PMF) and MS/MS protein identification. Abundant wheat low molecular weight glutenin subunit (LMW-GS) like proteins and a few high molecular weight glutenin subunits (HMW-GS) with low expression level were detected in B. distachyon. A total of 18 storage proteins and 15 albumin proteins were identified through PMF and MS/MS. The results demonstrated that the major seed storage proteins in B. distachyon are wheat LMW-GS like proteins and globulins. The identified albumins and globulins were mostly various enzymes that were classified into five groups according to their functions. The 2-DE spot distribution and MS results suggested that post-translational modifications (PTMs) such as phosphorylations and glycosylations are common phenomena in B. distachyon seed proteome.     

Co-current introgression of economically important traits in a potato-breeding program

Wild potato species contain many traits of economic importance. Late blight (LB) resistance and cold chipping are traits desired in potato cultivars. These traits could be co-currently introgressed if they occurred together in wild potato species. Our research objectives were (1) to determine if variation for cold chipping exists between potato species, accessions within species, and plants within accessions all having foliar LB resistance, and (2) to identify wild potato genotypes combining LB resistance and cold chipping. Materials include 665 genotypes from 43 LB-resistant accessions of 12 potato species having Endosperm Balance Numbers (EBN) of 1, 2, and 4, and 59 LB-resistant genotypes retained from these accessions for breeding. Potato chips were made from greenhouse-grown tubers following storage at 4 C for 6 months. Chip color was scored 1–10, ≤ 4 is acceptable by industry standards. Most of the variation for chip color was due to differences between species. Species ranged in the percentage of acceptably chipping genotypes (0% – 67%) with nine of 12 species having cold-chipping genotypes. Appreciable variation was present within accessions as well. The best chipping accessions wereS. verrucosum plant introduction (PI) 161173 – 4.33 / 0.67 (mean / proportion acceptable genotypes),S. stoloniferum PI 250510 -4.36 / 0.64,S. pinnatisectum PI 347766 -4.65 / 0.35 and 275233 -4.73 / 0.44, andS. megistacrolobum PI 195210 -5.14 / 0.29. Eleven 1EBN genotypes fromS. pinnatisectum andS. trifidum and five 2EBN genotypes fromS. verrucosum, S. fendleri,S. stoloniferum, andS. microdontum were identified that combined LB resistance and cold chipping. Co-current introgression would require fewer breeding cycles than other breeding methods to identify hybrid genotypes possessing both traits.