38–48 kDa subunits of buckwheat 13S globulins are controlled by a single locus

The inheritance of buckwheat (Fagopyrum esculentum Moench) seed storage proteins was investigated by control crosses between accessions containing different SDS‐PAGE protein patterns in the range of 38–48 kDa. Analysis of segregation of individual F₁ and F₂ seeds obtained by 17 crosses showed segregation of eight electrophoretic bands, from which 12‐banding variants were inferred. Two variants contained one band, nine contained two bands and one variant contained three bands and behaved as a single co‐dominant unit. The segregation of banding variants fits the expected ratios and thus supports the hypothesis of single Mendelian gene inheritance and that alleles are co‐dominant. The results therefore suggest that the genes controlling globulin subunits are tightly linked and inherited as a single locus. The locus was designated Glob‐1 and the twelve segregated protein variants as alleles a‐l. This is the first buckwheat globulin locus identified with multiple alleles, phenotypically expressed as groups of protein bands and thus applicable as a functional marker in buckwheat genomic studies.     

荞麦种子蛋白质组分分析

研究了不同品种荞麦种子中蛋白质组分情况，结果表明：养麦种子蛋白质主要由清蛋白、球蛋白、谷蛋白和醇溶蛋白构成。而不同类型蛋白质在总蛋白质中所占比例有明显差异，清蛋白是荞麦种子中的主要蛋白质，占总蛋白质含量的71．4％左右，其中以平荞最高。醇溶蛋白在荞麦种子总蛋白中所占比例较小，平均约为7．4％，榆养2号最高。球蛋白和谷蛋白所占比例非常接近，其中球蛋白平均约占9．1％，最高为日本大粒养。谷蛋白大约占总蛋白的12．1％，以京养1号最高。     

不同粒型稻米碾磨特性及蛋白质分布的比较

以3个具不同品质和粒形特点的典型水稻品种,比较研究了稻米碾磨特性及蛋白质含量与组分在籽粒内部的分布情况。碾磨程度和达到相应碾磨程度所需碾磨时间之间的非线性关系显示糠层由外到内硬度不断增加;淀粉胚乳层硬度保持不变,且高于糠层硬度。不同品种达到相同碾磨程度所需时间差异较大,说明淀粉等物质的沉积密度在品种中存在较大差异。蛋白质含量在籽粒不同部位不是均匀分布的,3个品种糙米中均约85%的蛋白质分布在胚乳层中,清芦占11的蛋白质含量以外层胚乳最高,苏御糯和扬辐粳4901蛋白质含量均以糠层最高。随碾磨程度的提高,籽粒蛋白质含量由表及里呈降低趋势,核心层胚乳只有糠层的1/2左右,但不同类型水稻品种的降低趋势不同。SDS-PAGE分析表明,不同种子贮藏蛋白组分在籽粒内部的分布是相对均匀的,说明不同蛋白质组分在积累过程中,其遗传表达相对同步。     

荞麦籽粒蛋白质组分特性研究

采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)方法研究了荞麦的2个栽培种苦荞与甜荞的总蛋白、清蛋白、球蛋白及谷蛋白特性。试验结果表明,甜荞籽粒总蛋白及蛋白质各组分的谱带具有丰富的多态性,而苦荞籽粒总蛋白及蛋白质各组分谱带的多态性有限;荞麦清蛋白主要由低分子量的亚基构成;甜荞球蛋白组分包含由中等到低分子量范围的5~12种亚基,苦荞球蛋白主要由8种亚基组成;甜荞谷蛋白主要由分子量在43~66.2 kDa之间的3~5种亚基组成,苦荞谷蛋白主要由分子量在31~43 kDa间的2种亚基组成。     

Effects of cultivar and environment on phenolic acids in buckwheat

Summary Phenolic acid contents were determined in seeds from five buckwheat cultivars grown at three locations in western Canada for four years. Buckwheat contained 12–16 g/kg of total phenolic acids, about 3 g/kg of esterified phenolic acids, and 8–13 g/kg of etherified phenolic acids. The latter represented 70 to 79% of the total phenolics. Variation in phenolic acids was mainly due to cultivar, seasonal effects and their interaction, while growing location had no significant effect. Phenolic acid contents of buckwheat were independent of seed color and protein content. Esterified phenolic acids exhibited a negative significant association with etherified phenolic acids in buckwheat.     

Identification of lily cultivars using isoelectric focusing of proteins from bulb scales and tissue culture bulblets

An efficient procedure is described for cultivar identification in lily, based on polymorphism in bulb scale proteins analysed by isoelectric focusing (IEF). Seventy (84%) out of 83 cultivars from which in vivo bulbs were analysed showed a unique banding pattern. Analysis of in vitro bulblets from 62 commercial cultivars resulted in 47 (76%) unique banding patterns. Cultivars with identical protein patterns always belonged to the same cultivars classification group (i.e., Asiatic hybrids, Oriental hybrids or Lilium longiflorum). Protein bands characteristic for the cultivar groups have been found. The protein patterns appeared to be very consistent and were not affected by the soil-type on which the bulbs were grown, long-term bulb storage, different protein sample storage methods, in vitro growing conditions, or in vitro bulblet storage. However, in six out of 10 cultivars from which in vivo bulbs and in vitro bulblets were analysed, protein patterns from in vivo bulbs consisted of a greater number of, and/or more intensely stained bands than those from in vitro bulblets of the same cultivar. The protein patterns appeared to be unsuitable markers for selection of reference cultivars for distinctness, uniformity and stability (DUS) testing as no relation was found between morphological characters and protein patterns. Cultivars with identical or very similar protein patterns were generally clearly distinct on the basis of morphological characters.     

Endosperm starch properties in several wheat cultivars preferred for Japanese noodles

To characterize superior genotypes for the white Japanese noodle, endosperm starch properties including amylose content, flour peak viscosity and starch-granule bound Waxy (Wx) proteins were compared using several cultivars preferred for noodle manufacture. Amylose contents from three seasons trials and flour peak viscosity from two seasons trials varied among cultivars. Low amylose content was a common property in the noodle cultivars, whereas a high peak viscosity was not always the case. When the Wx proteins were analyzed by sodium dodecylsulfate polyacrylamide gel electrophoresis, a clear reduction in the amount of low molecular weight protein or a lack of the high molecular weight protein occurred in the noodle cultivars. Segregation of Wx proteins was detectable in a B₁F₁ population, indicating that the Wx protein analysis has a potential as a surrogate of selecting low amylose genotypes in early generations.     

Polymorphism and DNA markers for asparagus cultivars identified by random amplified polymorphic DNA

Summary DNA polymorphism among five Asparagus officinalis L. cultivars-Imperial, Snow, Steline, UC-157 and Larac, as detected by random amplified polymorphic DNA (RAPD), is reported. Thirty one decamer primers were tested. and twenty six of them yielded amplification products. Fourteen primers gave products with at least one polymorphic DNA fragment. Among a total of 119 amplified fragments 33 were polymorphic. These RAPD markers enabled the identification of asparagus cultivars. Unique markers for cultivars were: Snow-bands 475 bp, 772 bp, 412 bp, 935 bp and 820 bp amplified by primers D5, OPA-07, OPA-09, OPA-10 and OPA-18, respectively. Steline-bands 645 bp, 680 bp and 997 bp amplified by primers A32, OPA-03 and OPA-09, respectively. A band 903 bp, amplitied by primer OPA-12, is a marker for Imperial, and a band 420 bp, amplified by primer D52, is a marker for Larac. Cultivar UC-157 could be identified by a combination of shared polymorphic bands. The pairwise marker difference between cultivars ranged from 0.08 to 0.17. A phenogram of the genetic relationship based on RAPD fits with the known origin of the cultivars.     

Stability of esterases and total proteins during seed storage of perennial ryegrass (Lolium perenne L.) and their use for cultivar discrimination

Polymorphisms in electrophoretic patterns obtained by isoelectric focusing (IEF) were examined to evaluate their suitability for cultivar identification in perennial ryegrass (Lolium perenne L.). It was possible to discriminate 64 (94%) of 68 cultivars by combining results from esterase and total protein analysis. Discrimination was based on quantitative differences (relative band intensity) rather than on qualitative differences (presence or absence of bands). Esterase patterns from different recent (fresh to seven years old) seed lots of the same perennial ryegrass cultivars were very stable. Occasionally, minor differences in band intensity were observed between recent and old (up to 30 years old) seed lots of a cultivar. Storage of meal samples up to two years at −20 ^°C had no effect on the total protein patterns. No correlation was found between esterase patterns and ploidy level, cultivar type (pasture or turf), heading date or breeding company. Esterase patterns appeared to be unsuitable markers for the selection of reference cultivars for distinctiveness, uniformity, and stability (DUS) testing, because no correlation was found between cultivars on the basis of esterase banding patterns and morphological characteristics. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic variation among cultivars of red clover (Trifolium pratense L.) detected by RAPD markers amplified from bulk genomic DNA

Summary The use of random amplified polymorphic DNA (RAPD) markers obtained from bulked samples was investigated for cultivar identification in red clover. Pooled samples were examined in order to minimize variation within cultivars. To determine the appropriate number of individuals to include in the bulked samples representing each cultivar, DNA samples from two, three, four, five, ten and twenty individuals were pooled. Twenty was found to be an appropriate number of red clover individuals per bulk in order to amplify only the DNA sequences shared among most individuals in each cultivar. Fourteen 10-mer primers were used to amplify genomic DNA from combined leaf samples of 15 red clover cultivars from European, Japanese and North American origins. A total of 79 amplified products, of which 55 were polymorphic, was obtained. Cultivar-specific bands were observed with 13 primers. The amplification patterns obtained from two primers could distinguish all 15 red clover cultivars. Rogers' genetic distances for all 105 pairwise comparisons were calculated to evaluate relationships among these cultivars. Cluster analysis based on these genetic distances separated these 15 cultivars into three groups, with two of the groups consisting of a single Japanese cultivar each, while the third group included cultivars from European, North American, and Japanese origins.     

Random amplified polymorphic DNA (RAPD) variation in Fagopyrum tataricum Gaertn. accessions from China and the Himalayan region

The diversity among 52 landraces and cultivars of tartary buckwheat (Fagopyrum tataricum Gaertn.) and one accession of its wild ancestor, F. tataricum ssp. potanini Batalin, from diverse geographic origins was examined using random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) markers. Eighteen primers produced a total of 240 fragments, of which 153 (63.75%) were monomorphic and 87 (36.25%) polymorphic bands. UPGMA-based pairwise Jaccard’s coefficient of similarity was used to deduce the relationships among 53 genetically diverse accessions. The similarity between cultivated tartary buckwheat accessions ranged from 0.61 to 1.00. Four distinct clusters were formed which corresponded well with the geographic distribution of the tartary buckwheat. Nepalese accessions showed maximum diversity followed by Chinese accessions. Tartary buckwheat accessions from the Himalayan region of northwestern India revealed a narrow gene pool. The wild buckwheat accession did not group with any of the three cultivated tartary buckwheat groups, and formed its own single-entry group. Genetic similarity (0.59) of Chinese buckwheat accessions with the wild ancestor reaffirmed that cultivated tartary buckwheat originated in the Yunnan province of northwestern China. Consistent with some earlier reports, our study demonstrated the usefulness of the RAPD technique for the characterization of plant genetic resources and assessment of diversity between species. This revised version was published online in August 2006 with corrections to the Cover Date.     

Comparison of phenotypic and molecular marker-based classifications of hard red winter wheat cultivars

Genetic diversity is the basis for successful crop improvement and can be estimated by different methods. The objectives of this study were to estimate the genetic diversity of 30 ancestral to modern hard red winter wheat (Triticum aestivum L.) cultivars adapted to the Northern Great Plains using pedigree information, morphological traits (agronomic measurements from six environments), end-use quality traits (micro-quality assays on 50 g grain or milled flour samples for the six environments), and molecular markers (seed storage proteins separated using SDS-PAGE, 51 SSRs, and 23 SRAP DNA markers), and to determine the relationships of genetic distance estimates obtained from these methods. Relationships among diversity estimates were determined using simple (Pearson) and rank (Spearman) correlation coefficients between distance estimates and by clustering cultivars using genetic-distances for different traits. All methods found a wide range in genetic diversity. The genetic distance estimates based on pedigree had the highest values due to possible over-estimation arising from model assumptions. The genetic diversity estimates based on seed storage protein were lowest because they were the major determinants of end-use quality, which is a highly selected trait. In general, the diversity estimates from each of the methods were positively correlated at a low level with the exceptions of SRAP diversity estimates being independent of morphologic traits (simple correlation), SDS-PAGE, and SSR diversity estimates (rank correlation). However, SSR markers, thought to be among the most efficient markers for estimating genetic diversity, were most highly correlated with seed storage proteins. The procedures used to accurately estimate genetic diversity will depend largely upon the tools available to the researcher and their application to the breeding scheme.     

Utilization of isozyme polymorphism for cultivar identification of 45 commercial peas (Pisum sativum L.)

Forty five Pisum sativum cultivars were analysed by isozyme electrophoresis with the objective to find protein markers for exact and reproducible discrimination of individual genotypes. The combination of six enzyme systems (acid phosphatase, amylase, esterase, leucine aminopeptidase, shikimate dehydrogenase and phosphoglucomutase) with two electrophoretic techniques (NATIVE-PAGE, isoelectric focusing) and use of seed and leaf tissue enabled to identify all 45 studied cultivars. Critical factors which may affect utilization of isozyme electrophoresis for commercial applications in pea breeding and seed production and testing are discussed. This revised version was published online in July 2006 with corrections to the Cover Date.     

不同颜色小麦籽粒的发芽特性和生理特征的研究

用３个不同籽粒颜色的小麦品种进行发芽特性的研究。结果表明，红皮小麦品种的发芽率和发芽指数低于白皮小麦品种，不同颜色籽粒的吸水速率没有明显差异，呼吸强度、电导率和α淀粉酶活性随色级的增加面降低。３个小麦品种的淀粉同工酶有８条共显带，而扬麦５号有一特殊的小分子酶带。ＧＡ３处理可明显提高发芽速度、显著增加ＰＰ紫色和扬麦５号的苗高和芽鞘长度。     

Relationship Between Gluten Strength and Glutenin Proteins in Durum Wheat Cultivars

An electrophoretic study of gliadin and glutenin proteins, mainly low-molecular-weight (LMW) Glutenin Subunits, was undertaken to investigate possible assoeiations between these proteins and gluten strength. Thirty-eight durum wheat cultivars having different origins and currently grown in Spam were analysed. Different electrophoretic methods were used to analyse the seed storage proteins. Protein grain content was estimated and gluten strength was measured by the SDS-sedimentation test. New patterns for LMW glutenins were observed. Besides the known patterns of LMW-1, associated with γ-gliadin 42, and LMW-2 associated with γ-gliadin 45, six cultivars had LMW-2^? associated with γ-gliadin 43, one cultivar showed LMW-2* associated with γ-gliadin 44, and another cultivar, null for γ-42 and γ-45, had LMW-1^?. Significant differences for gluten strength were found among groups of cultivars with different LMW patterns. High molecular weight glutenins were found in general to be poor indicators of viscoelastic properties, although band 20 showed a negative influence on quality. The results are discussed in relation to development ol cultivars with good gluten strength.     

不同浓度PEG-6000对苦荞种子萌发的影响及抗旱性评价

选取4个苦荞材料,以不同浓度(0%、5%、15%、25%)的PEG-6000溶液胁迫萌发,研究不同浓度PEG-6000对苦荞种子萌发影响,以期筛选出苦荞抗旱性鉴定指标并对不同苦荞材料的抗旱性进行综合评价。结果显示,5%PEG-6000胁迫促进了云南花苦和晋荞2号的种子萌发,提高了黔苦4号的幼苗活力指数、胚芽含水量、胚根干物重和储藏物质转运率。15%PEG-6000胁迫抑制了苦荞的种子萌发,降低了苦荞的胚芽长、幼苗活力指数和储藏物质转运率,提高了苦荞的根茎比、黔苦4号的胚根长和胚根干物重。25%PEG-6000胁迫下4个材料均未发芽,复水后川荞1号多倍体恢复发芽率仅为16.7%。通过灰色关联分析筛选出萌发指数、发芽率、储存物质转运率作为抗旱鉴定的初级指标,发芽势、胚芽干物重、胚芽含水量、胚根含水量作为抗旱鉴定的次级指标。通过隶属函数对苦荞材料抗旱性进行评价,发现不同苦荞材料在5%PEG-6000、15%PEG-6000胁迫下种子的抗旱性不同。     

Expression of heritable biochemical markers from various pecan tissues

Summary Isozyme expression of malate dehydrogenase (MDH), phosphoglucomutase (PGM), and phosphoglucose isomerase (PGI) were examined from 12 tissues of pecan by starch gel electrophoresis. Tissue type, stage of development, and extraction method had little effect on isozyme expression of MDH or PGM but did influence PGI expression. Additional PGI bands were observed from seed tissue and soaked pollen. A direct relationship was observed between PGI expression from seed tissue and eventual expression from the germinated seedling. Additional PGI bands from soaked pollen were thought to be produced from structural changes and not due to more efficient extraction. The genotype of immature pecan seeds could be determined for Mdh-1, Pgi-2, and Pgm-1 provided cellular endosperm had developed. Watery endosperm did not contain detectable levels of the study enzymes. The advantages of uniform isozyme expression among various tissues and ability to determine the genotype of immature pecan seed are discussed.     

Bulked AFLP analysis for the assessment of genetic diversity in white clover (Trifolium repens L.)

The use of bulked leaf samples from individual plants for amplified fragment length polymorphism (AFLP) analysis was evaluated as a tool for assessment of genetic diversity in white clover (Trifolium repens L.). Bulking of leaf samples produced slightly simpler AFLP profiles compared to the combined profiles of individual plants from the same cultivar. Approximately 90% of bands which were present in individual plants were present in bulked samples of the same cultivar. The majority of those absent were rare bands, shared by less than 25% of individual plants. Replicate bulk samples gave almost identical banding patterns, demonstrating the robustness of the bulked AFLP technique. Cluster analysis of AFLP data derived from individual plants resulted in a phenogram similar to that produced from data derived from bulked samples of the same plants. AFLP analysis of bulked samples detected significant amounts of genetic variability among 52 cultivars and accessions with genetic similarity values ranging from 0.42 to 0.92. However, cluster analysis of AFLP data only partially reflected the geographic origin of cultivars and accessions and was not congruent with cluster analysis based on variation for morphophysiological characters. Bulked AFLP analysis provides a powerful tool for rapid assessment of genetic variability in white clover and may also be used for cultivar identification. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic and chemical analyses of six cowpea and two <Emphasis Type="Italic">Phaseolus</Emphasis> bean species differing in resistance to weevil pest

The objective of this work was to evaluate the genetic variability among six cowpea (Vigna unguiculata) cultivars differing in their resistance to Callosobruchus maculatus (F.) weevil. Two resistant bean cultivars were used to compare between the sensitive, moderate tolerant, and high tolerant cowpea cultivars. The differentiations were performed by using random amplified polymorphic DNA (RAPD) fingerprinting, protein concentration and organic and non-organic components in seed coat. Six polymorphic primers were identified, resulting in different informative bands. Based on polymorphic profiles, three clusters were formed. Clustering was mainly affected by the resistance to weevil pest. The sensitive cowpea cultivars were separated in one group, the moderate tolerant and high tolerant cultivars came in separate groups, and finally, the resistant bean cultivars separated clearly in one distinct group. The most interesting result was represented by concentration of total protein in the seed coat. The protein concentration in the resistant bean cultivars were approximately 50% less than concentration in each of the moderate tolerant and sensitive cultivars of cowpea. Ferric ions were about 25% less than the moderate tolerant and sensitive cultivars. The concentrations of calcium and potassium in seed coats were higher in the resistant beans than in cowpea cultivars. Cobalt was about four times higher in resistant bean than in the sensitive and moderate tolerant cowpea cultivars, which may play a major role in seed resistance to weevil.     

Stylar ribonucleases in almond: correlation with and prediction of incompatibility genotypes

To clarify incompatibility relationships among almond cultivars, 35 were analysed for stylar ribonucleases, which have previously been shown to correlate with incompatibility S alleles. Stylar proteins were extracted and separated electrophoretically and the zymograms compared with ladders of ribonucleases corresponding to the 12 S alleles previously reported. Sixteen cultivars showed a band corresponding to two of the known ribonucleases, 17 showed one known ribonuclease and one ‘new’ band, and two showed two new bands. Twelve new ribonucleases were detected; 11 were attributed to new S alleles (S₁₃ to S23) and a mutant form of S₇ was attributed to S_7A. Genotypes were proposed for nine cultivars of five incompatibility groups that had not been genotyped previously, VII, X, XI, XII and XIII. Twenty‐four cultivars of unknown incompatibility relationships were provisionally genotyped: six of these could be assigned to existing groups and two new groups were established, XIV and XV, along with group O of cultivars with unique genotypes. Test crosses confirmed that eight pairs of cultivars showing similar zymograms were indeed cross‐incompatible, including the two representatives of each of the two new groups. Virtually all self‐incompatible cultivars of known genotype are listed in a table. The data should be useful for planning cultivar combinations for orchards and for designing crosses for breeding programmes.