四川小麦品种贮藏蛋白位点及SSR分析*

利用种子贮藏蛋白和SSR标记研究了四川近20多年育成的小麦(Triticum aestivum)品种第1和第6同源群染色体的蛋白基因位点及遗传多样性。结果表明：供试的47个小麦品种共检测出47条醇溶蛋白条带，其中39条具有多态性，占82．98％；高分子量(HMW)谷蛋白亚基共出现7种亚基和11种亚基组合，表明四川主栽小麦品种在醇溶蛋白位点上存在广泛的变异，而HMW谷蛋白亚基遗传基础较狭窄。SSR结果表明，在21个位点中有8个(38．1％)位点具多态性，共检测到19个等位变异，供试品种在：DNA水平上存在一定的变异。聚类分析表明：两种标记均可将供试品种(包括中国春)分为三大类。通过Mantel检测，两种标记的分析结果间有显著的相关性，反映了在蛋白质和DNA水平上的结果可以互相补充，增加可靠性。     

低能N+离子注入诱导小麦种子高分子量谷蛋白亚基和醇溶蛋白的变异

应用SDSPAGE和APAGE电泳技术,对不同剂量低能(25keV)N+离子注入小麦稳定品系CH3286的M3代种子储藏蛋白高分子量谷蛋白亚基和醇溶蛋白变异进行了系统的分析。结果表明低能N+离子束注入能有效地诱导小麦种子高分子量谷蛋白亚基的变异。高剂量(10.8×1016N+cm2)N+注入的诱变频率高于中剂量(7.2×1016N+cm2),其亚基总变异频率分别是13.7%和4.2%。不同位点的高分子量谷蛋白亚基对N+离子的敏感程度不同,其中以Glu1D最敏感,变异频率由大至小分别是Glu1D>Glu1B>Glu1A。低能N+离子束注入诱导的醇溶蛋白变异与高分子量谷蛋白亚基的变异有相似的规律。醇溶蛋白遗传区对N+离子的敏感程度也不同,其中ω醇溶蛋白最敏感,能产生较多的变异,其次是γ和β醇溶蛋白,最不敏感的是α醇溶蛋白。在M3代植株群体中筛选到一些农艺性状较稳定的高分子量谷蛋白亚基和醇溶蛋白变异株。     

青藏高原野生老芒麦种质醇溶蛋白遗传多样性分析

采用酸性聚丙烯酰胺凝胶电泳(A-PAGE) 对采集自青藏高原的54份野生老芒麦（Elymus sibiricus L.）种质进行遗传多样性分析,获得下述结果:（1）供试材料共分离出42条带纹,多态率达92.86%。4个电泳分区（α、β、γ、ω）的平均Shannon指数为0.4627,Nei-Li遗传相似系数（GS）变异范围为0.2424~0.9767,平均值为0.5822。说明供试野生老芒麦材料具有较为丰富的醇溶蛋白遗传多样性。（2）对所有材料的聚类分析和主成分分析发现,在GS值为0.562的水平上供试材料可聚成4个大类,绝大部分来自于相同或相似生态地理环境的材料聚成一类,主成分分析显示了相似的结果,即醇溶蛋白图谱类型与材料的生态地理环境具有一定的相关性。（3）基于Shannon 多样性指数估算了5个老芒麦地理类群内和类群间的遗传分化,发现类群内遗传变异占总变异的68.17%,而类群间的遗传变异占总变异的31.83 %。（4）对各地理类群基于Nei氏无偏估计的遗传一致度的聚类分析表明,各地理类群间的遗传分化与其所处的地理生态环境具有较高的相关性。     

中国小麦品种醇溶蛋白Gli-1和Gli-2编码位点等位基因组成分析

利用酸性聚丙烯酰胺凝胶电泳(A-PAGE)方法,鉴定分析了我国部分重要小麦品种或种质醇溶蛋白Gli-1和Gli-2编码位点等位基因组成特点.36个品种的研究结果表明,我国小麦品种在醇溶蛋白几个主要位点上均存在较大的变异度,Nei氏遗传变异系数(H)达0.775.6个位点一共检测到59个不同的醇溶蛋白等位基因,其中出现频率较高的有8个等位基因,即Gli-B2g(55.56 %)、Gli-D1k(50 %)、Gli-A1a(33.33 %)、Gli-A2f(30.56 %)、Gli-B1b(22.22 %)、Gli-D1f(22.22 %)、Gli-B2b(22.22 %)和Gli-D2g(22.22 %).国外品种中很少的4个等位基因(Gli-D1f、Gli-A2f、Gli-B2g和Gli-D2g)在我国具有较高的频率.可作为1BL/1RS易位标记的Gli-B1l等位基因在中国品种中的频率较高.另外,还发现一些优质醇溶蛋白等位基因(如Gli-B1b、Gli-B2c、Gli-A2b等)在我国品种中出现的频率很低,这可能是中国小麦品种品质普遍较差的一个原因.     

小麦醇溶蛋白基因的HPCE和A—PAGE染色体定位及其比较分析

利用高效毛细管电泳（ＨＰＣＥ）技术分离中国春醇溶蛋白获得了４５个不同组分（峰或峰肩），通过过中国春第１，４，６，７部分同源群染色体ＮＴ系和ＤＴ系的ＨＰＣＥ分析，确定了各个醇溶蛋白组分的编码基因所在染色本臂，结果显示，全部蛋白组分均由第１，部６部分同源染色体短臂上的基因编码（即Ｇｌｉ－１和Ｇｌｉ－２位点），其中Ｇｌｉ－Ａｌ，Ｇｌｉ－Ｂ１和Ｇｌｉ－Ｄ１分别编码５，１０和１２个组分Ｇｌｉ－Ａ２，Ｇｌｉ－     

小麦体细胞无性系籽粒胚乳醇溶蛋白的变异

聚丙烯酰胺凝胶电泳法测定表明,小麦体细胞无性系籽粒胚乳醇溶蛋白发生了变异,这种变异在SC_1代就能表达。测定的两个基因型间存在显著差异,农大139的变异频率明显高于衣大142;同一基因型不同株系间也有差别。SC_1代的变异有的直至SC_3代仍发生分离,但绝大部分变异能稳定地遗传到后代。     

水稻10kD醇溶蛋白基因表达双元质粒的构建及其对烟草的遗传转化

将高含硫水稻１０ｋＤ醇溶蛋白基因从质粒ｐＦＭ１８亚克隆到植物高效表达载体卡盒ｐＲＳ１００中，得到双元表达质粒ｐＲＳ１０Ｋ；对其进行ＰＣＲ扩增及ＰＣＲ产物酶切分析，表明ｐＲＳ１０Ｋ包含来自ｐＲＳ１００的ｒｂｃＳ高效表达启动子、终止子和Ｍ１３通用引物互补序列及水稻１０ｋＤ醇溶蛋白基因。将ｐＲＳ１０Ｋ通过农杆菌感染法转化烟草，并对烟草转化植株进行ＰＣＲ检测和Ｓｏｕｔｈｅｒｎ杂交，结果证明ｐＲＳ１０Ｋ的Ｒ     

(英文)

用SDS-PAGE方法测定了我国10个小麦主产省份171份小麦品种和高代品系的高分子量麦谷蛋白亚基（HMW-GS）组成。鉴定出18种HMW-GS，40种HMW-GS组成形式，其中20种亚基其组成形式只在一个品种（系）中出现。Glu-A1位点亚基1和Null出现最多，Glu-B1位点7＋8和7＋9亚基对占绝对优势，Glu-D1位点2＋12亚基对出现频率最高。Null、7＋9、2＋12、Null，7＋8，2＋12，1、7＋8，2＋12，1、7＋9、2＋12等亚基组成形成出现频率最高，占分析品种的49.71％。与前人研究相比，新育成品种HMW-GS亚基组成发生了明显变化，面包优质亚基（对）1、5＋10出现的频率显著升高，亚基多态性增加，组成形式明显改善，这些对于品质改良和品种选育是非常有利的，新育成品种Glu-1品质评分已超过7。尽管个别品种亚基组成好，品质优良，但总体上看，我国小麦品种与其它国家相比品质还存在一定差距，提高5＋10、17＋18等优质亚基的频率是改善我国小麦面包品质的重要措施。     

山西小麦品种资源醇溶蛋白组成的遗传变异

利用A-PAGE技术,比较分析了近40年来山西小麦育成品种、外引品种与地方品种在醇溶蛋白组成上的差异,从而揭示山西小麦品种醇溶蛋白的遗传变异。结果表明：山西育成品种与外引品种比地方品种醇溶蛋白谱带数目分别增加了2.1和1.5条。在70条迁移率不同的谱带中,26条从地方品种到育成品种出现频率明显增加,23条明显降低,其余的21条变化规律不明显;分区来看,与地方品种比较,育成品种ω区谱带数目及谱带出现频率的增减变化最大,其次是γ区,β和α区变化最小。同时发现16.5和19.1为双联带,12.9、15.7、17.8为三联带,二者都在ω区但总不同时出现。对3种类型品种的遗传差异分析发现,地方品种间的平均遗传距离明显大于其他2类品种;聚类分析也发现,大多数地方品种、山西育成品种或外引品种总是分别聚在同一亚类,表明地方品种与山西育成品种和外引品种之间编码醇溶蛋白的基因位点存在较大差异,同时也说明近40年来山西育成品种之间、外引品种之间醇溶蛋白的遗传变异水平不高。     

Genetic Diversity of Glutenin Protein Subunits Composition in Durum Wheat Landraces [Triticum turgidum ssp. turgidum Convar. durum (Desf.) MacKey] from the Mediterranean Basin

The genetic diversity of high and low molecular weight glutenin subunits of 63 durum wheat landraces from different geographical regions in the Mediterranean Basin was studied using SDS-PAGE. Great variability in glutenin composition was found, with 42 high and low molecular weight glutenin haplotypes, 20 allele combinations at the HMW-GS loci, and 18 at the LMW-GS. All five possible LMW models were detected in all Mediterranean regions. Rare alleles were found at Glu-B1 locus in high frequencies and a priori related alleles to grain quality were also observed. Global genetic diversity index was relatively high (0.67); it ranged from 0.33 to 0.66. Cluster analysis on the frequency patterns of origins grouped genotypes following a geographical structure. Rogers’ distance coefficient on frequency pattern for each region of origin showed two germplasm pools with distinct quality profiles, where South West Asian landraces were very different from the landraces of other Mediterranean areas. The relationship between different regions of origin is discussed and two possible ways of introduction of wheat in the Iberian Peninsula (N Africa and SE Europe) are hypothesized. The use of Mediterranean durum wheat landraces as source of genetic variability for grain quality improvement is highly recommended.     

Assessment of genetic variability in hexaploid wheat landraces of Pakistan based on polymorphism for HMW glutenin subunits

Summary Sixty hexaploid wheat landraces collected from five regions of Pakistan were assessed for genetic variability in terms of high molecular weight (HMW) glutenin subunits as revealed by SDS-PAGE. The germplasm appeared to be diverse and unique on the basis of HMW glutenin subunit compositions. Out of 24 alleles detected at all the Glu-1 loci, four belonged to Glu-A1, 12 to Glu-B1 and eight to Glu-D1 locus. The number of novel HMW glutenin subunits detected were 1, 4 and 6 at the three loci (Glu-A1, Glu-B1, Glu-D1), respectively. The frequency distribution patterns of 24 allelic variants detected at the three Glu-1 loci in 1080 samples analysed for 60 accessions were determined both on the basis of individual accessions and on the basis of regions (accessions pooled across the regions). One allele (null) at the Glu-A1 locus, three alleles (17+18, 7+8, 14) at the Glu-B1 locus and, two alleles (2+12 and 2^**+12) at the Glu-D1 locus were found most frequently distributed in the 60 populations. Maximum variation was observed in the Baluchistan and Gilgit regions of Pakistan in terms of distribution of novel Glu-1 alleles. A higher gene diversity was observed between the populations as compared to the gene diversity within the populations while, a reverse pattern of gene diversity was observed when populations were pooled across the regions (higher within the regions than between the regions). A data base has been generated in this study which could be expanded and usefully exploited for cultivar development or management of gene bank accessions.     

Effects of Salinity and Drought Stress on Grain Quality of Durum Wheat

This work aimed to assess the influences of soil salinity and drought stresses on grain quality characteristics of selected salt-tolerant genotypes differing in salinity tolerance in durum wheat. This study was conducted under control, drought, and saline field conditions in separate experiments during 2 years. A randomized complete block design with three replications was used for each experiment. The results showed significant effects of genotype and environmental conditions on all grain-quality related traits. Salt and drought stress caused the significant increment of grain protein content, wet and dry gluten contents, and sodium dodecyl sulfate (SDS) sedimentation volume. Thousand-grain weight, grain protein yield, and test weight reduced significantly under both salinity and drought stress conditions. Protein content showed positive correlation with wet gluten, dry gluten, SDS sedimentation, and volume and strong negative correlation with other traits. It is concluded that influence of salinity stress was greater than drought stress on grain protein yield and some other grain-quality-related traits.     

Genetic diversity of HMW glutenin subunits in diploid,tetraploid and hexaploid <Emphasis Type="Italic">Triticum</Emphasis> species

The genetic variations of high-molecular-weight (HMW) glutenin subunits in 1051 accessions of 13 Triticum subspecies were investigated using sodium dodecyl sulfate polyacrylamide-gel electrophoresis. A total of 37 alleles were detected, resulting in 117 different allele combinations, among which 20, 68 and 29 combinations were observed in diploid, tetraploid and hexaploid wheats, respectively. Abundance and frequency of allele and combinations in tetraploid wheats were higher than these in hexaploid wheats. Allele Glu-A1c was the most frequent subunit at Glu-A1 locus in tetraploid and hexaploid wheats. Consequently, the results also suggested that the higher variations occurred at Glu-B1 locus compared to Glu-A1 and Glu-D1. Therefore, carthlicum wheat possessing the allele 1Ay could be presumed a special evolutional approach distinguished from other tetraploid species. Furthermore, this provides a convenient approach of induction of the 1Ay to common wheat through direct cross with carthlicum wheat. Alleles Glu-B1c and Glu-B1i generally absent in tetraploid wheats were also found in tetraploid wheats. Our results implied that tetraploid and hexaploid wheats were distinguished in dendrogram, whereas carthlicum and spelta wheats and however displayed the unique performance. In addition, founder effect, no-randomness of diploidization, mutation and artificial selection could cause allele distribution of HMW-GS in Triticum. All alleles of HMW-GS in Triticum could be further utilized through hybrid in the quality improvement of common wheat.     

新疆小麦高分子量麦谷蛋白亚基组成及主效亚基分析

为探索新疆强筋小麦的主效亚基,本文采用SDS-PAGE方法对新疆主栽的21种中筋小麦和17种强筋小麦的高分子量麦谷蛋白亚基进行了分析。结果表明:供试材料中共出现12种类型的亚基和12种亚基组合。中筋小麦Glu-A1位点亚基主要为N,Glu-B1位点亚基主要为7+8,Glu-D1位点亚基主要为2+12。强筋小麦Glu-A1位点亚基主要为1和N,Glu-B1位点亚基主要为7+8和7+9,Glu-D1位点亚基主要为5+10。聚类分析结果显示:在相似系数为0.43时可将供试小麦分为2类:第1类包括28个品种,均含有2+12亚基,其中既有强筋小麦也有中筋小麦;第2类包括12个品种,均含有5+10亚基,全部为强筋小麦。5+10亚基对小麦品质贡献明显大于其他亚基,是新疆强筋小麦的主效亚基。本研究可为提高新疆小麦面筋强度的分子育种提供理论依据。     

Effects of Nitrogen and Sulfur Fertilizers on Gliadin Composition of Several Cultivars of Durum Wheat

Four fertilizer treatments were applied to 10 cultivars of durum wheat (Triticum turgidum var. durum L.) in three seasons of designed field trials. The fertilizer treatments were N0S0 ‐ neither N nor S applied; N1S0 ‐ only N applied; N1S1 ‐ both N and S applied; N0S1 ‐ only S applied. It was found, through densitometric SDS‐PAGE, that the relative proportion of different gliadin components was influenced by the distinct regimes, even though our region in the center of the Argentinean Pampa is not considered to be deficient in soil S‐availability. The effects observed were more marked in one particular season of the three studied, were principally associated with the treatment expected to produce the maximum degree of grain S‐deficiency (N1S0), and were consistent with the hypothesis that the relative content of S‐poor gliadins increases under conditions of S‐deficiency, as has been found in related crops. Multivariate reverse‐phase high‐performance liquid chromatography (RP‐HPLC) analysis of samples from the season most affected generally supported these observations, but refined them in identifying quantitative differences between all four fertilizer treatments for the cultivars studied. These findings imply that differences in industrial quality might also be expected between the fertilizer treatments. RP‐HPLC also identified numerous genetically controlled qualitative differences in the gliadins between the cultivars studied, adding to previous published work on these cultivars based on SDS‐PAGE. These observations confirmed that one particular cultivar of very low gluten strength showed a gliadin profile markedly distinct from the remaining cultivars studied.     

Analysis of Genetic Variability in a Sample of the Durum Wheat (Triticum durum Desf.) Spanish Collection Based on Gliadin Markers

In this work gliadin proteins were used to analyse the genetic variability in a sample of the durum wheat Spanish collection conserved at the CRF-INIA. In total 38 different alleles were identified at the loci Gli-A1, Gli-A3, Gli-B5, Gli-B1, Gli-A2 and Gli-B2. All the gliadin loci were polymorphic, possessed large genetic diversity and small and large differentiation within and between varieties, respectively. The Gli-A2 and Gli-B2 loci were the most polymorphic, the most fixed within varieties and the most useful to distinguish among varieties. Alternatively, Gli-B1 locus presented the least genetic variability out of the four main loci Gli-A1, Gli-B1, Gli-A2 and Gli-B2. The Gli-B1 alleles coding for the gliadin γ-45, associated with good quality, had an accumulated frequency of 69.7%, showing that the Spanish germplasm could be a good source for breeding quality. The Spanish landraces studied showed new gliadin alleles not catalogued so far. These new alleles might be associated with specific Spanish environment factors. The large number of new alleles identified also indicates that durum wheat Spanish germplasm is rather unique.     

Relationship Between the Normalized SPAD Index and the Nitrogen Nutrition Index: Application to Durum Wheat

In a three-year field experiment in Toulouse (in Southwest France), two indicators of plant nitrogen (N) status were compared on five durum wheat cultivars: the normalized SPAD index and the nitrogen nutrition index (NNI). SPAD value is a non-destructive measurement of chlorophyll content from the last expanded leaf. The normalized SPAD index is expressed relative to SPAD reading on a fully fertilized crop. The NNI is calculated from the crop biomass and total plant N content using a universal N-dilution curve for wheat. The normalized SPAD index and NNI were closely related irrespective of year, cultivar, and growth stage. When N was a limiting factor, the SPAD index measured at anthesis predicted grain yield and protein content accurately. Unlike NNI, SPAD index cannot be used to predict these variables when wheat is over-fertilized.     

Influence of High Molecular Weight (HMW) and Low Molecular Weight (LMW) Glutenin Subunits Controlled by Glu‐1 and Glu‐3 Loci on Durum Wheat Quality

The progenies of four intervarietal durum wheat crosses were used to determine the effects of glutenin variants coded at Glu‐1 and Glu‐3 loci on durum wheat quality properties. The F₂ lines were analyzed for high molecular weight (HMW) and low molecular weight (LMW) glutenin composition by electrophoresis. Whole grain derived F₃ and F₄ samples were analyzed for vitreousness, protein, and dry gluten contents, gluten index, SDS sedimentation volume, mixograph, and alveograph properties. Allelic variation at the Glu‐B1 and Glu‐B3 loci affected gluten quality significantly. Comparisons among the Glu‐B3 and Glu‐B1 loci indicated that the LMW glutenin subunits controlled by Glu‐B3 c and j made the largest positive contribution, followed by the alleles a, k, and b. HMW glutenin subunits 14+15 gave larger SDS values and higher mixing development times than subunits 7+8 and 20. The positive effects of the glutenin subunits LMW c and HMW 14+15 were additive. Flour protein content, vitreousness, and mixograph peak height values were positively correlated with each other as well as with Dglut values, whereas the SDS sedimentation highly correlated with mixing development time, alveograph strength, and extensibility but was not correlated with the other parameters. The results of quality analysis, together with the results of the genetic analysis, led to the conclusion that SDS sedimentation, mixograph mixing development time, and peak breakdown are the tests more influenced by allelic variation of prolamin. The uses of the results in durum wheat quality breeding programs are discussed.