小麦族四个属模式种的醇溶蛋白分析

利用酸性聚丙烯酰胺凝胶电泳 (APAGE)对小麦族披碱草属、鹅观草属、猬草属和仲彬草属 4个属的模式种进行了醇溶蛋白电泳分析 ,结果表明 :(1 ) 4个模式种具有明显的醇溶蛋白遗传多样性 ,其种间醇溶蛋白多态性高达 92 .3 % ;(2 ) Elymus sibiricus和 H ystrix patula具有相似的醇溶蛋白带型 ,而 Roegneria caucasica和Kengyilia gobicola的带型基本相似 ,其醇溶蛋白图谱能够反映一定的系统关系 ;(3 )不同收集地的 E. sibiricus材料间也存在明显的醇溶蛋白遗传差异 ,新疆的 E. sibiricus具有较丰富的醇溶蛋白带纹 ,而甘肃的 E. sibiricus的醇溶蛋白带纹较少。     

赖草属植物醇溶蛋白的遗传多态性

用酸性聚丙烯酰胺凝胶电泳 (A PAGE)对小麦族赖草属 2 0种和 1亚种的 45份材料进行醇溶蛋白遗传分析 ,结果表明 :(1 ) 4 5份材料共出现 43种不同的醇溶蛋白图谱 ,从中分离出的 38条带纹多态性高达 1 0 0 % ;(2 )赖草属植物具有丰富的醇溶蛋白遗传多态性 ,其种间和种内不同居群间均存在明显的醇溶蛋白遗传变异 ,其种间变异大于种内变异 ,醇溶蛋白图谱可以作为鉴定赖草属植物的指纹图谱 ;(3)醇溶蛋白图谱的聚类结果与形态学结果基本一致 ,表明醇溶蛋白资料可以运用于赖草属植物种间、种内遗传差异及亲缘关系研究     

种子醇溶蛋白提取及检测条件探索

以大麦、小麦、玉米、高粱和苏丹草的种子为材料提取种子的醇溶蛋白,分析了不同提取剂及不同固液比[种子粉末样品质量(g)与提取剂的体积(mL)的比例]对种子醇溶蛋白提取效果的影响,并对SDS-PAGE检测醇溶蛋白中的不同胶浓度、厚度以及样品上样量等的影响进行了研究.结果表明,60%的正丙醇、乙二醇、异丙醇和叔丁醇分别是小麦、大麦、玉米以及高粱和苏丹草的最佳提取剂,将1∶6比例提取的种子醇溶蛋白以15μL上样,0.5mm厚度的15%分离胶电泳可以得到清晰的电泳检测图谱.     

八倍体小冰麦及其亲本种子醇溶蛋白和高分子量麦谷蛋白亚基的研究

对５个八倍体小冰麦种子醇溶蛋白和高分子量麦谷蛋白亚基的电泳谱带进行了分析,结果表明：八倍体小冰麦中１和中２的电泳谱带基本相同,中３、中４、中５的电泳谱带基本相同,但完全不同于中１和中２的类型。八倍体小冰麦中１和中２同天蓝冰草（Ａｇｒｏｐｙｒｏｎｉｎｔｅｒｍｅｄｉｕｍ（Ｈｏｓｔ）Ｐ．Ｂ．＝Ｅｌｙｔｒｉｇｉａｉｎｔｅｒｍｅｄｉａ（Ｈｏｓｔ）Ｎｅｖｓｋｉ＝Ｔｈｉｎｏｐｙｒｕｍｉｎｔｅｒｍｅｄｉｕｍ（Ｈｏｓｔ）ＢａｒｋｗａｒｔｈａｎｄＤｅｗｅｙ）在高分子量麦谷蛋白亚基上存在一条相同的谱带,在醇溶蛋白谱带上出现了小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．）和冰草均没有的带型。中３、中４、中５在醇溶蛋白谱带上具有一条冰草×染色体组的特征谱带,其基因表达程度同冰草类似。从５个八倍体小冰麦种子醇溶蛋白和高分子量麦谷蛋白亚基的电泳图谱结果,分析了八倍体小冰麦染色体组构成及亲本来源,并探讨了八倍体小冰麦在优质麦育种过程中的价值。     

南瓜属4个栽培种种子蛋白质电泳分析

通过聚丙烯酰胺凝胶电泳(PAGE)对南瓜属4个栽培种中国南瓜、印度南瓜、美洲南瓜及黑籽南瓜的12个品种种子蛋白组分进行了分析。结果显示,黑籽南瓜种子以盐溶蛋白条带居多;其它3个栽培种以水溶蛋白条带居多,其次为盐溶蛋白;4个栽培种酸溶蛋白条带均较少;醇溶蛋白未分离得到蛋白谱带。水溶、盐溶、酸溶蛋白电泳图谱显示各栽培种间蛋白谱带差异明显,同一栽培种品种间蛋白谱带较为一致。     

鹅观草属三个物种及其居群间的醇溶蛋白分析

利用酸性聚丙烯酰胺凝胶电泳技术(APAGE)对小麦族鹅观草属3个物种:毛叶鹅观草、纤毛鹅观草和竖立鹅观草进行了醇溶蛋白电泳分析,23份材料电泳分离出22条相对迁移率不同的谱带,其中3条(16.6%)共同带,19条(83.4%)具多态性,每个材料可分离出9～16条谱带。结果表明:(1)三个物种具有相似的醇溶蛋白带型,但存在明显的种间差异;(2)同种不同居群间也存在遗传差异;(3)种间差异大于种内差异。     

鹅观草种质资源醇溶蛋白遗传多样性研究

以8个地理类群的90份鹅观草野生种质材料为研究对象,采用A-PAGE(酸性聚丙烯酰胺)凝胶电泳技术进行蛋白质水平遗传多样性检测.研究结果表明,来源于不同居群的鹅观草共分离出26条谱带,每个材料可以分离出5～26条迁移率不同的谱带, 平均谱带数为16.39条,其中平均多态性谱带为12.65条,多态性比例为77.22%;基于供试材料醇溶蛋白每个位点谱带出现的频率,分别计算了地理类群内多样性指数(0.345)和总多样性指数(0.471),类群间的遗传分化系数为26.8%,表明鹅观草变异的73.2% 来源于类群内;90份供试材料醇溶蛋白的Jaccard遗传相似系数变异范围为0.133 3～1.000,平均遗传相似系数为0.395 7;利用种子醇溶蛋白可将90份材料分为12类,鹅观草种质资源之间的亲缘关系呈现出一定的地域性规律;不同地理类群间的遗传多样性指数从高到低的排列顺序依次为云南＞四川＞内蒙古＞新疆＞山西＞甘肃＞宁夏＞河北.因此在进行鹅观草种质资源收集和原地保护时,建议对云南和四川地区的鹅观草种质资源应给予极大关注.     

人工合成六倍体小麦醇溶蛋白遗传多样性分析

运用酸性聚丙烯酰胺凝胶电泳(A-PAGE)技术,对96份人工合成六倍体小麦的醇溶蛋白多样性进行了分析。结果显示,96份人工合成小麦中,共分离出65条不同的醇溶蛋白谱带,其中ω区22条,β和γ区各17条,α区9条,但各醇溶蛋白在电泳图谱中出现的频率差异较大,其变化范围为1.04%~91.67%。醇溶蛋白遗传多样性指数(H′)及多态性信息含量(PIC)分析结果显示,β、ω两个谱带区醇溶蛋白组成最为丰富,而α区最低;聚类分析结果显示,材料间的平均遗传距离为0.86,在遗传距离为0.83水平上,96份材料被划分为4个主要类群,类群间的关系基本反映了合成双二倍体的亲缘关系。研究结果表明,人工合成六倍体小麦醇溶蛋白基因位点表现出广泛的遗传变异,具有丰富的遗传多样性。     

南方红豆杉不同居群遗传多样性的RAPD研究

用随机扩增多态性方法对广东、湖南、江西等3省的12个南方红豆杉自然居群进行了基因组DNA多态性分析,从100条引物中共筛选出10个引物,获得RAPD谱带86条,多态性谱带占51％。聚类分析结果表明：南方红豆杉居群间的遗传距离与这些居群的地理分布相关,即相同或相邻产地的居群间的遗传距离较小,不同产地个体间的遗传距离较大。粤北南方红豆杉的9个居群的遗传多样性较低,可能与近年来资源遭到严重破坏,及其生长缓慢、种子萌发率低、成活率不高等原因有关。     

半夏居群遗传分化的同工酶和可溶性蛋白电泳的聚类分析

用聚丙烯酰胺凝胶电泳（PAGE）技术分析了安徽产7个半夏（PinelliaternataBreit）居群的4种同工酶和可溶性蛋白谱带。根据其中的PER、EST和可溶性蛋白谱带的分布特征,用Forguson和Aquado＆Avise的公式分别计算其谱带的相似性系数和遗传距离,以类平均法（UPGMA）进行了聚类分析。结果表明：半夏居群间表现出一定的遗传分化;鹞落坪半夏与半夏的黄山、九华山、大别山万佛寺居群间的遗传距离较小（D≤0.214）,结合形态变异,笔者认为鹞落坪半夏只能是半夏居群中形态上较为特殊的地方宗,而不是一新种;狭叶半夏与其它6个居群的遗传距离较大（D=0.416）,是半夏居群中种系分化较为强烈的一个居群。文中还讨论了半夏的遗传分化与形态变异、生境等的相关性。     

Quantitative genetic diversity and conservation strategies for an allogamous annual species,Dasypyrum villosum (L.) Candargy (Poaceae)

Dasypyrum villosum (L.) Candargy is a weedy annual diploid (2n = 14, VV genomes) allogamous grass species (Poaceae, Triticeae). Genetic variation for 12 traits was studied in 43 natural populations (31 from Italy and 12 from Croatia and Montenegro of former Yugoslavia) grown in a common field environment in California. Although 7 of 12 traits followed the theoretical prediction that a larger proportion of genetic variation was distributed within populations than among populations, exceptions were found for spike length, plant height, and days to flag-leaf emergence, heading, and anthesis. Covariate analysis showed that developmentally closely related characters were more likely correlated at both population and family within population levels. Geographically closer populations shared more genetic similarity than distant populations as indicated by mean coefficients of variation and cluster analysis of the Euclidean distances among populations. As few as five populations, each population with five or more half-sib seeds taken randomly from 5 plants, is expected to capture more than 95% of the total genetic variation of this species in the region sampled, but sampling a much larger number of seeds per population (> 1000) for long-term storage would supply research and plant breeding needs for several decades. If seed regeneration is required, populations can be sampled from clusters having similar genetic variation, and grown in reproductive isolation or bulked seed samples from all populations of each cluster group can be grown in isolation. The former is recommended if population integrity is desired while the latter is sufficient to provide genetic resources for plant-breeding purposes.     

A Rapid DNA Extraction Method for RFLP and PCR Analysis from a Single Dry Seed

A single-seed DNA extraction method was developed for rapid identification of plant genotype. The method was applied to 12 plant species, including the oil seeds sesame and soybean. The results were comparable to those obtained for oil-less seeds such as rice. This method will be useful for genotypic selection which requires rapid screening of large populations. It can also be used to identify varietal purity of seed stocks by PCR and RFLP analysis. The method includes two major steps, (i) treatment by proteinase K in an SDS extraction buffer, and (ii) grinding of a single half seed in the buffer after incubation. About 1.5–2 µg of DNA per half seed (the endosperm part) of rice was obtained and more than 200 half seed samples could be handled by one person in a day. The DNA could be used for fingerprinting and detection of target genes in a transgenic plant by PCR. The amplified PCR products from the half seed DNA exhibited the same banding patterns as those from leaf DNA. Yield and quality of DNA extracted from half seeds of rice was also sufficient for RFLP analysis. The remnant half seeds containing the embryo can be maintained for later germination of selected genotypes.     

中国小麦地方品种内和品种间醇溶蛋白遗传多样性分析

为了揭示中国小麦地方品种内遗传异质性和品种间的遗传多样性,采用A-PAGE方法,对72份来自不同生态区的地方品种进行醇溶蛋白构成分析。结果发现,全部供试地方品种共观察到101条迁移率不同的务带,构成229种醇溶蛋白构型,每个品种醇溶蛋白条带数目为14—24。63份（87．5％）地方品种在品种内具有2种以上醇溶蛋白变异类型,其中,变异类型最多的品种二红皮小麦（ZM004659）30个子粒中有14种之多,多数品种具有2—3种变异类型。品种内醇溶蛋白构型一致的品种共有9个,占12．5％。这表明供试的大多数小麦地方品种内个体间在醇溶蛋白构成上具有遗传异质性。聚类分析表明,相同生态区的地方品种没有整齐地聚为一类。     

Long-term variation in seed mass and seed production in populations of Paris quadrifolia

Seed production of the perennial herb Paris quadrifolia L. (Liliaceae) was investigated in five populations in northern Poland. The long-term seed production per square metre differed significantly among populations and years. Moreover, throughout the 7-year study period, 30% of both whole ripe fruits and seeds alone were predated. Variation in seed mass per fruit in both space and time was significant. Throughout the 7-year study, nearly all the marked individuals produced fruits once every 2 years. The most frequent break between fruiting was 2 years and the longest was 5 years. Only the mean seed mass in fruits of the same individual varied significantly over subsequent years. In the five populations, the number of ovules, number of seeds in the fruit and seed mass varied significantly between populations. However seed/ovule ratio did not differ in fruits in the five populations. The seed mass/number trade-off in fruits was strongly partially correlated when the effect of total seed mass was considered. Breeding experiments suggest that P. quadrifolia has a substantial capacity for both inbreeding and outbreeding. There were no significant differences in the seed/ovule ratio, seed number or seed mass in fruits produced from bagged or control flowers. However, both seed/ovule ratio and number of seeds were significantly lower in fruits from emasculated flowers.     

Somaclonal Variation in the Gliadin Patterns of Grains of Regenerated Wheat Plants

Maddock, S. E., Risiott, R., Parmar, S., Jones, M. G. K. andShewry, P. R. 1985. Somaclonal variation in the gliadin patternsof grains of regenerated wheat plants.—J. exp. Bot. 36:1976–1984. The banding patterns of the gliadin storageproteins of the grains of 590 regenerated plants from six wheatcultivars were examined by polyacrylamide gel electrophoresisusing lactate buffer. Variation additional to that present incontrol material was observed at a low frequency (1%). Two variantlines showed extensive changes in banding patterns which wereaccompanied by morphological variation of the plants. More limitedvariation in the form of an extra –gliadin band was observedin a third line. Differences in the seed gliadins were not foundin four lines which had shown stable phenotypic changes in heightin field trials. Key words: Wheat, somaclonal variation, gliadins, tissue culture, seed proteins     

Rapid identification of lettuce seed germination mutants by bulked segregant analysis and whole genome sequencing

Lettuce (Lactuca sativa) seeds exhibit thermoinhibition, or failure to complete germination when imbibed at warm temperatures. Chemical mutagenesis was employed to develop lettuce lines that exhibit germination thermotolerance. Two independent thermotolerant lettuce seed mutant lines, TG01 and TG10, were generated through ethyl methanesulfonate mutagenesis. Genetic and physiological analyses indicated that these two mutations were allelic and recessive. To identify the causal gene(s), we applied bulked segregant analysis by whole genome sequencing. For each mutant, bulked DNA samples of segregating thermotolerant (mutant) seeds were sequenced and analyzed for homozygous single‐nucleotide polymorphisms. Two independent candidate mutations were identified at different physical positions in the zeaxanthin epoxidase gene (ABSCISIC ACID DEFICIENT 1/ZEAXANTHIN EPOXIDASE, or ABA1/ZEP) in TG01 and TG10. The mutation in TG01 caused an amino acid replacement, whereas the mutation in TG10 resulted in alternative mRNA splicing. Endogenous abscisic acid contents were reduced in both mutants, and expression of the ABA1 gene from wild‐type lettuce under its own promoter fully complemented the TG01 mutant. Conventional genetic mapping confirmed that the causal mutations were located near the ZEP/ABA1 gene, but the bulked segregant whole genome sequencing approach more efficiently identified the specific gene responsible for the phenotype.     

Intra-inflorescence variation in floral traits and reproductive success of the hermaphrodite Silene acutifolia

BACKGROUND AND AIMS: Intraspecific variation in floral components and reproductive success is often located at the intra-individual level. The arrangement of flowers within inflorescences may explain a great deal of this variation. The variation in number of ovules, fruit set, number of seeds per fruit, seed set, seed weight and seed germination is investigated at different positions within the inflorescence of Silene acutifolia. METHODS: Data were obtained in natural populations, and germination experiments were conducted in a germination chamber. KEY RESULTS: The number of ovules, fruit set, number of seeds, seed set and seed weight, showed a significant decline from early (primary) position to later (tertiary) position. The patterns of intra-inflorescence variation were consistent in different populations and years of study. Seed germination showed an opposite pattern, seeds from primary position showed the lowest germination percentages and seeds from tertiary position the highest, although the effect of position on germination was only marginally significant. There was significant among-population variation in number of ovules per flower. Fruit set also varied significantly among populations, with lower fruit set in the smaller and more isolated population. No significant among-population differences were detected in number of seeds per fruit and seed set. Seeds from the smallest and more isolated population (Arnado) were the lightest. Seed germination showed strong differences between populations, seeds from Arnado started to germinate later, and showed the lowest final germination percentages. CONCLUSIONS: Architectural effects or resource competition are the most commonly proposed hypothesis to explain these patterns. Data suggest that there is less pollen available to pollinate tertiary flowers, and that there is not enough outcross pollen in Arnado. The germination percentages suggest that there is variation in the source of pollen within inflorescences, with high probability of receiving outcross pollen in flowers from primary position, and higher probability of geitonogamous crosses in tertiary flowers.     

Patterns of pollen flow in a dense population of the insect-pollinated canopy tree species Castanopsis sieboldii

Insect pollinations of tree species with high-density populations have rarely been studied. Since the density of adults can affect effective pollen dispersal, short-distance pollination, even by insects, may frequently occur in high-density populations. To test this prediction, we investigated pollination patterns in a high-density population of the insect-pollinated canopy tree species Castanopsis sieboldii by paternity analysis using genotypes at 8 microsatellite loci of 145 adult trees and 439 seeds from 11 seed parents in a 4-ha plot. We then explored their genetic effects on the population by calculating other population genetics parameters. Although C. sieboldii has high potential for long-distance dispersal of pollen (as indicated by a fat-tailed dispersal kernel), the cumulative pollination at the local scale was spatially limited and strongly dependent on the distance between parents due to the high density of adults. Genetic diversity estimates for pollen pools accepted by each seed parent converged on a maximum as the effective number of pollen parents increased. The genetic diversity of pollen pool bulked over all the seed parents from inside the plot did not differ from that of the total pollen pools. Therefore, although pollen flow from distant pollen parents may help to maintain the genetic diversity of offspring, pollen parents neighboring seed parents may be the main contributors to the genetic diversity of the offspring at the seed stage.     

Wheat Seed Carboxypeptidase and Joint Action on Gliadin of Proteases from Dry and Germinating Seeds

A carboxypeptidase preparation, homogeneous according to polyacrylamidegel electrophoresis and ultracentrifugation, was obtained fromwheat seeds. The isolation procedure included (NH₄)₂SO₄ fractionation,gel-filtration on Sepharose-6B and affinity chromatography onCABS-Sepharose. Mr of the enzyme determined by gel-filtrationwas 126 000. The enzyme consisted of two non-identical subunitsof Mr 60 000 and 63 000. The pl of the carboxypeptidase was5.7. The inhibitory analysis revealed that the isolated enzymeis a serine carboxypeptidase. The carboxypeptidase preferentiallyhydrolysed N-substituted dipeptides with aromatic amino acidresidues at the C-terminus and showed weak hydrolysing activitytowards gliadin. The combined action of carboxypeptidase andaspartic proteinase from dry wheat seeds led to an increasein the degree of proteolysis of the storage protein comparedto that resulting from the sum of the action of individual enzymes.The cysteine proteinase from germinated wheat seeds caused completedegradation both of gliadin, which had first been treated withproteases of dry seeds (aspartic proteinase plus carboxypeptidase),and of untreated gliadin. However, when gliadin had first beenhydrolysed with dry seed proteases, the rate of its proteolysiswith cysteine proteinase increased 3–4 times comparedto the non-hydrolysed gliadin. The data indicate the importanceof preliminary modification of gliadin with dry wheat proteases,which, apparently, enhances the supply of nutritive substancesto the embryo in the course of seed germination. Key words: Wheat, gliadin, carboxypeptidase, proteinases, proteolysis     

Disc electrophoretic investigation of Coffea arabica and C. canephora: General protein and malate dehydrogenase of mature seeds

A preliminary disc electrophoretic investigation utilizing malate dehydrogenase and general protein banding patterns obtained from mature seeds revealed detectable differences among Coffea species, varieties and cultivars. Protein differences among seed samples from individual plants of the same variety were also implied. The analyzed protein data were presented as dendrograms and compared with published genetical and taxonomical data.