小麦-黑麦代换系间杂交后代染色体易位的研究

主要利用小麦-黑麦5R/5A二体代换系与6R/6A二体代换系杂交,在其杂种自交F3中未利用辐射、杀配子染色体、Ph基因等方法,鉴定出易位系,同时,首次观察到有丝分裂中的染色体断裂现象.发现在小麦遗传中,没对Ph基因进行特殊处理的情况下,小麦同祖染色体间可能发生部分同源配对并交换,产生易位.     

小麦-黑麦代换系5A/5R与6A/6R杂交诱导同祖染色体配对与易位的研究

利用两个小麦-黑麦异源双代换系DS 5A/5R与DS 6A/6R杂交,探讨同祖染色体配对的可能性与创制小麦黑麦异源易位系.在方法上对杂种F1的减数分裂行为进行研究,观察5R与5A、6R与6A配对频率,探讨同祖染色体配对规律.实验结果看到杂交F1减数分裂中有22.91%的花粉母细胞有小麦染色体(ABD组)与黑麦染色体(R组)发生同祖配对.在F2及以后世代,通过染色体C分带、原位杂交检测,选择小麦-黑麦易位系.在F2代的45株中检测到9株有易位,易位频率为20%,是目前小麦-黑麦染色体易位频率最高的.染色体易位有的来源于同祖配对的交换,有的来源于单价体错分裂或断裂的重建.     

应用原位杂交技术对小麦—黑麦代换系间杂交的细胞遗传研究

应用原位杂交技术结合染色体组型分析方法,对两个小麦-黑麦异源双代换系5R/5A和6R/6A杂交后代的遗传进行了研究,探讨同祖染色体配对的可能性并获得小麦-黑麦易位系。实验中对杂种F1代植株减数分裂各时期的花粉母细胞染色体行为进行分析,结果发现有22.91%的花粉母细胞中黑麦染色体与小麦染色体发生同祖配对。F2代通过C-分带、原位杂交鉴定,在45株中检测到9株易位,易位频率为20%,是目前报道易位频率最高的。染色体易位有的来源于同祖配对交换,有的来源于单价体错分裂或断裂的重建。     

GC1代换系与小麦杂交减数分裂行为的研究

用来自拟斯卑尔脱山羊草（Ae.speltoides）的杀配子染色体2S(GC基因),诱导普通小麦（Triticum aestivum L.）“宁农”、普通小麦—黑麦（Secale cereale L.）二体代换系（5R/5A）的染色体断裂,观察杂种F₁的减数分裂行为,在减数分裂中期I和中期II均出现较高频率的单价体、多价体,后期I和后期II出现大量的落后染色体、染色体断片和桥等异常现象,在二分孢子和四分孢子中出现较多的微核。在本实验中,GC1代换系与5R/5A代换系杂交F₁代减数分裂行为比GC1代换系与普通小麦“宁农”杂交F₁代减数分裂行为复杂,经统计学分析,含杀配子染色体的代换系间杂交比单一的杀配子染色体作用对单价体、多价体、微核的产生具有显著差异。因此,利用带有杀配子染色体的代换系间杂交来诱导染色体易位是一条有效的途径。     

7个大穗型小麦-黑麦代换系间杂交后代的形态学与SSR分析

对从小麦-黑麦代换系5R/5A与1R/1D杂交后代中选育的大穗型品系1-5、1-6、1-7、1-8、1-9、1-10、1-11进行形态学与SSR分析。结果表明,7个品系田间表现遗传性稳定,且具有黑麦大穗、抗病等优良性状;利用黑麦特异引物PSC119.1确定7个品系均导入了黑麦染色体片段,引物SCM138扩增结果表明,在1-6、1-7、1-8、1-9、1-10中导入了黑麦染色体5RS片段,引物SCM120、TSM604可以在7个品系中稳定扩增出黑麦5R长臂和1R短臂特异片段。以上结果可为小麦遗传育种与品质改良提供基础材料。     

小麦——黑麦易位系的研究

为了创制在小麦生产上有经济价值和在小麦育种中有利用价值的小麦—黑麦易位系,用小麦—黑麦代换系5R/5A与6R/6A杂交,在杂种后代中选择带有某些黑麦性状的普通小麦类型植株,并按性状继代选择直至稳定为止,至F6代共选出9个品系即6-26、06-60-11、06-6-24、06-6-15、06-6-35、6-34、6-28、06-6-17、06-6-14。本文对这9个品系及其亲本进行了田间观察和遗传分析,结果表明：它们田间表现生长整齐一致、遗传稳定、育性正常,具有大穗、多小穗、多分蘖、抗病、抗旱等优良性状;选择分布在黑麦5R和6R染色体上的微卫星引物共20对,结果表明引物SCM268在6-26、06-60-11、06-6-15、06-6-35、6-34、6-28、06-6-17、06-6-14这8个品系中扩增出黑麦特异产物,初步确定这8个品系是易位系,是有经济价值和利用价值的遗传材料。     

一个小麦—黑麦染色体代换的细胞学鉴定

对从六倍体小黑麦与普通小麦的杂种后代中获得的矮秆抗病选系84056-1-36-1进行体细胞C-分带鉴定,结果表明,它的21对染色体中,有1对短臂带型与1R相似的黑麦染色体代换了小麦的1D。观察“中国春”双端二体1A、1B与该选系杂种F_1的花粉母细胞染色体配对,发现分别有82.56%和65.71%的细胞出现异型三价体,所有细胞至少有两个形态不同的单价体;而在“中国春”端二体IDL与84056-1-36-1的杂种中,端体不配对的花粉母细胞占100%,经C-分带后,另外1条单价体显示明显的端带。从上述这些结果推断84056-1-36-1为1R(1D)代换系。     

应用基因组原位杂交及RFLP标记鉴定小麦中的大麦染色体

用生物素（Biotin-6-dUTP)标记的大麦Betzes基因组DNA作探针,以普通小麦中国春总DNA作封阻进行基因组原位杂交（Genomeinsituhybridization,简称GISH）,从13株小麦-大麦杂交后代中鉴定出2个含有3条大麦Betzes2H染色体的材料（2n＝43）;2个2H单体异代换系（2n＝42）;7个2H二体异代换系（2n＝42）。用已定位在小麦第2部分同源群短臂上的探针psr131进行RFLP分析,结果表明大麦Betzes、代换系A5有1条区别于小麦中国春的特异带,A     

组织培养诱导的普通小麦─黑麦代换系和附加系分子细胞遗传学检测

通过组织培养从普通小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．）与八倍体小黑麦（×ＴｒｉｔｉｃｏｓｅｃａｌｅＷｉｔｍａｃｋ）杂种Ｆ０幼胚再生植株后代中获得２个代换系９３０４９８、９３０４８３和１个附加系９３００２９。以黑麦（ＳｅｃａｌｅｃｅｒｅａｌｅＬ．）基因组ＤＮＡ为探针,采用荧光原位杂交（ＦＩＳＨ）证实了黑麦染色体的存在。在有丝分裂中期,经ＦＩＳＨ处理的黑麦染色体为黄绿色,明显区别于红色的小麦染色体。染色体配对、Ｃ分带、麦谷蛋白电泳分析,证明两个代换系为１Ｄ／１Ｒ代换,附加的也是黑麦１Ｒ染色体     

小麦——黑麦染色体易位系的细胞学鉴定

用C－带技术分析了普通小麦“中国春”、黑麦“胜利”及小黑麦与普通小麦经辐射处理的后代中产生的并经多代纯化的5个带有黑麦某些性状的普通小麦品系的根尖染色体,结果表明：品系98－5－1为1A／1R纯合易位系,具有抗锈病、抗白粉病等基因,可作为诱导小片段易位的资源。作者提出在小麦－黑麦易位系鉴定中应用更高分辩率的G－带技术识别黑麦染色体片段或小片段易位。     

昆虫生态学试验设计中时空转换的技巧

时空转换最初是研究植物群落演替的方法,目前作为一种理论推断在社会科学、地理信息和地质分析等领域应用,在昆虫生态学中的应用很少,本文介绍了空间换取时间与时间换取空间两种方法,能够巧妙地缩短很多昆虫生态学的试验时间,或增加研究范围内的样本数,在大尺度景观研究昆虫迁移、预测预报、栖境恢复等方面具有重要意义。这种时空转换的技巧在昆虫生态学的研究的有广阔的应用前景。     

Variation in the pattern of synonymous and nonsynonymous difference between two fungal genomes

The proportion of synonymous nucleotide differences per synonymous site (p(S)) and the proportion of nonsynonymous differences per nonsynonymous site (p(N)) were computed at 1,993,217 individual codons in 4,133 protein-coding genes between the two yeast species Saccharomyces cerevisiae and Saccharomyces paradoxus. When the modified Nei-Gojobori method was used, significantly more codons with p(N) > p(S) were observed than expected, based on random pairing of observed p(S) and p(N) values. However, this finding was most likely explained by the presence of a strong negative correlation between the number of synonymous differences and the number of nonsynonymous differences at codons with at least one difference. As a result of this correlation, codons with p(N) > p(S) were characterized not only by unusually high p(N) but also by unusually low p(S). On the other hand, the number of codons with p(N)>p(S) (where p(S) is the mean p(S) for all codons) was very similar to the random expectation, and the observed number of 30-codon windows with p(N) > p(S) was significantly lower than the random expectation. These results imply that the occurrence of a certain number of codons or codon windows with p(N) > p(S) is expected given the nature of nucleotide substitution and need not imply the action of positive Darwinian selection.     

Discrimination between adaptive and neutral amino acid substitutions in vertebrate hemoglobins

Summary A discriminant analysis on the basis of the physicochemical properties of amino acid residues is developed to investigate the accumulation pattern of amino acid substitutions in a family of proteins. The application of this analysis to vertebrate hemoglobins reveals the following new results. (1) The major components of teleost fish and amphibian hemoglobins showing the Root effect are sharply discriminated from mammalian hemoglobins in several regions of the and chains, whereas shark, minor components of teleost fish and amphibian, reptile, and bird hemoglobins showing no Root effect exhibit a gradual change to mammalian hemoglobin in a straightforward way. This result suggests at least two lines of molecular evolution in vertebrate hemoglobins. (2) The nonadult hemoglobin chains are allocated to the latter line, i.e., tadpole, , and chains are similar to shark and trout I chains, and and chains are similar to some of the reptile chains. (3) In any case, most of the amino acid residues causing the discrimination are located near the sites that carry the amino acid residues conserved well throughout all classes of vertebrates, suggesting that modifications adapting to the respective living conditions or respiratory organs have taken place effectively near the amino acid residues essential for the manifestation of cooperative oxygen binding. (4) The amino acid residues at other sites are changed from one to another species even within the same class, showing a constant substitution rate as a whole. These amino acid substitutions may be nearly neutral, being under a weak functional constraint. The number of sites allowing such neutral substitutions is rather small, less than one-half of all the sites in the adult hemoglobins of bird and mammal, whereas it amounts to two-thirds in teleost fish hemoglobins.     

Synonymous and nonsynonymous substitutions in mammalian genes and the nearly neutral theory

The nearly neutral theory of molecular evolution predicts larger generation-time effects for synonymous than for nonsynonymous substitutions. This prediction is tested using the sequences of 49 single-copy genes by calculating the average and variance of synonymous and nonsynonymous substitutions in mammalian star phylogenies (rodentia, artiodactyla, and primates). The average pattern of the 49 genes supports the prediction of the nearly neutral theory, with some notable exceptions.The nearly neutral theory also predicts that the variance of the evolutionary rate is larger than the value predicted by the completely neutral theory. This prediction is tested by examining the dispersion index (ratio of the variance to the mean), which is positively correlated with the average substitution number. After weighting by the lineage effects, this correlation almost disappears for nonsynonymous substitutions, but not quite so for synonymous substitutions. After weighting, the dispersion indices of both synonymous and nonsynonymous substitutions still exceed values expected under the simple Poisson process. The results indicate that both the systematic bias in evolutionary rate among the lineages and the episodic type of rate variation are contributing to the large variance. The former is more significant to synonymous substitutions than to nonsynonymous substitutions. Isochore evolution may be similar to synonymous substitutions. The rate and pattern found here are consistent with the nearly neutral theory, such that the relative contributions of drift and selection differ between the two types of substitutions. The results are also consistent with Gillespie's episodic selection theory.     

Avoiding Gas Bubble Formation During Freeze Substitution

Gas bubbles frequently are formed during freeze substitution, especially when tissues are warmed to room temperature. The problem arises largely from the extreme solubility of CO₂ in the freeze substitution solvent. Gas bubbles may be minimized by briefly transferring the tissue to freshly chilled solvent before warming to room temperature.     

Staining of Acid Mucopolysaccharides after Chromatography on Filter Paper

Acid mucopolysaccharides obtained both from commercial sources and by isolation from human urine have been chromatographed on Whatman No. 1 filter paper, using propanol or ethanol in pH 6.5 M/15 phosphate buffer as solvent systems. The chromatograms are then fixed by immersion in 95% alcohol and in diethyl ether. After drying, they are stained in 0.06% toluidine blue O in 0.5% aqueous acetic acid. A final rinsing in 2% aqueous acetic acid removes the excess dye from the paper and exposes the stained mucopolysaccharide to a pH favoring orthochromasia.     

Mutation-selection models of codon substitution and their use to estimate selective strengths on codon usage

Current models of codon substitution are formulated at the levels of nucleotide substitution and do not explicitly consider the separate effects of mutation and selection. They are thus incapable of inferring whether mutation or selection is responsible for evolution at silent sites. Here we implement a few population genetics models of codon substitution that explicitly consider mutation bias and natural selection at the DNA level. Selection on codon usage is modeled by introducing codon-fitness parameters, which together with mutation-bias parameters, predict optimal codon frequencies for the gene. The selective pressure may be for translational efficiency and accuracy or for fine-tuning translational kinetics to produce correct protein folding. We apply the models to compare mitochondrial and nuclear genes from several mammalian species. Model assumptions concerning codon usage are found to affect the estimation of sequence distances (such as the synonymous rate d(S), the nonsynonymous rate d(N), and the rate at the 4-fold degenerate sites d(4)), as found in previous studies, but the new models produced very similar estimates to some old ones. We also develop a likelihood ratio test to examine the null hypothesis that codon usage is due to mutation bias alone, not influenced by natural selection. Application of the test to the mammalian data led to rejection of the null hypothesis in most genes, suggesting that natural selection may be a driving force in the evolution of synonymous codon usage in mammals. Estimates of selection coefficients nevertheless suggest that selection on codon usage is weak and most mutations are nearly neutral. The sensitivity of the analysis on the assumed mutation model is discussed.     

Der substitutionsspezifische einfluss von chalkonen auf die anthocyanbiosynthese bei Petunia hybrida

The effect of two chalcones, 3,4,2′,4′,6′-pentahydroxy- and the 4, 2 ′,4′,6′ -tetrahydroxy- 3-methoxy-chalcone- 4′-glucoside, on the synthesis of different flower anthocyanins in isolated petals of Petunia hybrida has been investigated. The results show that the synthesis of those anthocyanins, which have the same substitution pattern as the chalcone used was increased. This suggests that the chalcones are incorporated into the anthocyanins concerned. When the chalcones were fed together with acetic acid-[1-¹⁴C], this specific substitution effect was detectable only for the 3,4,2′,4′,6′-pentahydroxy-chalcone-4′-glucoside.     

Not so different after all: a comparison of methods for detecting amino acid sites under selection

We consider three approaches for estimating the rates of nonsynonymous and synonymous changes at each site in a sequence alignment in order to identify sites under positive or negative selection: (1) a suite of fast likelihood-based "counting methods" that employ either a single most likely ancestral reconstruction, weighting across all possible ancestral reconstructions, or sampling from ancestral reconstructions; (2) a random effects likelihood (REL) approach, which models variation in nonsynonymous and synonymous rates across sites according to a predefined distribution, with the selection pressure at an individual site inferred using an empirical Bayes approach; and (3) a fixed effects likelihood (FEL) method that directly estimates nonsynonymous and synonymous substitution rates at each site. All three methods incorporate flexible models of nucleotide substitution bias and variation in both nonsynonymous and synonymous substitution rates across sites, facilitating the comparison between the methods. We demonstrate that the results obtained using these approaches show broad agreement in levels of Type I and Type II error and in estimates of substitution rates. Counting methods are well suited for large alignments, for which there is high power to detect positive and negative selection, but appear to underestimate the substitution rate. A REL approach, which is more computationally intensive than counting methods, has higher power than counting methods to detect selection in data sets of intermediate size but may suffer from higher rates of false positives for small data sets. A FEL approach appears to capture the pattern of rate variation better than counting methods or random effects models, does not suffer from as many false positives as random effects models for data sets comprising few sequences, and can be efficiently parallelized. Our results suggest that previously reported differences between results obtained by counting methods and random effects models arise due to a combination of the conservative nature of counting-based methods, the failure of current random effects models to allow for variation in synonymous substitution rates, and the naive application of random effects models to extremely sparse data sets. We demonstrate our methods on sequence data from the human immunodeficiency virus type 1 env and pol genes and simulated alignments.     

Goodman et al.'s method for augmenting the number of nucleotide substitutions

Summary Statistical properties of Goodman et al.'s (1974) method of compensating for undetected nucleotide substitutions in evolution are investigated by using computer simulation. It is found that the method tends to overcompensate when the stochastic error of the number of nucleotide substitutions is large. Furthermore, the estimate of the number of nucleotide substitutions obtained by this method has a large variance. However, in order to see whether this method gives overcompensation when applied together with the maximum parsimony method, a much larger scale of simulation seems to be necessary.