Isozyme Analysis of F 5 and BC 1F 4 from Cultivated Barley (Hordeum vulgare)× Roegneria ciliaris

栽培大麦,纤毛鹅观草,属间杂种,酯酶,过氧化物酶 ISOZYME ANALYSIS OF F5 AND BCiF4 FROM CULTIVATED BARLEY ( HORDEUM VULGARE ) ~ ROEGNERIA CILIARIS LI Wan-Ji LI Yi-Ping L1U Fang Abstract Esterase and peroxidase isozymes were analysed in the variants including 4 types, 16 lines of Fs, BC1F4 and the parents derived from cultivated barley ( Hordeum vulgare cv. Arupo) x Roegneria ciliaris (Trin.) Nevski in young roots, shoots, spikes and seeds. The zymogram patterns of esterase and peroxidase demonstrated that the 16 lines of F5 and BC1F4 had all or most bands of the cultivated barley parent cv. "Arupo", 1 to 3 bands from the male R. ciliaris, and new hybrid isozyme bands in various amount. Some bands of parent "Arupo" were lost. It suggested that the genetic substances come from R. ciliaris were stably inherited to the progenies of selfing and backcrossing, and there were some variations among the lines. There was certain relationship between isozyme variance and plant characters. Thus, in identifying the translocation lines by isozyme analysis, it would be preferable to study the various organ-specific isozymes or to trace one type of isozyme pattern in consequence.     

Active expression of Gγ globin gene on chromosome 11 with Yunnanese (Ayγδβ)~0-thalasseinia deletion in MEL cells

A permanent lymphocyte cell line of a heterozygote with Yunnanese (Aγδβ)0-thalassemia deletion, associated with an increased production of Cry globin in adult, was founded using Epstein-Barr virus transformation. The hybrids of the lymphocyte cell and mouse erythroleukemia cell (MEL) were achieved and the hybrids containing human chromosome 11 were selected with the monoclonal antibody 53/6. The subclones containing only either the normal or the abnormal human chromosome 11 were separated and the expression of the human globin genes was studied. Expression of the β-globin gene, but not the Cγ and Aγ, was observed in the hybrids containing only the normal human chromosome 11, while active expression of the Cγ globin gene was observed in the hybrids containing only the abnormal human chromosome 11. These results have confirmed that the DNA deletion in the β-globin gene cluster is the cause of persistent active expression of the Cγ globin gene in the Yunnanese mutant.     

Expression of PIN Genes in Rice （Oryza sativa L.）： Tissue Specificity and Regulation by Hormones

Twelve genes of the PIN family in rice were analyzed for gene and protein structures and an evolutionary relationship with reported AtPINs in Arabidopsis. Four members of PIN1 （designated as OsPINla-d）, one gene paired with AtPIN2 （OsPIN2）, three members of PIN5 （OsPIN5a-c）, one gene paired with AtPIN8 （OsPIN8）, and three monocot-specific PINs （OsPIN9, OsPINIOa, and b） were identified from the phylogenetic analysis. Tissue-specific expression patterns of nine PIN genes among them were investigated using RT-PCR and GUS reporter. The wide variations in the expression domain in different tissues of the PIN genes were observed. In general, PIN genes are up-regulated by exogenous auxin, while different responses of different PIN genes to other hormones were found.     

Genetic analysis and gene mapping of a rice few-tillering mutant in early backcross populations (Oryza sativa L.)

A rice mutant, G069, characteristic of few tiller numbers, was found in anther culture progeny from the F1 hybrid between an indica-japonica cross, Gui630×02428. The mutant has another two major features: delayed tillering development and yellowing apex and margin on the mature leaves. As a donor parent, G069 was further backcrossed with the recurrent parent, 02428, for two turns to develop a BC2F2 population. Genetic analysis in the BC2F2 population showed that the traits of few-tillering and yellowing apex and margin on the mature leaves were controlled by one recessive gene. A pool of equally mixed genomic DNA, from few-tillering individual plants in BC2F2, was constructed to screen polymorphism with simple sequence repeat (SSR) markers in comparison with the 02428 genome. One SSR marker and three restriction fragment length polymorphism (RFLP) markers were found possibly linked with the recessive gene. By using these markers, the gene of few-tillering was mapped on chromosome 2 between RFLP marker C     

Allele-specific DNA methylation analyses associated with siRNAs in Arabidopsis hybrids

Accumulating evidence has suggested that epigenetic marks including DNA methylation,small RNA and histone modification may involve hybrid vigor in plants.However,knowledge about how epigenetic marks in hybrids regulate gene expression is still limited.Based on genome-wide DNA methylation landscapes of Arabidopsis thaliana Ler and C24 ecotypes and their reciprocal F1 hybrids which were obtained in our previous work,we analyzed allele-specific DNA methylation and distinguished cis-and trans-regulated DNA methylation in hybrids.Our study indicated that both cis-and trans-regulated DNA methylation played roles in hybrids,when cis-regulation played a major role in CG methylation and trans-regulation played major roles in CHG and CHH methylation.In addition,we observed correlations between trans-regulated DNA methylation and siRNA densities.Enriched siRNA regions were significantly concurrent with highly trans-regulated DNA methylation regions.Our results illustrated DNA methylation regulation patterns integrated with siRNAs in Arabidopsis hybrids,and shed light on understanding the mechanism of epigenetic reprogramming for hybrid vigor.     

Behavior of meiotic chromosomes in Pinus wallichiana, P. strobus and their hybrid and nrDNA localization in pollen mother cells of the hybrid by using FISH

The complete process of meiosis was investigated in Pinus wallichiana, P. strobus and their artificial hybrid （F1） using microsporocytes. It is revealed that there were slightly lower chiasma frequency, lower ring bivalent frequency, lower meiotic index and distinctly higher frequency of aberrance （chromosomal bridges, fragments or micronuclei） in pollen mother cells （PMCs） of the hybrid （F1） than those of the parental species, which showed a certain degree of differentiation between homologous chromosomes of the two parents. However, relatively higher frequency of ring bivalents and higher meiotic index in all the three entities indicate the great stability of genomes of parental species, and the differentiation of genomes between the two parents must have been slight. Total nineteen signal loci of 18S rDNA were observed in nine bivalents of the hybrid （F1）, among which one bivalent bears two loci, while the others have only one. It is suggested that distinct differentiation at genetic level existed in homologous chromosomes of the two parental species, whereas only slight differentiation at karyotypic and genomic levels take place between the parent species.     

Molecular characterization and expression analysis of Triticum aestivum squamosa-promoter binding protein-box genes involved in ear development

Wheat(Triticum aestivum L.) is one of the most important crops in the world. Squamosa-promoter binding protein(SBP)-box genes play a critical role in regulating flower and fruit development. In this study, 10 novel SBP-box genes(TaSPL genes) were isolated from wheat((Triticum aestivum L.) cultivar Yanzhan 4110). Phylogenetic analysis classified the TaSPL genes into five groups(G1–G5). The motif combinations and expression patterns of the TaSPL genes varied among the five groups with each having own distinctive characteristics: TaSPL20/21 in G1 and TaSPL17 in G2 mainly expressed in the shoot apical meristem and the young ear, and their expression levels responded to development of the ear; TaSPL6/15 belonging to G3 were upregulated and TaSPL1/23 in G4 were downregulated during grain development; the gene in G5(TaSPL3) expressed constitutively. Thus, the consistency of the phylogenetic analysis, motif compositions, and expression patterns of the TaSPL genes revealed specific gene structures and functions.On the other hand, the diverse gene structures and different expression patterns suggested that wheat SBP-box genes have a wide range of functions. The results also suggest a potential role for wheat SBP-box genes in ear development.This study provides a significant beginning of functional analysis of SBP-box genes in wheat.     

小麦农家品种大籽糙抗条锈性的遗传分析

以抗条锈病的农家品种大籽糙作父本、感病品种铭贤169作母本杂交获得F1代杂交种,F1代植株自交获得F2代种子,F1代植株与铭贤169回交获得 BC1代种子。在人工控制条件下,用我国小麦条锈菌优势小种条中28号和条中32号,分别对F1、F2、BC1代及其亲本的幼苗进行人工接种,研究了它们的抗性表现和杂交后代中抗条锈性的分离情况。结果表明,大籽糙对条中32号小种的抗性由一对隐性基因控制;对条中28号小种的抗性由一对显性基因和一对隐性基因的互补作用控制。 Abstract:Dazicao,a native wheat variety with stripe rust resistance from Henan,China,was crossed with susceptible cultivar Mingxian 169 as the female parent.The F1 progeny was selfed to produce F2 progeny and backcrossed with Mingxian 169 to produce BC1 progeny.In air-conditioned greenhouse,seedlings of the F1,F2,BC1 progenies and their parents were inoculated with the prevalent races CY28 and CY32 of Puccinia striiformis respectively.The phenotypes of the F1,F2 and BC1 plants were analyzed for resistance to the two races.The results indicated that the resistance in the Dazicao to race CY32 was controlled by one recessive gene,and the resistance to race CY28 by complementary action of one dominant gene and one recessive gene.     

The Role of Viral Infection in Inducing Variability in Virus-Free Progeny in Tomato

The effect of virus-host interactions on subsequent generations is poorly understood. The evaluation of the effects of viral infection on inheritance of quantitative traits in the progeny of infected plants and elucidation of a possible relationship between chiasma frequency in the infected plants and variability of traits in the progeny were investigated. The current study involved genotypes of four intraspecific hybrids of tomato （Solanum lycopersicum L.）, their parental forms and two additional cultivars. Used as infection were the tobacco mosaic virus （TMV） and potato virus X （PVX）. The consequences of the effect of viral infection were evaluated based on chromosome pairing in diakinesis and/or by examining quantitative and qualitative traits in the progeny of the infected tomato plants. Tomato plants infected with TMV ＋ PVX were found to differ in chiasma frequency per pollen mother cell or per bivalent. Deviations have been observed for genotypes of both F1 hybrids and cultivars. At the same time, differences in mean values of the traits under study have only been found for progeny populations （F2-F4） derived from virus-infected F1 hybrids, but not in the case of progeny of the infected cultivars. The rate of recombinants combining traits of both parents increased significantly （2.22-8.24 times） in progeny populations of hybrids infected with TMV＋PVX. The above suggests that the observed effects could be the result of modification of recombination frequencies that can be manifested in heterozygous hybrids and make small contributions to variability in cases of ＇homozygous＇ tomato genotypes （i.e. cultivars）.     

小麦杂交种与亲本发育早期种子的基因表达差异及其与杂种优势关系的初步研究

为深入探讨小麦杂种优势形成的分子机理,选取3个冬小麦品种(系)为一组亲本,3个为另一组亲本,配制了正反交18个杂交组合,以授粉后6d的杂交和自交种子为材料,应用mRNA差异显示技术(DDRT—PCR)研究了小麦杂交当代种子与其亲本自交种子基因的表达差异,并与杂种优势进行相关分析。为降低DDRT—PCR技术假阳性的不利影响,对每个引物组合均作了两次PCR扩增,在处理数据时,仅统计能重复出现的条带。结果发现：杂交种和亲本之间的基因表达模式有8类共15种：(1)单亲沉默型(2种),(2)单亲一致型(2种),(3)正交或反交沉默型(2种),(4)正交或反交特异型(2种),(5)正交或反交单亲一致型(4种),(6)杂交种特异型(1种),(7)双亲共沉默型(1种),(8)表达一致型(1种)。分析发现,小麦杂交种和亲本间存在显著的表达差异。在差异表达类型中,杂交种特异型和双亲共沉默型比例最低。对上述15种表达模式与杂种优势进行相关分析,结果表明,表达一致型与各产量性状杂种优势之间的相关均不显著,说明杂种优势是由某些有表达差异的基因造成。9个产量性状均能检测到一种以上与其显著或极显著相关的基因表达模式,有些性状受正负相关效应的共同影响;沉默型(包括单亲沉默型、正交或反交沉默型和双亲共沉默型)和正交或反交单亲一致型在杂种优势形成中发挥重要作用。这些研究表明,在种子发育早期,基因的差异表达与杂种优势形成之间可能存在较为复杂的关系。     

玉米杂种与亲本穗分化期功能叶基因差异表达与杂种优势

为探讨玉米杂种优势的分子机理,以10个玉米自交系及其组配的38个杂交种为材料,利用cDNA-AFLP技术,分析杂种与亲本在玉米雌穗小穗分化期功能叶片的基因差异表达类型与主要农艺性状的杂种表现及杂种优势的关系。研究表明：（1）杂种的基因相对于其双亲,存在质和量的表达差异,其中质的差异表达类型包括：单亲沉默表达,双亲沉默表达,亲本显性表达和杂种特异表达等类型。（2）在雌穗小穗分化期,同一差异表达类型中不同杂交组合间差异很大;从总体平均看,杂种特异表达类型占25.22%,亲本显性表达类型占21.46%,双亲沉默表达类型占8.27%,单亲沉默表达类型占33.49%。（3）单亲沉默表达与株高的杂种表现呈显著正相关;双亲沉默表达与穗粗的杂种优势呈显著负相关,显性表达与行粒数和单株粒重的杂种优势呈显著负相关,其余表达类型与所有农艺性状杂种表现及杂种优势均不相关,并对结果进行了讨论。     

小麦杂种及其亲本苗期叶片家族基因差异表达及其与杂种优势关系的初步研究

为探讨小麦杂种优势形成的分子机理,以一套双列杂交组合的苗期叶片为材料,利用mRNA差异显示技术分析了杂种及其亲本间MADS－box、G－ box、Ser/Thr蛋白激酶、EIF－4A、ARF1基因家族共5类家族基因在杂交种和亲本之间的表达差异。并与杂种性状表现和杂种优势进行了相关分析。结果发现,除ARF1家族基因外,其余家族基因在杂种和亲本间存在显著的表达差异,差异表达类型可概括为4种：（1）双亲共沉默;（20单亲表达沉默;（3）杂种特异表达;（4）单亲表达一致。分析发现,MADS－box、G－box和EIF－4A家族基因在杂种和亲本间的差异表达模式相似,均以单亲特异表达和种特异表达类型所占比例最高。相关分析结果表明,以上所有家族基因的总体差异表达程度与所有性状的杂种表现均不相关,MADS－box家族基因中杂种特异表达类型与小穗数、单株产量和单穗产量杂种优势呈显著正相关,双亲共沉默类型与小穗数、千粒重和单穗产量杂种优势呈显著负相关,另外,EIF－4A家族基因中单亲表达一致型与单穗产量杂种优势呈显著正相,但双亲共沉默类型与小穗数和单穗产量杂种优势呈显著负相关,对于G－box基因家族而言,仅小穗数杂种优势和双亲共沉默类型成显著负相关,而蛋白激酶家族基因的各种差异类型与性状杂种优势的相关分析均不显著。这些研究表明,调控基因的差异表达与杂种优势形成有密切关系。     

Relationships of differential gene expression in leaves with heterosis and heterozygosity in a rice diallel cross

Using differential display analysis, we assessed the patterns of differential gene expression in hybrids relative to their parents in a diallel cross involving 8 elite rice lines. The analysis revealed several patterns of differential expression including: (1) bands present in one parent and F₁ but absent in the other parent, (2) bands observed in both parents but not in the F₁, (3) bands occurring in only one parent but not in the F₁ or the other parent, and, (4) bands detected only in the F₁ but in neither of the parents. Relationships between differential gene expression and heterosis and marker heterozygosity were evaluated using data for RFLPs, SSRs and a number of agronomic characters. The analysis showed that there was very little correlation between patterns of differential expression and the F₁ means for all six agronomic traits. Differentially expressed fragments that occurred only in one parent but not in the other parent or in F₁ in each of the respective crosses were positively correlated with heterosis and heterozygosity. And conversely, fragments that were detected in F₁s but in neither of the respective parents were negatively correlated with heterosis and heterozygosity. The remaining patterns of differential expression were not correlated with heterosis or heterozygosity. The relationships between the patterns of differential expression and heterosis observed in this study were not consistent with expectations based on dominance or overdominance hypotheses.     

Differential gene expression in liver of inbred chickens and their hybrid offspring

Differential display of mRNA was used to analyse the differences of gene expression in liver between chicken hybrids and their parents in a 4 x 4 diallel crosses in order to study the molecular basis of heterosis in chickens. The results indicated that patterns of gene expression in hybrids differ significantly from their parents. Four patterns of differential gene expression were revealed, which included: (i) bands only detected in the hybrid F1s (UNF1); (ii) bands only absent in the hybrid F1s (ABF1); (iii) bands only detected in the parental P1 or P2 lines (UNP1 and UNP2) and (iv) bands absent in the parental P1 or P2 lines (ABP1 and ABP2). In addition, correlations between patterns of gene expression and heterosis percentages of nine carcass traits of 8-week-old chickens were evaluated. Statistical results showed that negative correlations between heterosis percentages and the percentage of F1-specific bands (UNF1) were significant at P < 0.01 for breast muscle yield, leg muscle yield, wing weight, eviscerated weight and eviscerated weight with giblet of 8-week-old chickens, and at P < 0.05 for intermuscular fat width. Heterosis percentage was negatively correlated with ABP (bands present in the hybrid F1s and one parental line but absent in the other parental line, ABP1 and ABP2) for breast muscle yield, leg muscle yield, wing weight, eviscerated weight and eviscerated weight with giblet of 8-week-old chickens (P < 0.01). Bands detected only in the hybrid F1s but not in either of the parental lines (UNF1) and bands absent in parental P1 or P2 lines (which includes ABP1 and ABP2) may play important roles in chicken heterosis.     

Cis-transcriptional variation in maize inbred lines B73 and Mo17 leads to additive expression patterns in the F1 hybrid

Microarray analysis of gene expression patterns in immature ear, seedling, and embryo tissues from the maize inbred lines B73 and Mo17 identified numerous genes with variable expression. Some genes had detectable expression in only one of the two inbreds; most of these genes were detected in the genomic DNA of both inbreds, indicating that the expression differences are likely caused by differential regulation rather than by differences in gene content. Gene expression was also monitored in the reciprocal F1 hybrids B73xMo17 and Mo17xB73. The reciprocal F1 hybrid lines did not display parental effects on gene expression levels. Approximately 80% of the differentially expressed genes displayed additive expression patterns in the hybrids relative to the inbred parents. The approximately 20% of genes that display nonadditive expression patterns tend to be expressed at levels within the parental range, with minimal evidence for novel expression levels greater than the high parent or less than the low parent. Analysis of allele-specific expression patterns in the hybrid suggested that intraspecific variation in gene expression levels is largely attributable to cis-regulatory variation in maize. Collectively, our data suggest that allelic cis-regulatory variation between B73 and Mo17 dictates maintenance of inbred allelic expression levels in the F1 hybrid, resulting in additive expression patterns.     

香菇杂交子农艺性状与差异基因表达类型的相关性研究

以香菇(Lentinula edodes)4个菌株为亲本组成3个杂交组合,采用单孢菌株配对获得杂交子,观察锁状联合鉴别出真正的杂交子,测定杂交子及其亲本的农艺性状。在每个杂交组合中分别各选取3个杂交子,研究杂交子和亲本在子实体发育阶段差异基因表达情况。结果表明:杂交子共有4种基因表达类型:双亲沉默型(W1型),单亲沉默型(W2型),杂交子特异表达型(W3型),单亲表达型(W4型)。香菇杂交子农艺性状与基因差异表达类型的相关性分析表明:菌盖厚与双亲沉默型(W1型)呈极显著负相关,而菌柄长与单亲沉默型(W2型)呈显著负相关。     

Central cell nuclear-cytoplasmic incongruity:a mechanism for segregation distortion in advanced backcross and selfed generations of (Allium cepa L. x Allium fistulosum L.) x A. cepa interspecific hybrid derivatives

A model is presented as an explanation for an anomaly observed in germination and establishment and isozyme segregation patterns in Allium cepa x A. fistulosum F2BC3 populations generated in an introgression-breeding program. The F1BC3 parent of these populations was selected for its heterozygous PGI phenotype, Pgi-1(2/3); Pgi-1(2) was inherited from an A. cepa (Ac) seed parent and Pgi-1(3) from an A. fistulosum (Af) pollen parent. Germination and establishment was recorded for the F2BC3 progeny population. Segregation of Ac and Af Pgi-1 alleles was investigated in F2BC3 seeds and embryo and endosperm tissue was isolated and tested for isozyme expression. A pooled goodness-of-fit test of the segregation of Pgi-1 alleles in the populations to the expected Mendelian 1:2:1 ratio using the chi-square statistic gave a chi2 = 185.9, well beyond the accepted limits at 2 degrees of freedom. The 1:2:1 ratio expected for simple Mendelian inheritance was rejected, while a pooled chi-square goodness-of-fit test of the segregation of Pgi-1 alleles in the populations fit a 1:1 ratio with a chi2 = 0.203, based on the incongruity model. We present here the central cell nuclear-cytoplasmic incongruity hypothesis to explain the observed anomalies.     

盛花期抗虫杂交棉及其亲本叶片基因表达差异与杂种优势

采用DDRT-PCR技术,以棉花盛花期项尖叶片cDNA为材料,对上海生物工程公司合成的专用于基因差异显示分析的3个锚定引物和全套26个随机引物进行筛选,最后选择了15个扩增差异带丰富的随机引物。采用3个锚定引物和这15个随机引物组成的45对引物组合对24个抗虫棉杂交组合及其10亲本盛花期叶片cDNA进行扩增和差显,2次扩增重复率达70．1％,表明在扩增过程中存在较高的假阳性,通过重复PCR扩增,统计稳定扩增的条带,可减少假阳性干扰。根据基因表达方式,将其划分5种模式：MI为双亲表达沉默,双亲出现条带而杂种没有条带;M2为单亲表达沉默,带仅出现在亲本之一,包括仅母本有带而父本和杂种无带和仅父本有带而母本和杂种无带2种表达方式：M3为杂种特异表达,带仅出现在杂种,双亲无带;M4为单亲表达一致,带在双亲之一和杂种中出现,而在另一亲本中不出现,包括母本、杂种中有带而父本无带和父本、杂种中有带而母本无带2种方式;M5为基因表达一致,带在双亲和杂种中均出现。差显表达模式比例与产量性状和杂种优势分析表明：M4与所有产量性状均呈正相关,并且与单位面积铃数相关达显著水平,其他各种模式与杂种产量性状表型值均未达到显著水平;M2与单位面积铃数杂种优势呈显著负相关,M3与皮棉产量杂种优势呈显著正相关。上述结果表明,盛花期叶片中的基因显性表达和杂种特异表达有利于产量形成和杂种优势发挥。     

应用过氧化物酶同工酶鉴定山茶属植物杂种F1代的研究

对4个杂交组合的12个杂种F1代植株与亲本进行了形态学比较和过氧化物酶同工酶分析,结果表明以金花茶为父本,云南山茶或云南野山茶为母本杂交产生的11个杂种的幼枝、叶的特征均与母本相似,H-86-1-1的花与母本一致,H-87-2-2的花兼有父母本的特征;H-78-1-1的幼枝、叶的特征与父本相似,其花也兼有父母本的特征。有9株的酶谱为“互补型酶带”,且都出现了杂种带,为真正的杂种;其余2株的酶谱与母本一致,与其形态特征表现出一定的相关性,可能为非真正的杂种。同一个杂交组合产生的杂种,酶谱却有差异,金花茶最为特征的一条带(Rf=0.813)在其所有11个杂种F1代植株中都未表达。