Analysis of dominant spring mutations in rye

The dominant mutant genes responsible for the spring habit were studied in seven rye plants according to the developed scheme of two-step crosses and analysis of the F₂ progeny. The genotypes with a particular genetic formula (heterozygote) were obtained by crossing the studied plants with the winter rye Korotkostebel’naya 69 carrying the recessive genes that control the winter habit of plants. Heterozygotes yielded by different combinations were crossed with each other. The F₁ hybrids were either self-pollinated to obtain F₂ progeny or crossed with the winter rye. Analysis of the progeny suggests that all seven plants carry the same gene.     

Effect of the 5R(5A) alien chromosome substitution on the growth habit and winter hardiness of wheat

The growth habit, ear emergence time, and frost tolerance of wheat/rye substitution lines have been studied in cultivars Rang and Mironovskaya Krupnozernaya whose chromosome 5A is substituted with chromosome 5R of Onkhoyskaya rye. Hybrid analysis has demonstrated that the spring habit of the recipient cultivars Rang and Mironovskaya Krupnozernaya is controlled by dominant gene Vrn-A1 located in chromosome 5A. Onokhoyskaya rye has a dominant gene for the spring habit (Sp1) located in chromosome 5R. It has been found that the resultant 5R(5A) alien-substitution lines have a winter type of development and ears do not emerge during summer in plants sown in spring. The change in growth habit has been shown to be related to the absence of the rye Spl gene expression in the substitution lines. The winter hardiness of winter 5R(5A) alien-substitution lines has been studied under the environmental conditions of Novosibirsk. Testing the lines in the first winter demonstrated that their winter survival is 20-27%. The possible presence of the frost resistance gene homeoallelic to the known genes Fr1 and Fr2 of the common wheat located on chromosomes 5A and 5D, respectively, is discussed.     

Effect of the 5R(5A) Alien Chromosome Substitution on the Growth Habit and Winter Hardiness of Wheat

The growth habit, ear emergence time, and frost tolerance of wheat/rye substitution lines have been studied in cultivars Rang and Mironovskaya Krupnozernaya whose chromosome 5A is substituted with chromosome 5R of Onkhoyskaya rye. Hybrid analysis has demonstrated that the spring habit of the recipient cultivars Rang and Mironovskaya Krupnozernaya is controlled by dominant gene Vrn-A1 located in chromosome 5A. Onokhoyskaya rye has a dominant gene for the spring habit (Sp1) located in chromosome 5R. It has been found that the resultant 5R(5A) alien-substitution lines have a winter type of development and ears do not emerge during summer in plants sown in spring. The change in growth habit has been shown to be related to the absence of the rye Sp1 gene expression in the substitution lines. The winter hardiness of winter 5R(5A) alien-substitution lines has been studied under the environmental conditions of Novosibirsk. Testing the lines in the first winter demonstrated that their winter survival is 20–27%. The possible presence of the frost resistance gene homeoallelic to the known genes Fr1 and Fr2 of the common wheat located on chromosomes 5A and 5D, respectively, is discussed.     

Genome sequencing reveals agronomically important loci in rice using MutMap

The majority of agronomic traits are controlled by multiple genes that cause minor phenotypic effects, making the identification of these genes difficult. Here we introduce MutMap, a method based on whole-genome resequencing of pooled DNA from a segregating population of plants that show a useful phenotype. In MutMap, a mutant is crossed directly to the original wild-type line and then selfed, allowing unequivocal segregation in second filial generation (F(2)) progeny of subtle phenotypic differences. This approach is particularly amenable to crop species because it minimizes the number of genetic crosses (n = 1 or 0) and mutant F(2) progeny that are required. We applied MutMap to seven mutants of a Japanese elite rice cultivar and identified the unique genomic positions most probable to harbor mutations causing pale green leaves and semidwarfism, an agronomically relevant trait. These results show that MutMap can accelerate the genetic improvement of rice and other crop plants.     

Introgression of the reduced height gene Rht10 from the wheat cultivar Ai-bian 1 into the triticale genotype

A gene determining reduced height, Rht10, from the wheat cultivar Ai-Bian 1 was introgressed into the triticale genotype. Initially, Ai-Bian 1 was crossed with the wheat cultivar Chinese Spring (CS), a carrier of Kr genes, to overcome the uncrossability of this cultivar with rye. Amphidiploids were produced by hybridizing the F2 (CS x Ai-Bian 1) plants displaying reduced height (at the level of Ai-Bian 1) with rye. Free pollination of F1 (F2 of CS x Ai-Bian 1) x Saratovskaya 7 with triticale pollen gave fertile viable hybrids; the majority of hybrids were phenotypically closer to octoploid triticale; however, the variants intermediate between octo- and hexaploids were also present. The height of amphidiploids varied from 40 to 90 cm, and the grain yield per spike amounted on the average to 11.7--24.7 grains, which exceeded essentially this value in F1 plants.     

Epistatic interaction between vernalization genes Vrn-Am1 and Vrn-Am2 in diploid wheat

Genes Vrn-A(m)1 and Vrn-A(m)2 control the vernalization requirement in diploid wheat (Triticum monococcum). The epistatic interaction between these two genes on flowering date was studied here using a factorial analysis of variance. One hundred and two F2 plants were classified according to their genotypes for molecular markers tightly linked to Vrn-A(m)1 and Vrn-A(m)2. Mean comparisons showed that the VrnA(m)2 allele for winter growth habit was dominant to the vrn-A(m)2 allele for spring growth habit and that the Vrn-A(m)1 allele for spring growth habit was dominant to the vrn-A(m)1 allele for winter growth habit. A significant interaction was found between these two genes, suggesting that they work in the same developmental pathway. Plants homozygous for the recessive vrn-A(m)2 allele for spring growth habit flowered earlier than plants from the Vrn-A(m)2 class independently of the alleles present at Vrn-A(m)1. However, differences in heading date between plants with the Vrn-A(m)1 allele and those with the vrn-A(m)1 allele were significant only when the dominant Vrn-A(m)2 allele was present. A genetic model for the action of these two vernalization genes is proposed in which the role of Vrn-A(m)1 is to counteract the Vrn-A(m)2-mediated delay of flowering.     

Meiotic pairing in hybrids between tetraploid Triticale and related species: new elements concerning the chromosome constitution of tetraploid Triticale

Summary Two F5 strains of tetraploid triticale (2n= 4x=28), obtained from 6x triticaleX2 rye progenies, were crossed with diploid and tetraploid rye, some durum and bread wheats, and various 8x and 6x triticale lines. Meiosis in the different hybrid combinations was studied. The results showed that the haploid complement of these triticales consists of seven chromosomes from rye and seven chromosomes from wheat. High frequencies of PMCs showing trivalents were observed in hybrids involving the reference genotypes of wheat and triticale. These findings proved that several chromosomes from the wheat component have chromosome segments coming from two parental wheat chromosomes. The origin of these heterogeneous chromosomes probably lies in homoeologous pairing occurring at meiosis in the 6x triticaleX2x rye hybrids from which 4x triticale lines were isolated. A comparison among different hybrids combinations indicated that the involvement of D-genome chromosomes in homoeologous pairing is quite limited. In contrast, meiotic patterns in 4x triticale X 2x rye hybrids showed a quite high pairing frequency between some R chromosomes and their A and B homoeologues.     

The effects of low temperature acclimation of winter rye on catalytic properties of its ribulose bisphosphate carboxylase-oxygenase.

A comparison was made of the kinetics of the carboxylation reaction of bicarbonate-magnesium-activated ribulose biphosphate carboxylase-oxygenase purified from cold-hardened and unhardened winter rye (Secale cereale L. cv. Puma). The activity of the (NH4)2SO4-precipitated enzyme from hardened plants was stable at -20 degrees C for a month, whereas the form from unhardened plants was reversibly cold inactivated. The KmCO2 of the unhardened form increased more rapidly with decreasing pH below 8.2, but the estimated pKa of chemical groups associated with the active site was not affected by the cold hardening. The temperature dependencies of the KmCO2 of the two forms of the enzyme crossed at 10 degrees C with the effect that the catalysis of carboxylation by ribulose biphosphate carboxylase-oxygenase from Puma rye was most efficient in the temperature range to which the plants had been adapted.     

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.     

Inheritance of apospory in bahiagrass,Paspalum notatum

Previous studies on the inheritance of aposporous apomixis in bahiagrass showed a wide range of segregation ratios in crosses involving sexual and aposporous apomictic plants. The F1 progenies were classified through a visual progeny test carried out on few F2 plants. The number of sexual F1s highly exceeded the apomictics leading to the conclusion that apomixis was controlled by a few recessive genes. The present study examines the inheritance of apospory in bahiagrass. A sexual plant was self-pollinated and crossed with an aposporous apomictic plant as pollen donor. Backcross and F2 progenies were obtained in several combinations. All self-pollinated sexual plants or sexual x sexual crosses produced progenies free of apospory. All crosses involving a sexual and an apomictic plant produced approximately three times more apospory-free plants than plants with apospory. Bahiagrass is of autotetraploid origin and hence is expected to display tetrasomic inheritance. The most widely accepted genetic model for inheritance of apospory in tropical grasses is a single dominant gene with tetrasomic inheritance. In the present experiments none of the apospory-free F1s segregated for the apospory trait indicating that it is most likely a dominant character. However, the observed results fit better a modified model: tetrasomic inheritance of a single dominant gene with pleiotropic effect and incomplete penetrance. The excess of apospory-free plants in the F1 progeny could be ascribed to some distortion in the segregation pattern due to a pleiotropic lethal effect of the dominant A allele with incomplete penetrance. Alternatively, partial lethality of factors linked to aposporous gene may account for segregation distortion against apospory.     

Validation of the <Emphasis Type="Italic">VRN-H2</Emphasis>/<Emphasis Type="Italic">VRN-H1</Emphasis> epistatic model in barley reveals that intron length variation in <Emphasis Type="Italic">VRN-H1</Emphasis> may account for a continuum of vernalization sensitivity

The epistatic interaction of alleles at the VRN-H1 and VRN-H2 loci determines vernalization sensitivity in barley. To validate the current molecular model for the two-locus epistasis, we crossed homozygous vernalization-insensitive plants harboring a predicted “winter type” allele at either VRN-H1 (Dicktoo) or VRN-H2 (Oregon Wolfe Barley Dominant), or at both VRN-H (Calicuchima-sib) loci and measured the flowering time of unvernalized F₂ progeny under long-day photoperiod. We assessed whether the spring growth habit of Calicuchima-sib is an exception to the two-locus epistatic model or contains novel “spring” alleles at VRN-H1 (HvBM5A) and/or VRN-H2 (ZCCT-H) by determining allele sequence variants at these loci and their effects relative to growth habit. We found that (a) progeny with predicted “winter type” alleles at both VRN-H1 and VRN-H2 alleles exhibited an extremely delayed flowering (i.e. vernalization-sensitive) phenotype in two out of the three F₂ populations, (b) sequence flanking the vernalization critical region of HvBM5A intron 1 likely influences degree of vernalization sensitivity, (c) a winter habit is retained when ZCCT-Ha has been deleted, and (d) the ZCCT-H genes have higher levels of allelic polymorphism than other winterhardiness regulatory genes. Our results validate the model explaining the epistatic interaction of VRN-H2 and VRN-H1 under long-day conditions, demonstrate recovery of vernalization-sensitive progeny from crosses of vernalization-insensitive genotypes, show that intron length variation in VRN-H1 may account for a continuum of vernalization sensitivity, and provide molecular markers that are accurate predictors of “winter vs spring type” alleles at the VRN-H loci.     

Ethylene Induces Antifreeze Activity in Winter Rye Leaves

Antifreeze activity is induced by cold temperatures in winter rye (Secale cereale) leaves. The activity arises from six antifreeze proteins that accumulate in the apoplast of winter rye leaves during cold acclimation. The individual antifreeze proteins are similar to pathogenesis-related proteins, including glucanases, chitinases, and thaumatin-like proteins. The objective of this study was to study the regulation of antifreeze activity in response to ethylene and salicyclic acid, which are known regulators of pathogenesis-related proteins induced by pathogens. Nonacclimated plants treated with salicylic acid accumulated apoplastic proteins with no antifreeze activity. In contrast, when nonacclimated plants were exposed to ethylene, both antifreeze activity and the concentration of apoplastic protein increased in rye leaves. Immunoblotting revealed that six of the seven accumulated apoplastic proteins consisted of two glucanases, two chitinases, and two thaumatin-like proteins. The ethylene-releasing agent ethephon and the ethylene precursor 1-aminocyclopropane-1-carboxylate also induced high levels of antifreeze activity at 20 degrees C, and this effect could be blocked by the ethylene inhibitor AgNO(3). When intact rye plants were exposed to 5 degrees C, endogenous ethylene production and antifreeze activity were detected within 12 and 48 h of exposure to cold, respectively. Rye plants exposed to drought produced both ethylene and antifreeze activity within 24 h. We conclude that ethylene is involved in regulating antifreeze activity in winter rye in response to cold and drought.     

普通小麦远缘杂交F1代表现型研究

小麦远缘杂交试验采用常规杂交方法,不进行胚培养,在自然条件下,实现了普通小麦与小黑麦(6X,AABBRR)、柱穗山羊草(2X,DD)、人工合成六倍体小麦(6X,AABBDD)、CIMMYT人工合成小麦(6X,AABBDD)的杂交.发现在F1代中,出现了2～7种植株表现型、7种性状分离现象;小麦颖壳颜色变化可能受多个加性基因控制;杂交F2代结实率较F1代有数倍或十余倍的增长.     

A Cre::FLP fusion protein recombines FRT or loxP sites in transgenic maize plants

SUMMARY: The coding sequences of Cre (site-specific recombinase from bacteriophage P1) and FLP (yeast 2-microm plasmid site-specific recombinase) were fused in frame to produce a novel, dual-function, site-specific recombinase gene. Transgenic maize plants containing the Cre::FLP fusion expression vector were crossed to transgenic plants containing either the loxP or FRT excision substrate. Complete and precise excisions of chromosomal fragments flanked by the respective target sites were observed in the F1 and F2 progeny plants. The episomal DNA recombination products were frequently lost. Non-recombined FRT substrates found in the F1 plants were recovered in the F2 generation after the Cre::FLP gene segregated out. They produced the recombination products in the F3 generation when crossed back to the FLP-expressing plants. These observations may indicate that the efficiency of site-specific recombination is affected by the plant developmental stage, with site-specific recombination being more prevalent in developing embryos. The Cre::FLP fusion protein was also tested for excisions catalysed by Cre. Excisions were identified in the F1 plants and verified in the F2 plants by polymerase chain reaction and Southern blotting. Both components of the fusion protein (FLP and Cre) were functional and acted with similar efficiency. The crossing strategy proved to be suitable for the genetic engineering of maize using the FLP or Cre site-specific recombination system.     

Comparative molecular-genetic mapping of genomes of rye (Secale cereale L.) and other cereals

The genetic map of rye consisting of 149 RFLP, 20 isozyme and 12 microsatellite markers was developed. Using the collection of cross-hybridizing probes, the presence of multiple translocations in rye genome with respect to wheat and barley genomes was shown. However, within large regions of genome a strict collinearity of marker order was observed that allow us to use the method of comparative mapping for an introduction of new genes. In the developed genetic map 18 morphological and breeding-valuable genes mapped in different rye populations were integrated. The comparative analysis of homeological loci in genomes of Triticeae species as well as in genomes of rice and maize was carried out. The genes controlling a number of morphological traits, plant height, photoperiodic response and winter/spring growth habit were shown to be conserve among cereals and to form clear homoeologous rows.     

Bt水稻中crylAb基因的遗传分析

用PCR、GUS染色和Western点杂交技术检测了Bt水稻杂交后代群体,在394株GUS阳性株中,有392株表达Bt蛋白,协同表达株率达99.49%。由此表明,在杂交后代中报告基因gus和目的基因crylAb紧密连锁遗传与表达。GUS组织染色和Southern杂交检测表明Bt水稻中的crylAb基因呈单位点显性基因遗传,且在有性世代中能稳定传递,还发现,在BC1、BC1F2和粳粳交F2群体中crylAb基因呈单位点显性基因遗传,而在籼粳交F2群体中偏离3：1分离。     

Genetics of resistance to phosphine in Rhyzopertha dominica (Coleoptera: Bostrichidae)

The inheritance of resistance to phosphine was studied in two strains of the lesser grain borer, Rhyzopertha dominica (F.), labeled 'Weak-R' and 'Strong-R'. These strains were purified versions of field-selected populations collected in Queensland, Australia. Weak-R and Strong-R were, respectively, 23.4 times (20-h exposure) and 600 times (48-h exposure) resistant to phosphine compared with a reference susceptible strain (S-strain). Each -R strain was crossed with the S-strain and the response to phosphine was measured in their respective F1, F2, and F1-backcross (F1-BC) progenies. Data from testing of reciprocal F1 progeny indicated that resistance in Weak-R was autosomal and incompletely recessive with a degree of dominance -0.96. Modified chi-square analysis and contingency analysis of the observed response to phosphine of F1-BC and F2 progenies rejected the hypothesis of single gene inheritance of resistance. Analysis of the response of the F1, F2, and F1-BC progeny from the Strong-R x S-strain cross also rejected the null hypothesis for single gene resistance. Resistance in the Strong-R strain was autosomal and incompletely recessive with a degree of dominance of -0.64. The Weak-R and Strong-R strains were then crossed. Analysis ofthe F1 and F2 progenies of this reciprocal cross revealed that the strong resistance phenotype was coded by a combination of the genes already present in the Weak-R genotype plus an extra major, incompletely recessive gene. There was also evidence of a minor dominant gene present in approximately 5% of Strong-R individuals.     

The Characterization and Geographical Distribution of the Genes Responsible for Vernalization Requirement in Chinese Bread Wheat

The frequency and distribution of the major vernalization requirement genes and their effects on growth habits were studied.Of the 551 bread wheat genotypes tested,seven allelic combinations of the three Vrn.1 genes were found to be responsible for the spring habit,three for the facultative habit and one for the winter habit.The three Vrn-1 genes behaved additively with the dominant allele of Vrn-A1 exerting the strongest effect.The allele combinations of the facultative genotypes and the discovery of spring genotypes with "winter" allele of Vrn-1 implied the presence of as yet unidentified alleles/genes for vernalization response.The dominant alleles of the three Vrn-1 genes were found in all ten ecological regions where wheat Is cultivated in China,with Vrn-D1 as the most common allele in nine and Vrn-A1 in one.The combination of vrn-A 1vrnB 1Vrn-D1 was the predominant genotype in seven of the regions.Compared with landraces,improved varieties contain a higher proportion of the spring type.This was attributed by a higher frequency of the dominant Vrn-A1 and Vrn-B1 alleles in the latter.Correlations between Vrn-1 allelic constitutions and heading date,spike length,plant type as well as cold tolerance were established.     

Fecundity, phenology, and seed dormancy of F1 wild-crop hybrids in Sunflower (Helianthus annuus, Asteraceae)

Crop-to-wild hybridization has the potential to introduce beneficial traits into wild populations. Gene flow from genetically engineered crops, in particular, can transfer genes coding for traits such as resistance to herbicides, insect herbivores, disease, and environmental stress into wild plants. Cultivated sunflower (Helianthus annuus) hybridizes spontaneously with wild/weedy populations (also H. annuus), but little is known about the relative fitness of F1 hybrids. In order to assess the ease with which crop-to-wild introgression can proceed, we compared characteristics of F1 wild-crop progeny with those of purely wild genotypes. Two nontransgenic, cultivated varieties were crossed with wild plants from three different regions-Texas, Kansas, and North Dakota. Seed burial experiments in the region of origin showed that wild-crop seeds had somewhat higher germination rates (less dormancy) than wild seeds from Kansas and North Dakota, while no differences were seen in seeds from Texas. Progeny from each type of cross were grown in outdoor pots in Ohio and in a weedy field in Kansas to quantify lifetime fecundity and flowering phenology. Flowering periods of hybrid and wild progeny overlapped considerably, especially in plants from North Dakota and Texas, suggesting that these hybrids are very likely to backcross with wild plants. In general, hybrid plants had fewer branches, flower heads, and seeds than wild plants, but in two crosses the fecundity of hybrids was not significantly different from that of purely wild plants. In Ohio, wild-crop hybrids from North Dakota appeared to be resistant to a rust that infected 53% of the purely wild progeny, indicating a possible benefit of "traditional" crop genes. In summary, our results suggest that F1 wild-crop hybrids had lower fitness than wild genotypes, especially when grown under favorable conditions, but the F1 barrier to the introgression of crop genes is quite permeable.     

Genetic determination of stem-rust resistance in rye

The harmful effect of stem rust on the crops of short-stem diploid winter rye was studied. If stem rust affected the plants by 70-100%, this decreased the mass of 1000 grains by about 35.8%. The genes that control the stemrust resistance of rye might originate from the following cultivars and forms: Ilmen, Orlovskii Gibrid, Kharkovskaya 55, Kharkovskaya 60, Kustovka, Kombaininyai, Kazanskaya, Krupnozernaya, Novozybkovskaya 4, Alfa, Derzhavinskaya 29, Chulpan, and Rossul, as well as wild populations of the perennial rye Secale montanum. This study was first to demonstrate that the resistance of the Kharkovskaya 55 and Rossul rye cultivars to the population of stem rust was controlled by a single dominant gene, which was designated Sr1.