Assessment of the Combining Ability of Triticale and Secalotriticum with Respect to in Vitro Androgenesis Characteristics

The combining ability of cultivars and strains of triticale and secalotriticum with respect to the characteristics of androgenesis in anther culture has been analyzed (the 6 × 3 polytester design and parental forms). Nonadditive effects of gene interaction have been found to be prevalent in the genetic control of the characters analyzed in plants with the given genotypes. However, the characteristics of androgenesis were the highest in those hybrid combinations in which genotypes with high values of general combining ability (GCA) and positive effects of specific combining ability (SCA) participated. A program of the analysis of combining ability in triticale and secalotriticum is proposed. The program is aimed at detecting hybrid combinations with the best androgenetic capacities and takes into account both additive and nonadditive effects.     

Analysis of meiotic genome stabilization in the rye-wheat amphidiploid secalotriticum (×Secalotriticum,S/RRAABB, 2n = 42)

The results of cytogenetic studies of genome stabilization in secalotriticum are presented. Comparative analysis of microsporogenesis in hexaploid secalotriticum, triticale, and reciprocal hybrids of them (^S/RRAABB, ^T/AABBRR, 2n = 6x = 42) was carried out. The cytogenetic properties and genotypic factors of genome stabilization in heteroplasmic triticale were found. The inheritance by the secalotriticum of the genotypically caused first meiotic division, the so-called reduction division, of univalents from original combinations of rye-triticale hybrids was demonstrated for the first time.     

籼型杂交稻光合特性的配合力分析

以6个籼型杂交稻三系不育系和5个恢复系按不完全双列杂交设计配制的30个杂交稻组合及其亲本品种为材料,对其光合性状进行了测定和分析,结果表明:(1)杂交稻组合的光合特性存在显著或极显著的组合间遗传差异,光合特性的遗传变异主要来自基因的非加性效应;(2)胞间CO2浓度、蒸腾速率、叶绿素a、叶绿素b和类胡萝卜素含量受不育系的影响大于恢复系,而气孔导度、叶绿素a+b含量受恢复系的影响大于不育系;(3)杂交稻光合性状的广义遗传力均大于狭义遗传力,各性状主要受基因互作及环境的影响。狭义遗传力的大小依次为叶绿素b、叶绿素a+b、叶绿素a、气孔导度、胞间CO2浓度、类胡萝卜素和蒸腾速率,这些性状具有中等遗传力;(4)9个光合性状杂交稻F1表型值与父母本一般配合力效应值之和的相关系数均达极显著水平。因此,可以根据父母本一般配合力效应值之和来预测杂交稻组合光合性状的表现,有利于高效选育高光效杂交稻组合。     

Combining ability for the traits of stem branching and plant height in linseed lines

Combining ability for the traits of stem branching and plant height has been studied in ten pure lines of flax under complete and incomplete diallel crosses. High heritability of the traits "plant height", "the number of lateral stems" and "the number of lateral shoots" and essential role of genes with dominant effects of interaction in genetic control of the traits of stem branching and plant height have been shown. On the basis of combining ability indexes the ways for usage of certain genotypes and crossing combinations in flax breeding were defined. As a result of individual selection from hybrid combinations some new complexes of habit traits and agriculturally valuable plant characteristics were obtained.     

Combining ability according to the traits of stem branching and plant height in linseed lines

The combining ability according to the traits of stem branching and plant height has been studied for ten pure lines of oil flax under the conditions of complete and incomplete diallel crosses. The high heritability of traits, such as plant height, the number of lateral stems and shoots, and the essential role of genes with dominant effects of interaction in genetic control of the traits of stem branching and plant height have been shown. The methods of application of certain genotypes and crossed combinations in flax breeding based on combining-ability indexes were identified. Some new complexes of habitus traits and agriculturally valuable plant characteristics were obtained as a result of individual selection from hybrid combinations.     

Quantitative trait loci associated with androgenic responsiveness in triticale (×Triticosecale Wittm.) anther culture

Quantitative trait loci (QTLs) associated with androgenic responsiveness in triticale were analyzed using a population of 90 DH lines derived from the F1 cross between inbred line ‘Saka 3006’ and cv. ‘Modus’, which was used in a number of earlier studies on molecular mapping in this crop. Using Windows QTL Cartographer and MapQTL 5.0, composite interval mapping (CIM) and association studies (Kruskal–Wallis test; K–W) for five androgenesis parameters (androgenic embryo induction, total regeneration and green plant regeneration ability, and two characteristics describing final androgenesis efficiency) were conducted. For the studied components of androgenic response, CIM detected in total 28 QTLs which were localized on 5 chromosomes from A and R genomes. Effects of all QTLs that were identified at 2.0 or above of the LOD score explained 5.1–21.7?% of the phenotypic variation. Androgenesis induction was associated with seven QTLs (LOD between 2.0 and 5.8) detected on chromosomes 5A, 4R, 5R and 7R, all of them confirmed by K–W test as regions containing the markers significantly linked to the studied trait. What is more, K–W test revealed additional markers on chromosomes: 5A, 2BL, 7B and 5R. Both total and green regeneration ability were controlled by genes localized on chromosome 4A. Some of the QTLs that affected final androgenesis efficiency were identical with those associated with androgenic embryo induction efficiency, suggesting that the observed correlation may be either due to tight linkage or to pleiotropy. Key message Five regions of the triticale genome were indicated as revealing significant marker/trait association. Markers located in these regions are potentially useful for triticale breeding through marker-assisted selection.     

Aneuploidy among androgenic progeny of hexaploid triticale (XTriticosecale Wittmack)

Doubled haploids are an established tool in plant breeding and research. Of several methods for their production, androgenesis is technically simple and can efficiently produce substantial numbers of lines. It is well suited to such crops as hexaploid triticale. Owing to meiotic irregularities of triticale hybrids, aneuploidy may affect the efficiency of androgenesis more severely than in meiotically stable crops. This study addresses the issue of aneuploidy among androgenic regenerants of triticale. Plant morphology, seed set and seed quality were better predictors of aneuploidy, as determined cytologically, than flow cytometry. Most aneuploids were hypoploids and these included nullisomics, telosomics, and translocation lines; among 42 chromosome plants were nulli-tetrasomics. Rye chromosomes involved in aneuploidy greatly outnumbered wheat chromosomes; in C₀ rye chromosomes 2R and 5R were most frequently involved. While the frequency of nullisomy 2R was fairly constant in most cross combinations, nullisomy 5R was more frequent in the most recalcitrant combination, and its frequency increased with time spent in culture with up to 70% of green plants recovered late being nullisomic 5R. Given that 5R was not involved in meiotic aberrations with an above-average frequency, it is possible that its absence promotes androgenesis or green plant regeneration. Overall, aneuploidy among tested combinations reduced the average efficiency of double haploid production by 35% and by 69% in one recalcitrant combination, seriously reducing the yield of useful lines.     

In vitro androgenesis of triticale in isolated microspore culture

Culture conditions for triticale (X Triticosecale Wittmack) androgenesis were studied using microspore culture. Sporophytic development of isolated triticale microspores in culture is described in five winter hexaploid triticale genotypes. Microspores were isolated using a microblendor, and embryogenesis was induced in modified 190-2 medium both in the presence and absence of growth regulators. The highest induction of microspore embryogenesis was obtained in a growth regulator-free medium. Adventitious embryogenesis was observed during in vitro development of triticale microspores. Albino and green plantlets were regenerated from embryo-like structures. More than 50% of regenerants were albino. In total, 126 green plantlets were produced, transplanted and established in soil. Cytological evidence revealed that 90% of the transplanted regenerants were haploid. This revised version was published online in June 2006 with corrections to the Cover Date.     

Effect of induction medium pH and maltose concentration on in vitro androgenesis of hexaploid winter triticale and wheat

Low levels of in vitro androgenesis limit the utility of anther culture as a routine tool for the improvement of triticale. The objectives of this research were to determine the effect of induction medium modifications on the embryoid induction (EI) and green plant regeneration (GPR) of three hexaploid winter triticales and a hexaploid winter wheat. Medium modifications were factorial combinations of gelling agents, pH and maltose concentration. There was a significant difference in the response of wheat and triticales. An induction medium pH of 4.8 vs. 5.8 led to higher EI percentages in all cases and in Ficoll containing medium to a significantly higher GPR in the wheat, but it had no effect on the response of the triticales. Maltose at 0.26 M vs. 0.21 M increased the EI of both the triticales and the wheat but led to higher GPR only in the case of wheat. Averaged over genotypes, induction media with agar and Ficoll 400 were superior to the liquid form. In terms of GPR the wheat gave the highest response: 9.1%. The GPR percentages for the triticales ranged from 1.4 to 7.1. Genotype specificity must be overcome if anther culture is to be routinely used in diverse arrays of triticale germplasm.     

Formation of a new original material based on genetic heterogeneity of disomal wheat plants from offspring of aneuploids

A series of disomic lines of spring wheat Opal selected on the basis of monosomic lines of this cultivar has been studied. The lines have been tested for combining ability, and the heterosis effect has been studied in disomic lines of F1 hybrids obtained by topcrossing. The line have been demonstrated to differ both from one another and from cultivar Opal in the expression of quantitative traits, combining ability, and the degree of heterosis in F1. These data suggest that recombinations accompanying the formation of the monosomic series have changed their genetic program. To test this suggestion, intramolecular heterogeneity of 42-chromosome plants has been analyzed using polymerase chain reaction (PCR) and isoenzyme analysis. The results confirmed the differences at the DNA and protein levels. According to the results of molecular analyses, A-genome lines are the most polymorphic. Strong heterosis effects have been detected in hybrid combinations contributed by D- and B-genome lines, which are characterized by medium and low degrees of molecular genetic polymorphism. Lines that are promising in terms of breeding programs have been identified.     

Hybrid necrosis in triticale and the expression of necrosis genes in allopolyploids

Summary The occurrence in triticale of four different genes causing hybrid necrosis is described: Ne1 and Ne2 in the B genome of wheat and Ner1 and Ner2 in the rye genome. Hybrid necrosis develops due to dominant complementary interaction of two genes. This interaction in triticale, however, may take place not only between genes belonging to the same genome but also between genes of different genomes. In triticale, these genes can cause hybrid necrosis in four different combinations. The inheritance of the phenomenon in triticale is, therefore, more complicated than it is in wheat or rye. To avoid hybrid necrosis in triticale, attention should be paid that no necrosis genes are introduced into the primary triticale stocks from the wheat and rye parents. The expression of necrosis genes is influenced by the level of ploidy. Any additional genome — A, B, D, or R — may exert a suppressing effect on the expression of necrosis genes. Therefore, when identifying genotypes of triticale with regard to their necrosis genes, the level of ploidy has to be accounted for. Moreover, the present results illustrate that gene expression in polyploids is not only determined by interactions with other single genes but that it may also be modified by the total genotype of the respective individual.     

Biolistic Transfer of the Gene uidA and Its Expression in Haploid Embryo-like Structures of Triticale (×Triticosecale Wittmack)

Haploid embryo-like structures (ELS) of triticale were obtained by in vitro androgenesis. These structures were bombarded with gold microparticles 1 µm in diameter coated with the plasmid pAHGUS, using the Dupont PDS He/1000 apparatus. Analysis was made of the influence of genotype, duration of ELS pre-culture, helium pressure and shooting distance on the transfer and expression of the gene uidA. No significant differences were seen between genotypes or duration of pre-culture; differences were found, however, with respect to helium pressure and shooting distance. The combination of 1100 psi helium and a 6 cm shooting distance led to the greatest mean number of uidA expression foci in the ELS (1.63), and the greatest mean number of ELS showing at least one focus (0.38). With a pressure of 1800 psi and a 9 cm distance, only 0.35 foci were obtained per ELS, and only 0.18 ELS had one or more foci. These results are being used as references in a program for obtaining double haploid and transgenic triticale plants.     

Formation of New Original Material Based on the Genetic Heterogeneity of the Disomic Offspring of Aneuploid Wheat Plants

A series of disomic lines of spring wheat Opal developed on the basis of monosomic lines of this cultivar has been studied. The lines have been tested for combining ability, and the heterosis effect has been studied in disomic lines of F₁ hybrids obtained by topcrossing. The line have been demonstrated to differ both from one another and from cultivar Opal in the expression of quantitative traits, combining ability, and the degree of heterosis in F₁. These data suggest that recombinations accompanying the formation of the monosomic series have changed their genetic program. To test this suggestion, intramolecular heterogeneity of 42-chromosome plants has been analyzed using polymerase chain reaction (PCR) and isoenzyme analysis. The results confirmed the differences at the DNA and protein levels. According to the results of molecular analyses, A-genome lines are the most polymorphic. Strong heterosis effects have been detected in hybrid combinations contributed by D- and B-genome lines, which are characterized by medium and low degrees of molecular genetic polymorphism. Lines that are promising in terms of breeding programs have been identified.     

Evaluation of genetic behavior of some Egyption Cotton genotypes for tolerance to water stress conditions

Water stress is a critical abiotic stress for plant reduction in arid and semiarid zones and, has been discovered to be detrimental to the development of seedlings as well as the growth and physiological characteristics of many crops such as cotton. The objectives of our study were to determine the combining ability and genetic components for five quantitative traits [(leaf area (LA), leaf dry weight (LDW), plant height (PH), fiber length (2.5 percent SL), and lint cotton yield/plant (LCY/P)] under water shortage stress, a half diallel cross between six cotton genotypes representing a wide range of cotton characteristics was evaluated in RCBD with four replications. The genotype mean squares were significant for all traits studied, demonstrating significant variation among genotypes for all characters under water shortage stress. LCY/P had the highest phenotypic and genotypic correlation co-efficient with PH, LDW, and LA shortage. The highest direct effect on lint cotton yield was exhibited by leaf area (3.905), and the highest indirect effects of all traits were through LA, with the exception of 2.5 percent SL, which was through LDW. The highest dissimilarity (Euclidean Distance) between parental genotypes was between G.87 and G.94, followed by G.87 and Menoufi. G.94 was also a well-adapted genotype, and the combinations G.87 x G.94 and G.87 x Menoufi may outperform their parents. The combining ability analysis revealed highly significant differences between parental GCA effects and F1 crosses SCA effects. The variation of GCA and SCA demonstrated the assurance of additive and non- additive gene action in the inheritance of all traits studied. In terms of general combining ability (GCA) effects, parental genotype G.94 demonstrated the highest significant and positive GCA effects for all traits studied, with the exception of 2.5 percent SL, where G.87 revealed the highest significant and positive GCA effects. The effects of specific combining ability (SCA) revealed that the cross (G.87 x2G.94) revealed stable, positive, and significant SCA for all of the studied traits.     

Assessing genetic variation to predict the breeding value of winter triticale cultivars and lines

The objective of this study was to evaluate the usefulness of amplified fragment length polymorphism (AFLP) analyses for selection of the best parents for breeding of hybrid winter triticale. Phenotypic diversity was measured for 8 agronomic traits in 10 parents and 27 F1 hybrids. Genotypic diversity was measured by 91 AFLP markers. Coefficients of correlation of genetic similarity (AFLP-GS) with both Euclidean distances and mean values of the traits were generally not significant. A correction of the preliminary binary matrix into trait-specific derived matrices increased the values of 5 correlation coefficients to a significant level. The correlation of AFLP-GS with mid-parent heterosis of grain weight per plant was low but significant (r = -0.452). Our study confirms the effectiveness of marker preselection for obtaining AFLP-GS better correlated with heterosis. The use of derived matrices is promising for reducing the number of cross combinations tested for specific combining ability.     

Genotypic features of morphogenesis in spring barley anther culture

The variability of characteristics in haploid production was assessed depending on the growth conditions for donor plants, ear pretreatment technique, and nutrient medium composition. The ranks of genotypes included in the experiment remained unchanged in terms of their in vitro ability toward androgenesis, which evidences the significant contribution of a genetic component to the general variability and demonstrates the need to investigate how genetic control of androgenesis works.     

Effect of rye chromosomes on features of androgenesis in wheat-rye substituted lines of Triticum aestivum L. sort Saratovskaya 29/Secale cerale L. sort Onokhoiskaia and Triticale]

The characteristic features of androgenesis in six wheat-rye substitution lines Triticum aestivum L. (cv. Saratovskaya 29)/Secale cereale L. (cv. Onokhoiskaya) and triticale (2n = 56) using anther culture at different concentrations of 2,4-D in the growth medium were studied. Under variable cultivation conditions, the significant effect of genotypic diversity on the variability of such androgenesis parameters as the frequency of productive anthers, the frequency of embryoid formation, and the frequency of total regenerated plantlets, was shown. It was demonstrated that chromosomes 1R, 3R, and 7R stimulated the formation of androgenous embryoids, while chromosome 5R produced an opposite effect. In triticale and substitution lines, the regeneration ability of androgenous embryoids induced by elevated 2,4-D concentrations was inhibited. Chromosome 1R of the Onokhoiskaya cultivar was suggested to contain genes suppressing regeneration of green plantlets, while chromosome 3R, conversely, stimulated their formation. Chromosomes 1R, 2R, 3R, and 7R of the Onokhoiskaya cultivar did not inhibit the spontaneous formation of androgenous hexaploids in the substitution lines.     

Doubled haploids in production of male sterility maintaining triticale (<Emphasis Type="Italic">Triticosecale</Emphasis> Wittmack) lines

The androgenetic response of several selected male sterility-maintainer genotypes of triticale was investigated. Androgenesis induction was obtained in all cultivars, but a large genotypic variation in green plant regeneration was observed. The number of regenerated triticale plants varied from 0.1 to 4.7 green plants per spike, depending on genotype. Spontaneous doubling of chromosomes varied from 14 to 60% for particular genotypes and, on average, reached the value of 34% for all genotypes. Fertile DH lines obtained in this study will find practical application in the development of triticale male sterile lines that are desirable in hybrid breeding.     

Spike morphology alternations in androgenic progeny of hexaploid triticale (× Triticosecale Wittmack) caused by nullisomy of 2R and 5R chromosomes

Induction of androgenesis, followed by chromosome doubling, is a crucial method to obtain complete homozygosity in one-generation route. However, in vitro androgenesis can result in various genetic and epigenetic changes in derived triticale plants. In this study, we evaluated chromosome alternations and we associated them with the changes of spike morphology in androgenic progeny of triticale. We karyotyped offspring plants that derived from double haploid plants using fluorescence in situ hybridization techniques. We distinguished four major groups of karyotypes: double ditelosomics, nullisomics N2R, nullisomics N5R, and triticale plants with a complete set of chromosomes. It is known that more than half of QTLs connected with androgenic response are located in R-genome of triticale but 2R, 5R, and 6R chromosomes are not included. We hypothesized that the reason why only aberrations of chromosomes 2R and 5R appear during androgenesis of triticale is that because these chromosomes are not involved in the stimulation of androgenic response and the following regeneration of plants is not disrupted. Concerning the established groups, we evaluated following quantitative traits: spike length, number of spikes per plant, number of spikelets per spike, and number of grains per spike. The nullisomy of chromosome 2R and 5R resulted in vast changes in spike architecture of triticale plants, which can be correlated with the location of major QTLs for spike morphology traits on these chromosomes. The spikes of nullisomic plants had significantly decreased spike length which correlated with the reduction of number of spikelets per spike and number of grains per spike.

     

DNA methylation changes in triticale due to in vitro culture plant regeneration and consecutive reproduction

Doubled haploids of triticale are of interest for plant breeders due to hybrid breeding programs based on cytoplasmic male sterility Tt phenomenon. However, (epi)mutations appearing during in vitro culture regeneration may lead to a phenotypic variation that makes the uniformity of plant materials questionable. Using RP-HPLC genomic DNA methylation of donor doubled haploid plants utilized as a source of tissues for the in vitro regeneration (via androgenesis and somatic embryogenesis) of triticale cv. Bogo and their consecutive generative progeny was evaluated. It was demonstrated that in vitro cultures induced a decrease of the DNA methylation of the regenerants independently of the approach used for plant regeneration. The decrease in DNA methylation of genomic DNA proceeded up to the first/second successive generations followed by the beginning of its reestablishment. Moreover, somatic embryogenesis resulted in a higher level of genomic DNA demethylation in regenerants than androgenesis and the process of methylation seems to be affected by donor plant. It is being speculated that long term changes in genomic DNA methylation may be a source of off-type individuals that may spontaneously arise during plant breeding.