Chromosome substitutions of Triticum timopheevii in common wheat and some observations on the evolution of polyploid wheat species

Whether the two tetraploid wheat species, the well known Triticum turgidum L. (macaroni wheat, AABB genomes) and the obscure T. timopheevii Zhuk. (A^tA^tGG), have monophyletic or diphyletic origin from the same or different diploid species presents an interesting evolutionary problem. Moreover, T. timopheevii and its wild form T. araraticum are an important genetic resource for macaroni and bread-wheat improvement. To study these objectives, the substitution and genetic compensation abilities of individual T. timopheevii chromosomes for missing chromosomes of T. aestivum Chinese Spring (AABBDD) were analyzed. Chinese Spring aneuploids (nullisomic-tetrasomics) were crossed with a T. timopheevii x Aegilops tauschii amphiploid to isolate T. timopheevii chromosomes in a monosomic condition. The F₁ hybrids were backcrossed one to four times to Chinese Spring aneuploids without selection for the T. timopheevii chromosome of interest. While spontaneous substitutions involving all A^t- and G-genome chromosomes were identified, the targeted T. timopheevii chromosome was not always recovered. Lines with spontaneous substitutions from T. timopheevii were chosen for further backcrossing. Six T. timopheevii chromosome substitutions were isolated: 6A^t (6A), 2G (2B), 3G (3B), 4G (4B), 5G (5B) and 6G (6B). The substitution lines had normal morphology and fertility. The 6A^t of T. timopheevii was involved in a translocation with chromosome 1G, resulting in the transfer of the group-1 gliadin locus to 6A^t. Chromosome 2G substituted for 2B at a frequency higher than expected and may carry putative homoeoalleles of gametocidal genes present on group-2 chromosomes of several alien species. Our data indicate a common origin for tetraploid wheat species, but from separate hybridization events because of the presence of a different spectrum of intergenomic translocations.     

Identification of the G-genome donor to Triticum timopheevii by DNA:DNA hybridizations

In vitro DNA:DNA hybridizations and hydroxyapatite thermal-elution chromatography were employed to identify the diploid Triticum species ancestral to the G genome of Triticum timopheevii. Total genomic, unique-sequence, and repeated-sequence fractions of ³H-T. timopheevii DNA were hybridized to the corresponding fractions of unlabeled DNAs of T. searsii, T. speltoides, T. sharonensis, T. longissimum, and T. bicorne. The heteroduplex thermal stabilities indicated that, of the five species examined, T. speltoides was the most closely related to the G genome of T. timopheevii. Thus, T. spelotides appears to be the G-genome donor to T. timopheevii. The thermal stability profiles further indicated that the repeated DNA fractions from the five diploid species and the tetraploid T. timopheevii are more similar than the unique DNA fractions. This indicates that all of these species are closely related and that the sequences which comprise the current repeated fractions in the various species have not undergone any significant change since the formation of various species.Published with the approval of the Director of the West Virginia Agriculture and Forestry Experiment Station as Scientific Paper No. 1850.     

Wheat Microsatellites: The Prospects of Application for Gene Mapping and Analysis of the Reconstructed Genomes

Microsatellite markers Xgwmand Xgdmwere used to map the S1, S2, and S3genes of the induced sphaerococcoid mutants of Triticum aestivumL. and to analyze the introgressive lines of common wheat, obtained by crossing several common wheat cultivars to T. timopheeviiZhuk.; these lines carry the Lrgenes of resistance to leaf rust. All sphaerococcoid genes were linked to centromeric markers of the short and long arms of chromosomes of homoeologous group 3 of T. aestivum: the S1locus was located between the markers Xgdm72and Xgwm456; the S2gene, betweenXgwm845and Xgwm566; and the S3was located between Xgwm2and Xgwm720. The introgressive lines of common wheat carry the following substitutions from T. timopheevii, most of 2A and 2B and telomeric region of the 5AL chromosome in the line 821, the same introgression and also the completely substituted chromosome 4B in line 837, and the partially substituted chromosomes 2A and 2B in line 842. The introgression of the genomic material fromT. timopheeviiinto the chromosomes of homoeologous group 2 was the common trait of all three lines resistant to leaf rust. The authors discuss the feasibility of using microsatellite-derived data for analyzing nonmapped wheat species, linking new genes to wheat molecular genetic maps, and analyzing wheat genomes of diverse hybrid origins.     

Activity of trypsin inhibitors in wheat seedlings exposed to pathogenic fungus Tilletia caries and phytohormones

We compared the effect of common bunt (caused by fungus Tilletia caries Tul.) and treatment with phytohormones IAA, ABA, and cytokinins (CK) on the activity of trypsin inhibitors (TI) in wheat seedlings. The experiments were conducted with pathogen-susceptible species of wheat Triticum aestivum L. (cv. Zhnitsa) and resistant species T. timopheevii Zhuk. (accession k-58666 from the collection of All-Russian Institute of Plant Industry). In the resistant wheat, the fungus elevated the activity of TI and the content of CK, whereas in the susceptible wheat, it induced accumulation of IAA. In the seedlings of wheat T. aestivum, TI activity increased under the effect of CK, same as upon the action of pathogen. ABA briefly increased the activity of TI, whereas IAA did not considerably affect it. It was concluded that among the investigated hormones, CK play a leading role in the regulation of defense responses of wheat plants involving TI.     

Hormonal Status in Wheat and Rice Seedlings under Anoxia

A comparative analysis of the effects of anoxia on growth, fresh weight gain, and phytohormones in wheat (Triticum aestivum L.) and rice (Oryza sativa L.) seedlings was performed. In both plant species, a total cessation of root growth occurred during the initial hours of anoxia. In an anaerobic environment, the fresh weight of wheat seedlings decreased. An increase in the shoot length and weight under the stress conditions was found only in rice seedlings. During the initial hours of anoxia, the level of free ABA in wheat and rice tissues increased manifold, and the accumulation of a free ABA form occurred at the expense of the hydrolysis of its bound forms. The IAA content in plant tissues also increased. In wheat, the accumulation of IAA was short, but in rice, a high hormone level was retained during the entire experiment, and, as a result, its concentration exceeded that of ABA. A level of cytokinins in the tissues of both plant species was affected by anoxia to a lesser extent than that of other phytohormones. This level somewhat decreased under anoxia similarly to the level in darkness under aeration. It is suggested that IAA accumulation in hypoxia-tolerant rice seedlings under anoxia favors maintenance of shoot growth and simultaneous inhibition of root growth. At the same time, in the hypoxia-sensitive wheat, an increase in the ABA level resulted in growth cessation.     

Wheat genome structure: translocations during the course of polyploidization

The genomic organization of Triticum timopheevii (2n=28, A^tA^tGG) was compared with hexaploid wheat T. aestivum (2n=42, AABBDD) by comparative mapping using microsatellites derived from bread wheat. Genetic maps for the two crosses T. timopheevii var. timopheevii × T. timopheevii var. typica and T. timopheevii K-38555×T. militinae were constructed. On the first population, 121 loci were mapped, and on the second population 103 loci. The transferability of the wheat markers to T. timopheevii was generally better for the A genome-specific markers (76–78% produced amplification products; 26 and 29% were polymorphic) than for B genome-specific markers (54% produced amplification products; 14 and 16% were polymorphic). Of the D genome-specific markers, one third produced amplification products in T. timopheevii, but only 5 and 2% were polymorphic in the corresponding mapping populations. The maps constructed confirmed the previously described translocation between chromosome arms 6A^tS and 1GS and revealed at least two yet unknown rearrangements on chromosomes 4A^t and 6A^t. The presence of other translocations and rearrangements between T. timopheevii and T. aestivum was demonstrated by a variety of markers mapping to nonhomoeologous positions.     

Cytogenetic investigation of Triticum timopheevii (Zhuk.) Zhuk. and related species using the C-banding technique

Triticum timopheevii and related species T. militinae (2n=28, A^tG) and T. zhukovskyi (2n=42, A^mA^tG), hybrids T. kiharae, T. miguschovae, the amphidiploid T. timopheevii x T. tauschii (all 2n=42, A^tGD), T. fungicidum (ABA^tG) and T. timonovum (2n=56, A^tA^tGG) were analyzed using the C-banding technique. Chromosomes of the A^m and A^t genomes in the karyotype of T. zhukovskyi differed in their C-banding pattern. Partial substitutions of A^t-genome chromosomes and a complete substitution of the G-genome chromosomes by homoeologous chromosomes of an unidentified tetraploid wheat species with an AB genome composition were found in the T. timonovum karyotype. A^t- and G-genome chromosomes in the karyotypes of all studied species had similar C-banding patterns and were characterized by a low level of polymorphism. The comparative stability of the A^t and G genomes is determined by the origin and specifity of cultivation of studied species.     

Physiological response of riparian plants to watering in hyper-arid areas of Tarim River, China

The physiological responses and adaptive strategies of Populus euphratica Oliv. (arbor species), Tamarix ramosissima Ldb. (bush species), and Apocynum venetum L. (herb species) to variations in water and salinity stress were studied in the hyper-arid environment of the Tarim River in China. The groundwater table, the saline content of the groundwater, as well as the content of free proline, soluble sugars, plant endogenous hormones (abscisic acid (ABA), and cytokinins (CTK)) of the leaves of the three species were monitored and analyzed at the lower reaches of the Tarim River in the study area where five transects were fixed at 100 m intervals along a vertical sampling line before and after water release. Saline stress dramatically increased soluble sugar concentration of the three species. Differences in sugar accumulation were determined among the species at different transects. The free proline concentration of the leaves of T. ramosissima and P. euphratica showed a proportional decrease with various degrees of elevation of the groundwater table after water release. There was a least correlation between the soluble sugars and proline stimulation in T. ramosissima. It was strongly suggested that T. ramosissima developed a different strategy to accumulate organic solutes to adapt to the stress environment. The soluble sugars and proline accumulation responded to the changes of groundwater table independently: the former occurred under salt stress, whereas the latter was more significant under drought stress. The concentration and the increase in concentration of ABA and CTK involved in stress resistance of the three species were also determined. This increase in the hormone concentration in P. euphratica was different from that of the other two species. Expressed as a function of increase of ABA concentration in leaves, A. venetum and T. ramosissima showed a different solute accumulation in response to groundwater table. There was a significant correlation between ABA accumulation and Δ [proline] in A. venetum as well as between ABA accumulation and Δ [sugar] in T. ramosissima. Translated from Acta Ecologica Sinica, 2005, 25(8): 1966–1973 [译自: 生态学报]     

Molecular Analysis of Leaf Rust-Resistant Introgression Lines Obtained by Crossing of Hexaploid Wheat Triticum aestivum with Tetraploid Wheat Triticum timopheevii

Twenty-four Triticum aestivum×T. timopheevii hybrid lines developed on the basis of five varieties of common wheat and resistant to leaf rust were analyzed by the use of microsatellite markers specific for hexaploid wheat T. aestivum. Investigation of intervarietal polymorphism of the markers showed that the number of alleles per locus ranged from 1 to 4, depending on the marker (2.5 on average). InT. timopheevii, amplification fragments are produced by 80, 55, and 30% of primers specific to the A, B, and D common wheat genomes, respectively. Microsatellite analysis revealed two major areas of introgression of the T. timopheevii genome: chromosomes of homoeological groups 2 and 5. Translocations were detected in the 2A and 2B chromosomes simultaneously in 11 lines of 24. The length of the translocated fragment in the 2B chromosome was virtually identical in all hybrid lines and did not depend on the parental wheat variety. In 15 lines developed on the basis of the Saratovskaya-29, Irtyshanka, and Tselinnaya-20, changes occurred in the telomeric region of the long arm of the 5A chromosome. Analysis with markers specific to the D genome suggested that introgressions of the T. timopheevii genome occurred in chromosomes of the D genome. However, the location of these markers on T. timopheevii chromosomes is unknown. Our data suggest that the genes for leaf rust resistance transferred from T. timopheevii to T. aestivum are located on chromosomes of homoeological group 2.     

Assessment of the Respiratory Costs of Adaptation in Plant Species That Differ in Their Responses to Insufficient and Excessive Mineral Nutrition

The effects of insufficient and excessive mineral nutrition on the relative growth rate (RGR); the rate of total dark respiration (R); gross photosynthesis (Pg); and the contents of endogenous IAA, ABA, and cytokinins (CK) were studied in 20–60-day-old seedlings of plant species that differed in their responses to these factors: beet root (Amaranthus retroflexusL.), orchard-grass (Dactylis glomerataL.), and common wheat (Triticum aestivumL.). The changes in the level of mineral nutrition lowered the RGR and Pgvalues and the IAA/ABA and CK/ABA ratios and increased the Rand R/Pgvalues and ABA content, especially in A. retroflexusand T. aestivum. The authors propose a new way to assess the respiratory cost of adaptation to stress based on the relation between the RGR and the R/Pgvalues. The adaptation component of Ris shown to relate to the ABA content.     

Rearrangement of the Cytoskeleton in Triticum aestivum Cells during Cold Hardening and after Treatment with Abscisic Acid

Immunocytochemical study of the basic characteristics of the tubulin and actin cytoskeleton (total content, orientation, structure, and stability) was performed for various root zones of the seedlings of winter wheat cultivars contrasting in their freezing tolerance. Plant cold hardening (3°C, 7 days) and ABA treatment (30 M, 3 days) increased the stability of tubulin microtubules (MT), that is, reduced the depolymerizing action of oryzalin in vivo. However, the mechanisms of hardening and ABA stabilizing action on the cytoskeleton were different: low temperature enhanced spatial MT aggregation and resulted in the formation of a dense network of thick MT bundles, whereas ABA reduced the content of tubulin components and induced microfilament (MF) depolymerization. Most pronounced temperature- and ABA-induced cytoskeleton changes were observed in the differentiation zone, which indicates an important role of this root zone in plant adaptation and development of root freezing tolerance. Low temperatures reduced the hormonal effect on the structural arrangement and stability of MT and MF in wheat cultivars of high and moderate freezing tolerance but increased hormonal effects in the slightly tolerant cultivar. MF depolymerization and an increase in the proportion of stable MT are supposed to be a necessary condition for seedling growth retardation after their treatment with ABA and for seedlings at the initial phase of their adaptation to low temperature. At the final phase of cold hardening, some growth acceleration is evidently determined by the accumulation of highly labile MT and greater actin polymerization.     

Comparative cytological and molecular analysis of common wheat introgression lines containing genetic material of <Emphasis Type="Italic">Triticum timopheevii</Emphasis> Zhuk

A total of 40 introgression lines of common wheat (2 n = 42) Triticum aestivum L × T. timopheevii Zhuk., resistant to leaf rust and partly to powdery mildew, were examined. Based on cytological analysis of meiosis in pollen mother cells (PMC), hybrid lines were subdivided into two groups characterized by either stable or unstable meiosis. In cytologically stable lines, chromosome configuration at the MI stage of meiosis was mostly bivalent (21II) with small proportion of defect cells (almost 10%), which at most contained two univalents (20II + 2I). Cytologically unstable group was comprised of the lines, containing high proportions of cells with abnormal chromosome pairing in meiotic PMC, as well as the cells with multivalents, and the lines containing aneuploid plants. Localization of the T. timopheevii fragments performed with the use of SSR markers showed that the lines with unstable meiosis were characterized by higher numbers of introgressions compared to stable lines. The influence of certain chromosomes of T. timopheevii on chromosome pairing stability was also demonstrated. In cytologically unstable lines, the increased frequency of 2A substitutions along with the high frequency of introgression of T. timopheevii genetic material into chromosome 7A was observed. Multivalents were scored in all cases of introgression in chromosome 7A. It was suggested that the reason for the genome instability in hybrid forms lied in insufficient compensating ability of certain T. timopheevii chromosomes and/or their parts, involved into recombination processes.     

Abscisic acid concentrations are correlated with leaf area reductions in two salt-stressed rapid-cycling Brassica species

The greater sensitivity of B. carinata to salinity in comparison to B. napus has been linked to a greater reduction in net assimilation rate. Apparently this is not due to ion toxicity; the cause is unknown. In this report, we test the hypothesis that increases in abscisic acid (ABA) are involved in the reduction of growth by salinity. Salinity (8 dS m^–1) caused an increase of ABA concentrations in the shoot, root and callus of both species. ABA concentrations were lower in the salt-tolerant species, B. napus, than the salt-sensitive species, B. carinata, both in the whole plant and callus. Leaf expansion for both species was equally sensitive to ABA; salt stress did not significantly alter sensitivity to applied ABA. The growth inhibition increased in a hyperbolic manner with an increase in endogenous ABA concentration. These results indicate that ABA in salt-stressed plants may play a role in the inhibition of growth. The photosynthesis of salt-sensitive species, B. carinata, was also decreased by salinity, corresponding to the reduction in growth. The decreased photosynthesis does not appear to be the cause of the growth reduction, because photosynthesis was not inhibited by short-term exposure to salinity and photosynthesis was poorly correlated with endogenous ABA concentrations.     

Effect of increased irradiance on the hormone content,water relations,and leaf elongation in wheat seedlings

In wheat (Triticum durum Desf., cv. Bezenchukskaya 139) seedlings, an increase in irradiance from 20 to 400 μmol/(m² s) PAR enhanced transpiration and increased stomatal conductance by three times on the background of reduced relative water content (RWC). After this treatment, leaves quickly ceased to grow and became even shrunk later. In 40 or 50 min, leaf growth was resumed. At this period, we observed an increase in hydraulic conductivity and RWC and also in leaf extensibility. As soon as 10 min after treatment, some changes in hormone content were noted. In the zones of leaf growth and its mature part, zeatin and zeatin riboside were accumulated, whereas ABA accumulation was observed in the zone of leaf growth and in the roots. The results obtained indicate that leaf expansion at increased irradiance was related to changes in cell-wall extensibility and hydraulic conductivity. The first effect could be due to cytokinin accumulation, whereas the second one, to ABA accumulation.     

The relationship between leaf water potential ψ leaf and the levels of abscisic acid and ethylene in excised wheat leaves

The amount of diffusible ethylene from excised wheat leaves (Triticum aestivum L. cv. Eclipse) increased when they were subjected to water stress. The quantity of ethylene produced was related to the severity of the stress, reaching a maximum at a leaf water potential _leaf of approximately-12 bars. Irrespective of the severity of the stress, the maximum rate of ethylene production usually occurred between 135–270 min after applying the stress and then the rate declined. Part of the decline may have been due to an oxygen deficiency in the leaf chambers. In excised water-stressed leaves there was a sigmoid relationship between increasing ethylene and abscisic acid (ABA) levels and decreasing leaf water potential values. The two curves were displaced from each other by approximately 1 bar, with ethylene evolution leading that of ABA accumulation. The maximum rate of increase in ethylene occurred between-8 and-9 bars and for ABA between-9 and-10 bars. A significant increase in the levels of these two plant growth regulators was found when the _leaf decreased outside the normal diurnal _leaf range by 1 bar for ethylene and 2 bars for ABA. Because of the sigmoid nature of the curves there was no distinct threshold _leaf value triggering-off an increase in ethylene or ABA, but with ABA the curve became very steep at a _leaf value of-9.3 bars and this could be looked upon as a kind of threshold value.It seems unlikely that the stress-induced ethylene evolution in excised wheat leaves stimulated the accumulation of ABA, because when the leaves were subjected to a substantial water stress (e.g. _leaf bars) ABA increased immediately and at a faster rate than ethylene.Abbreviations ABA abscisic acid - GLC gas-liquid chromatography - RWC relative water content - TLC thin-layer chromatography - _leaf leaf water potential     

The use of RAPD analysis to classify Triticum accessions

 Crop germplasm collections contain a considerable percentage of misclassified accessions which may affect the use of germplasm for agricultural crop improvement. The objective of this study was to determine if random amplified polymorphic DNA (RAPD) analysis could be used to reclassify misclassified Triticum accessions. Twelve accessions suspected to be misclassified, based on morphological characters, as either macha or vavilovii wheat were studied using RAPD and cytological analyses. In the RAPD analysis, a dendrogram, based on Jaccard genetic similarity coefficients, grouped 5 dicoccum-like, 1 timopheevii-like, and 6 monococcum-like accessions with Triticum dicoccum, T. timopheevii, and T. monococcum accessions, respectively. These results were confirmed by the cytological analysis. A RAPD marker specific to the D genome was also detected. This study suggests that RAPD analysis can be used to classify germplasm and to distinguish some species in Triticum. Received: 12 June 1998 / Accepted: 18 August 1998     

Molecular characterization of vernalization loci VRN1 in wild and cultivated wheats

Background  

Variability of the VRN1 promoter region of the unique collection of spring polyploid and wild diploid wheat species together with diploid goatgrasses (donor of B and D genomes of polyploid wheats) were investigated. Accessions of wild diploid (T. boeoticum, T. urartu) and tetraploid (T. araraticum, T. timopheevii) species were studied for the first time.     

GA,ABA, phenol interaction in the control of growth: Phenolic compounds as effective modulators of GA-ABA interaction in radish seedlings

Abscisic acid, a potent growth inhibitor inhibits hypocotyl growth ofRaphanus sativus seedlings. Phenolic compounds,viz., trans-cinnamic acid, chlorogenic acid, ferulic acid, salicylic acid, tannic acid and quercetin when applied with ABA, antagonize ABA action and restore normal seedling growth. Gibberellic acid promotes hypocotyl growth and on combined application with ABA, the ratio of their concentrations determines the course of the resultant growth. This interaction can be modulated by phenolic compounds. Phenolic compounds in low concentrations when present together with GA and ABA, favour GA-induced growth by antagonizing the inhibitory influence of ABA. The inhibitory action of abscisic acid on a wide range of growth processes is so far known to be reversed only by growth promoting hormones,viz., IAA, GA and cytokinins. Antagonistic action of phenolic compounds towards ABA, and increasing the action of GA when present together with GA and ABA, establishes a dual role to this class of compounds; balancing the effect of both growth promoting and growth inhibiting hormones. Part I.