Chromosomal control of the tolerance of gradually and suddenly imposed salt stress in the Lophopyrum elongatum and wheat,Triticum aestivum L. genomes

The facultatively halophytic Lophopyrum elongatum, closely related wheat, Triticum aestivum, and their amphiploid tolerate salt stress better if they are gradually exposed to it than if they are suddenly stressed. Lophopyrum elongatum has greater tolerance of both forms of salt stress than wheat, and its genome partially confers this tolerance on their amphiploid. Chromosomal control of the tolerance of both stress regimes in the L. elongatum and wheat genomes was investigated with disomic and ditelosomic addition lines and disomic substitution lines of L. elongatum chromosomes in wheat and with wheat tetrasomics. The tolerance of the sudden salt stress is principally controlled by L. elongatum chromosomes 3E and 5E and less by 1E, 2E, 6E, and 7E and the tolerance of gradually imposed salt stress principally by chromosomes 3E, 4E, and 5E, and less by chromosome 1E and 7E. Ditelosomic analysis indicated that genes conferring the tolerance of sudden stress are on chromosome arms 1EL, 5ES, 5EL, 6EL, 7ES and 7EL and those controlling the gradual stress regime are on 1ES, 1EL, 5ES, 5EL, 6ES, 7ES, and 7EL. In wheat, chromosomes in homoeologous groups 1, 3, and 7 and chromosomes in homoeologous groups 1, 4, and 6 were shown to enhance the tolerance of suddenly and gradually imposed stress, respectively. The arms of chromosome 3E individually conferred tolerance to neither stress regime. Chromosome 2E and wheat chromosomes 2B and 2D reduce the tolerance of both stress regimes in a hyperploid state. In 2E this effect was associated with arm 2EL. A potential relationship between the tolerance of these stress regimes and the expression of the early-salt induced genes is examined.     

Nucleolus organizer competition in Triticum aestivum — Aegilops umbellulata chromosome addition lines

Summary The nucleolar organizer activity of wheat (Triticum aestivum, AABBDD) and Aegilops umbellulata (UU) chromosomes have been analyzed in the complete set of the chromosome addition lines by using a highly reproducible silver-staining procedure. Chromosomes 1U and 5U produce the partial inactivation of wheat nucleolar organizer chromosomes 6B, 1B and 5D. The chromosomes D and G from Ae. umbellulata, which are not SAT-chromosomes, seem to specifically influence the activity of wheat NORs. The predominant status of the U genome with respect to nucleolar competition in the Triticeae is confirmed.     

Effects on Growth and Development of Individual Chromosomes from Slow-growing Lophopyrum elongatum Love when Incorporated into Bread Wheat (Triticum aestivum L.)

Aspects of growth and development were evaluated in the fast-developingannual Triticum aestivum L. ‘Chinese Spring’, theslow-developing perennial Lophopyrum elongatum Löve, theiramphiploid, and chromosome addition and substitution lines ofL. elongatum into ‘Chinese Spring’. Relative growthrates (RGR) of shoots of L. elongatum and the amphiploid werelower than those of ‘Chinese Spring’ (34 and 13%respectively) and main stem development was also slower. Therewas no difference in shoot RGR of any of the chromosome additionor substitution lines and that of ‘Chinese Spring’when assessed between Haun stages 2.0 and 5.0. In contrast,several aspects of plant development were observed to differin the chromosome addition and substitution lines. SubstitutingE genome chromosomes (with the exceptions of 3E and 4E) forD genome chromosomes, or adding E genome chromosomes, slowedthe rate of main stem development, at least up to Haun stage5.0. Despite these differences in the rate of main stem development,the appearance of adventitious roots commenced at approximatelyHaun stage 2.0 in all genotypes. However, the numbers of adventitiousroots and tillers at the 5.0 Haun stage differed between someof the lines when compared to ‘Chinese Spring’.Although incorporation of some L. elongatum chromosomes alteredaspects of plant development, all lines showed more similarityto bread wheat than to L. elongatum, reflecting, in part, thegreater genetic contribution made by bread wheat to these lines.Copyright 2001 Annals of Botany Company Adventitious roots, chromosome addition and substitution lines, Haun stage, Lophopyrum elongatum, relative growth rate (RGR), Triticum aestivum(wheat)     

C-banding polymorphism and the analysis of nucleolar activity in Dasypyrum villosum (L.) Candargy,its added chromosomes to hexaploid wheat and the amphiploid Triticum dicoccum — D. villosum

Summary C-banding patterns and nucleolar activity were analyzed in Dasypyrum villosum, its added chromosomes to hexaploid wheat and the hexaploid amphiploid Triticum dicoccum-D. villosum. Two different populations of the allogamous species D. villosum (2n= 14, VV) from Greece and Italy were analyzed showing a similar polymorphism for C-banding pattern. Six of the seven addition lines were identified by their characteristic C-banding pattern. No polymorphism between both members of each added alien chromosome was found. Furthermore, nucleolar activity and competition were studied by using silver staining procedure. In D. villosum only one chromosome pair, A, was found to be responsible for organizing nucleoli. The results obtained in the amphiploid and in the addition lines demonstrate that nucleolar activity is restricted to SAT-chromosomes 1B and 6B of wheat, while those of D. villosum remain inactive.     

<Emphasis Type="Italic">Agropyron elongatum</Emphasis> chromatin localization on the wheat chromosomes in an introgression line

The introgressed small-chromosome segment of Agropyron elongatum (Host.) Neviski (Thinopyrum ponticum Podp.) in F₅ line II-1-3 of somatic hybrid between common wheat (Triticum aestivum L.) and A. elongatum was localized by sequential fluorescence in situ hybridization (FISH), genomic in situ hybridization (GISH) and karyotype data. Karyotype analysis offered basic data of arm ratios and relative lengths of 21 pairs of chromosomes in parent wheat Jinan177 and hybrid II-1–3. Using special high repetitive sequences pSc119.2 and pAs1 for FISH, the entire B- and D-genome chromosomes were detected. The FISH pattern of hybrid II-1-3 was the same as that of parent wheat. GISH using whole genomic DNA from A. elongatum as probe determined the alien chromatin. Sequential GISH and FISH, in combination with some of the karyotype data, localized the small chromosome segments of A. elongatum on the specific sites of wheat chromosomes 2AL, 1BL, 5BS, 1DL, 2DL and 6DS. FISH with probe OPF-03₁₂₉₆ from randomly amplified polymorphic DNA (RAPD) detected E-genome chromatin of A. elongatum, which existed in all of the small chromosome segments introgressed. Microsatellite primers characteristic for the chromosome arms above were used to check the localization and reveal the genetic identity. These methods are complementary and provide comprehensive information about the genomic constitution of the hybrid. The relationship between hybrid traits and alien chromatin was discussed.     

C-banding and in-situ hybridization analyses of Agropyron intermedium,a partial wheat x Ag. intermedium amphiploid,and six derived chromosome addition lines

Summary C-banded karyotypes of Agropyron intermedium (2n=6x=42, E₁E₂X), a partial amphiploid Triticum aestivum —Ag. intermedium (2n=8x=56, TAF46), and six derived chromosome addition lines, were analyzed. In Ag. intermedium, diagnostic C-bands were present on 14 pairs of chromosomes, designated from A to N, while the remaining seven pairs, designated O to U, either lacked, or had only faint, C-bands and were not always identified unambiguously. All seven Ag. intermedium chromosome pairs of the partial amphiploid TAF46, and the added Ag. intermedium chromosomes present in the six derived addition lines, were identified by their characteristic C-banding patterns. Chromosome morphology and banding patterns were similar to those of the corresponding chromosomes present in the parent Ag. intermedium accession, suggesting that these chromosomes were not structurally rearranged. In-situ hybridization, using a 18s.265s rDNA probe, showed that the Ag. intermedium chromosomes 1Ai-1 and 5Ai-l present in the addition lines L3 and L5 were carrying actively transcribed nucleolus organizer regions. The results are discussed with respect to the genomic relationships of these chromosomes.Contribution no. 91-561-J from the Wheat Genetics Resource Center and Kansas Agricultural Experiment Station, Kansas State University, Manhatten, USA     

Modification of the salinity response of wheat by the genome ofElytrigia elongatum

Summary Triticum aestivum cv. Chinese Spring wheat,Elytrigia elongatum (tall wheatgrass), and theTriticum-Elytrigia amphiploid were grown in complete nutrient culture containing, in addition, 0, 40, 80 and 120 mM NaCl. The 3 genotypes responded quite differently to increasing salinity; the Na concentration of wheat shoots increased in direct proportion to the increase in salinity of the external medium whereas the Elytrigia response was interpreted as showing high affinity for Na at low external Na (40 mM) but comparative exclusion of Na at high salinities (120 mM). In contrast, Na levels of the amphiploid were less than those of either wheat or Elytrigia under both low and high salinities. Thus the amphiploid behaved like wheat at 40 mM NaCl but more like Elytrigia at 120 mM NaCl because Na transport to the amphiploid shoot was restricted over the whole salinity range. The K concentration of the amphiploid shoot at high salinities was significantly greater than the K concentrations of either wheat or Elytrigia.     

Combined effect of salinity and hypoxia in wheat (Triticum aestivum L.) and wheat-Thinopyrum amphiploids

The effects of sodium chloride salinity and hypoxia were studied in eight wheat lines and three wheat-Thinopyrum amphiploids in vermiculite-gravel culture. The lines were treated with either 100 or 150 mol m^–3 NaCl with and without hypoxia. Saline hypoxic conditions significantly reduced the vegetative growth, water use, grain and straw yields for all wheat varieties except the amphiploids, whereas NaCl or hypoxia alone had less pronounced effects. In addition, saline hypoxic stress reduced K⁺ concentration and increased significantly the Na⁺ and Cl^– concentrations in cell sap expressed from leaves. There was more Na⁺ and Cl^– accumulation in wheats than the amphiploids in hypoxic conditions at 150 mol m^–3 NaCl. Of the wheats, Pato was the most sensitive at all stress levels while aTriticum aestivum cv. Chinese Spring ×Thinopyrum elongatum amphiploid was the most tolerant of the three amphiploids.     

C-banding pattern and polymorphism of Aegilops caudata and chromosomal constitutions of the amphiploid T. aestivum — Ae. caudata and six derived chromosome addition lines

Summary C-banding patterns were analysed in 19 different accessions of Aegilops caudata (= Ae. markgrafii, = Triticum dichasians) (2n = 14, genomically CC) from Turkey, Greece and the USSR, and a generalized C-banded karyotype was established. Chromosome specific C-bands are present in all C-genome chromosomes, allowing the identification of each of the seven chromosome pairs. While only minor variations in the C-banding pattern was observed within the accessions, a large amount of polymorphic variation was found between different accessions. C-banding analysis was carried out to identify Ae. caudata chromosomes in the amphiploid Triticum aestivum cv Alcedo — Ae. caudata and in six derived chromosome addition lines. The results show that the amphiploid carries the complete Ae. Caudate chromosome complement and that the addition lines I, II, III, IV, V and VIII carry the Ae. caudata chromosome pairs B, C, D, F, E and G, respectively. One of the two SAT chromosome pairs (A) is missing from the set. C-banding patterns of the added Ae. caudata chromosomes are identical to those present in the ancestor species, indicating that these chromosomes are not structurally rearranged. The results are discussed with respect to the homoeologous relationships of the Ae. caudata chromosomes.     

Chromosomal location of genes controlling seed proteins in species related to wheat

Summary The seed proteins of Chinese Spring wheat stocks which possess single chromosomes from other plant species related to wheat have been separated by gel electrophoresis in the presence of sodium dodecyl sulphate. Marker protein bands have been detected for both arms of barley chromosome 5, chromosome E (= 1R) and B (= 2R) of rye, chromosomes A,B (= 1C^u) and C (= 5C^u) of Aegilops umbellulata and chromosomes I and III of Agropyron elongatum. These studies, and previous findings, indicate that chromosome 5 of barley, chromosome 1R of rye, chromosome I of Ag. elongatum and possibly chromosome 1C^u of Ae. umbellulata are similar to chromosomes 1A, 1B and 1D in hexaploid wheat in that they carry genes controlling prolamins on their short arms and genes controlling high-molecular-weight (apparent molecular weight greater than 86,000) seed protein species on their long arms. These findings support the idea that all these chromosomes are derived from a common ancestral chromosome and that they have maintained their integrity since their derivation from that ancestral chromosome.     

Nucleolar organizer activity in wheat,rye and derivatives analyzed by a silver-staining procedure

Nucleolar activity was analyzed in wheat (Triticum sp.), rye (Secale cereale) and several types of wheat-rye derivatives using a modified, highly reproducible, silver staining procedure (Lacadena et al. 1984). A comparative analysis of the nucleolar organizer regions (NORs) of somatic metaphase chromosomes was made by phase contrast, C-banding, and silver staining. The frequency distribution of the number of nucleoli visualized at interphase by silver staining was also used to infer the activity of NORs. The results agree quite well with data from in situ hybridization reported by other authors. The behavior of euploid, ditelosomic and nulli-tetrasomic plants of common wheat showed the relative nucleolar activity of the four organizer chromosomes to be: 6B > 1B > 5D > 1A. — Several types of wheat-rye derivatives were analyzed: interspecific hybrid, triticale, addition and substitution lines, and plants with the genome constitutions, AABBDR, ABDR + 5D, ABRR, and ABRRR. In all cases the nucleolar organizer chromosome 1R of rye was suppressed by the presence of wheat chromosomes.     

Wheat Neocentromeres Found in F1 Triticale × Tritordeum Hybrids (AABBRH<Superscript>ch</Superscript>) After 5-Azacytidine Treatment

The DNA hypomethylation effect of 5-azacytine (5-AC; a cytosine analog) is widely known. This agent has been used for rRNA gene expression studies of Triticeae amphiploids and hybrids regarding rye rRNA genes suppression caused by the wheat nucleolar dominance phenomenon. However, this situation is reverted by 5-AC treatment which activates rye rRNA gene expression as it has been intensively observed in triticale. For nucleolar dominance studies, we produced F1 multigeneric hybrids (AABBRH^ch; 2n = 6x = 42) from crosses between the triticale cultivar ‘Corgo’ (AABBRR; 2n = 6x = 42) and the tritordeum cultivars HT9 and HT31 (AABBH^chH^ch; 2n = 6x = 42). The hybrid seeds were germinated in a low concentration of 5-AC (treatment) and in distilled water (nontreated control plants). Silver nitrate staining performed in one 5-AC-treated F1 hybrid revealed a reduced number of interphase cells with seven nucleoli, metaphases with eight Ag-NORs, and neocentromeres in the long arm of three wheat chromosomes. Nontreated hybrids presented six Ag-NORs per mitotic metaphase cell and a maximum of six nucleoli per interphase because of the 1R Ag-NOR suppression. No neocentromere was found in the control F1 hybrid plants. Both treated and nontreated seedlings were subsequently evaluated by fluorescent in situ hybridization performed with genomic and repetitive DNA probes to identify H^ch and rye genomes, to confirm Ag-NORs location, and to detect inactive rDNA loci. DAPI counterstaining was also helpful for the detection of neocentromeres in the long arm of three wheat chromosomes. This study allowed us to suggest that 5-AC treatment specifically induced wheat neocentromeres in the F1 multigeneric triticale × tritordeum hybrids.     

Meiotic pairing of the amphiploid Hordeum chilense X Triticum turgidum conv. durum studied by means of Giemsa C-banding technique

Summary The meiotic behaviour of the amphiploid Hordeum chilense X Triticum turgidum conv. durum using a C-banding staining method is studied. Nine pairs of chromosomes at metaphase-1 (4A, 7A and the seven of the B genome) were identified and the remaining wheat chromosomes (1A, 2A, 3A, 5A and 6A) and seven of the chilense (1 to 7 H ^ch chromosomes) were assigned to its particular genome. A similar mean number of univalents from parental genomes (wheat and wild barley) were found. No meiotic pairing between chilense and turgidum chromosomes was detected. Differences in the meiotic behaviour per chromosome and amongst genomes are explained on the basis of cytomorphological and heterochromatin characteristics.     

Cytologically based physical maps of the group 3 chromosomes of wheat

Cytologically based physical maps for the group 3 chromosomes of wheat were constructed by mapping 25 Triticum aestivum deletion lines with 29 T. tauschii and T. aestivum RFLP probes. The deletion lines divide chromosomes 3A, 3B, and 3D into 31 discrete intervals, of which 18 were tagged by marker loci. The comparison of the consensus physical map with a consensus RFLP linkage map of the group 3 chromosomes of wheat revealed a fairly even distribution of marker loci on the long arm, and higher recombination in the distal region.     

Meiosis Aspects and Nucleolar Activity in Triatoma vitticeps (Triatominae, Heteroptera)

Some aspects of both the nucleolar organizer activity and meiosis were studied in the testes of Triatoma vitticeps (Heteroptera, Triatominae). The techniques used included squashing followed by lacto-acetic orcein staining, silver-ion impregnation, fluorescent banding (CMA₃, Quinacrine mustard and DAPI) and fluorescent in situ hybridization (FISH). A close relationship between heterochromatin and nucleolus in testicular cells was observed. During meiosis, the silver-ion impregnation pattern varied. At metaphase plate, a small body appeared apart from the chromosomes. In the spermatids this small body was seen in preparations stained with orcein and silver- ion impregnation but not with fluorochromes or FISH. These characteristics combined suggest that these corpuscles represent a source of ribonucleoproteins (RNP) – RNA and specific nucleolar proteins. Silver-ion impregnation and (FISH) revealed nucleolar organizer activity in two metaphase sex chromosomes (X). These results indicate that, in these species, nucleolar organizer regions (NORs) are located in the sex chromosomes, X chromosomes were CMA₃⁺ and Y chromosome was DAPI⁺.     

Fusarium head blight resistance in hexaploid wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis>)-<Emphasis Type="Italic">Lophopyrum</Emphasis> genetic lines and tagging of the alien chromatin by PCR markers

The objective of this research was to identify Fusarium head blight (FHB) resistance in wheat (Triticum aestivum)-Lophopyrum genetic lines that might complement FHB resistance in common wheat; and to identify DNA markers that can be used to tag the resistance gene in the alien chromatin (E or el₂ genome) for the development of improved wheat cultivars. FHB resistance was evaluated in 19 Chinese Spring-Lophopyrum elongatum (EE) substitution lines, two Thatcher-L. ponticum (el₁ and el₂) substitution lines, and four Thatcher-L. ponticum translocation lines. Significant resistance was identified in the substitution lines 7E(7A), 7E(7B), and 7E(7D). The homoeologous chromosome, 7el₂,also showed resistance in the Thatcher genetic background. Both the Thatcher-7el₁ substitution and translocation lines were susceptible, like Thatcher, indicating that there is no resistance gene on the 7el₁ chromosome. Simple sequence repeat (SSR) and cleaved amplified polymorphic sequences (CAPS) in homoeologous group 7 chromosomes were used to identify DNA markers located on 7E and 7el₂. As expected, the transferability of wheat SSR markers to Lophopyrum is low. Of the 52 SSR markers that we tested, only five were found to be co-dominant on 7E of L. elongatum versus 7A, 7B, and 7D, one of which is also positive on 7el₂. A CAPS marker, derived from the RFLP probe PSR129, can serve as a dominant marker for 7el₂ chromatin.Communicated by J. Dvorak     

Tolerance to high concentrations of boron for the amphiploid of Triticum aestivum × Agropyron elongatum

The response to high concentrations of B in soil was compared for Triticum aestivum L. (vars. Chinese Spring and Halberd) and the amphiploid of Chinese Spring × Agropyron elongatum (Host). The tolerance of the three genotypes, with respect to yield at the high B treatments, was amphipolid>Halberd>Chinese Spring. The concentration of B in whole shoots was similar for the amphiploid and Halberd and significantly higher for Chinese Spring. The mechanism for tolerance to B appears to be the same for the amphiploid and wheat and is related to reduced accumulation of B in shoots.     

Standard karyotype of Triticum umbellulatum and the characterization of derived chromosome addition and translocation lines in common wheat

A standard karyotype and a generalized idiogram of Triticum umbellulatum (syn. Aegilops umbellulata, 2n = 2x = 14) was established based on C-banding analysis of ten accessions of different geographic origin and individual T. umbellulatum chromosomes in T. aestivum — T. umbellulatum chromosome addition lines. Monosomic (MA) and disomic (DA) T. aestivum — T. umbellulatum chromosome addition lines (DA1U = B, DA2U = D, MA4U = F, DA5U = C, DA6U = A, DA7U = E = G) and telosomic addition lines (DA1US, DA1UL, DA2US, DA2UL, DA4UL, MA5US, (+ iso 5US), DA5UL, DA7US, DA7UL) were analyzed. Line H was established as a disomic addition line for the translocated wheat — T. umbellulatum chromosome T2DS·4US. Radiation-induced wheat — T. umbellulatum translocation lines resistant to leaf rust (Lr9) were identified as T40 = T6BL·6BS-6UL, T41 = T4BL·4BS-6UL, T44 = T2DS·2DL-6UL, T47 = Transfer = T6BS·6BL-6UL and T52 = T7BL·7BS-6UL. Breakpoints and sizes of the transferred T. umbellulatum segments in these translocations were determined by in situ hybridization analysis using total genomic T. umbellulatum DNA as a probeContribution no. 94-349-J from the Kansas Agricultural Experiment Station, Kansas State University, Manhattan, KS 66506-5502, USA     

Comparison of the genetic organization of the early salt-stress-response gene system in salt-tolerant Lophopyrum elongatum and salt-sensitive wheat

Lophopyrum elongatum is a facultative halophyte related to wheat. Eleven unique clones corresponding to genes showing enhanced mRNA accumulation in the early stages of salt stress were previously isolated from a L. elongatum salt-stressed-root cDNA library. The chromosomal distribution of genes complementary to these clones in several genomes of the tribe Triticeae and their copy number in the L. elongatum and wheat genomes are reported. Genes complementary to clones pESI4, pESI14, pESI15, pESI28, and pESI32 were found in homoeologous group 5, those complementary to pESI18 and pESI35 in homoeologous group 6, and those complementary to pESI47, pESI48, pESI3, and pESI2 in homoeologous groups 1, 3, 4, and 7, respectively. The genes are present in a single copy per genome in L. elongatum with the exception of those complementary to pESI2 and pESI18 which are present in at least two and five copies, respectively. Since similar copy numbers per genome were found in wheat (except for pESI2), the ability of L. elongatum to accumulate higher mRNA levels than wheat in response to salt shock apears to have evolved by changes in the regulation of these genes.