Effects of loci on chromosomes 2 (2H) and 7 (5H) on developmental patterns in barley (Hordeum vulgare L.) under different photoperiod regimes

 Heading-date in cereals is the final result of a number of interacting characters that include vernalization requirement, photoperiod sensitivity, and earliness per se. Progress in developing adapted varieties may be achieved by determining the chromosomal locations of genes controlling these characters. Nineteen doubled-haploid (DH) lines from the Dicktoo×Morex mapping population were phenotyped in controlled- environment photoperiod experiments to determine the role of two previously detected QTLs on the developmental patterns of barley. The QTLs are hypothesised to represent the effects of the Ppd and Sh2 loci on chromosomes 2 (2H) and 7 (5H), respectively. Alleles at the Ppd locus were found to be vary in response to photoperiod duration. Vernalization had some effect on alleles at both loci. The presence of early and late- flowering transgressive segregants in this mapping population can be explained by interactions between the Ppd and Sh2 loci. The Ppd and Sh2 loci are hypothesised to be homoeologous with the Ppd and Vrn1 loci of wheat. Received: 1 August 1996 / Accepted: 15 November 1996     

Acrotrisomic analysis in linkage mapping in barley (Hordeum vulgare L.)

Summary Three acrotrisomic lines, Triplo IL^1S, 3L^3S, and 4L^4S, each carrying an extra acrocentric chromosome, were used for cytogenetic linkage mapping of barley chromosomes. The cytological structures of the acrocentric chromosome of the three acrotrisomic lines were studied with an improved Giemsa N-banding technique. The long (1L) and short arm (1S) of chromosome 1 had deficiencies of approximately 38% and 65%, respectively. The percentages of deficiencies were 0 and 77.8% for 3L and 3S, and 31.7 and 59.3% for 4L and 4S, respectively. All three genes tested (br, f _c , gs3) in 1S and all three genes tested, f8, n and 1k2 in 1L showed a disomic ratio indicating that they are located in the deficient segments. Two genes (a _c , yst2) located in the middle segment of 3S in linkage map showed a trisomic ratio, and two others a _n , x _s showed a disomic ratio. The only gene(f9) tested in 4L showed a trisomic ratio. Two genes (1g4, g1) located in the proximal segment of 4S in the linkage map showed a trisomic ratio, whereas two genes (br2, g13) located distally in 4S showed a disomic ratio, indicating that the breakage occurred between g1 and br2. This experiment demonstrates a new method for physical localization of genes on chromosome segments in material such as barley in which pachytene analysis can not be effectively used for accurate determination of break points in structural changes. Problems associated with this new technique are discussed.Contribution from the Department of Agronomy and published with the approval of the Director of Colorado State University Experiment Station as Scientific Series Paper No. 2823. Supported by USDA/SEA Competitive Research Grant Nos. 5901-0410-9-0334-0 and 82-CRCR-1-1020 and USDA-CSU Cooperative Research Grant 58-9AHZ-2-265     

Detection of quantitative trait loci for agronomic,yield, grain and disease characters in spring barley (Hordeum vulgare L.)

Quantitative trait loci (QTLs) have been revealed for characters in a segregating population from a spring barley cross between genotypes adapted to North-West Europe. Transgressive segregation was found for all the characters, which was confirmed by the regular detection of positive and negative QTLs from both parents. A QTL for all the agronomic, yield and grain characters measured except thousand grain weight was found in the region of the denso dwarfing gene locus. There were considerable differences between the location of QTLs found in the present study and those found in previous studies of North American germ plasm, revealing the diversity between the two gene pools. Thirty-one QTLs were detected in more than one environment for the 13 characters studied, although many more were detected in just one environment. Whilst biometrical analyses suggested the presence of epistasis in the genetic control of some characters, there was little evidence of interactions between the QTLs apart from those associated with yield. QTLs of large effect sometimes masked the presence of QTLs of smaller effect.     

Microarray analysis of gene expression in germinating barley embryos (Hordeum vulgare L.)

A cDNA library containing approximately 5,000 clones from germinating barley embryos was constructed and used to examine the variation in gene expression patterns during the first 4 days postimbibition. The expression profiles of embryos (including scutellum) from 4 to 96 h postimbibition were compared to a reference profile from 24 h postimbibition using microarray analysis. A subset of clones exhibiting tenfold or greater differential expression patterns was sequenced to elucidate function. All of the sequenced clones could be identified to at least EST level with 64% exhibiting homology to published protein sequences. Almost 95% of the library exhibited similar expression levels at the 4 h time point as at the 24 h reference point. From 24 to 96 h, however, considerable fluctuations in gene expression occurred. The observed patterns of gene expression for the classified genes are consistent with the expected genetic changes required to prepare an embryo for germinative development. A replicate set of clones for the 23-kDa jasmonate-induced protein was identified. The current data not only provides conclusive evidence for the expression patterns of this abundant stress-response protein in germinating embryos, but also serves to validate previous research into JIP-23 isoforms, function and the relationship between timing of mRNA upregulation and protein abundance.     

Quantitative trait loci on barley (Hordeum vulgare L.) chromosome 7 associated with components of winterhardiness.

Quantitative trait loci (QTL) controlling traits associated with winterhardiness in barley (field survival, LT50, growth habit, and crown fructan content) were mapped to chromosome 7 in a population of 100 F1-derived doubled haploid lines. The largest QTL effects for all traits were detected in a 21% recombination interval on the long arm of chromosome 7. QTL in this region accounted for 37-68% of the variation for three measures of cold tolerance, 47% of the variation for growth habit, and 28% of the variation in crown fructan content. Trait association may be due to linkage rather than pleiotropy.     

Triple hybridization with cultivated barley (Hordeum vulgare L.)

Summary A crossing programme for trispecific hybridization including cultivated barley (Hordeum vulgare L.) as the third parent was carried out. The primary hybrids comprised 11 interspecific combinations, each of which had either H. jubatum or H. lechleri as one of the parents. The second parent represented species closely or distantly related to H. jubatum and H. lechleri. In trispecific crosses with diploid barley, the seed set was 5.7%. Crosses with tetraploid barley were highly unsuccessful (0.2% seed set). Three lines of diploid barley were used in the crosses, i.e. Gull, Golden Promise and Vada. Generally, cv Gull had high crossability in crosses with related species in the primary hybrid. It is suggested that Gull has a genetic factor for crossability not present in cv Vada and cv Golden Promise. One accession of H. brachyantherum used in the primary hybrid had a very high crossability (seed set 54.7%) in combination with cv Vada but no viable offspring was produced. In all, two trispecific hybrids were raised, viz. (H. lechleri x H. brevisubulatum) x Gull (2n=7–30) and (H. jubatum x H. lechleri) x Gull (2n=20–22). The first combination invariably had a full complement of seven barley chromosomes plus an additional chromosome no. 7, but a varying number of chromosomes (19–22) of the wild-species hybrid. The second combination had a full set of barley chromosomes. The meiotic pairing was low in both combinations.     

Studies of soil moisture stress in barley (Hordeum vulgare L.)

Summary The objective was to find the optimum range of water contents for inducing better growth, physiological efficiency and yield potential of barley plants (Hordeum vulgare L. var. K18). A pot culture experiment was conducted in the Division of Crop Physiology and Biochemistry Kanpur-2. The plants were subjected to various soil moisture stresses,i.e., 0.15, 0.30, 0.45, 0.60 and 0.75 atm tension throughout the crop growth period measured by irrometers.Plants maintained at 0.45 soil moisture tension required 19.07 litre of water and had the best water use efficiency (1765 mg dm/litre of water) which favourably influenced the leaf water balance (85.9%), plant growth as measured by plant height (85.4 cm) and tiller production (35.6) per hill, photosynthetic efficiency (2.185 mg CO₂/g dm/h), grain number (722) and grain yield (33.7 g) per hill while plants irrigated at a tension greater than 0.45 SMT did not develop as well. However, protein and gluten percentage increased gradually with the subsequent increase in soil moisture tension. On the other hand respiration rate (2.090 mg CO₂/g dm/hr) and leaf area (4375 cm²) were recorded to be the highest at 0.60 and 0.30 atm SMT respectively.Thus it is suggested that for reaping high harvest of barley crop, the physiological need of water (19.07 litre) in total of plant life should be made available through scheduled irrigation based on maintenance of plant at 0.45 SMT from seeding to maturity.     

QTL analysis of tolerance to a German strain of BYDV-PAV in barley (Hordeum vulgare L.)

One hundred and forty six barley doubled-haploid lines (DH lines) were tested for variation in grain yield, yield components, plant height, and heading date after artificial infection with a German isolate of barley yellow dwarf virus (BYDV-PAV-Braunschweig). Of these 146 lines 76 were derived from the cross of the barley yellow dwarf virus (BYDV) tolerant cultivar ’Post’ to cv ’Vixen’ (Ryd2) and 70 from the cross of Post to cv ’Nixe’. Phenotypic measurements were gathered on both non-infected plants and plants artificially inoculated with BYDV-PAV by viruliferous aphids in pot and field experiments for three years at two locations. For all traits a continuous variation was observed suggesting a quantitative mode of inheritance for tolerance against BYDV-PAV. Using skeleton maps constructed using SSRs, AFLPs and RAPDs, two QTLs for relative grain yield per plant after BYDV infection, explaining about 47% of the phenotypic variance, were identified in Post × Vixen at the telomeric region of chromosome 2HL and at a region containing the Ryd2 gene on chromosome 3HL. In Post × Nixe, a QTL was found in exactly the same chromosome 2HL marker interval. In this cross, additional QTL were mapped on chromosomes 7H and 4H and together these explained about 40% of the phenotypic variance. QTL for effects of BYDV infection on yield components, plant height, and heading date generally mapped to the same marker intervals, or in the vicinity of the QTL for relative grain yield, on chromosomes 2HL and 3HL, suggesting that these regions are of special importance for tolerance to the Braunschweig isolate of BYDV-PAV. Possible applications of marker-assisted selection for BYDV tolerance based on these results are discussed. Received: 1 December 2000 / Accepted: 9 March 2001     

Phenotypic variation in barley (Hordeum vulgare L.) landraces from north Shewa in Ethiopia

An experiment was conducted in a greenhouse to estimate the level of morphological variability within 44 barley landraces (14 farmers' cultivars and 30 accessions) from north Shewa in Ethiopia. Four qualitative traits (spike type, kernel color, caryopsis type and spike density) were recorded. Variation in spike type (two-rowed, irregular or six-rowed) was high in many landraces. Variability in spike type within landraces from Debre Libanose, Kuyu and Girar Jarso was high. The highest mean frequencies of the six-row type were recorded for Ankober-Mezezo (85%) and Kimbibit (80.9%). The two-row types were dominant in Wuchale (53.3%), while they were absent at the Ankober-Mezezo, Debre Libanose and Kimbibit localities. A larger proportion of the irregular types occurred in Debre Libanose (71.3%), followed by Kuyu (62.6%). Diversity for kernel color was generally low except in landraces from Kuyu, but it was very high among landraces. White kernel color was predominant. The mean diversity index pooled over characters ranged from 0.12 ± 0.08 to 0.57 ± 0.11, and 11 landraces had a mean diversity larger than 0.50. Differences among landraces within localities were mostly highly significant for all characters except that of caryopsis type (covered or naked). Landraces from Girar Jarso, Wuchale, and Kuyu in particular differed significantly in spike type and kernel color. Unlike farmers' cultivars, the accessions were found to be more variable in spike type and kernel color. Moreover, the mean diversity indices pooled over characters were relatively higher in the accessions than within farmers' cultivars.     

Protoplast preparation without centrifugation: plant regeneration of barley (Hordeum vulgare L.)

Summary Using a modification of the alginate film culture technique we show that it is possible to prepare and culture tobacco mesophyll and barley cell suspension protoplasts without centrifugation. Comparable division frequencies and colony development were observed from protoplasts embedded with enzyme and protoplasts purified by centrifugation. A 3 × 30 min washing regime was found to be the minimum time necessary to remove the enzyme from the gelled alginate matrix. The procedure provides a more gentle method for isolating protoplasts. It has the additional benefit of recovering all of the cells released from the starting tissue. In particular, the smaller protoplasts that are frequently lost during conventional isolation, are maintained. In barley, we illustrate the use of the system for recovering plants from embryogenic protoplast-derived calli from the cultivars Dissa and Igri. Finally, using small volumes of enzyme (50 l) single cell aggregates were used to isolate and culture protoplasts.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - BAP 6-benzyl aminopurine - MES 2-(N-morpholino)ethanesulfonic acid - NAA -naphthaleneacetic acid     

Strong extracellular nuclease activity displayed by barley (Hordeum vulgare L.) uninucleate microspores

 Electroporation is becoming an increasingly important technique for plant transformation. Nevertheless, no positive results were achieved in barley when uninucleate microspores were used as target cells. Since it was previously demonstrated that electric shocks create pores in the microspore cell wall, experiments were designed to verify the presence of nucleases in the electroporation mix. Aliquots of all the solutions used for microspore extraction, purification and transformation were collected and analysed using supercoiled pBI 221 as a substrate; a nuclease activity was detected in all samples. Though microspore rinsing removed most nucleolytic activity in the supernatants, DNA preservation in the electroporation buffer was difficult to achieve, because microspores appeared capable of synthesising and releasing endonucleases at any time. Microspore chilling at 0°C was fairly effective in reducing nuclease secretion in the mix, whereas 1%PEG or 10 mM EDTA maintained most of the DNA in a supercoiled or circular relaxed form. EDTA effects were counterbalanced by Mg²⁺, but not Ca²⁺ or Zn²⁺, and enhanced by Mn²⁺. Barley microspore nucleases actively degraded different DNAs as well as TMV RNA, and apparently had a molecular weight above 30 kDa. Nuclease inactivation with EDTA did not alter microspore viability and allowed a transient expression of the uidA gene in electroporated barley microspores. Received: 13 January 1997 / Accepted: 28 February 1997     

Advanced backcross QTL analysis in barley (Hordeum vulgare L.)

This paper reports on the first advanced backcross-QTL (quantitative trait locus) project which utilizes spring barley as a model. A BC(2)F(2) population was derived from the initial cross Apex ( Hordeum vulgare ssp. vulgare, hereafter abbreviated with Hv) x ISR101-23 ( H. v. ssp. spontaneum, hereafter abbreviated with Hsp). Altogether 136 BC(2)F(2) individuals were genotyped with 45 SSR (simple sequence repeat) markers. Subsequently, field data for 136 BC(2)F(2) families were collected for 13 quantitative traits measured in a maximum of six environments. QTLs were detected by means of a two-factorial ANOVA with a significance level of P < 0.01 for a marker main effect and a marker x environment (M x E) interaction, respectively. Among 585 marker x trait combinations tested, 86 putative QTLs were identified. At 64 putative QTLs, the marker main effect and at 27 putative QTLs, the M x E interaction were significant. In five cases, both effects were significant. Among the putative QTLs, 29 (34%) favorable effects were identified from the exotic parent. At these marker loci the homozygous Hsp genotype was associated with an improvement of the trait compared to the homozygous Hv genotype. In one case, the Hsp allele was associated with a yield increase of 7.7% averaged across the six environments tested. A yield QTL in the same chromosomal region was already reported in earlier barley QTL studies.     

Several QTLs involved in osmotic-adjustment trait variation in barley (Hordeum vulgare L.)

 Osmotic adjustment (OA) was previously demonstrated to be an important adaptive mechanism of drought tolerance in cereals. In order to determine which genomic regions are involved in OA variation, 187 barley (Hordeum vulgare L.) recombinant inbred lines (RILs) derived from a cross between Tadmor (drought tolerant) and Er/Apm (susceptible) were studied in a growth chamber for their OA capacity (through correlated traits and by calculation), at an early growth stage and under two water treatments (soil moisture of 14% and 100% of field capacity). The continuous distribution of the traits and their broad-sense line heritabilities, ranging from 0.04 to 0.44, indicated that OA and related traits should have a polygenic nature. A subset of 167 RILs were also genotyped using 78 RFLP, 32 RAPD and three morphological markers and a linkage map was constructed. Despite strong environmental effects acting on the traits, interval mapping and single-marker ANOVA allowed the detection of three QTLs for relative water content (RWC), four QTLs for osmotic potential (ψ_π), two QTLs of osmotic potential at full turgor (ψ_π100) and one QTL for osmotic adjustment at a soil moisture of 14% field capacity. For the irrigated treatment, only two QTLs were detected: one for RWC and one for ψ_π100. Two chromosomal regions were involved in several OA-related trait variations and could be considered as regions controlling OA; these were present on chromosome 1 (7H) and chromosome 6 (6H), whereas other regions were specific for one trait. No major QTL was found. However, the genomic region involved in OA-related traits on chromosome 1 (7H) in barley seemed to be conserved for OA variation among cereals. Epistatic effects, with or without additive effects, acted on the traits. Received: 15 July 1997 / Accepted: 29 October 1997     

Mapping of chromosome regions conferring boron toxicity tolerance in barley (Hordeum vulgare L.)

 Boron toxicity has been recognised as an important problem limiting production in the low-rainfall regions of southern Australia, West Asia and North Africa. Genetic variation for boron toxicity tolerance in barley has been characterised but the mode of inheritance and the location of genes controlling tolerance were not previously known. A population of 150 doubled-haploid lines from a cross between a boron toxicity tolerant Algerian landrace, Sahara 3771, and the intolerant Australian cultivar Clipper was screened in four tolerance assays. An RFLP linkage map of the Clipper×Sahara population was used to identify chromosomal regions associated with boron tolerance in barley. Interval regression-mapping allowed the detection of four chromosomal regions involved in the boron tolerance traits measured. A region on chromosome 2H was associated with leaf-symptom expression, a region on chromosome 3H was associated with a reduction of the affect of boron toxicity on root growth suppression, a region on chromosome 6H was associated with reduced boron uptake, and a region on chromosome 4H was also associated with the control of boron uptake as well as being associated with root-length response, dry matter production and symptom expression. The benefits and potential of marker-assisted selection for boron toxicity tolerance are discussed. Received: 18 December 1997 / Accepted: 28 November 1998     

Ring chromosomes in barley (Hordeum vulgare L.)

A plant with 2n = 14 + 1 ring chromosomes was obtained in the progeny of a primary trisomie for chromosome 7 of a two-rowed cultivar, Shin Ebisu 16. The morphological characteristics of the trisomic plants with an extra ring chromosome were similar to the primary trisomic for chromosome 7 (Semierect), which suggests that it originated from this chromosome. The ring chromosomes were not completely stable in mitotic cells because of abnormal behavior. Chromosome complements varied in different plants and in different roots within a plant. Root tip cells and spikes with 2n = 14 and 14 + 2 ring chromosomes were observed on plants with 14 + 1 ring chromosomes. Breakage-fusion-bridge cycle was inferred. The ring chromosome was associated with two normal homologues forming a trivalent in 17.6% sporocytes at metaphase I. The transmission of the extra ring chromosome was 23.1% in the progeny of the plant with 14 + 1 ring chromosomes. Trivalent formation may have been much higher at early prophase stages which were difficult to analyze in barley; only 4 of 120 sporocytes analyzed showed an isolated ring at pachytene. The ring chromosome moved to one pole without separation in 24.7% of the sporocytes at AI, and divided in 27.1% sporocytes giving rise to 8-8 separation. Only 10% of the sporocytes showed bridge formation at AI.     

Nuclear genes affecting percentage of green plants in barley (Hordeum vulgare L.) anther culture

Summary The genetics behind response in barley anther culture was studied with 22 reciprocal and one single: cross between three varieties with high and four varieties with low capacity for green plant formation. Effects of genotypes dominated embryo formation and percentages of green plants, accounting for 62 and 76% of total variation, respectively, with almost no genetic effect on the ability to regenerate plants from pollen embryos. Nuclear genes could explain all genotype effects in this plant material, since no reciprocal effects were indicated. The three parents with high and the four parents with low capacity for green plant formation formed two phenotypically homogeneous groups, producing 27–52% and 0–7% green plants, respectively. Genetic variation within hybrids for both embryo and green plant formation could be explained completely by general combining ability (GCA). The results are discussed with respect to a previous similar study in hexaploid wheat and the reported existence of DNA deletions in the plastid genomes in albino plants from anther culture of wheat and barley.     

Cytogenetics of ditelotetrasomics for short arms of four chromosomes of barley (Hordeum vulgare L.)

Summary Plants with a pair of extra homologous telocentric chromosomes in addition to the normal chromosome complement are called ditelotetrasomics. Six types of ditelotetrasomics of barley have been obtained. Four types obtained in the selfed progenies of telotrisomics (Triplo 2S, Triplo 5S, Triplo 6S, and Triplo 7S) are reported in this paper. The ditelotetra 2S showed a stronger expression of the diagnostic characteristics of Triplo 2S. It was weak and small, with narrow, short, dark-green leaves, and was almost completely pollen and seed sterile. However, three other ditelotetrasomics (ditelotetra 5S, 6S, and 7S) did not show specific diagnostic characteristics and were similar to normal diploid plants, with the exception of ditelotetrasomic 5S, which showed some effects. At meiotic diakinesis and metaphase I, these ditelotetrasomic plants showed chromosome configurations of 6_II+1_IV, 7_II+1 telo_II, 6_II+1_III+1telo_I, or 7_II+2 telo_I. Most of the sporocytes at anaphase and telophase in the first and second meiotic divisions showed almost normal chromosome behavior. Quartets were mostly normal with no micronuclei. Approximately 30% of the selfed progenies of these three ditelotetrasomics were ditelotetrasomics and almost 50% were telotrisomics, indicating a high percentage of male and female transmission of the extra telocentric chromosomes.     

Development of STS markers closely linked to the Ppd-H1 photoperiod response gene of barley (Hordeum vulgare L.)

A BC₂ population of 353 plants segregating for the Ppd-H1 photoperiod response gene was developed from a cross between the winter barley ’Igri’ and the spring barley ’Triumph.’ Bulk segregant analysis identified six AFLP markers closely linked to the Ppd–H1 gene and three strongly amplified AFLP bands that mapped 0.8-cM distal, 0.6-cM proximal and 2.3-cm proximal to Ppd-H1 were cloned and sequenced. Southern-blot analysis showed that the cloned fragments were single-copy sequences in ’Igri’, the variety from which they were derived. Two of the sequences were absent from ’Triumph’ while the third detected a single-copy sequence. The cloned fragments were used to design specific sequence tagged site (STS) primer pairs to assist in the construction of a high-resolution map of the Ppd-H1 region. Received: 22 March 2000 / Accepted: 10 April 2000