Plant critical levels for the evaluation of boron toxicity in spring barley (Hordeum vulgare L.)

Summary Pot experiments on B toxicity in spring barley (Hordeum vulgare L., cv. Trumpf) using sandy soils indicated that there are significant relationships between B content of the leaves and of the shoots respectively in the toxic range and degree of damage of the leaves at stage 7–8 of the Feekes scale, which may be used to derive plant critical levels of B toxicity. Symptoms due to B excess begin to develop on the leaves (leaf tip necroses) relatively independently of the ontogenetical stage of development as soon as the B content of the leaf tissue reaches 60–80 mg/kg DM.The corresponding symptom-related toxic plant critical level of shoots (i.e. the B content of the whole shoot at which the first damage in leaves begins to occur) ranges from 30 mg/kg shoot DM (related to older leaves) to 80 mg/kg shoot DM (related to younger leaves). Grain yield is significantly reduced only when the B content of shoots at Feekes stages 7–8 exceeds the yield-related toxic plant critical level (yield reduction to 90% of the optimum yield) of 120–130 mg/kg shoot DM.B contents of the shoots at Feekes stages 7–8 from 80–120 mg/kg shoot DM define the range at which plants have marked toxicity symptoms, but at which there are no yield reductions.     

Aquaporin-facilitated water uptake in barley (Hordeum vulgare L.) roots

It is not known to what degree aquaporin-facilitated water uptake differs between root developmental regions and types of root. The aim of this study was to measure aquaporin-dependent water flow in the main types of root and root developmental regions of 14- to 17-d-old barley plants and to identify candidate aquaporins which mediate this flow. Water flow at root level was related to flow at cell and plant level. Plants were grown hydroponically. Hydraulic conductivity of cells and roots was determined with a pressure probe and through exudation, respectively, and whole-plant water flow (transpiration) determined gravimetrically in response to the commonly used aquaporin inhibitor HgCl(2). Expression of aquaporins was analysed by real-time PCR and in situ hybridization. Hydraulic conductivity of cortical cells in seminal roots was largest in lateral roots; it was smallest in the fully mature zone and intermediate in the not fully mature 'transition' zone along the main root axis. Adventitious roots displayed an even higher (3- to 4-fold) cortical cell hydraulic conductivity in the transition zone. This coincided with 3- to 4-fold higher expression of three aquaporins (HvPIP2;2, HvPIP2;5, HvTIP1:1). These were expressed (also) in cortical tissue. The largest inhibition of water flow (83-95%) in response to HgCl(2) was observed in cortical cells. Water flow through roots and plants was reduced less (40-74%). It is concluded that aquaporins contribute substantially to root water uptake in 14- to 17-d-old barley plants. Most water uptake occurs through lateral roots. HvPIP2;5, HvPIP2;2, and HvTIP1;1 are prime candidates to mediate water flow in cortical tissue.     

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.     

Salinity tolerance in different cultivars of barley (Hordeum vulgare L.)

Seven barley(Hordeum vulgäre L.) cultivars tested varied greatly in their responses to root medium salinity (electrical conductivity of 3, 5, 10, 15 and 20 dS nr^-1)-lant growth was relatively more adversely affected than seed germination. Dry/fresh mass ratio increased at higher salinity levels in all barley cultivars indicating reduced water uptake. Higher K/Na ratio in plant shoots compared to that in the root medium solution indicated selective uptake of K that seems to be among processes involved in tolerance of cultivars to salinity stress.     

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.     

Polymorphism at the Hor 1 locus of barley (Hordeum vulgare L.)

The Hor 1 locus of barley encodes a group of seed storage polypeptides called C hordein. Two-dimensional electrophoretic analysis of C-hordein fractions from six cultivars with different alleles at the Hor 1 locus showed extensive polymorphism. A total of 34 major polypeptides was mapped, with between 4 and 18 present in each cultivar. There was less variation among the same cultivars in the numbers (6 to 10) of restriction fragments of genomic DNA which hybridized to a cDNA clone related to C hordein. The total number of restriction fragments was also lower (22), and most pairs of cultivars had more restriction fragments than polypeptides in common. A total number of about 20–30 C-hordein genes per haploid genome was estimated. The results indicate that cultivars differ mainly in the extent of gene and polypeptide divergence, rather than in the degree of gene reiteration. They are consistent with the proposed origin of the multiple structural genes at the Hor 1 locus by the duplication and divergence of a single ancestral gene.NACB was supported by a grant from the Home Grown Cereals Authority.     

Chlorophyll a fluorescence--A useful tool for the early detection of temperature stress in spring barley (Hordeum vulgare L.)

The photosynthetic activity of two Syrian barley (Hordeum vulgare L.) landraces, Arabi Abiad (A. Abiad) and Arabi Aswad (A. Aswad), grown under low- and high- temperature stresses, were studied by analyzing the measured chlorophyll fluorescence signals. Both the applied stresses influenced photosystem II (PSII) activity. However, the effects depend on the stress type and the duration of its application. Phenomenological parameters were shifted shortly after the application of both stresses, whereas fluorescence ratios and yield values were altered most significantly after 7 days of stress imposition. The earliest changes in PSII activity of both cultivars were observed in the case of high temperature treatment. The maximal quantum efficiency of the photosynthetic apparatus (F(v)/F(M)) did not alter after stress application. Therefore, we could not recommend this parameter for early detection of such stress. In contrast, the results from the present investigation strongly indicate that the most significantly changed chlorophyll a fluorescence parameters could be used as an efficient tool for the early diagnosis of temperature stress in barley.     

Effect of seed nitrogen on response to applied nitrogen in six spring barley (Hordeum vulgare L.) cultivars in a glasshouse

Summary Variation in the nitrogen content of seed of six barley cultivars was brought about by growing parent plants at four nitrogen levels. Shoot dry weight of plants grown for 23 days from these seeds was generally enhanced by an increase in seed nitrogen content. The most responsive cultivar was a primitive type of barley from Ethiopia. Cultivars with a longer breeding history were less responsive. Risø 1508 apparently had physiological and biochemical limitations in responding to extra seed nitrogen. In the barley cultivars studied extra seed nitrogen seems to supplement, rather than substitute for, nitrogen fertilizer in the seed bed.     

Expression of hydrogenase activity in barley (Hordeum vulgare L.) after anaerobic stress

Germinated seeds of barley, after anaerobic treatment, produce considerable amounts of hydrogen. The anaerobic stress induces hydrogenase activity, as measured by the ability to evolve hydrogen from reduced methyl viologen. Several pieces of evidence, including bacteriological controls and induction in axenic calluses, indicate that the hydrogenase activity is induced by the plant itself and not by contaminant microorganisms. The hydrogenase is selectively induced in roots and to a lower extent in hypocotyls, but no hydrogenase activity can be detected in leaves.     

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.     

Accumulation of phenolics and growth rate of barley seedlings (Hordeum vulgare L.)

Accumulation of phenolics in barley seedlings was examined in relation to elongation; the seedlings were cultivated at 5 °C or 26 °C in light or in darkness. It was found that a higher accumulation of phenolics (mainly saponarin) was accompanied by slower elongation. This relation was repeatedly observed regardless of whether growth retardation or stimulation was obtained by light and temperature conditions of growth or treatment with CCO orp-fluorophenylalanine (p-FPA), respectively. It is proposed that PAL and peroxidase activities are responsible for maintaining the level of phenolics in seedlings. These enzyme activities are differently influenced by temperature conditions of growth. It is also suggested that accumulation of saponarin may lead to slowing down the growth by stimulating IAA oxidase and lowering the auxin level in the tissues. Thus, phenolics may belong to the factors through which environmental conditions influence elongation of seedlings.     

The effects of priming treatments using seaweed suspensions on the water sensitivity of barley (Hordeum vulgare L.) caryopses

Two seaweed suspensions, one prepared from Ascophyllum nodosum (Linnaeus) Le Jolis (ANS), another from Laminaria hyperborea (Gunn.) Foslie (LHS), were evaluated for their effects on the water sensitivity of barley (Hordeum vulgare L.) seeds. Priming in either seaweed suspension did not reduce viability. Priming in ANS was beneficial to seeds germinated in elevated water levels. The suspension of A. nodosum reduced the water sensitivity of barley seeds better than either water or polyethylene glycol (PEG) priming treatments. ANS also reduced the microbial population on the seeds by 86%. The hygroscopic properties and the antibiotic effect of the suspension of A. nodosum resulted in greater oxygen availability to the embryo, enabling more seeds to germinate under oxygen deficient conditions.     

Transgenic barley (Hordeum vulgare L.) by electroporation of protoplasts

Summary Protoplasts isolated from calli derived from cultured microspores of barley (Hordeum vulgare L. cv. Kymppi, an elite cultivar) were transformed with the neomycin phosphotransferase marker gene (nptII) by electroporation. Screening of the regenerated plants for the NPTII activity by gel assay resulted in three positive signals. Southern blot analysis and NPTII assays of second and third generation plants confirmed the genomic integration of the transferred gene and that the new trait was inherited by the progeny.     

A flavonoid mutant of barley (Hordeum vulgare L.) exhibits increased sensitivity to UV-B radiation in the primary leaf

The aim of the present investigation was to define the role of soluble flavonoids as UV-B protectants in the primary leaf of barley (Hordeum vulgare L.). For this purpose we used a mutant line (Ant 287) from the Carlsberg collection of proanthocyanidin-free barley containing only 7% of total extractable flavonoids in the primary leaf as compared to the mother variety (Hiege 550/75). Seven-day-old leaves from plants grown under high visible light with or without supplementary UV-B radiation were used for the determination of UV-B sensitivity. UV-B-induced changes were assessed from parameters of chlorophyll fluorescence of photosystem II, including initial and maximum fluorescence, apparent quantum yield, and photochemical and non-photochemical quenching. A quartz fibre-optic microprobe was used to evaluate the amount of potentially harmful UV-B (310 nm radiation) penetrating into the leaf as a direct consequence of flavonoid deficiency. Our data indicate an essential role of flavonoids in UV-B protection of barley primary leaves. In leaves of the mutant line grown under supplementary UV-B, an increase in 310nm radiation in the mesophyll and a strong decrease in the quantum yield of photosynthesis were observed as compared to the corresponding mother variety. Primary leaves of liege responded to supplementary UV-B radiation with a 30% increase in the major flavonoid saponarin and a 500% increase in the minor compound lutonarin. This is assumed to be an efficient protective response since no changes in variable chlorophyll fluorescence were apparent. In addition, a further reduction in UV-B penetration into the mesophyll was recorded in these leaves.     

Estimation of genetic parameters in barley (Hordeum vulgare L.)

Summary Four different sets of partial diallels were analysed for their relative efficiencies for estimating the genetic parameters in barley: (1) partial diallel with 12 parents, each involved in only 5 crosses; (2) partial diallel with 12 parents, each involved in only 3 crosses; (3) partial diallel with 8 parents, each involved in only 5 crosses; and (4) partial diallel with 8 parents, each involved in only 3 crosses. In partial diallel experiments, the estimates of gca effects were higher than in those of full diallel. Ranking pattern of the parents on the basis of gca effects in partial diallels deviated considerably from the ranking in full diallel. With decreasing s per parent, the deviation in ranking was also more. This clearly suggests the unsuitability of partial diallel analysis for screening high general combiners. Selection of best cross combinations is also not possible because only a sample of crosses (s out of n) is analysed under partial diallel so that there is every possibility of the best cross being excluded from the sample. In general, overdominance was exhibited, indicating that there is ample scope for heterosis breeding in barley.     

Estimation of genetic parameters in barley (Hordeum vulgare L.)

Summary Four exotic and four indigenous strains of barley were used for making diallel crosses. The sets of parents and crosses making full, half and quarter diallel were analysed in a randomized block design for plant height, number of effective tillers, ear length, grain yield per plant, 100 grain weight and number of grains per ear.The three alternatives of diallel were similar with respect to the estimates of degree of dominance, general combining ability and specific combining ability, indicating that all these three methods of diallel were equally efficient. However, as the number of entries are minimum in quarter diallel, it would be economical in terms of cost, time and labour to estimate genetic parameters by this method. Average degree of dominance was found in the range of overdominance. The ranking of parents on the basis of their array mean was similar to the ranking based on gca effects. Similarly, the ranking of crosses on the basis of per se performance was similar to the ranking based on sca effects. This suggests that the selection of best general combiner or best cross combinations may be easier and more effective through array mean for per se performance rather than through high gca and sea effects, respectively. From among 56 crosses, IB-226 X X C-164 was the one which showed superiority for maximum number of characters followed by AB-12/59 X PTS-57. High sea effect for plant height, ear length, grain yield, 100 grain weight and number of grains per ear was the result of cross between parents having high X low general combining ability, indicating additive X dominance type of gene interaction. For number of effective tillers, high sca was produced by low x low general combiners, indicating dominance x dominance gene interaction.Part of a Ph. D. thesis submitted by senior author to Haryana Agricultural University, Hissar.     

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.     

Root contribution to nitrate reduction in barley seedlings (Hordeum vulgare L.)

Summary The leaf and root nitrate reductase activities were measured in 7 day-old barley seedlings by anoxic nitrite accumulation in darkness, during 48h after the transfer from a N-starved medium to a 1.5 mM K¹⁵NO₃ medium. Thisin situ nitrate reduction was compared with the¹⁵N incorporation in the reduced N fraction of the whole seedlings.The nitrate reduction integrated fromin situ measurements was lower than the reduced¹⁵N accumulation. The rootin situ nitrate reductase activity seemed to account for only the third of the real root nitrate reduction, which may have been responsible for the overall underestimation. This discrepancy was partly explained by the ability of the root to reduce nitrite in an anoxic environment.These results suggest that, after correction of thein situ estimation of the nitrate reduction. the roots contribute to about 50% of the total assimilation.     

Polymorphism of the cultivated barley (Hordeum vulgare L.) of South Arabia at hordein-coding loci

Electrophoresis in starch gel was used to study the polymorphism of hordeins controlled by loci Hrd A, Hrd B, and Hrd F in 89 samples of the local barleys from South Arabia (Yemen). Overall, 36 alleles were detected for locus Hrd A; 48 alleles, for Hrd B; and 5 alleles, for Hrd F. The existence of the blocks of hordein components controlled by loci Hrd A and Hrd B was demonstrated. Calculation of genetic distances allows us to conclude that the barley populations from Yemen and Ethiopia are more similar compared with the populations from Egypt. This confirms the hypothesis of Bakhteev on the origin of Ethiopian barleys.