SESSION 9 ABIOTIC STRESS

Session 9 Abiotic Stress

Oral Presentation

Genetic Bases of Barley Response to Abiotic Stresses

L. Cattivelli1, C. Crosatti1, C. Mare1, S. Barilli1, S. Belloni1, E. Mazzucotelli2
and A. M. Stanca1

1Experimental Institute for Cereal Research, Section of Fiorenzuola dArda (PC) and 2Section of Foggia, 29017 Fiorenzuola dArda, Italy, E-mail: michele@stanca.it

The ability of barley to withstand stress situations is mediated by specific sets of genes which modify the cell metabolism making cells able to cope with adverse conditions. Fluctuations in water availability, changes of temperature and solute concentrations are common in all climates. All these stresses can damage plants, causing changing in cell volume and membrane shape, disruption of water potential gradients, physical damages to the membranes and protein degradation. Although in the recent years a number of genes whose expression is induced or enhanced by environmental changes have been cloned, little is known about the regulatory mechanism controlling the stress responses. This is mainly due to the fact that all stress responses are multigenic traits involving many genes that may have either redundant or additive effects and may interact with each other. Improvement of stress resistance will be a crucial factor in the next future to increase yield stability, grain quality and to expand the growing area of barley. This objective can be achieved either exploiting the genetic variability naturally present in the H. vulgare and H. spontaneum gene pools, or introducing new or modified genes in the barley genome either via molecular marker assisted selection or plant transformation.

Drought Tolerance in Barley

S. Grando, M. Baum and S. Ceccarelli

Germplasm Program, International Center for Agricultural Research in the Dry Areas (ICARDA),
Aleppo, Syria, E-mail: s.grando@cgiar.org

Since agriculture began, drought has been one of the major plagues limiting crop production worldwide. Although it reaches the front pages of the media only when it causes famine and death, drought is a permanent constraint to agricultural production in many areas of the world. Drought causes marginalization and poverty, and its negative effects are likely to increase as water resources decline globally. Climate changes will increase the frequency of droughts, particularly in Southeast Asia and Central America; by 2,050 water shortages are expected to affect 67% of the worlds population. Drought resistance has been always a challenge to plant breeders. Physiologists, biochemists, geneticists and breeders have dissected drought resistance into individual components, with the aim of finding simple associated traits. Examples include proline accumulation, osmotic adjustment, stomatal conductance, canopy temperature, and various root characteristics. Today, most scientists agree that a drought resistance gene does not exist, and that differences in drought resistance are due to the effects of several genes, affecting different characters that interact with each other. This is because drought is unpredictable in its occurrence, severity, timing and duration; and because drought seldom occurs in isolation and often interacts with other stresses, mostly temperature extremes thus determining many combinations of stresses. Often cultivars successful in one dry year may fail in another, thus eluding the need of farmers for consistency of performance. Therefore, while drought is a global issue, its effects need to be addressed locally because every dry area has its own type of drought. Future research to increase the level of drought tolerance in barley needs to address the interaction between a number of traits and assemble those combinations of traits that maximize economic yields per unit of water utilized, possibly with the use of molecular tools.

Freezing Injury and Breeding on the Frost Resistance
in Continental Russia

V. M. Shevtsov

Barley Breeding Department, Krasnodar Lukyanenko Research Institute of Agriculture,
Krasnodar, 350012, Russia,
E-mail: vshevtsov@inbox.ru

Winter cereals have a significant yield advantage in comparison with spring ones in Russia and some other countries. On the North Caucasus winter barley is higher in yield than spring barley 2050% and even more. During the last three seasons in Krasnodar region winter barley two times out-yielded spring barley. Winter hardness is an important agronomic character, facilitating barley yield stability. Resistance to freezing is a principal component of the trait. The deeper formation of crown node can significantly improve over-wintering. It was shown that there are reserves for barley improvement on cold tolerance by methods of the international germplasm exchange, mutagenesis and hybridization. Methods and approaches for evaluation and screening of breeding material on cold tolerance in the field and controlled conditions have been described. The method of experimental mutagenesis proved to be useful to induce a genetic variation on cold tolerance. A selection from the class of visible mutations was more effective. Some accessions from the world collection, mutants and breeding lines, having a high level of winter hardness, were identified to combine these characters with high productivity. The evaluation on cold tolerance should be regular and started from early generations by testing of big volume of breeding material in locations with sever winters and also in freezing chambers. Results of many years research have proved that the recombination of adaptive and productive characters is possible. High yielding and cold tolerant varieties Radical, Bastion and Dobrynia-3 were developed and introduced on farmers fields. The new winter barley varieties Fakir and Almaz carry the highest level of winter hardness in combination with the high yield potential.

Versatile Barley Varieties for the Mexican Highlands

F. Capettini

ICARDA/CIMMYT Latin American Regional Program, 06600, Mexico D.F., Mexico,
E-mail: f.capettini@cgiar.org

Barley production in the highlands of Mexico dates from the time of the Spanish conquest. Today, all the barley planted in the region is used for malting to produce beer. Globalization, most notably the North American Free Trade Agreement, is prompting farmers to look for new crops, such as forage barley, which appears to offer an economically viable alternative to malt barley production.

Screening Barley Germplasm for Waterlogging Tolerance

M. X. Zhou1, N. J. Mendham1, G. P. Zhang2, Q. Q. Shen3, R. G. Xu4, G. R. Zhang5 and H. Chen6

1Tasmanian Institute of Agricultural Research, University of Tasmania, Kings Meadows, TAS 7250, Australia, E-mail: meixue.zhou@dpiwe.tas.gov.au;
2Agronomy Department, Zhejiang University, Hangzhou, 310029, P. R. China; 3Zhejiang Academy
of Agricultural Sciences, Hangzhou, 310021, P. R. China; 4Agronomy Department, Yangzhou University, Yangzhou, 225009, P. R. China; 5Shanghai Academy of Agricultural Sciences, Shanghai,
201106, P. R. China; 6Yancheng Institute of Agricultural Research, Yangcheng, P. R. China

Field experiments were conducted to screen barley germplasm for waterlogging tolerance at four Chinese Institutes/Universities during 2001/02 and 2002/03 growing seasons. More than 500 barley varieties or breeding lines from different Chinese breeding programs were selected. Death rate of seedlings and leaf chlorophyll content were measured after waterlogging treatment. Other agronomic traits and yield components were also recorded. Varieties showed diverse responses to waterlogging. Based on the differences between the treated and controls, such as the reduction of yield component, more than 10% varieties were found to be tolerant to waterlogging. Most of these tolerant varieties were from the lower Yangtze River area of China. This paper will detail the response of different barley genotypes to waterlogging and the distribution of waterlogging tolerance varieties.

Poster Presentation

Combining of Drought Tolerance and Yield Potential in Crosses Developed from three Different Parents in Barley

S. Abbas1, G. Backes1, M. Baum2, S. Grando2, S. Ceccarelli2, H. Kayal3
and A. Jahoor1

1Plant Research Department, Risoe National Laboratory, DK-4000 Roskilde, Denmark,
E-mail: shahinaz.abbas@risoe.dk;
2International Center for Agricultural Research in the Dry Areas (ICARDA), Germplasme Program, Aleppo, Syria; 3Crops Production Department, Faculty of Agriculture, Damascus University, Damascus, Syria

Four crosses involving three different parents have been developed to conduct QTL analysis in diverse genetic background and under various environmental conditions in close so-cooperation with ICARDA. To develop these crosses, F1 plants produced from crosses of Syrian landrace with European high yielding variety were backcrossed with the third parents, again a Syrian landrace or a wild barley (Hordeum spuntanum) line. Several hundreds of BCF1 seeds were made by hand. Each BCF1 plant has been multiplied and planted at ICARDA. Two of these crosses in which either tow landrace from Syria and one European variety or one landrace and a wild barley line from Syria and a European variety, respectively are involved have been selected to be screened with a large number of microsatellite markers distributed evenly on all barley chromosomes. Theses crosses have been grown and observed at ICARDA at two locations for two seasons, to eventually identify effective QTLs for drought tolerance and to identify the QTLs that have been detected in other crosses for dry land agriculture. The results of QTLs analysis for drought tolerance and other agricultural traits will be presented.

Improving High Yielding Malting Barley Cultivars
for Supplementary Irrigated Areas in Turkish Highlands

T. Akar1, M. Avci1, F. Duesuenceli1, I. Sayim1, A. Atli1, H. Tosun1, A. Ottekin1 and A. Engin2

1Central Research Institute for Field Crops, 06042 Ulus, Ankara, Turkey,
E-mail: taner_akar@ankara.tagem.gov.tr;
2Anadolu Efes Malting and Beer Company, Cumra, Konya, Turkey

Some of the supplementary irrigated areas devoted for barley cultivation after sugar beet production were mainly dominated by six-rowed feed barley cultivars in Turkish Highlands. In order to meet good malting quality cultivars demand of malting industry located in the highlands a collaborative breeding program supported by the private company was commensed in 1994. The aims of the project are to improve new malting cultivars having high yield level and desirable good quality when compared to six-rowed feed cultivars in short term. These joint breeding efforts in collobaration with cereal quality, plant pathology and agronomy disciplines during the last ten years resulted in developing new winter and facultative two rowed malting cultivars namely Aydanhanim and Zeynelaga, that outyielded six-rowed feed cultivars interms of both yield and grain quality over locations in target areas.

Sequencing, Annotation and Transcriptional Analysis
of Molybdenum Cofactor Sulfurase Gene from Wild Barley,
Hordeum spontaneum

A. Altinkut1,2, S. Weining2, A. Distelfeld2 and E. Nevo2

1Tubitak, Research Institute for Genetic Engineering and Biotechnology, 41470, Gebze, Kocaeli, Turkey; 2Institute of Evolution, University of Haifa, Mount Carmel, Haifa, 31905, Israel,
E-mail: altinkut@research.haifa.ac.il

Plant Aldehyde Oxidases (AOs) form a multigene family whose members catalyse the final step in the biosynthesis of the phytohormone Abscisic Acid (ABA). Molybdenum cofactor (MoCo) sulfurase catalyzes the generation of the sulfurylated form of MoCo, a cofactor required by AO functioning in the last step of ABA biosynthesis in plants. This function is important in plant development and adaptation to environmental stresses such as drought. The coding region of MoCo sulfurase gene has been cloned and sequenced in the genotypes of wild barley, Hordeum spontaneum from a northern mesic Mediterranean area, semi-xeric steppes, and a southern xeric desert in Israel. Southern blot analysis of wild barley DNAs revealed evidence that MoCo sulfurase is a single copy gene in the genome of wild barley. After hybridization of the MoCo sulfurase gene probe with Morex barley BAC filter set, positive BAC clones containing flanking regions, promoters of the corresponding gene were identified by DNA Walking Speed Up Strategy. Chromosomal mapping using Chinese Spring Betzes addition lines revealed that the MoCo sulfurase gene is located on the barley chromosome 1 (7H). Using RT-PCR and Real-time PCR to study the gene expression analysis of this gene under water stress is currently under investigation.

QTLs for Drought Tolerance in Barley Grown in a Mediterranean Environment

H. Bahri1, D. This2, M. Mellouk1, M. El Ouahabi1, M. Bernokh1 and B. P. Forster3

1Ecole Nationale dAgriculture de Meknés, PBS40 Meknés, Morocco, E-mail: hbahri@enameknes.ac.ma; 2UMR 1096, INRA-ENSAM-CIDAR, 34060 Montpellier, France; 3Scottish Crop Research Institute, Invergowrie, Dundee, DD2 5DA, UK

Breeding for drought tolerance is a challenging task for many researchers. The ability to identify and select genotypes with desirables traits is influenced by the environment. In Mediterranean environments, grain yield is limited largely by drought, which is becoming more of a constraint in the last decade. In this regard, and in the framework of a INCO-DC programme, a population of 167 barley Recombinant Inbred Lines (RIL) and the two parents Tadmor and ER/Apm were grown under field conditions for two consecutive years in Meknés, Morocco. The aim was to evaluate the population for agronomic characteristics in the field, then to perform QTL analysis on the data obtained using a genetic map developed for the same population using different molecular markers. Grain yield, yield components, relative water content, and grain carbon isotope were evaluated during the two seasons. Simple interval mapping was used to identify QTL for the traits studied. QTL s were obtained for each environment separately and for both environments combined. The results show co-localisation of QTLs for some agronomic traits, wile others seem specific. The largest number of QTLs was detected on chromosomes 2(2H) and 7(5H). Four QTLs were identified for grain yield on chromosomes 3(3H) and 5(1H) for season 1, and two QTLs on chromosomes 1(7H) and 5(1H) for season 2. QTLs were also identified for relative water content, and grain carbon isotope. A significant negative association was obtained between carbon isotope discrimination and grain yield, and between carbon isotope discrimination and water use efficiency.

The Genetic Basis of Adaptation to Low Rainfall Environments
in Australia

S. J. Coventry1, M. Baum2, H. Sayed2, S. Grando2, S. Ceccarelli2, A. R. Barr3
and J. K. Eglinton4

1Molecular Plant Breeding CRC, School of Agriculture and Wine, University of Adelaide, Glen Osmond, SA 5064, Australia, E-mail: stewart.coventry@adelaide.edu.au;
2International Center for Agricultural Research in the Dry Areas (ICARDA), Aleppo, Syria; 3Australian Grain Technologies, University of Adelaide, Glen Osmond, SA 5064, Australia; 4School of Agriculture
and Wine, University of Adelaide, Glen Osmond, SA 5064, Australia

Improving the adaptation of barley to low rainfall environments is a challenge faced by many barley breeding programs. Constraints to productivity in these environments include drought and high temperature stress, subsoil constraints, and biotic stresses. Specific mapping populations have been grown in Australian low rainfall environments to characterise the genetic basis of adaptation. The mapping populations include the Australian advanced backcross QTL (H. spontaneum × Barque × 73) mapping population and ICARDA mapping populations Arta × Harmal and Tadmor × ER/Apm. QTL mapping of yield and grain quality traits, agronomic traits, and biotic/abiotic stress related traits has identified genetic locations different to those identified in previous Australian mapping populations. Mapping results from the Arta × Harmal population evaluated in Australian field trials will be compared with results from trials conducted in Syria and Jordan to compare the nature of low rainfall environments between the two regions.

Boron Toxicity in Barley: Prospects for Delivering a Yield Advantage

J. K. Eglinton1, R. J. Reid2, J. E. Hayes2, P. Davies3, D. Jha3 and G. McDonald1

1School of Agriculture and Wine, Waite Campus, and 2School of Earth and Environmental Sciences,
University of Adelaide, Glen Osmond, SA 5064, Australia, E-mail: jason.eglinton@adelaide.edu.au;
3South Australian Research and Development Institute, Glen Osmond, SA 5064, Australia

The physiological role of boron is not clearly understood, however it has long been established that it is essential for the growth of higher plants (Aghulon 1910). Boron can be phytotoxic if present in soils at high concentrations and has been recognised as a production constraint in regions of southern Australia, West Asia and North Africa. The genetic basis of tolerance to boron toxicity has been examined in a mapping population derived from Sahara 3771 and Clipper (Jefferies et al. 1999). This study identified four significant QTL involved in aspects of boron tolerance, however subsequent validation experiments have shown modest or inconsistent improvements in grain size and yield in boron toxic environments. The current paper presents results from studies aimed at further characterising yield responses associated with Sahara derived boron tolerance alleles, and examining the basis for potential yield penalties associated with these loci. The backcross lines were evaluated in further field trials that included analysis of rooting depth and water use in the soil profile. The sizes of introgression segments derived from Sahara were determined in this germplasm, highlighting linkage problems that potentially confound any yield advantage. Recent advances in our understanding of the mechanisms of boron toxicity tolerance are considered in relation to the genetic control of the trait. A novel method of selecting for boron tolerance is presented and discussed in the context of developing breeding strategies to maximise recombination.

QTLs and Candidate Genes in the Nure × Tremois Mapping Population

E. Francia1, D. Barabaschi1, A. Tondelli1, G. Laido1, C. Caffagni1, A. Aprile1,
A. Gianinetti1, A. M. Stanca1, G. Galiba2, P. M. Hayes3 and N. Pecchioni4

1Experimental Institute for Cereal Research, 29017 Fiorenzuola dArda (PC), Italy;
E-mail: enryfrancia@libero.it; 2Agricultural Research Institute of the Hungarian Academy of Sciences,
H-2462 Martonvásár, Hungary; 3Department of Crop and Soil Science, Oregon State University, Corvallis, OR 97331, USA; 4Universita di Modena e Reggio, Facolta di Agraria, I-42100 Reggio Emilia, Italy

In order to study in an unique genetic system cold and drought stresses, together with malting quality, a new genetic map based on the Nure × Tremois doubled-haploid population has been developed. Two low temperature tolerance QTLs were mapped on chromosome 5H. A first, distal QTL for cold tolerance and vernalization requirement was identified at the VrnH1/FrH1 region. The second proximal QTL was coincident with two QTLs regulating the accumulation of two different COR proteins (COR14b and TMC-Ap3); its peak position, named FrH2, is coincident with the HvCBF4 locus, that is in turn the candidate gene underlying this QTL. The Nure × Tremois population is also being tested for malting quality traits. Two separate replicated yield trials have been set up, the first in one location (2001/2002), and the second one for validation in different locations in Italy (2002/2003). Moreover, a multilocational irrigated vs. non-irrigated field trial in Mediterranean Europe, North Africa and West Asia is being carried out in 2004 for mapping QTLs and candidate genes of drought tolerance.

The Changes of Aluminum and Mineral Nutrient Contents
in Response to Al and Cd Toxicity and Al-Induced Organic Acid Exudation in Two Barley Varieties Differing in Al-Tolerance

T. R. Guo1, G. P. Zhang1, M. X. Zhou2, F. B. Wu1 and J. X. Chen1

1Agronomy Department, Huajia Chi Campus, Zhejiang University, Hangzhou, 310029, P. R. China,
E-mail: zhanggp@zju.edu.cn;
2Tasmanian Institute of Agricultural Research, University of Tasmania, Kinds Meadows, TAS 7249, Australia

A hydroponic experiment was carried out to study varietal difference in effect of Al and Cd on Al and mineral nutrient contents in various plant parts and Al-induced organic acid exudation by using two barley varieties differing in Al tolerance. The results showed that Al sensitive cv. Shang70-119 had significantly higher Al content and accumulation in various plant parts than Al tolerant cv. Gebeina, especially in root, when subjected to low pH (4.0) and Al treatments (100M Al and 100M Al + 1.0M Cd). Without Al addition, 1.0M Cd treatment caused a slight decrease in Al contents of both cultivars, however at the presence of Al in the solution, Cd addition further increased Al contents in plant parts compared with 100M Al treatment alone. Both low pH and two Al treatments caused significant reductions in Ca and Mg contents in all plant parts, P and K contents in shoot and leaf, Fe, Zn and Mo contents in leaf, Zn and B contents in shoot, Mn contents both in root and leaf. Moreover, the greater change in nutrient content was detected in the plants exposed to Al and Cd combination treatment than in those exposed to Al treatment alone. A dramatic enhancement of malate, citrate and succinate was found in the plants exposed to 100M treatment over the whole duration of experiment relative to the control, and Al-tolerant cultivar had a considerable higher exudation of these organic acids than Al-sensitive one, indicating that more Al-induced enhancement of these organic acids is very likely to be associated with Al-tolerance. The exudation of oxalate in response to Al exposure was not consistent during the treatment and showed less difference between the two cultivars, suggesting that oxalate is not likely to act a mechanism of Al tolerance in barley.

Black Point of Barley: A Possible Enzymatic Browning Reaction

T. K. Hadaway, H. M. Collins and A. J. Able

Discipline of Plant and Pest Science, School of Agriculture and Wine,
University of Adelaide, Waite Campus, Glen Osmond, SA 5064, Australia,
E-mail: tamara.hadaway@adelaide.edu.au

The possibility of an enzymatic browning reaction as the cause of black point in barley was investigated. Black point discolouration appears to be similar to the enzymatic browning characteristic of plant tissues subjected to physical damage, which involves the oxidation of phenolic compounds present in plant tissues by polyphenol oxidase (PPO) or peroxidase (POX) and the transformation of the oxidation products to black or brown pigments. Particularly high levels of oxidative enzymes such as POX and PPO are of particular importance in barley grain, as they are implicated to promote haze, off-flavour and staling of the finished beer. Lipoxygenase (LOX) is widely known to impart cardboard flavour to beer. Activity of the oxidative enzymes PPO, POX and LOX was measured during development of barley varieties differing in susceptibility to black point. The key enzyme for the production of phenolics in plant tissues, phenylalanine ammonia-lyase (PAL), was also measured. In order to determine differences in enzymatic activity of clean and black pointed grains, and hence the suitability of black pointed barley grain for use in malting and brewing, POX, PPO, LOX and PAL was measured during grain development using enzymatic activity assays, isoelectric focusing and Northern blotting for mRNA expression.

Effects of Cultivar and Provenance on Vigour of Barley Seed

P. Hrstkova, O. Chloupek and J. Bebarova

Mendel University of Agriculture and Forestry Brno, 613 00 Brno, Czech Republic;
e-mail: hrstkova@mendelu.cz

High vigour of barley seed is a precondition for rapid and homogenous field emergence and good malting quality. Seed quality has traditionally been associated with germination and purity. Another important seed quality component is seed vigour. Symptoms of vigour loss include reduction in germination rate and uniformity and consequently inferior seedling emergence and growth. The study was aimed to determine potential differences in seed quality of spring and winter barley in relation to cultivars and growing conditions. Several tests were performed on each seed lot, i.e. the standard germination test, the low temperature and water stress germination test and germination in a salt solution. Some of the seed samples were also evaluated using the controlled deterioration test described in the Handbook of Vigour Test Methods, ISTA 1995. CD test was initially developed for evaluating quality of small seeds of vegetable species as brassicas, carrot and lettuce, but seems to be also suitable for barley. The results were compared and seed lots with different values were identified. The effects of main factors as cultivar, provenance and year on seed vigour were estimated using analysis of variance. Statistically significant differences were particularly found among cultivars.

Supported by the Grant Agency of the Czech Republic (Grant No. 521/02/153).

Differences in Carbon Isotope Discrimination between Two-Rowed and Six-Rowed Barley Lines

Q. z. Jiang, D. Roche and D. Hole

Plants, Soils and Biometeorology Department, Utah State University, Logan, UT 84322-4820, USA,
E-mail: droche@mendel.usu.edu

For the last twenty years carbon isotope discrimination () has been extensively used in order to estimate the water use efficiency of C3 cereals species. Genetic lines of barley belong to two phenotypic classes for ear morphology, two-rowed (2R) or six-rowed (6R), a simply inherited trait. No experimental work has been attempted to test if values for differ between these two barley types for flag leaf, awn, and seeds. Here, we present data indicating that among breeding lines/varieties of both ear types grown in different conditions of water availability within line source experiments at two location years, values for 2R are usually lower than those found in 6R types. In an independent experiment conducted on double haploid progenies, issued from a cross between Harrington (2R) and Morex (6R), that segregate for ear type, we found that values for penultimate leaves and awns in field-collected materials are also lower in 2R progenies. Some of these observed differences may be due to differences in precocity as indicated by heading dates. This working hypothesis would confirm the importance of earlier maturity in C3 cereals in order to avoid terminal water stress during the grain filling period. Another contributing hypothesis would be, considering the individual fertile tiller as a physiological unit, that the carbon metabolism of each tiller may be intrinsically different between the two types since size of flag leaf and grain load per tiller are known to differ. Testing the segregation of values for in several plant tissues among unselected (Harrington × Morex) doubled haploid lines may provide further unexpected insights on the influence of the vrs-1 and int-c loci on carbon metabolism.

Gas Exchanges and Chlorophyll Fluorescence Parameters
of Fourteen Barley Lines in Response to Salinity

Q. z. Jiang, D. Roche, S. Durham and T. Monaco

Plants, Soils and Biometeorology Department, Utah State University, Logan, UT 84322-4820, USA,
E-mail: droche@mendel.usu.edu

Barley is one of the few economically significant crops relatively tolerant to salinity. Differences in salinity tolerance also exist between germplasm lines/varieties of barley; however, the physiological mechanisms that enable barley plants to tolerate salt stress are not clearly understood. Simultaneous measurements of leaf gas exchange (CO2) and H2O) and chlorophyll fluorescence parameters (Fv/Fm and PSII) were made on control plants and plants grown in saline conditions. Five days after emergence, the salinity treatment was conducted by soaking individual plants in a saline solution of NaCl and CaCl2 (EC: 30dS/m). Gas exchange and chlorophyll fluorescence measurements were determined for attached leaves at the fourth-fifth leaf stage. We will discuss the value of these measurements as a possible tool for salinity tolerance screening of genotypes.

Allelic Variation of Dehydrin Genes in Barley

S. Lababidi1,2, W. Choumane1,3, S. Ceccarelli1, S. Grando1, W. Al-Said2 and M. Baum1

1International Center for Agricultural Research in the Dry Areas (ICARDA), Aleppo, Syria,
E-mail: s.lababidi@cgiar.org; 2Department of Botany, Faculty of Science, Aleppo University, Aleppo, Syria; 3Faculty of Agriculture, Tishreen University, Lattakia, Syria

Dehydrins are proteins that accumulate during water deficient conditions during the maturing drying phase of seed development, in seedlings or more mature plants and are thought to play an important role in drought resistance in plants. In barley, 12 of the 14 possible dehydrin genes have been localized on almost all of the 7 chromosomes. The aim of our study was to identify the presence and the variation of dehydrin genes in different barley genotypes and to assess the dehydrin genes expression induced by drought conditions in the barley genotypes. Fourteen barley genotypes were used in this study. Plants were subjected to drought conditions and leave samples were collected every 3 to 5 days. DNA and RNA was extracted from plants subjected to water deficiency and from control plants. Genomic DNA and reverse transcribed cDNA was analyzed with specific dehydrin primers by RT-PCR. Variation of the length of the dehydrin gene alleles was detected in the different barley genotypes in genomic and cDNA. The result of using labeled dehydrin genes probes for northern hybridization demonstrated differences in dehydrin gene expression depending on the genes and genotypes used.

Characterisation of Traits Associated with Improved Growth
and Grain Yield of Barley on Sandy Soils of Low Fertility

N. R. Long1, R. E. Holloway3, S. J. Jefferies2 and A. R. Barr2

1School of Agriculture and Wine, University of Adelaide, Glen Osmond, SA 5064, Australia,
E-mail: nigel.long@adelaide.edu.au; 2Australian Grain Technologies, Roseworthy Campus, University
of Adelaide, Roseworthy, SA 5371, Australia; 3Minnipa Agricultural Centre, South Australian Research and Development Institute, Minnipa, SA 5654, Australia

Sandy soils of low fertility constitute approximately 30% of the total area (968,600 ha) sown to barley in South Australia (SA). With such a significant proportion of the barley sown on sandy soils, the provision of specifically adapted cultivars for this soil type is an integral component of feed barley breeding objectives in the SA Barley Improvement Program (SABIP). While potential genetic variation for sand adaptation has been observed in some Australian germplasm, there has been no concerted effort to characterise specific traits associated with improved performance on sand. Superior performance and grain yield on sandy soils was associated with improved establishment, early vigour, phosphorus utilisation efficiency, the reallocation of carbohydrates from the stem to the developing grain post-anthesis, and a deep root system. Besides the genotypic differences, establishment, early vigour and grain yield could be improved by selecting the large seed size fraction for sowing. Large seed size also increased coleoptile length and seed nutrient content. Superior varieties also exhibited an erect growth habit and an earlier flowering phenology. Evidence also suggests that adaptation response on sand requires drought tolerance to accommodate prevailing environmental conditions that easily override the sand effect. To elucidate chromosome regions associated with traits conferring improved adaptation on sandy soils a linkage map was constructed from a RIL population of a cross between the Australian cultivars Mundah (very good sand adaptation) and Keel (moderately poor sand adaptation). Significant QTL-marker associations for grain yield, grain weight, low screenings and early vigour were established that identified broad chromosome regions associated with sand adaptation.

Two Mutants Affecting Adaptative Responses to Abiotic Stresses
in Barley Seedlings

A. E. Martinez1,2, A. Landau1,3, P. T. Garcia1, G. Polenta1, M. C. Arias1, R. Murray1,
N. Pensel1 and A. R. Prina1

1Instituto Nacional de Tecnología Agropecuaria (INTA), Argentina; 2Consejo Nacional
de Investigaciones Científicas y Técnicas (CONICET), Argentina; 3Agencia Nacional
de Promoción Científica y Tecnológica (ANPCyT), Argentina,
E-mail: aprina@cnia.inta.gov.ar

We want to describe here two novel mutants which have affected the adaptative responses of barley seedlings to different abiotic stresses. They allow us to explore some aspects of adaptative phenomena which are little known in higher plants. One of those mutants corresponds to a plastid encoded gene which is involved in photosystems I and II stability and, probably indirectly, affects the acclimation of the seedlings to higher temperatures, a fact which seems to occur through the control of unsaturation/saturation levels of the thylakoid membrane fatty acids. The other mutant corresponds to a nuclear gene which, under certain circumstances, induces an additional ethylene production in the seedling roots. This mechanism seems to be involved in eliciting a negative hydrotropic growth of the roots, a phenomenon that we interpret as a waterlogging avoiding response.

AB-QTL Analysis for Osmotic Adjustment and Relative Water Content of Drought Tolerance in Spring Barley

K. A. H. Mohammed1, M. v. Korff1, H. Wang1, G. A. R. El-Sherbeny 2, K. Pillen1 and J. Leon1

1Department of Crop Sciences and Plant Breeding, University of Bonn, D-53115 Bonn,
E-mail: khalafhamam@yahoo.com, j.leon@uni-bonn.de; Germany;
2Department of Genetics, Sohag Faculty of Agriculture, South Valley University, Egypt

The advanced backcross quantitative trait loci (AB-QTL) analysis was used to identify QTLs for osmotic adjustment and relative water content (RWC) in a BC2DH population derived from a cross between the spring barley variety Scarlett and the wild barley ISR42-8. Ninety-seven markers were employed to genotype 323 BC2DH lines and phenotypic data were collected for two parameters related to drought tolerance. Two treatments were tested for the drought experiment, 50% field capacity (FC) level for drought stress and 100% FC level as a control. QTLs were detected by means of ANOVA with a significance level of P < 0.01 for a marker main effect and a marker × drought treatment (M × D) interaction, respectively. A total of 10 putative QTLs were detected, of which seven were for osmotic adjustment and three for relative water content. Altogether, 5 (50%) favorable effects of exotic alleles were detected for drought tolerance, despite the fact that the exotic barley donor had an overall inferior phenotype with respect to agronomic performance. The present study indicated that favorable QTL alleles for drought tolerance could be transferred from wild barley into an elite barley variety by the advanced backcross QTL strategy and molecular breeding.

Thermal Requirements for Barley Leaf Development
during Tillering

V. Momcilovic1, N. Przulj1 and D. Knezevic2

1Institute of Field and Vegetable Crops, Novi Sad, Serbia and Montenegro,
E-mail: przulj@ifvcns.ns.ac.yu;
2ARI SERBIA Small Grains Research Centre, 34000 Kragujevac, Serbia and Montenegro

Time interval between appearances of successive leaves is designated as phyllochron interval (PI). The objective of this study was to evaluate PI of mainstem leaf (MSL) development during tillering of spring barley. Six spring barley varieties were grown at Novi Sad, former Yugoslavia (45°20 N, 15°51 E, 86 m a.s.l.) from 1999 to 2001. Phyllochron was estimated using Haun scale where growing-degree-days (GDD) have been used as time scale. PI ranged from 60.5 GDD in fifth to 75.8 GDD in third leaf. All three factors, i.e., year, variety, and their interaction were included in phyllochron determination of first leaf. The highest percentage of variance component for first leaf phyllochron belonged to year. Variation in phyllochron of second leaf was controlled by variety and an interaction year × variety, variation in phyllochron of third, fourth, and fifth leaf by year and the interaction, and variation in sixth leaf by the interaction only. The linearity suggests that MSL stage can be used as a predictive measure of plant development and can retroactive shows quality of the preemergent seedbed environment.

The Use of Real-Time PCR for Barley Gene Expression Analysis: Cold Stress Response

J. Ovesna, A. Heldova, L. Kucera, I. T. Prasil and K. Polakova

Research Institute of Crop Production, 161 06 Prague-Ruzyne, Czech Republic,
E-mail: ovesna@vurv.cz

A two-step real-time quantitative reverse transcription-PCR (RT Q-PCR) RNA samples with a SYBR Green I detection chemistry is the technique of choice for sensitive, reproducible and large-scale measurements of gene expression levels. To investigate the changes in inducible gene expression measurement of endogenous standards is critical. A non-regulated house keeping gene ubiquitin has been used as an endogenous standard. Some studies in animal system have already shown that house keeping genes are regulated and differ under experimental conditions. For high-throughput and reliable quantification model of RT Q-PCR in barley plant, this study enters into the normalization of ubiquitin genes for the relative quantification of a target gene(s) with 2-fold. Expression levels of ubiquitin was correlated with the spectroscopic measurement of total RNA and amplification efficiencies during RT Q-PCR. To show applicability of RT Q-PCR cold stressed barley cultivars differing in cold stress response were evaluated by a set of primers specific for different dehydrin genes and quantitative data were correlated with LT50 values. Resulting data are discussed.

Growth, Morphological and Anatomical Responses of Two Barley Genotypes to Hypoxia

J. Pang1, M. X. Zhou2, N. J. Mendham1 and S. Shabala1

1Tasmanian Institute of Agricultural Research, University of Tasmania, Hobart, TAS 7001,
Australia; 2Tasmanian Institute of Agricultural Research, University of Tasmania,
Kings Meadows, TAS 7250, Australia,
E-mail: meixue.zhou@dpiwe.tas.gov.au

TX9425 (Chinese variety, waterlogging tolerant) and Naso Nijo (Japanese variety, waterlogging susceptible) were used to investigate the growth, morphological and anatomical responses of barley genotypes to hypoxia, produced experimentally by growing seedlings de-oxygenated nutrient solution containing 0.1% (w/v) agar with controls in the aerated nutrient solution. Both varieties were subjected to hypoxia for 3 weeks starting from the four-unfolded-leaf stage. After the treatment, TX9425 showed less reduction in the length of the longest seminal root and dry weight of plants than Naso Nijo. Both varieties had an increase in growth of adventitious roots after hypoxia treatment with TX9425 showing greater increase. Anatomical study of the adventitious root showed that aerenchyma was formed in the cortex in both genotypes after 3 weeks hypoxia treatment, and the percentage of aerenchyma area in the root cross-section area was larger for TX9425 than the corresponding value for Naso-Nijo. Hypoxia decreased the ratio of stele to root cross-Session area in TX9425, while it had little effect in Naso-Nijo. In plants suffering from hypoxia the ratio of the total xylem area to root cross-Session area increased for TX9425 but decreased for Naso-Nijo compared with the controls. This ratio should be an indicator of ability to transport water from roots to stems and leaves.

QTL Mapping of Reproductive Frost Tolerance in Barley

J. L. Reinheimer1, A. R. Barr1 and J. K. Eglinton2

1Australian Grain Technology, Plant Breeding Unit, Roseworthy Campus, Rosevorthy, SA 5371, Australia; 2School of Agriculture and Wine, Waite Institute, University of Adelaide, SA 5064, Australia,
E-mail: jason.eglinton@adelaide.edu.au

Spring radiation frost is a major abiotic stress affecting broad acre grain production in Australia. Low temperatures during reproductive stages of cereal development can cause floret and spike abortion as well as damage to developing grain, significantly reducing yield and quality. No previous study has been successful in identifying genetic variation for reproductive frost tolerance in barley. A survey of diverse germplasm collected from throughout the world was undertaken to identify genetic variation for reproductive frost tolerance in barley. Field based screening nurseries were developed to discriminate between reproductive frost tolerant and susceptible genotypes. Compared to Australian commercial varieties, two genotypes from Japan and one from Ethiopia were identified to have significantly less frost induced sterility and grain damage during a discriminating frost event. Both Japanese lines were used to develop mapping populations from a cross with Australian frost susceptible varieties. Frost induced sterility mapped to chromosome 2HL and 5HL in both populations. The QTL on chromosome 2HL mapped distal to the earliness per se Eps2 locus. The QTL on chromosome 5HL mapped close to Vrn-H1 and the previously identified vegetative frost tolerance locus. This region is also homoeologous to the Vrn-A1, Fr-A1 region on wheat chromosome 5AL. The results suggest that this region on chromosome 5H conditions frost tolerance of both reproductive and vegetative plant tissue challenging the traditional view that these traits are independent.

Biodiversity for Yield Performance and Associated Morpho-Physiological Traits in Barley Genotypes under Rainfed and Irrigated Conditions

F. Rizza1, F. W. Badeck2, L. Cattivelli1, J. Ghashghaie3, N. Di Fonzo4, O. Lidestri 4
and A. M. Stanca1

1Experimental Institute for Cereal Research, I-29017 Fiorenzuola dArda (PC), Italy,
E-mail: fulvia.rizza@libero.it;
2Potsdam Institute for Climate Impact Research (PIK), D-14412 Potsdam, Germany;
3Laboratoire dEcophysologie Végétale, Université Paris Sud, Orsay Cedex, 91405 France;
4Experimental Institute for Cereal Research, I-71100 Foggia, Italy

The biodiversity for yield performance under rainfed (NI) and irrigated (I) conditions was studied for three years at Foggia (South Italy) in a set of 100 barley genotypes. For each trial a Water Stress Index (WSI) was calculated based on the daily potential and actual evapotranspiration occurred during each growing season, estimated by the Thorntwaites method. As the WSI explained a high proportion of the yield variation among years and treatments, the yield vs WSI regression was employed to describe the behaviour of a given genotype across trials. The biodiversity among genotypes was also analysed for morpho-physiological traits as: heading date, plant height, kernel number, kernel weight and 13C isotopic signature. The high yield potential of the genotypes played a pre-eminent role in assuring high yield in both NI and I trials. High yield were achieved throughout different strategies associated to genotypic traits (higher kernel weight or number per m2, reduced plant height, early heading) conserved under favourable or stress conditions.

Root-Specific O-Methyltransferase Gene Expressed
in Salt-Tolerant Barley

M. Sugimoto1, Y. Okada2, K. Sato3, K. Ito2 and K. Takeda3

1Laboratory of Cytomolecular Biochemistry, Research Institute for Bioresources, Okayama University, Kurashiki, Okayama 710-0046, Japan, E-mail: manabus@rib.okayama-u.ac.jp;
2Plant Bioengineering Research Laboratories, Sapporo Breweries Ltd., Nitta, Gunma 370-0393, Japan;
3Barley Germplasm Center, Research Institute for Bioresources, Okayama University, Kurashiki,
Okayama 710-0046, Japan

O-Methylated flavonoids and derivatives play an important role not only in plant growth but also in interactions with environmental factors as regulators of auxin transport, signal molecules in plant-symbiont interactions, and precursors of phytoalexins. Enzymatic O-methylation is catalyzed by o-methyltranserase (OMT), which transfers the methyl group of S-adenosyl-L-methionine to the hydroxyl group of an acceptor molecule. The OMT gene from barley was induced by fungal pathogens and UV light and the gene from ice plant is highly expressed under the exposure to salt stress. These results led us to expect that the OMT would function in protecting cells from environmental stresses in barley. We identified an OMT gene expressed preferentially in salt-tolerant barley roots. The structure of the OMT encoded by the gene was different from those isolated from barley as reported previously. The OMT gene was a single copy and positioned on chromosome 1H between Ica1 and ABG500A. Northern blot analysis revealed that the OMT gene was expressed constitutively in the salt-tolerant barley roots and the expression level was increased 1.5 times by salt stress, whereas the salt-sensitive barley showed no expression of the gene in roots and leaves. Our results suggest that the OMT gene would be a candidate component for an anti-salt stress system in barley.

Differential Expression of Dehydrin Genes under Water Stress
in Wild Barley, Hordeum spontaneum

T. Suprunova, T. Krugman, T. Fahima, G. Chen, I. Shams, A. B. Korol and E. Nevo

Institute of Evolution, University of Haifa, Haifa 31905, Israel,
E-mail: fahima@research.haifa.ac.il

Dehydrin gene (Dhn) expression is associated with plant response to dehydration. Our aim was to study the association of Dhn gene (Dhn 1, 3, 5, 6, and 9) expression with drought tolerance found in wild barley (Hordeum spontaneum). Tolerant and sensitive genotypes were identified from Israeli (Tabigha microsite) and Jordanian (Jarash and Waddi Hassa) populations (based on scoring of water loss rate of 400 genotypes). The expression of the five Dhn genes was up-regulated by dehydration in both resistant and sensitive wild barley genotypes. Notably, differences between resistant and sensitive genotypes were detected, mainly in the expression of Dhn1 and Dhn6 genes, depending on the duration of dehydration stress. Resistant genotypes showed earlier expression of Dhn1 (after 3 h) and higher level of expression (12 h and 24 h) as compared to sensitive genotypes. The level of expression of Dhn6 was significantly higher in the resistant genotypes at the earlier stages after stress. However, after 12 and 24 h Dhn6 expression level was relatively higher in sensitive genotypes. Our results may indicate that these genes have some functional role in the dehydration tolerance of wild barley. We suggest that the observed differences of Dhn expression in wild barley, originating from different micro- and macro-ecogeographic locations, may be the result of edaphic and climatic selective pressures.

Effect of Cadmium on Free Amino Acids, Glutathione and Ascorbic Acid Content in Two Barley Genotypes (Hordeum vulgare L.) Differing in Cadmium Tolerance

F. B. Wu and G. P. Zhang

Agronomy Department, Huajia Chi Campus, Zhejiang University, Hangzhou, 310029, P. R. China,
E-mail: zhanggp@zju.edu.cn

A hydroponic experiment was carried out to study the effect of three Cd levels on glutathione (GSH), free amino acids, and ascorbic acid (ASA) content in the different organs of 2 barley cultivars with different Cd tolerance. Cadmium concentration in both roots and shoots increased with external Cd levels, while biomass and ASA content declined, and Wumaoliuling, a Cd sensitive genotype was more affected than ZAU 3, a Cd tolerant genotype. The effect of Cd on GSH content was dose- and time-dependent. In the 5 day exposure, root GSH content increased in 0.5 M Cd treatment compared with control, but decreased significantly in 5 M Cd treatment, irrespective of genotypes. However, in the 10 d exposure, GSH content in leaves, stems and roots decreased with increasing Cd levels in the culture medium, and Wumaoliuling was much more affected than ZAU 3. Cadmium treatment also modified free amino acid content and composition in plant organs. The effect of Cd on Glu content in roots showed a genotype-dependent change. ZAU 3 showed a steady increase in root Glu content in both 0.5 and 5 M Cd treatments, while Wumaoliuling showed 38.0% decrease in 5 M Cd treatment, compared with control. The results indicate that GSH and ASA may attribute to Cd-detoxicity, and the relative less reduction in GSH content at 5 M Cd in ZAU 3 relative to the control may account for its higher tolerance to Cd toxicity.

Progress in Malting Barley Breeding in a Family Owned Company

J. Breun and M. Breun

Saatzucht Josef Breun GdbR, D-91074 Herzogenaurach, Germany
E-mail: saatzucht@breun.de

The Saatzucht Josef Breun GdbR is a family company in the third generation and exists since 1906. The crops, we breed, are spring and winter barley, winter wheat and triticale. Because of the self fertilization uses the breeder the linebreeding method for spring barley. According to a definite time the method cuts in bulk and pedigree method. Often a combination of both methods succeeds. We use single seed descent to get earlier homozygous lines, for fastening the breeding process. The breeding of a new malting barley, which is on the one hand homogeneous and on the other a progress on the permitted varieties is a long process, that takes sometimes 10 to 12 years. An important breeding principal object for malting barley is the enhancement of the quality: higher full barley, less protein, progression of the malt- and beer quality. The selection for resistance against plant diseases has a central role in barley breeding, for example against mildew, netblotch, Rhynchosporium (leafblotch), dwarfrust as well as against new mycosis like Ramularia oder virus disease especially at winter barley. But thoughts are also given to new kinds of damage, caused by nonparasitic reasons like atmospheric strains, to assure high yields. The cultivation of mycosis resistant varieties should abate the costs of production by reducing fungizides, the products are nonhazardous and last but not least conserve the environment. There has been made an explicit progress. Additionally through the increasing demands of the consumers the breeding aims must be constant redefined and the utilization of new methods of molecular biology in practical breeding is continuously evaluated and verified.

Progress in Malting Barley Breeding in Secobra

M. Madre

SECOBRA Recherches, 78580 Maule, France,
E-mail: michel.madre@secobra.com

Established in 1902 by the French Brewing industry, the first mission of cv. Secobra has been to promote, improve and develop the barley crop in France. For this purpose, cv. Secobra has implemented breeding and trialling programs, carried out work on varietal identification, developed tools for quality evaluation. Since the seventies, with the impulse of its main industrial shareholders from the Malting and Brewing industry (Soufflet, Malteurop, Kronenbourg ). Cv. Secobra has broadened and intensified its breeding programs and consequently participated in the evolution of the barley crop in France and in the development of malting barley varieties during the last 30 years: (1) Maintenance of the area cultivated with spring barley since the nineties thanks to the development of suitable productive varieties (Nevada, Astoria); (2) Development of 6 rowed winter malting barley with mosaic resistant varieties (Esterel);.(3) Development of the two-rowed winter barley crop: malting and feeding varieties (Clarine, Pastoral). Leader in France with more than 40% of barley cultivated areas, cv. Secobra is also present in other countries with six-rowed winter barleys (cv. Theresa, Franziska), as well as malting spring barley like cv. Josefin. Today, cv. Secobra is developing important barley breeding programs (winter and spring) in 4 countries (France, Germany, Great Britain and Spain) and uses modern breeding methods and tools: nearly 2,500 crosses per year, winter generations in New Zealand and dihaploidisation, many predictive technological analyses, molecular marking, specific programs. Constantly led by its aims and thanks to a strong specialisation and to the continuous support of the industrial chain, cv. Secobra is a unique model in Europe of collaboration between a breeding company and its downstream industry.