The Stress of Suicide: Temporal and Spatial Expression of Putative Heat Shock Protein 70 Protect the Cells from Heat Injury in Wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis>)

Heat stress adversely affects growth, development, and yield of winter wheat (Triticum aestivum). Plants have, however, evolved mechanisms to adapt to such conditions mainly by the expression of stress-associated chaperones, the heat shock proteins (HSPs), for modulating the tolerance level. Here, we report cloning of cytosolic putative HSP70 of 1678 bp from a thermotolerant cultivar (C306) of wheat (T. aestivum). A BLASTn search showed maximum homology with the predicted HSP70 protein reported from Hordeum vulgare (accession no AK354795.1). In silico characterization showed the presence of a nucleotide-binding domain of the sugar kinase/HSP70/actin superfamily in the sequence. Putative HSP70 showed temporal and spatial variations in the expression under heat stress (HS). We observed the abundance of HSP70 protein, H₂O₂, proline, and guaiacol peroxidase activity during the seed-hardening stage under HS; accumulation was, however, higher in the thermotolerant C306 than in thermosusceptible HD2329 cultivar. A gradual decrease in cell membrane stability (CMS) and an increase in total antioxidant capacity (TAC) were observed in both the cultivars at the different stages of growth. The expression of HSP70 showed a negative correlation with CMS and a positive correlation with TAC under HS; changes were less pronounced in C306 than in HD2329 at all the stages of growth studied. HSP70 seems to play diverse roles associated with thermotolerance, and partially protect wheat from terminal HS. Being the important member of family of the HSPs, HSP70 needs to be studied in detail, to be used for developing climate-smart wheat crops, through genetic engineering/breeding approaches.     

Genetic mapping in pea. 1. RAPD-based genetic linkage map of Pisum sativum

 A genetic linkage map of Pisum sativum L. was constructed based primarily on RAPD markers that were carefully selected for their reproducibility and scored in a population of 139 recombinant inbred lines (RILs). The mapping population was derived from a cross between a protein-rich dry-seed cultivar ‘Térèse’ and an increased branching mutant (K586) obtained from the pea cultivar ‘Torsdag’. The map currently comprises nine linkage groups with two groups comprising only 6 markers (n=7 in pea) and covers 1139 cM. This RAPD-based map has been aligned with the map based on the (JI281×JI399) RILs population that currently includes 355 markers in seven linkage groups covering 1881 cM. The difference in map lengths is discussed. For this alignment 7 RFLPs, 23 RAPD markers, the morphological marker le and the PCR marker corresponding to the gene Uni were used as common markers and scored in both populations. Received: 13 March 1998 / Accepted: 29 April 1998     

Chromosomal location of genes for resistance to powdery mildew in common wheat (Triticum aestivum L. em. Thell.) 4. Gene Pm 24 in Chinese landrace Chiyacao

 Chinese wheat landrace Chiyacao exhibited a response pattern different from that of the cultivars/lines possessing documented Pm genes after inoculation with 106 isolates of Erysiphe graminis f. sp. tritici. To characterize this resistance and to determine the chromosomal location of the gene or genes present, we crossed the landrace to susceptible cultivar ‘Chinese Spring’ and also to a set of 21 ‘Chinese Spring’ monosomic lines. Monosomic F₁ plants were allowed to self-pollinate and to produce F₂ seeds. Seedlings of F₂ plants and their parents were inoculated with isolates nos. 5 and 12 of Erysiphe graminis f. sp. tritici. The results revealed that one major dominant gene is located on chromosome 6D of Chinese common wheat landrace Chiyacao. The new gene is designated Pm 24. Received: 12 May 1997 / Accepted: 23 May 1997     

Genetic analysis of the dwarfing gene (Rht8) in wheat. Part I. Molecular mapping of Rht8 on the short arm of chromosome 2D of bread wheat (Triticum aestivum L.)

 Two sets of single chromosome recombinant lines comparing 2D chromosomes from the wheat varieties ‘Ciano 67’ and ‘Mara’ with the common 2D chromosome of ‘Cappelle-Desprez’ in a ‘Cappelle-Desprez’ background were used to detect a diagnostic wheat microsatellite marker for the dwarfing gene Rht8. The genetic linkage maps place the wheat microsatellite marker WMS 261 0.6 cM distal to Rht8 on the short arm of chromosome 2D. By PCR analysis the WMS 261 alleles of ‘Mara’, ‘Cappelle-Desprez’ and ‘Ciano 67’ could be distinguished by different fragment sizes of 192 bp, 174 bp and 165 bp, respectively. A screen of over 100 international varieties of wheat showed that the three allelic variants were all widespread. It also demonstrated that a limited number of varieties carried novel WMS 261 variants of over 200 bp. Following classification of the individual recombinant lines for allelic variants at the WMS 261 locus it was possible to attribute a 7- to 8-cm reduction in plant height with the WMS 261-192-bp allele compared to the WMS 261-174-bp allele in the set of recombinant lines comparing 2D chromosomes of ‘Mara’ and ‘Cappelle-Desprez’. A height reduction of around 3 cm was detected between the WMS 261-174-bp allele and the WMS 261-165-bp allele in the recombinant lines comparing 2D chromosomes of ‘Cappelle-Desprez’ and ‘Ciano 67’. Received: 17 October 1997 / Accepted: 12 November 1997     

A linkage map of the pea (Pisum sativum L.) genome containing cloned sequences of known function and expressed sequence tags (ESTs)

 A linkage map of the pea (Pisum sativum L.) genome is presented which is based on F₂ plants produced by crossing the marrowfat cultivar ‘Primo’ and the blue-pea breeding line ‘OSU442-15’. This linkage map consists of 209 markers and covers 1330 cM (Kosambi units) and includes RFLP, RAPD and AFLP markers. By mapping a number of anchor loci, the ‘Primo’בOSU442-15’ map has been related to other pea linkage maps. A feature of the map is the incorporation of 29 loci representing genes of known function, obtained from other laboratories. The map also contains RFLP loci detected using sequence-characterized cDNA clones developed in our laboratory. The putative identities of 38 of these cDNA clones were assigned by examining public-sequence databases for protein or nucleotide-sequence similarities. The conversion of sequence-characterized pea cDNAs into PCR-amplifiable and polymorphic sequence-tagged sites (STSs) was investigated using 18 pairs of primers designed for single-copy sequences. Eleven polymorphic STSs were developed. Received: 18 June 1997 / Accepted: 11 August 1997     

Use of the multi-allelic self-incompatibility gene in apple to assess homozygocity in shoots obtained through haploid induction

 To obtain homozygous genotypes of apple, we have induced haploid development of either the female or the male gametes by parthenogenesis in situ and anther culture, respectively. Of the shoots obtained, which were mainly of a non-haploid nature, some could be derived from fertilised egg cells or from sporophytic anther tissue. In order to select the shoots having a true haploid origin, and thus homozygotes, we decided to use the single multi-allelic self-incompatibility gene as a molecular marker to discriminate homozygous from heterozygous individuals. The rationale behind this approach was that diploid apple cultivars contain 2 different alleles of the S-gene and therefore the haploid induced shoots obtained from them should have only one of the alleles of the single parent. The parental cultivars used were ‘Idared’ (parthenogenesis in situ) and ‘Braeburn’ (androgenesis), and their S-genotypes were known, except for 1 of the ‘Braeburn’S-alleles. To stimulate parthenogenetic development ‘Idared’ styles were pollinated with irradiated ‘Baskatong’ pollen, the S-alleles of the latter (2n) cultivar were also unknown. The cloning and sequence analysis of these 3 unidentified S-alleles, 1 from ‘Braeburn’ and 2 from ‘Baskatong’ is described, and we show that they correspond to the S ₂₄ -, S ₂₆ - and S ₂₇ -alleles. We have optimised a method for analysis of the S-alleles of ‘Idared/Baskatong’- or ‘Braeburn’-derived in vitro plant tissues and have shown that this approach can be applied for the screening of the in vitro shoots for their haploid origin. Received: 18 August 1997 / Accepted: 10 September 1997     

Heat shock protein synthesis and thermal tolerance in wheat

Plants respond to high temperature stress by the synthesis of an assortment of heat shock proteins that have been correlated with an acquired thermal tolerance to otherwise lethal temperatures. This study was conducted to determine whether genotypic differences in acquired thermal tolerance were associated with changes in the pattern of heat shock protein synthesis. The pattern of heat shock protein synthesis was analyzed by ³⁵S-methionine incorporation in wheat (Triticum aestivum L.) varieties exhibiting distinct levels of acquired thermal tolerance. Significant quantitative differences between the cultivars Mustang and Sturdy were observed in the HSP exhibiting apparent molecular weights of 16, 17, 22, 26, 33, and 42 Kilodaltons. Genotypic differences in the synthesis of the small subunit of ribulose 1,5-bisphosphate carboxylase/oxygenase were observed at 34°C. Two-dimensional electrophoretic analysis revealed unique proteins (16, 17, and 26 kilodaltons) in the thermal tolerant variety Mustang that were absent in the more thermal sensitive variety Sturdy. These results provide a correlation between the synthesis of specific low molecular weight heat shock proteins and the degree of thermal tolerance expressed following exposure to elevated temperatures.     

Mapping QTLs for pre-harvest sprouting tolerance on chromosome 2D in a synthetic hexaploid wheat×common wheat cross

Based on segregation distortion of simple sequence repeat (SSR) molecular markers, we detected a significant quantitative trait loci (QTL) for pre-harvest sprouting (PHS) tolerance on the short arm of chromosome 2D (2DS) in the extremely susceptible population of F₂ progeny generated from the cross of PHS tolerant synthetic hexaploid wheat cultivar ‘RSP’ and PHS susceptible bread wheat cultivar ‘88–1643’. To identify the QTL of PHS tolerance, we constructed two SSR-based genetic maps of 2DS in 2004 and 2005. One putative QTL associated with PHS tolerance, designatedQphs.sau-2D, was identified within the marker intervalsXgwm261-Xgwm484 in 2004 and in the next year, nearly in the same position, between markerswmc112 andXgwm484. Confidence intervals based on the LOD-drop-off method ranged from 9 cM to 15.4 cM and almost completely overlapped with marker intervalXgwm261-Xgwm484. Flanking markers near this QTL could be assigned to the C-2DS1-0.33 chromosome bin, suggesting that the gene(s) controlling PHS tolerance is located in that chromosome region. The phenotypic variation explained by this QTL was about 25.73–27.50%. Genotyping of 48 F₆ PHS tolerant plants derived from the cross between PHS tolerant wheat cultivar ‘RSP’ and PHS susceptible bread wheat cultivar ‘MY11’ showed that the allele ofQphs.sau-2D found in the ‘RSP’ genome may prove useful for the improvement of PHS tolerance.     

Hybrid origin of some ornamentals of Allium subgenus Melanocrommyum verified with GISH and RAPD

 Random amplified polymorphic DNA (RAPD) and genomic in situ hybridization (GISH) methods have been used to verify the hybridogenic origin and to identify the parental species of some ornamental cultivars in the subgenus Melanocrommyum of the genus Allium. The cultivars had been selected from seed obtained after uncontrolled pollination in breeders’ fields. The combination of GISH analysis with RAPD markers is very suitable for testing the hybridogenic origin of plants and to ascertain the parental species of the hybrids in such cases. As suspected, A. macleanii and A. cristophii are the parental species of ‘Globemaster’. The parental species of cultivar ‘Globus’ are A. karataviense and A. stipitatum, and not A. cristophii and A. giganteum as has been assumed on morphological grounds. Cultivars ‘Lucy Ball’ and ‘Gladiator’ are of hybrid origin, though only one of the parental species, A. hollandicum, could be confirmed. The cultivars ‘Purple Sensation’, ‘Mount Everest’, ‘White Giant’, ‘Michael H. Hoog’ and ‘Mars’ are not hybrids since neither GISH nor RAPD suggest the presence of a second genome. ‘Purple Sensation’ belongs to A. hollandicum, ‘Mount Everest’, ‘White Giant’ and ‘Mars’ to A. stipitatum,‘Michael H. Hoog’ to A. rosenorum. Received: 3 July 1997 / Accepted: 9 October 1997     

Mapping of resistance to spot blotch disease caused by Bipolaris sorokiniana in spring wheat

Spot blotch caused by Bipolaris sorokiniana is a destructive disease of wheat in warm and humid wheat growing regions of the world. The development of disease resistant cultivars is considered as the most effective control strategy for spot blotch. An intervarietal mapping population in the form of recombinant inbred lines (RILs) was developed from a cross ‘Yangmai 6’ (a Chinese source of resistance) × ‘Sonalika’ (a spot blotch susceptible cultivar). The 139 single seed descent (SSD) derived F₆, F₇, F₈ lines of ‘Yangmai 6’ × ‘Sonalika’ were evaluated for resistance to spot blotch in three blocks in each of the 3 years. Joint and/or single year analysis by composite interval mapping (CIM) and likelihood of odd ratio (LOD) >2.2, identified four quantitative trait loci (QTL) on the chromosomes 2AL, 2BS, 5BL and 6DL. These QTLs were designated as QSb.bhu-2A, QSb.bhu-2B, QSb.bhu-5B and QSb.bhu-6D, respectively. A total of 63.10% of phenotypic variation was explained by these QTLs based on the mean over years. Two QTLs on chromosomes 2B and 5B with major effects were consistent over 3 years. All QTL alleles for resistance were derived from the resistant parent ‘Yangmai 6’.     

b-D-glucosidase activity in pelargonium and poinsettia leaves infected by <Emphasis Type="Italic">Botrytis cinerea</Emphasis> Pers. ex Fr.

The involvement of β-D-glucosidase activity in grey mould was studied in two ornamental plant species attacked by Botrytis cinerea. β-D-glucosidase activity in the susceptible pelargonium cultivar ‘Shiva’ gradually increased with the disease development in the leaf spots and their surroundings. The endogenic level of the studied hydrolase in the resistant pelargonium ‘Cascade’ was several times higher than in the susceptible cultivar ‘Shiva’ and in principle underwent no changes after inoculation. The postinfection increase in the activity of β-D-glucosidase noted in the leaves of the susceptible poinsettia cultivar ‘Malibu Red’ was evidently weaker in the intensity, but its tendencies were similar to those of the susceptible pelargonium cultivar. In the leaves of the medium-resistant poinsettia ‘Coco White’ the constitutional level of β-D-glucosidase was 2-3-fold higher in that cultivar than in the susceptible cv. ‘Malibu Red’. In attacked leaves of ‘Coco White’, the enzyme activity continued to increase temporarily until the 3^rd h after inoculation. The process of healthy leaf senescence in both species had no significant influence on the change of the studied enzyme activity which was generally low. A high activity of β-D-glucosidase was also observed in the homogenate prepared from mycelium and in the fungal spores.     

Back-cross reciprocal monosomic analysis of Fusarium head blight resistance in wheat (Triticum aestivum L.)

 Fusarium head blight (FHB or scab) caused by Fusarium spp. is a widespread disease of cereals causing yield and quality losses and contaminating cereal products with mycotoxins. The breeding of resistant varieties is the method of choice for controlling the disease. Unfortunately, the genetic basis of scab resistance is still poorly understood. We present the results of a back-cross reciprocal monosomic analysis of FHB resistance using the highly resistant Hungarian winter wheat line ‘U-136.1’ and the highly susceptible cultivar ‘Hobbit-sib’. Resistance testing was performed in a field trial artificially inoculated with a Fusarium culmorum conidial suspension. Five hemizygous families containing ‘U-136.1’ chromosomes 6B, 5A, 6D, 1B, and 4B had a visually reduced spread of infection compared to lines having the ‘Hobbit-sib’ chromosome. Chromosome 2B from ‘U-136.1’ had an increased spread of infection. The critical chromosomes controlling seed weight were 6D, 3B, 5A, and 6B while those controlling deoxynivalenol (DON) content were homoeologous groups 2 and 6, although the latter effects were not significant due to a high coefficient of variation. Results from this and other studies show that chromosomes 6D, 6B, 5A, 4D, and 7A have frequently been associated with scab resistance in a number of wheat cultivars. Research groups now attempting to map scab resistance in wheat using markers should pay special attention to the above-mentioned chromosomes. Received: 31 March 1998 / Accepted: 14 July 1998     

Effects of drought and salinity on some growth-contributing parameters in wheat and barley

Summary This study is mainly concerned with some parameters contributing to growth as indicators of difference in drought resistance and salt tolerance of wheat and barley cultivars. Parameters made use of are: transpiration efficiency (‘dry matter/transpiration’ ratio), ‘leaf/root’ ratio, chlorophyll content, chlorophyll stability to heat and anatomical modifications. The results revealed that transpiration efficiency is much higher in mexican ‘super-x’ wheat than in the egyptian ‘Giza-155’ cultivar under reduced soil water matric- or osmotic potentials. Chlorophyll content increased in super-x with decreasing soil water potential while chlorophyll heat stability decreased. The reverse is true in Giza-155 cultivar. Decreased leaf/root ratio in super-x is interpretted in favour of more beneficial water balance in this cultivar. Development of more sclerenchyma in its stems supports this judgement. Of barley cultivars tested, Borg El-Arab is favoured for drought resistance and Giza-117 for salt tolerance. re]19751014     

Genetic studies of Triticeae dehydrins: assignment of seed proteins and a regulatory factor to map positions

 A collection of 200 wheat (Triticum aestivum L. cv ‘Chinese Spring’) cytogenetic stocks (nullisomic, tetrasomic, nulli-tetrasomic, ditelosomic and deletion lines, addition and substitution stocks from intra- and inter-specific crosses) was utilized to determine the proteins encoded by some of the wheat and barley dehydrin genes, using a western blot procedure. Proteins extracted from seeds were reacted with antibodies that recognize dehydrins in a wide range of plants, including wheat and barley. Proteins encoded by dehydrin loci in chromosome arms 4DS, 5BL and 6AL of ‘Chinese Spring’ wheat were assigned by this method. There was also evidence of a regulatory factor on 5B in the vicinity of the dhn genes, and on 5H in wheat-barley addition lines, that is required for a normal level of expression of seed dehydrins in hexaploid wheat. Further understanding of this putative regulatory factor would be helpful for the interpretation of linkage studies that may relate dehydrin gene expression to phenotypes such as dehydration, salinity or low-temperature tolerance. Received: 27 August 1997 / Accepted: 4 February 1998     

Comparative mapping of the wheat chromosome 5A Vrn-A1 region with rice and its relationship to QTL for flowering time

 The vernalization gene Vrn-A1 on chromosome 5A is the predominant gene determining the spring/winter habit difference in bread wheat. Vrn-A1 was physically mapped using a set of deletion lines which located it to the region of chromosome 5A flanked by deletion breakpoints 0.68 and 0.78. This interval was shown to be homoeologous to a region of rice chromosome 3 that contains the flowering-time QTL Hd-6, previously mapped in a Nipponbare×Kasalath cross, and FLTQ1, a novel QTL identified by analysis of 78 F₃ families derived from a cross of ‘IR20’ב63–83’. Possible relationships between Vrn-A1 and rice QTL are discussed. Analysis of the chromosome 5A deletion lines showed evidence for a second, more proximal flowering-time effect located between deletion breakpoints 0.56 and 0.64. The proximal part of chromosome 5A is homoeologous to rice chromosome 9, on which two QTL were detected in the ‘IR20ב63–83’ cross. The possible relationship between these effects is also discussed. Received: 23 December 1997 / Accepted: 12 January 1998     

Over-expression of the cloned rice thaumatin-like protein (PR-5) gene in transgenic rice plants enhances environmental friendly resistance to Rhizoctonia solani causing sheath blight disease

 A 1.1-kb DNA fragment containing the coding region of a thaumatin-like protein (TLP-D34), a member of the PR-5 group, was cloned into the rice transformation vector pGL2, under the control of the CaMV 35S promoter. The Indica rice cultivars, ‘Chinsurah Boro II’, ‘IR72’, and ‘IR51500’ were transformed with the tlp gene construct by PEG-mediated direct gene transfer to protoplasts and by biolistic transformation using immature embryos. The presence of the chimeric gene in T₀, T₁, and T₂ transgenic plants was detected by Southern blot analysis. The presence of the expected 23-kDa TLP in transgenic plants was confirmed by Western blot analysis and by staining with Coomassie Brilliant Blue. Bioassays of transgenic plants challenged with the sheath blight pathogen, Rhizoctonia solani, indicated that over-expression of TLP resulted in enhanced resistance compared to control plants. Received: 11 August 1998 / Accepted: 26 August 1998