Effects of light, temperature, gibberellin (GA3) and their interaction on coleoptile and leaf elongation of tall, semi-dwarf and dwarf wheat

Near-isogenic wheat lines differing in height-reducing (Rht) alleles, in each of two cultivars, were used to investigate the effects of light intensity and of their interaction with temperature and GA₃ application, on the elongation of the coleoptile and the first seedling leaf. Darkness caused a conspicuous increase in the lengths of the coleoptile and of the sheath and lamina of the first leaf, in GA₃ treated and untreated seedlings of all genotypes grown at 11 and 25°C. The genotype effects and the effects of light intensity and GA₃ application on leaf length were ascribed entirely to their effects on the rate of leaf elongation since the duration of leaf elongation was not affected by these factors. Temperature elevation from 11 to 25°C caused a 55% shortening of the duration of leaf elongation and a concomitant increase in elongation rate, which diminished with increased genotypic dwarfness. Accordingly, temperature elevation resulted in a significant reduction in leaf-length of the light-grown dwarf genotypes and the dark-grown dwarf and semi-dwarf genotypes. It is suggested that this temperature × light × genotype interaction effect is due to environmental dependent upper limits of elongation rate set by the Rht alleles.Abbreviations PAR Photosynthetic Active Radiation     

Carbon monoxide sensitivity of cytochrome oxidase in wheat leaves in relation to dwarfing genes (Rht) in wheat cultivars

Summary Leaves of young seedlings of a number of tall cultivars of wheat, lacking the dwarfing Rht genes, readily responded to a brief 2 min exposure to CO, as assessed by in vivo aerobic assay of nitrate reductase. This test depends on the inhibition of cytochrome c oxidase by CO, which in turn renders cytosolic NADH available for the reduction of nitrate to nitrite in vivo. Semi-dwarf cultivars of wheat (Rht present) did not respond to CO in this way. Since CO forms a complex only with reduced cytochrome a₃, the results indicate differences in the redox state of cytochrome a₃, during in situ respiration of leaves from tall and semi-dwarf plants which are likely to be under genetic control.     

Genetic analysis of the anther-culture response of three spring wheat crosses

Summary Anther-culture response was examined among three spring wheat (Triticum aestivum L.) cultivars to evaluate the genetic component of response and to determine whether androgenetic performance could be improved by selection. The three lines, the three possible F₁'s among the three lines, their F₂'s, and the backcrosses to the parents were evaluated for callus production and regeneration capacity. Significant variation was observed among the generations of the three crosses for callus formation. Genetic variation for regenerability was nonsignificant. Callus production was negatively correlated (-0.24) with regeneration capacity. The random variation in the study was too great to determine whether major-gene differences for antherculture response exist among the three lines by examining population distributions. When the material was evaluated for quantitative gene effects, the estimates for the additive gene effects were generally greater than the estimates for the dominance gene effects for callus formation. Only the Pavon x Chris cross, however, exhibited a significant narrow sense heritability estimate for callusing response (0.94). Due to the large component of random variation and the varying selection potential among crosses for androgenetic performance, improving anther-culture response in wheat by selection could prove difficult unless the anther-culture process itself selects for response traits at the gametic level.     

Influence of the gibberellin-sensitive Rht8 dwarfing gene on leaf epidermal cell dimensions and early vigour in wheat (Triticum aestivum L.)

BACKGROUND AND AIMS: The gibberellin-insensitive Rht-B1b and Rht-D1b dwarfing genes are known to reduce the size of cells in culms, leaves and coleoptiles of wheat. Resulting leaf area development of gibberellin-insensitive wheats is poor compared to standard height (Rht-B1a and Rht-D1a) genotypes. Alternative dwarfing genes to Rht-B1b and Rht-D1b are available that reduce plant height, such as the gibberellin-responsive Rht8 gene. This study aims to investigate if Rht8 has a similar dwarfing effect on the size of leaf cells to reduce leaf area. METHODS: The effect of Rht8 on cell size and leaf area was assessed in four types of epidermal cells (interstomatal, long, sister and bulliform) measured on leaf 2 of standard height (rht8) and semi-dwarf (Rht8) doubled-haploid lines (DHLs). The DHLs were derived from a cross between very vigorous, standard height (rht8) ('Vigour18') and less vigorous, semi-dwarf (Rht8) ('Chuan-Mai 18') parents. KEY RESULTS: Large differences were observed in seedling vigour between the parents, where 'Vigour18' had a much greater plant leaf area than 'Chuan-Mai 18'. Accordingly, 'Vigour18' had on average longer, wider and more epidermal cells and cell files than 'Chuan-Mai 18'. Although there was correspondingly large genotypic variation among DHLs for these traits, the contrast between semi-dwarf Rht8 and tall rht8 DHLs revealed no difference in the size of leaf 2 or average cell characteristics. Hence, these traits were independent of plant height and therefore Rht8 in the DHLs. Correlations for leaf and average cell size across DHLs revealed a strong and positive relationship between leaf width and cell files, while the relationships between leaf and cell width, and leaf and cell length were not statistically different. The relative contribution of the four cell types (long, sister, interstomatal and bulliform) to leaf size in the parents, comparative controls and DHLs is discussed. CONCLUSIONS: Despite a large range in early vigour among the DHLs, none of the DHLs attained the leaf area or epidermal cell size and numbers of the vigorous rht8 parent. Nonetheless, the potential exists to increase the early vigour of semi-dwarf wheats by using GA-sensitive dwarfing genes such as Rht8.     

Gibberellin status and responsiveness in shoots of tall and dwarf genotypes of diploid rye (Secale cereale)

The recessive dwarfing alleles of rye ( Secale cereale L.), ct1 and ct2 , caused a 35–55% reduction in the length of leaf 2 compared with corresponding tall lines grown at both 10°C and 20°C. The dwarf lines were 45–50% as responsive to applied GA₃ as the tall lines at 20°C but the absolute GA-responsiveness of the dwarfs was greater at 10°C than at 20°C. There was no significant difference in the contents of GA₁₉, GA₂₀, GA₂₉, GA₁, GA₃ and GA₈ in the leaf extension zone of tall and dwarf seedlings grown at 20°C. It was concluded that the mechanism whereby GA homeostasis is maintained is functional in both tall and dwarf lines despite marked differences in leaf extension rate. The recessive rye mutations may cause loss of function late in the GA-cell elongation pathway or, alternatively, indirectly affect GA-responsiveness in vegetative tissues. The genetic and physiological evidence indicates that ct1 and ct2 are unrelated to the GA-insensitive Rht genes in hexaploid bread wheat.     

Modelling potassium uptake by wheat (Triticum aestivum) crops

Spring wheat was grown in the field under deficient and sufficient levels of soil K and with high and low supplies of fertiliser nitrogen. Measurements were made of K uptake, soil nutrient supply parameters, root growth and, in solution culture, root influx parameters. Mechanistic models predicted uptake reasonably well under K-deficient conditions, but over-predicted uptake, by as much as 4 times, under K-sufficient conditions. The over-prediction was apparently due to poor characterisation of plant demand.     

Meiosis and fertility of F1 hybrids between hexaploid bread wheat and decaploid tall wheatgrass (Thinopyrum ponticum)

As the first step in the transfer of barely yellow dwarf virus resistance and salt tolerance from decaploid tall wheatgrass (Thinopyrum ponticum) into hexaploid bread wheat (Triticum aestivum L.), octoploid intergeneric hybrids (2n = 8x = 56) were synthesized by crossing the tall wheatgrass cultivar Alkar with wheat cvs. Fukuhokomugi (Fuko) and Chinese Spring. (Fuko x Alkar) F₁ hybrids were studied in detail. The F₁ hybrids were perennial and generally resembled the male wheatgrass parent with regard to morphological features and gliadin profile. Most hybrids were euploid with 56 chromosomes and showed high chromosome pairing. On an average, in 6 hybrids 83.6% of the complement showed chiasmatic association, some between wheat and wheatgrass chromosomes. Such a high homoeologous pairing would be obtained if Ph1, the major homoeologous pairing suppressor in wheat, was somehow inactivated. Some of the Fuko x Alkar hybrids had high pollen fertility (18.5–42.0% with a mean of 31.5%) and high seed fertility (3–29 seeds wtih a mean of 12.3 seeds per spike), offering excellent opportunities for their direct backcrossing onto the wheat parent.     

Genetic analysis of photosynthetic variation in hexaploid and tetraploid wheat and their interspecific hybrids

Intra- and inter-specific variation in CO₂ assimilation rate (A) in Triticum spp. is well documented for reproductive growth stages. Research was conducted to characterize early vegetative photosynthetic variation in a diverse set of cultivated hexaploid wheat (T. aestivum L.) germplasm and in wild tetraploid (T. dicoccoides Korn) and hexaploid x tetraploid populations. Choice of hexaploid genotypes was based on maximum genetic distance between cultivars within the HRW and SRW wheat classes of the USA. The tetraploid material was produced by hybridizing two accessions of T. dicoccoides previously shown to differ widely in A and A/Chl but with similar leaf morphology. Genetic variability in the HRW and SRW gene pools was attributed to more recently developed descendent lines and unrelated lines rather than parental lines. Phenotypic distributions for A, stomatal conductance (gs), and internal CO₂ concentration (Ci) in the F₂ tetraploid population were continuous and showed transgressive segregation, reflecting quantitative inheritance with intermediate heritability. Variability in A was not associated with chlorophyll content or CO₂ supply to the mesophyll measured as Ci. Genetic variability in A was also observed in the interspecific backcross population, 2*TAM W-101/PI 428109, thereby providing a germplasm pool to select for high A while restoring the D genome of hexaploid wheat. These results suggest that genetic improvement of vegetative assimilation rate is feasible in hexaploid wheat via homologous transfer from an alien source.Abbreviations HRW hard red winter - LA leaf area - r_G genotypic correlation - r_P phenotypic correlation - SRW soft red winter     

Inheritance of partial resistance to powdery mildew in spring wheat

Summary Four spring wheat (Triticum aestivum L.) cultivars exhibiting partial resistance to powdery mildew induced by Erysiphe graminis f.sp. tritici were crossed to a common susceptible cultivar to study the inheritance of resistance. The genetic parameters contributing to resistance were estimated by generation means analyses. Additive gene action was the most important genetic component of variation among generation means in all four crosses. Additive by additive effects were significant in one cross and both additive by additive and additive by dominance effects were significant in another. Dominance effects were not significant. The F2/F3 correlations in three crosses ranged from 0.27 to 0.43. Three additional crosses among resistant cultivars were employed to study the effectiveness of selection in improving resistance. By selecting the most resistant plants from the F2 and evaluating the progenies in the F4, increases in resistance ranging from 21% to 31% were obtained. In all crosses, there was transgressive segregation in both directions indicating that the genes conferring resistance to these cultivars differ and exhibit additive effects.     

Copper supply and the leaf emergence rate of spring wheat

Copper deficiency can delay flowering and plant maturity. However, the effect of copper deficiency on the rate of leaf emergence has not been quantified. We tested the hypothesis that low copper supply decreases the rate of leaf emergence of wheat (Triticum aestivum L. cv Gamenya). Copper foliar sprays are commonly applied to wheat. We examined the response of the rate of leaf emergence to a foliar application of copper sulphate.Wheat was grown in root cooling tanks (20°C) in the glasshouse. Soil copper treatments were applied as solutions of CuSO₄.5H₂O at three rates: Cu₀=no added Cu, Cu₄₀₀=400 g Cu per 3 kg pot, and Cu₁₆₀₀=1600 g Cu per pot. An additional treatment of a foliar spray of CuSO₄.5H₂O (0.4 mg Cu per plant) was applied to Cu₀ and Cu₄₀₀ plants 45 days after sowing (5.5 leaves on the main stem). Leaves on the main stem were counted and the rate of leaf emergence was estimated from the regression of number of emerged leaves against thermal time (base 0°C). The phyllochron was calculated as 1/rate of emergence.Leaves on Cu₀ and Cu₄₀₀ plants took longer to emerge than on Cu₁₆₀₀ plants, with the phyllochron of Cu₁₆₀₀ plants being 130 compared to 137 for the Cu₄₀₀ plants and 158 for the Cu₀ plants. The foliar application of CuSO₄ at the 5–6 leaf stage resulted in a decrease in the phyllochron of the Cu₀ plants to 127, but no change in that of the Cu₄₀₀ plants.     

Fertilizer nitrogen budgets of 15N-labelled sugarbeet (Beta vulgaris) tops and Na 15NO3 dressings split-applied to winter wheat (Triticum aestivum) in microplots on a loam soil

The fate of N from sugarbeet (Beta vulgaris L.) tops returned to the soil (50 T ha^-1) in autumn 1986 before sowing winter wheat (Triticum aestivum L.), and from NaNO₃ split-applied in 3 equal dressings (at tillering, stem elongation and flag leaf stages) was studied using isotopically labelled ¹⁵N in open stainless-steel cylinders pressed into the soil.At harvest, the percentage utilization (PU) of N from sugarbeet was very low (6.66%) and negatively influenced by fertilizer N (5.59%), while that of fertilizer N was rather high (69.64%) and unchanged by addition of tops. Residual N in soil represented 25.9% of the amount applied in tops and ranged from 33% for the tillering application to 21% for the flag leaf application. N losses (mainly denitrification) from sugar beet tops amounted to 67% and were very low for mineral fertilizer (less than 5%).     

Effect of Nitrogen and Water Stress on Photosynthesis and Nitrogen Content in Wheat

The relationships between photosynthesis, leaf nitrogen content and water stress were studied in ten genotypes of wheat differing in the presence of dwarfing genes. Net photosynthetic rate (P_N) was mostly higher at ear emergence stage than at anthesis stage. P_N decreased with water stress (leaf water potential from –2.0 to –2.5 MPa), and with reduced leaf N content in all genotypes studied. Among the various genotypes, single dwarf and wild types showed higher P_N rate and maintained higher leaf N content under different N doses and water supply as compared to the other types studied.     

Molecular characterization of the fate of transgenes in transformed wheat (Triticum aestivum L.)

Molecular analysis of the transgenes bar and gus was carried out over successive generations in six independent transgenic lines of wheat, until the plants attained homozygosity. Data on expression and integration of the transgenes is presented. Five of the lines were found to be stably transformed, duly transferring the transgenes to the next generation. The copy number of the transgenes varied from one to five in the different lines. One line was unstable, first losing expression of and then eliminating both the transgenes in R3 plants. Although the gus gene was detected in all the lines, GUS expression had been lost in R2 plants of all but one line. Rearrangement of transgene sequences was observed, but it had no effect on gene expression. All the stable lines were found to segregate for transgene activity in a Mendelian fashion.     

Genotypic differences in phosphorus efficiency of wheat

In an attempt to evaluate whether breeding and selection for high yielding capacity did change the P requirements of modern wheat cultivars, the response of two wheat cultivars to different levels of P supply was investigated. A traditional cultivar ("Peragis") and a modern cultivar ("Cosir") were cultivated in a C-loess low in available P and high in CaCO₃ in 120 cm high PVC pots. Shoot and root growth at different developmental stages was compared. The grain yield of the modern cultivar Cosir was higher at limiting and non-limiting P supply and, therefore, this cultivar can be considered as more P-efficient than the traditional cultivar. From the results it can be concluded that the main factors contributing to the higher P efficiency of the modern cultivar are (i) efficient use of assimilates for root growth characteristics which enhance P acquisition: smaller root diameter, and longer root hairs, (ii) efficient remobilization of P from vegetative plant organs to the grains, and (iii) lower P requirement for grain yield formation because of lower ear number per plant but higher grain number per ear.     

Genetically controlled differences in the effects of chlormequat on tetraploid wheat (Triticum turgidum)

Two crosses between Triticum turgidum wheat lines differing in their response to chlormequat (CCC) were tested. In the F₂ population of one cross, which was segregating for the Rht1 dwarfing allele, each plant was cloned by separation of two tillers, one of which was treated with CCC. The tall (rht1/rht1) and the intermediate (Rht1/rht1) genotypes showed a greater response to CCC than the semi-dwarf (Rht1/Rht1) genotype, as expressed by culm length and date of ear emergence. The F₃ families of another cross and their two semi-dwarf parents were grown in a three-replicated field test in paris of rows, one of which was treated with CCC. In one of the parents and in 1/4 of the F₃ families CCC induced a wide-angled tiller growth, suggesting a monogenic control of this growth habit in response to CCC.Based on an M.Sc. thesis presented by the senior author to the Faculty of Agriculture of The Hebrew University of Jerusalem.     

Accelerated production of transgenic wheat (Triticum aestivum L.) plants

We have developed a method for the accelerated production of fertile transgenic wheat (Triticum aestivum L.) that yields rooted plants ready for transfer to soil in 8–9 weeks (56–66 days) after the initiation of cultures. This was made possible by improvements in the procedures used for culture, bombardment, and selection. Cultured immature embryos were given a 4–6 h pre-and 16 h post-bombardment osmotic treatment. The most consistent and satisfactory results were obtained with 30 g of gold particles/bombardment. No clear correlation was found between the frequencies of transient expression and stable transformation. The highest rates of regeneration and transformation were obtained when callus formation after bombardment was limited to two weeks in the dark, with or without selection, followed by selection during regeneration under light. Selection with bialaphos, and not phosphinothricin, yielded more vigorously growing transformed plantlets. The elongation of dark green plantlets in the presence of 4–5 mg/l bialaphos was found to be reliable for identifying transformed plants. Eighty independent transgenic wheat lines were produced in this study. Under optimum conditions, 32 transformed wheat plants were obtained from 2100 immature embryos in 56–66 days, making it possible to obtain R3 homozygous plants in less than a year.     

Gibberellins and leaf expansion in near-isogenic wheat lines containing Rht1 and Rht3 dwarfing alleles

In near-isogenic lines of winter wheat (Triticum aestivum L. cv. Maris Huntsman) grown at 20° C under long days the reduced-height genes, Rht1 (semi-dwarf) and Rht3 (dwarf) reduced the rate of extension of leaf 2 by 12% and 52%, respectively, compared with corresponding rht (tall) lines. Lowering the growing temperature from 20° to 10° C reduced the rate of linear extension of leaf 2 by 2.5-fold (60% reduction) in the rht3 line but by only 1.6-fold (36% reduction) in the Rht3 line. For both genotypes, the duration of leaf expansion was greater at the lower temperature so that final leaf length was reduced by only 35% in the rht3 line and was similar in the Rht3 line at both temperatures. Seedlings of the rht3 (tall) line growing at 20° C responded positively to root-applied gibberellin A₁ (GA₁) in the range 1–10 μM GA₁; there was a linear increase in sheath length of leaf 1 whereas the Rht3 (dwarf) line remained unresponsive. Gibberellins A_{1, 3, 4, 8, 19, 20, 29, 34, 44} and ₅₃ were identified by full-scan gas chromatography-mass spectrometry in aseptically grown 4-d-old shoots of the Rht3 line. In 12-d-old seedlings grown at 20° C, there were fourfold and 24-fold increases in the concentration of GA₁ in the leaf expansion zone of Rht1 and Rht3 lines, respectively, compared with corresponding rht lines. Although GA₃ was present at a similar level to GA₁ in the rht3 (tall) line it accumulated only fivefold in the Rht3 (dwarf) line. The steady-state pool sizes of endogenous GAs were GA₁₉ ? GA₂₀ = GA₁ in the GA-responsive rht3 line whereas in the GA non-responsive Rht3 line the content of GA₁₉≈ GA₂₀ ? GA₁. It is proposed that one of the consequences of GA₁ action is suppression of GA₁₉-oxidase activity such that the conversion of GA₁₉ to GA₂₀ becomes a rate-limiting step on the pathway to GA₁ in GA-responsive lines. In the GA-non-responsive Rht lines it is suggested that GA₁₉ oxidase is not downregulated to the same extent and GA₁ accumulates before the next rate-limiting step on the pathway, its 2β-hydroxylation to GA₈. The steady-state pool sizes of GA_{19, 20, 1, 3} and ₈ were similar in developmentally equivalent tissues of the rht3 (tall) line growing at 10° C and 20° C despite a 2.5-fold difference in the rate of leaf expansion. In contrast, in the Rht3 (dwarf) line, the extent of accumulation of GA₁ reflected the severity of the phenotype at the two temperatures with slower growing tissues accumulating less, not more, GA₁. These results are interpreted as supporting the proposed model of regulation of the GA-biosynthetic pathway rather than previous suggestions that GA₁ accumulates in GA-insensitive dwarfs as a consequence of reduced growth rates.     

Epistatic,additive and dominance variation in a triple test cross of bread wheat (Triticum aestivum L.)

Summary Triple test cross progenies resulting from the crossing of three testers (Kloka, UP 368 and an f₁ intermediate between them) and 24 varieties of bread wheat have been studied for plant height (cm), peduncle length (cm), ear length (cm), number of spikelets per spike and harvest index (ratio between economic and total yield). Epistasis was not significant for any of the characters studied. The testers were inadequate for plant height and for peduncle length although the testers varied considerably for these traits. Additive variance played a significant role in the inheritance of all the characters except number of spikelets per spike. The dominance variance was important for plant height, ear length and harvest index. The degree of dominance was in the over-dominance range for plant height. Complete dominance was operative for ear length, number of spikelets per spike and harvest index whereas for peduncle length only partial dominance was observed. The possibility of the isolation of the recombinants with high harvest index has been stressed.