Growth and gibberellin-A1 metabolism in normal and gibberellin-insensitive (Rht3) wheat (Triticum aestivum L.) seedlings

Growth parameters were determined for tall (rht3) and dwarf (Rht3) seedlings of wheat (Triticum aestivum L.). Plant statures and leaf length were reduced by 50% in dwarfs but root and shoot dry weights were less affected. Leaves of dwarf seedlings had shorter epidermal cells and the numbers of cells per rank in talls and dwarfs matched the observed relationships in overall length. Talls grew at twice the rate of dwarfs (2.3 compared with 1.2 mm h^-1). [³H]Gibberellin A₁ ([³H]GA₁) was fed to seedlings via the third leaf and metabolism was followed over 12 h. Immature leaves of tall seedlings transferred radioactivity rapidly to compounds co-chromatographing with [³H]gibberellin A₈ ([³H]GA₈) and a conjugate of [³H]GA₈, whereas leaves of dwarf seedlings metabolised [³H]GA₁ more slowly. Roots of both genotypes produced [³H]GA₈-like material at similar rates. Isotopic dilution studies indicated a reduced 2-hydroxylation capacity in dwarfs, but parallel estimates of the endogenous GA pool size, obtained by radioimmunoassay, indicated a 12–15 times higher level of GA in the dwarf immature leaves. Dwarfing by the Rht3 gene does not appear to operate through enhanced, or abnormal metabolism of active gibberellins and the act of GA metabolism does not bear an obligate relationship to the growth response.Abbreviations GA_n gibberellin A_n - HPLC high-performance liquid chromatography     

Genotypic variation in the responsiveness to GA3 within tall (rht1) and semi-dwarf (Rht1) spring wheat

The effect of GA₃ on coleoptile-and first leaf elongation of tall (rht1) and semi-dwarf (Rht1) nearly-isogenic genotypes, within each of 25 random F₉ wheat families, was determined on seedlings grown in a growth room at 18 °C. Conspicuous and very significant inter-family variation in the response of the first leaf to GA₃ application was found in both the rht1 and Rht1 genotypes. The magnitudes of the response of the different families within genotypes to GA₃ were not related to the leaf length of their untreated seedlings. It is suggested that, under given environmental conditions, background genotypic effects, inducing inter-family variation in responsiveness to GA₃, regulate the elongation growth up to the limits set by the Rht alleles.     

Exogenous GA3 Application Can Compensate the Morphogenetic Effects of the GA-Responsive Dwarfing Gene Rht12 in Bread Wheat

The most common dwarfing genes in wheat, Rht-B1b and Rht-D1b, classified as gibberellin-insensitive (GAI) dwarfing genes due to their reduced response to exogenous GA, have been verified as encoding negative regulators of gibberellin signaling. In contrast, the response of gibberellin-responsive (GAR) dwarfing genes, such as Rht12, to exogenous GA is still unclear and the role of them, if any, in GA biosynthesis or signaling is unknown. The responses of Rht12 to exogenous GA₃ were investigated on seedling vigour, spike phenological development, plant height and other agronomic traits, using F_2∶3 and F_3∶4 lines derived from a cross between Ningchun45 and Karcagi-12 in three experiments. The application of exogenous GA₃ significantly increased coleoptile length and seedling leaf 1 length and area. While there was no significant difference between the dwarf and the tall lines at the seedling stage in the responsiveness to GA₃, plant height was significantly increased, by 41 cm (53%) averaged across the three experiments, in the GA₃-treated Rht12 dwarf lines. Plant height of the tall lines was not affected significantly by GA₃ treatment (<10 cm increased). Plant biomass and seed size of the GA₃-treated dwarf lines was significantly increased compared with untreated dwarf plants while there was no such difference in the tall lines. GA₃-treated Rht12 dwarf plants with the dominant Vrn-B1 developed faster than untreated plants and reached double ridge stage 57 days, 11 days and 50 days earlier and finally flowered earlier by almost 7 days while the GA₃-treated tall lines flowering only 1–2 days earlier than the untreated tall lines. Thus, it is clear that exogenous GA₃ can break the masking effect of Rht12 on Vrn-B1 and also restore other characters of Rht12 to normal. It suggested that Rht12 mutants may be deficient in GA biosynthesis rather than in GA signal transduction like the GA-insensitive dwarfs.     

Rht3基因及带有Rht3基因的4B染色体对普通小麦光合碳同化特性的影响

本文系统地研究了带有Ｒｈｔ３基因的４Ｂ染色体二体（宁矮１号）、单体（宁矮１号Ｍ４Ｂ）、缺体（宁矮１号Ｎ４Ｂ）材料的光合特性,发现带有Ｒｈｔ３基因的４Ｂ染色体对光合速率、叶绿素含量、ＲｕＢＰ羧化酶含量及活性、叶片导度均有正效应,并有累加作用,而且对叶绿素含量缓降期和光合速率高值持续期也有正效应,因此带有Ｒｈｔ３基因的４Ｂ染色体具有促进光合作用的效应。对具有不同Ｒｈｔ３基因剂量的矮秆系（宁矮１号,即苏麦３号的Ｒｈｔ３矮秆等基因系）、半矮秆系（ＭＤ苏麦３号）及其苏麦３号的光合碳同化特性的研究,发现Ｒｈｔ３基因对光合速率、叶绿素含量、叶片导度具有正效应,但对于ＲｕＢＰ羧化酶含量和活性、叶绿素含量缓降期、光合速率高值持续期有负效应。     

Shoot elongation in Lathyrus odoratus L.: Gibberellin levels in light- and dark-grown tall and dwarf seedlings

The levels of gibberellin A₁ (GA₁), GA₂₀, GA₁₉, GA₈, GA₂₉ and GA₈₁ (2-epiGA₂₉) were measured in tall (L-) and dwarf (ll) sweet-pea plants grown in darkness and in light. In both environments the apical portions of dwarf plants contained less GA₁; GA₈ and GA₁₉, but more GA₂₀, GA₂₉, and GA₈₁ than did those of tall plants. It is concluded that the partial block in 3β-hydroxylation of GA₂₀ to GA₁ is imposed by allele l in darkness as well as in the light. Furthermore, darkness does not appear to enhance elongation in sweet pea by increasing GA₁ levels. The reduction of the pool size of GA₁₉ in dwarf plants supports recent theories on the regulation of GA biosynthesis, formulated on the basis of observations in monocotyledonous species. Darkness results in decreased GA₂₀, GA₂₉, and GA₈₁ levels in the apical portions of tall and dwarf plants and possible reasons for this are discussed.     

Influence of Gibberellic Acid and the Rht3 Gene on Choline and Phospholipid Metabolism in Wheat Aleurone Tissue

The effect of gibberellic acid (GA3) on phospholipid metabolismand -amylase production was studied in aleurone tissue of twonear-isogenic lines of wheat (Triticum aesuvum L.). Incubationof embryoectomized seeds from a GA-responsive line (rht3, tall)with GA₃ caused the induction of -amylase activity after a lagphase of 30 h. In the case of embryoectomized seeds from a ‘GA-insensitive’line (Rh13, dwarf), however, the lag phase was extended up to50 h. During the first 14 h following imbibition, GA₃ inhibitedcholine uptake and its subsequent incorporation into phosphatidylcholine in the Rhr3 line but not in the rht3 line. GA₃ promotedphospholipid breakdown in both the lines during this period,however. GA₃ also terminated independent turnover of the cholineN-methyl groups in phosphatidyl choline and promoted turnoverof the whole choline headgroup. These results are discussedin relation to the possibility that phosphatidyl choline turnoveris an integral part of the GA₃ signal-transduction mechanismin aleurone tissue. Key words: GA3, Rht3 gene, choline, phospholipid     

Comparison of IAA-Induced and Low Temperature-Induced GA(3) Responsiveness and alpha-Amylase Production by GA(3) Insensitive Dwarf Wheat Aleurone

Rht3-containing gibberellic acid (GA₃) insensitive deembryonated wheat (Triticum aestivum L. var Cappelle Desprez × Minister Dwarf) aleurone, that can be made responsive to GA₃ by low temperature, can also be rendered GA₃ sensitive by preincubation with indoleacetic acid (IAA). The IAA-induced response of the dwarf selection is concentration-dependent, relatively sensitive, and similar in magnitude to that induced by low temperature. Other auxins also induce GA₃ responsiveness to a greater or lesser degree. IAA has no apparent effect on the wild type (rht, tall) selection.     

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.     

Effect of Temperature on Gibberellin (GA) Responsiveness and on Endogenous GA(1) Content of Tall and Dwarf Wheat Genotypes

Near-isogenic wheat (Triticum aestivum L.) lines differing in height-reducing (Rht) alleles were used to investigate the effects of temperature on endogenous gibberellin (GA) levels and seedling growth response to applied GA₃. Sheath and lamina lengths of the first leaf were measured in GA treated and control seedlings, grown at 11, 18, and 25°C, of six Rht genotypes in each of two varietal backgrounds, cv Maris Huntsman and cv April Bearded. Endogenous GA₁ levels in the leaf extension zone of untreated seedlings were determined by gas chromatography-mass spectrometry with a deuterated internal standard in the six Maris Huntsman Rht lines grown at 10 and 25°C. Higher temperature increased leaf length considerably in the tall genotype, less so in the Rht1 and Rht2 genotypes, and had no consistent effect on the Rht1+2, Rht3 and Rht2+3 genotypes. In all genotypes, endogenous GA₁ was higher at 25°C than at 10°C. At 10°C the endogenous GA₁ was at a similar level in all the genotypes (except Rht2+3). At 25°C it increased 1.6-fold in the tall genotype, 3-fold in Rht1 and Rht2, 6-fold in Rht3, and 9-fold in Rht1+2. Likewise, the genotypic differences in leaf length were very conspicuous at 25°C, but were only slight and often unsignificant at 11°C. The response of leaf length to applied GA₃ in the Rht1, Rht2, and Rht1+2 genotypes increased significantly with lowering of temperature. These results suggest the possibility that the temperature effect on leaf elongation is mediated through its effect on the level of endogenous GA₁ and that leaf elongation response to endogenous or applied GAs is restricted by the upper limits set by the different Rht alleles.     

The effects of temperature and the Rht3 dwarfing gene on growth, cell extension, and gibberellin content and responsiveness in the wheat leaf

The effects of low temperature and the Rht3 dwarfing gene onthe dynamics of cell extension in leaf 2 of wheat were examinedin relation to gibberellin (GA) content and GA-responsivenessof the extension zone. Leaf 2 of wild-type (rht3) wheat closelyresembled that of the Rht3 dwarf mutant when seedlings weregrown at 10C. The maximum relative elemental growth rate (REGR)within the extension zone in both genotypes was lower at 10Cthan at 20C, but the position with respect to the leaf basewas unaffected by temperature. The size of the extension zoneand epidermal cell lengths were similar in both genotypes at10C. Growth at 20C, instead of 10C, increased the lengthof the extension zone beyond the point of maximum REGR in thewild type, but not in the Rht3 mutant. Increasing temperatureresulted in longer epidermal cells in the wild type. Treatingwild-type plants at 10C with gibberellic acid (GA₃) also increasedthe length of the extension zone, but the Rht3 mutant was GA-non-responsive.However, the concentrations of endogenous GA₁ and GA₃ remainedsimilar across the extension zone of wild-type plants grownat both temperatures, despite large differences in leaf growthrates. The period of accelerating REGR as cells enter the extensionzone, and the maximum REGR attained, are apparently not affectedby GA. It is proposed that GA functions as a stimulus for continuedcell extension by preventing cell maturation in the region beyondmaximum REGR and that low temperature increases the sensitivitythreshold for GA action. Key words: Cell extension, gibberellin, Rht3 dwarfing gene, temperature, wheat leaf     

Effect of the Rht3 dwarfing gene on dynamics of cell extension in wheat leaves, and its modification by gibberellic acid and paclobutrazol

The second leaf of wheat was used as a model system to examinethe effects of the Rht3 dwarfing gene on leaf growth. Comparedto the rht3 wild type, the Rht3allele decreased final leaf length,surface area and dry mass by reducing the maximum growth rates,but without affecting growth duration. Gibberellic acid (GA₃)increased final leaf length and maximum growth rate in the rht3wild type, but was without effect on the Rht3 mutant, whichis generally regarded as being non-responsive to gibberellin(GA). Paclobutrazol, an inhibitor of GA biosynthesis, decreasedfinal leaf length and maximum growth rate in the rht3 wild typeto values similar to those in the untreated Rht3 mutant. NeitherGA₃ nor paclobutrazol affected the duration of leaf growth.The decrease in leaf length was produced by reduction of celllength rather than cell number. The maximum relative elementalgrowth rate (REGR) for cell extension was essentially the samein all treatments, as was the time between the cells leavingthe meristem and achieving maximum extension rate. The differencesbetween the genotypes and treatments were all almost entirelydue to differences in the time taken from the attainment ofmaximum REGR of cell extension to the cessation of extension.This was reflected in the length of the extension zone, whichwas approximately 6–8 per cent of final leaf length. Theeffects of the Rht3 allele, GA₃ and paclobutrazol all appearto be on the processes which promote the cessation of cell elongation. Key words: Cell extension, gibberellin, leaf growth, Rht3 gene, Triticum, wheat     

Ontogenetic variation in levels of gibberellin A1 in Pisum

The levels of the biologically active gibberellin (GA), GA₁, and of its precursor, GA₂₀, were monitored at several stages during ontogeny in the apical portions of isogenic tall (Le) and dwarf (le) peas (Pisum sativum L.) using deuterated internal standards and gas chromatography-selected ion monitoring. The levels of both GAs were relatively low on emergence and on impending apical arrest. At these early and late stages of development the internodes were substantially shorter than at intermediate stages, but were capable of large responses to applied GA₃. Tall plants generally contained 10–18 times more GA₁ and possessed internodes 2–3 times longer than dwarf plants. Further, dwarf plants contained 3–5 times more GA₂₀ than tall plants. No conclusive evidence for the presence of GA₃ or GA₅ could be obtained, even with the aid of [²H₂]GA₃ and [²H₂]GA₅ internal standards. If GA₃ and GA₅ were present in tall plants, their levels were less than 0.5% and 1.4% of the level of GA₁, respectively. Comparison of the effects of gene le on GA₁ levels and internode length with the effects of ontogeny on these variables shows that the ontogenetic variation in GA₁ content was sufficient to account for much of the observed variation in internode length within the wild-type. However, evidence was also obtained for substantial differences in the potential length of different internodes even when saturating levels of exogenous GA₃ were present.Abreviations GA_n gibberellin A_n We thank Noel Davies, Omar Hasan, Leigh Johnson, Katherine McPherson and Naomi Lawrence for technical help, Professor L. Mander (Australian National University, Canberra) for deuterated GA standards and the Australian Research Council for financial assistance.     

Gibberellins in immature seeds and dark-grown shoots of Pisum sativum

Gibberellins (GAs) A₁₇, A₁₉, A₂₀, A₂₉, A₄₄, 2OH-GA₄₄ (tentative) and GA₂₉-catabolite were identified in 21-day-old seeds of Pisum sativum cv. Alaska (tall). These GAs are qualitatively similar to those in the dwarf cultivar Progress No. 9 with the exception of GA₁₉ which does not accumulate in Progress seeds. There was no evidence for the presence of 3-hydroxylated GAs in 21 day-old Alaska seeds. Dark-grown shoots of the cultivar Alaska contein GA₁, GA₈, GA₂₀, GA₂₉, GA₈-catabolite and GA₂₉-catabolite. Dark-grown shoots of the cultivar Progress No.9 contain GA₈, GA₂₀, GA₂₉ and GA₂₉-catabolite, and the presence of GA₁ was strongly indicated. Quantitation using GAs labelled with stable isotope showed the level of GA₁ in dark-grown shoots of the two cultivars to be almost identical, whilst the levels of GA₂₀, GA₂₉ and GA₂₉-catabolite were significantly lower in Alaska than in Progress No. 9. The levels of these GAs in dark-grown shoots were 10²- to 10³-fold less than the levels in developing seeds. The 2-epimer of GA₂₉ is present in dark-grown-shoot extracts of both cultivars and is not thought to be an artefact.Abbreviations cv cultivar - GA_n gibberellin A_n - GC gas chromatography - GC-MS combined gas chromatographymass spectrometry - HPLC high-pressure liquid chromatography - KRI Kovats retention index - MeTMSi methyl ester trimethylsilyl ether     

GA-Responsive Dwarfing Gene Rht12 Affects the Developmental and Agronomic Traits in Common Bread Wheat

Opportunities exist for replacing reduced height (Rht) genes Rht-B1b and Rht-D1b with alternative dwarfing genes, such as the gibberellin-responsive gene Rht12, for bread wheat improvement. However, a comprehensive understanding of the effects and mode of action of Rht12 is lacking. In the present study, the effects of Rht12 were characterized by analyzing its effects on seeding vigour, seedling roots, leaf and stem morphology, spike development and carbohydrate assimilation and distribution. This was carried out in the four genotypes of F_2:3 lines derived from a cross between Ningchun45 and Karcagi (12) in two experiments of autumn sowing and spring sowing. Rht12 significantly decreased stem length (43%∼48% for peduncle) and leaf length (25%∼30% for flag leaf) while the thickness of the internode walls and width of the leaves were increased. Though the final plant stature was shortened (40%) by Rht12, the seedling vigour, especially coleoptile length and root traits at the seedling stage, were not affected adversely. Rht12 elongated the duration of the spike development phase, improved the proportion of spike dry weight at anthesis and significantly increased floret fertility (14%) in the autumn sowing experiment. However, Rht12 delayed anthesis date by around 5 days and even the dominant Vrn-B1 allele could not compensate this negative effect. Additionally, grain size was reduced with the ability to support spike development after anthesis decreased in Rht12 lines. Finally, grain yield was similar between the dwarf and tall lines in the autumn sowing experiment. Thus, Rht12 could substantially reduce plant height without altering seeding vigour and significantly increase spikelet fertility in the favourable autumn sowing environment. The successful utilization of Rht12 in breeding programs will require careful selection since it might delay ear emergence. Nonetheless, the potential exists for wheat improvement by using Rht12.     

Modification by gibberellin of the growth-temperature relationship in mutant and normal genotypes of several cereals

High-resolution growth measurements were conducted using a linear variable displacement transformer in conjunction with a temperature-programmed meristem-cooling collar. Chilling and rewarming profiles were determined for a range of Gramineae, in the presence and absence of varying concentrations of gibberellic acid (GA₃). In wheat (Triticum aestivum L.) seedlings, the growth-constraining temperature (P_e) was progressively lowered by increasing GA₃ concentration, with a difference of-4.8°C between controls and material treated with 10^–4 M GA₃. Dwarf-5 maize (Zea mays L.) seedlings had a higher P_e than tall segregates and the difference was markedly reduced by exposure to a saturating concentration of GA₃. A similar effect was observed with Tanginbozu dwarf rice (Oryza sativa L.). The growth ratetemperature responses of Rht3 gibberellin-insensitive dwarf wheat seedlings were unaffected by GA₃ and the P_e values for these segregates were around 5° C higher than for normals. Slender (s1) barley (Hordeum vulgare L.) genotypes had P_e values of-7° C, compared with +4° C for wild-type material, and did not show positive hysteresis for growth rate during the rewarming phase. These studies indicate that GA₃ modifies the thermal sensitivity of meristem function in Gramineae in a manner which enhances low-temperature growth.Abbreviations GA gibberellin - GA₃ gibberellic acid - LVDT linear variable displacement transducer     

Gibberellins in dark- and red-light-grown shoots of dwarf and tall cultivars of Pisum sativum: The quantification,metabolism and biological activity of gibberellins in Progress no. 9 and Alaska

The stem growth in darkness or in continuous red light of two pea cultivars, Alaska (Le Le, tall) and Progress No. 9 (le le, dwarf), was measured for 13 d. The lengths of the first three internodes in dark-grown seedlings of the two cultivars were similar, substantiating previous literature reports that Progress No. 9 has a tall phenotype in the dark. The biological activity of gibberellin A₂₀ (GA₂₀), which is normally inactive in le le geno-types, was compared in darkness and in red light. Alaska seedlings, regardless of growing conditions, responded to GA₂₀. Dark-grown seedlings of Progress No. 9 also responded to GA₂₀, although red-light-grown seedlings did not. Gibberellin A₁ was active in both cultivars, in both darkness and red light. The metabolism of [¹³C³H]GA₂₀ has also been studied. In dark-grown shoots of Alaska and Progress No. 9 [¹³C³H]GA₂₀ is converted to [¹³C³H]GA₁, [¹³C³H]GA₈, [¹³C]GA₂₉, its 2-epimer, and [¹³C³H]GA₂₉-catabolite. [¹³C³H] Gibberellin A₁ was a minor product which appeared to be rapidly turned over, so that in some feeds only its metabolite, [¹³C³H]GA₈, was detected. However results do indicate that the tall growth habit of Progress No. 9 in the dark, and its ability to respond to GA₂₀ in the dark may be related to its capacity to 3-hydroxylate GA₂₀ to give GA₁. In red light the overall metabolism of [¹³C³H]GA₂₀ was reduced in both cultivars. There is some evidence that 3-hydroxylation of [¹³C³H]GA₂₀ can occur in red light-grown Alaska seedlings, but no 3-hydroxylated metabolites of [¹³C³H]GA₂₀ were observed in red light-grown Progress. Thus the dwarf habit of Progress No. 9 in red light and its inability to respond to GA₂₀ may be related, as in other dwarf genotypes, to its inability to 3-hydroxylate GA₂₀ to GA₁. However identification and quantification of native GAs in both cultivars showed that red-light-grown Progress does contain native GA₁. Thus the inability of red light-grown Progress No. 9 seedlings to respond to, and to 3-hydroxylate, applied GA₂₀ may be due to an effect of red light on uptake and compartmentation of GAs.Abbreviations AMO-1618 2-isopropyl-4-(trimethylammonium chloride)-5-methylphenyl piperidine-1-carboxylate - cv. cultivar - GC-MS gas chromatography-mass spectrometry - GA_(n) gibberellin A_(n) - HPLC high-pressure liquid chromatography     

The involvement of gibberellin signalling in the effect of soil resistance to root penetration on leaf elongation and tiller number in wheat

Background and aims

The concept of root-sourced chemical signals that affect shoot growth in response to drought is widely reported; in particular the role of ABA in regulating stomatal conductance has received much attention. ABA, alone, does not fully explain all the effects of abiotic stresses in the root zone on shoot architecture. An increase in mechanical impedance, which can occur on even relatively modest soil drying, results in reduced root and shoot growth, processes that are potentially regulated by gibberellins (GAs).

Methods

In this study we explored the role of mechanical impedance and exogenous gibberellin (GA₃) on root and shoot architecture in wheat seedlings containing the Rht-B1a (tall), Rht-B1b (semi-dwarf) or Rht-B1c (dwarf) alleles in the April-Bearded or Mercia backgrounds. Our experiments were based on the use of the sand culture system which allows the mechanical impedance of the root growth environment to remain constant and independent of water and nutrient availability. We investigated the effects of the application of exogenous GA₃ to the root system.

Results

We found that impeding soil reduced leaf elongation in the tall and semi-dwarf lines, confirming the stunting effect of mechanical impedance which is widely reported. However, leaf elongation in the dwarf lines was not affected by root impedance. Application of GA₃ to the roots restored leaf elongation in the tall and semi-dwarf lines growing in impeding soil, with some growth response even in the dwarf line, the longest leaves being obtained when GA was applied to impeded roots of a tall line. Both exogenous GA and root impedance reduced the number of tillers, but there was no interaction with the Rht genotype. The genetic background did not affect the results.

Conclusion

We suggest that the GA signalling pathway has an unidentified role in the leaf elongation response to mechanical impedance to root growth.     

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.     

Distribution of endogenous gibberellins in dwarf and giant off-types banana (Musa AAA,cv.‘Grand nain’) plants from in vitro propagation

GA₃ and GA₂₀ were quantified in leaf extracts from true-to-type and somaclonal variants (dwarf and giant) of Musa AAA cv. Grand nain by GC-MS-SIM after purification on reverse- and normal-phase HPLC and detection by ELISA with GA₃ antibodies and by a dwarf rice bioassay. GA₃ concentration in dwarf plants was 811 ng g^–1 dry weight. For normal and giant plants, the endogeneous GA₃ levels were respectively 3.6 and 4.6 times higher. The GA₂₀ concentration in the giant plant was 68 ng g^–1 of dry weight. This concentration was, respectively, 4.6 and 7.3 times higher than those of normal and dwarf plants. These results suggest that the somaclonal variations affecting banana plant height are associated with modifications in GA metabolism.Abbreviations HPLC High Performance Liquid Chromatography - GC-MS Gas Chromatography-Mass Spectrometry - SIM Selected Ion Monitoring - GA Gibberellin - BSA Bovine Serum Albumin - PB Phosphate Buffer