Peroxidase Ontogeny in a Dwarf Pea Stem as Affected by Gibberellin and Decapitation

The ontogeny of peroxidase activity and isoenzyme pattern wasinvestigated in the stem of dwarf pea plants. Peroxidase activityper unit soluble protein was a given internode is highest inthe youngest growth stage, drops during elongation, remainsconstant upon cessation of growth, and increase at senescence.The lower the internode on the stem the higher is its peroxidaseactivity. These developmental differences are already apparentat the youngest growth stage of the internodes adn increaseduring elongation. Several anodic and five cathodic isoperoxidasesare apparent after starch gel electrophoresis. This patternis constant for all internodes at all growth stages, but therelative importance of particular isoenzymes changes with time. Gibberellic acid (GA₃) treatment causes greatly elongated internodes,decreased soluble protein, and inhibition of the rise in peroxidaseactivity within 4–8 h. Application of GA₃ to young internodesleads to a persistent depression in peroxidase activity, whiletreated older internodes suffer only a temporary depression.GA₃ causes no qualitative changes in the isoenzyme pattern butproduces some quantitative alterations in internodes in whichits influence on peroxidase activity is persistent. Decapitation of untreated and GA₃-treated dwarfs has littleinfluence on internode elongation, causes an increase in peroxidaseactivity, especially in the upper internodes, and alters therelative activity of particular isoenzymes. By contrast, decapitationinhibits elongation of young internodes in genetically tallpea plants.     

Peroxidase activities in relation to plant height and grain weight in bread wheat (Triticum aestivum L.)

Summary The relationship of peroxidase activity with plant height and grain weight has been studied in seven different varieties of bread wheat belonging to diverse genotypes, and their F₁ crosses. The association between plant height and peroxidase activity was highly significant and negative. Based on the similarity index values of peroxidase isoenzymes, the seven wheat genotypes could be classified into two groups: the first group consisting of triple and quadruple dwarf varieties and the other of tall, single and double dwarf. A negative correlation between peroxidase activity and grain weight was also observed. However, the results of this study indicate a possibility of developing a dwarf plant type with low peroxidase activity and well filled grains.     

Role of polyamines in gibberellin-induced internode growth in peas

To determine the requirement for polyamines in gibberellin (GA) induced internode growth polyamine content was measured in internodes of peas of various internode phenotypes (slender, tall, dwarf, nana) with and without applied gibberellin (GA₃) and polyamine synthesis inhibitors. Polyamines were assayed as dansyl derivatives which were separated by reverse phase high performance liquid chromatography and detected by fluorescence spectrophotometry. The amounts of polyamines in the different genetic lines of peas, which differed in internode lengths and extractable GA content, correlated with the extent of internode elongation. High polyamine concentrations were associated with young internodes and decreased with internode expansion. Extremely short internodes of nana plants without GA exhibited equal or higher amine concentrations relative to internodes of other lines of peas and GA-stimulated nana seedlings. The polyamine synthesis inhibitors, α-difluoromethylornithine and α-difluoromethylarginine, independently or in combination, inhibited polyamine accumulation and internode elongation of tall peas and GA-stimulated nana plants. Agmatine and putrescine restored growth and endogenous polyamine content to variable degrees. However, exogenous polyamines were not effective in promoting growth unless intracellular amines were partially depleted.

These results suggest that polyamines do not have a role in cell elongation, but may be required to support cell proliferation. Polyamines do not mediate the entire action of GA in internode growth of peas since GA induction of growth involves both cell division and cell elongation, whereas polyamines appear to affect cell division only.

     

ent-Kaurene Biosynthesis in Cell-Free Extracts of Excised Parts of Tall and Dwarf Pea Seedlings

Investigations on the sites of ent-kaur-16-ene (ent-kaurene) biosynthesis were conducted with cell-free extracts from several excised parts of 10-, 13-, and 16-d-old tall and dwarf pea (Pisum sativum L.) seedlings. [¹⁴C]Mevalonic acid was incorporated into ent-kaurene in cell-free extracts from young developing leaves and elongating internodes of tall (`Alaska') and dwarf (`Progress No.9') pea seedlings at all three stages of development. ent-Kaurene biosynthesis also occurred readily in cell-free extracts from shoot tips, petioles, and stipules near the young elongating internodes. The ent-kaurene-synthesizing activity found in young developing tissues declined as tissues matured. Little or no activity was detectable in enzyme extracts from cotyledons and root tips at different stages. In light grown tall pea internodes ent-kaurene-synthesizing activity was low as they began to elongate, reached a maximum when the internodes reached about 2 cm in length and declined as they matured. Activity in extracts of dwarf shoot tips and internodes was generally lower than in equivalent tall plants, but the activity in dwarf leaves and stipules was somewhat higher than in tall plants. With the exception of root tips, there is a strong correlation between growth potential of a tissue and the rate of ent-kaurene biosynthesis in extracts from that tissue.     

Peroxidase Isoenzymes in Development of Tall and Dwarf Cultivars of Rice

Four cultivars of rice, Hansraj (tall), Jamuna, Padma and Sabarmati(all dwarf) were analysed for variation in isoperoxidase patternsduring development by horizontal starch-gell electrophoresis.The samples were taken at weekly intervals, starting with soakedseeds (12 h) until the post panicle stage. There is a distinctpattern of peroxidase isoenzymes in the tall and dwarf cultivarsstudied. Some isoenzymes appear at a particular developmentalstage while others disappear while some remain fairly constantonce they are activated. Hansraj showed more qualitative fluctuationsin peroxidase than did the three dwarf varieties. A few isoperoxidaseswere specific to some varieties such as bands A₃ and A₇ forHansraj and band A₄ for Sabarmati. These specific bands of peroxidasemay have some role to play in development of Hansraj (tall)and Sabarmati (with aroma). Besides this, greater fluctuationsin peroxidase activity in the tall variety may be due to loosecontrol in the regulation of peroxidase activity. The significanceof controlling elements of enzyme activity is discussed.     

Xyloglucan endotransglycosylase activity in pea internodes. Effects of applied gibberellic acid.

Xyloglucan endotransglycosylase (XET) activity extractable from internodes of tall and dwarf varieties of pea (Pisum sativum L.) was assayed radiochemically using tamarind seed xyloglucan as donor substrate and an oligosaccharidyl-[3H]alditol as acceptor substrate. Internodes I and II showed little elongation during the period 15 to 21 d after sowing; XET activity remained relatively constant and was unaffected by exogenous gibberellic acid (GA3). A single application of GA3 to the dwarf genotype resulted in a small enhancement of elongation in internode III between d 17 and 21 and caused a small increase in XET activity in internode III. Repeated applications of GA3 caused internode V to elongate between d 20 and 26, to the same extent as in the tall variety, and concomitantly led to greatly elevated XET activity (expressed per unit fresh weight, per unit of extractable protein, and per internode). Thus, XET activity correlated with GA3-enhanced length in pea internodes; the possibility that this represents a causal relationship is discussed.     

Inheritance of a dwarf seed stalk character in onion

Results are reported of crossing a dwarf seed stalk line (mean height 44.8 cm) from Israel with five inbred lines of the Rijnsburger type each having normal tall seed stalks (mean heights 83.1–105.9 cm). The seed stalk height of F₁ generations from these crosses was slightly greater than that of the Rijnsburger parents but segregation occurred in the F₂ giving plants which could be considered ‘dwarf’ and ‘normal’ with respect to seed stalk height. Selfing individual F₂‘dwarf’ plants produced F₃ progenies whose mean heights were in the dwarf range, and were significantly correlated with those of their F₂ parent. The results obtained in the F₂ and F₃ families suggest that although a major recessive gene for dwarfness may be present, substantial minor gene and environmental variation is also involved.     

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.     

Role of Assimilates and Phosphates in the Control of Internode Elongation in Tall and Dwarf Indica Rice Varieties

Six indica rice varieties belonging to three duration groups,each group containing one tall and one dwarf variety, were grownunder irrigated field conditions during the wet season of 1986.At the time of culm elongation, the distal 5–6 internodesof a variety elongated. The pattern of extension was hierarchical,each successive internode dominating the one immediately belowit and hence the final internode lengths increased in acropetalfashion from the bottom to the top. Between the tall and dwarfvarieties, the lengths of the individual intemodes did not differmuch in the apical and basal positions, but significant differenceswere found between the lengths of the middle internodes. Withincrease in length, the internodes became visibly thinner anddry matter content per unit length decreased. Analyses of solublecarbohydrates, amino acids, and phosphate contents of the internodesindicated that the concentration of the metabolites increasedin acropetal succession from the base to the apex. In general,the dwarf varieties contained at least the same, if not a higherconcentration of the metabolites as the tall varieties. Thepossible role of the metabolites in internode extension is discussed. Key words: Rice, internode extension, assimilates, phosphates, sink     

Gibberellic acid and the light inhibition of stem elongation

Summary The ability of gibberellic acid (GA₃) to prevent the light inhibition of stem elongation in peas was examined for several varieties under a wide range of irradiation conditions.A saturating dose of GA₃ largely prevented the inhibitory effect of red light on total stem height in Duke of Albany (tall), Alaska (medium) and Meteor (dwarf) although a small, but statistically significant, effect persisted in all varieties after 3 days of light. The growth of the second internode was, however, markedly inhibited by red light even with a saturating dose of GA₃. With gibberellin there was no difference between the effects of continuous red light and 15 minutes per day on height but the second internode was much shorter in the former treatment. The number of internodes present was the same in both cases and, therefore, the upper internodes in continuous light were as long or longer than in the 15-minute treatment. The number of internodes was only slightly fewer in darkness than in light so that, with GA₃, the effect of red light was transient and only the growth of the lower internodes was inhibited. Without GA₃ overall height was less in both red light treatments than in darkness for all three varieties.In blue light, on the other hand, there was no difference depending on whether height or internode length is considered, and even with a saturating dose of GA₃ the growth rate remained depressed in continuous blue light. There was, however, some interaction between blue light and GA₃.Red/far-red reversal experiments showed that in the varieties Alska and Duke of Albany the far-red stimulation of elongation persisted in the presence of a saturating dose of GA₃ while for the dwarf variety Meteor there was a significant interaction between far-red and GA₃.At least a quantitative difference was found between tall and dwarf peas in their response to light. Tall varieties showed a much greater effect of a prolonged exposure to blue and a smaller effect of a short exposure to red than dwarf varieties. Increasing the duration of exposure to red increasingly inhibited the growth of tall varieties. The medium variety Alaska grew to approximately the same height in continuous red and blue light.     

New sources of dwarfing genes in pearl millet (Pennisetum americanum)

Summary Thirteen naturally occurring dwarf lines of pearl millet [Pennisetum americanum (L.) Leeke], identified from the world collection, varied for several morphological and agronomic characters. Extreme dwarfs were characterized by a tufted growth habit which could be distinguished from the time of germination, while the other dwarf lines could be distinguished only after anthesis. The F₁ hybrids between the tall and dwarf genotypes were tall, indicating that dwarfness is a recessive trait. In 10 out of the 13 crosses, the F₂ segregation ratio was three tall to one dwarf (31) suggesting that the dwarfness is controlled by a single recessive gene, while the height differences in 3 of the dwarfs (IP 8056, IP 8210 and IP 8214) were controlled by more than one gene as they showed continuous variation for plant height in F₂. When the remaining 10 single gene dwarfs were crossed to either d ₁ (Tift 238) or d ₂ (Tift 23 DB) dwarfs, only 2 crosses produced tall F₂ hybrids and they segregated for height in F₂ indicating that these 2 dwarfs are non-allelic to d ₁ and d ₂. Reciprocal crosses of these 2 dwarfs produced tall F₁ hybrids and showed a dihybrid segregation of 934 in F₂ indicating that the dwarfing genes of these 2 parents are non-allelic to each other. These non-allelic dwarfs were assigned the gene symbols d ₃ (IP 10401), and d ₄ (IP 10402).Submitted as J.A. No. 429 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT)     

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.     

Genes for dwarfing and photoperiod flowering response inPharbitis nil choisy

Dwarfing and sensitivity to the duration of a single inductive dark period for flowering ofPharbitis nil in F₂ progeny of a cross between the tall strain Tendan, and the dwarf, Kidachi appear to be controlled by the alleles at two independent loci. Progeny of a similar cross between the tall strain Violet and the dwarf Kidachi at F₂ and F₃ also showed single locus segregation for tall: dwarf plants. In this cross, differences in photoperiodic response could be identified in F₃ families but they were not simply inherited. There was some evidence of difficulties with classification of the F₂ plants, but also, the flowering of the F₁ between the two less sensitive strains Tendan and Violet indicated complex inheritance of their photoperiodic response. Complementary dominant alleles at three independent loci may be necessary for flowering in even shorter dark periods with the sensitive strain Kidachi. The dwarf strain Kidachi has a reduced gibberellin (GA) content (Barendse and Lang 1972), it flowers in a short dark period without terminal flowering, and it responds positively to GA application both for flowering and growth. However, since control of dwarfing and photoperiodic sensitivity can be separated genetically, there is no strick link between the gibberellin responsiveness of Kidachi for its growth and flowering. Despite the complexity of flowering genetics in Violet×Kidachi, a short-dark-period-sensitive, terminal flowering and tall F₇ line was obtained in a pedigree previously held heterozygous for the dwarf: tall character but not selected for flowering time. Thus, flowering in a short dark period can also be obtained in the presence of the non-dwarfing allele from strain Violet, again demonstrating genetic independence.     

Inheritance of height and maturity in crosses between pearl millet landraces and inbred Tift 85DB

Summary Over 300 landraces of pearl millet were collected in Burkina Faso and grown at the Coastal Plain Experiment Station in Tifton/GA. At Tifton, these landraces are predominantly tall and late-maturing. The photoperiod requirements of these landraces hinder evaluation of their performance in the field and their use in breeding programs. A conversion program has been initiated to transfer genes for dwarf stature and early flowering into the tall, late-maturing landraces. The inbred Tift 85DB is being used as a donor of genes for the dwarf and early characteristics, and was crossed to nine randomly selected landraces from Burkina Faso. The parents, F₁, F₂, and backcrosses to each parent were grown in the field and evaluated for plant height at anthesis and time in days from planting to anthesis. In general, plant height of F₁s was taller than the tallest parent, and in all crosses the maturity of F₁s was intermediate between the parents. Numbers of loci conferring height varied among crosses, ranging from 0 to 9.6, and averaged 1.6. Estimated numbers of loci conferring maturity ranged from 0 to 12.8 and averaged 3.4. Broad-sense heritability estimates for height and maturity averaged 60.2 and 65.7%, respectively. Corresponding narrow-sense estimates averaged 23.8 and 48.2%. Joint scaling tests revealed that additive-genetic effects were highly significant for both traits, but dominance and epistatic-genetic effects contributed to the inheritance of each trait in some crosses. The low gene numbers, high heritability estimates, and preponderance of additive-genetic effects suggest that selection for these traits should be effective.     

Isoperoxidase pattern and internode length genotype in Pisum

Inbred Pisum sativum lines of known constitution for the intemode length genes Le, La and Cry, and representing four height phenotypes, were grown to the 7-intemode stage in the light. Six cationic isoperoxidases, making up ca. 90% of the activity of stem extracts, were resolved by concave gradient elution from Dowex 50 columns and shown to run as single peroxidase bands on starch gel electrophoresis. They were all able to oxidise IAA in the presence of 2,4-dichlorophenol, but fell into two groups with widely differing IAA oxidase/peroxidase ratios. The isoperoxidase patterns were independent of both genotype and phenotype for internode length, thus making it unlikely that these loci exert their effect on internode extension via control of synthesis of a particular isoperoxidase. Amongst the lines screened polymorphism was detected involving two of the isoperoxidases, and limited F₂ data suggest that these two variants fire determined by alleles of a single gene. Isoperoxidase patterns of stem extracts of 6 other Pisum species did not differ significantly from the two found in P. sativum.     

Ontogenetic development in architecture and biomass allocation of 13 rattan species in Indonesia

Rattans are climbing, nonbranching palms with diverse growth forms ranging from stems that remain at the forest floor to stems that reach the canopy. We analyzed changes in architecture and biomass allocation during ontogenetic development of 13 Indonesian species in the genera Calamus, Ceratolobus, Daemonorops, Korthalsia, and Plectocomiopsis. Species included both nonclimbers (<5 m tall maximum) and climbers (10 to 50 m tall). Nonclimbers retain a rosette form—that is, thick, short internodes with length/internode diameter ratio <10 but with long petioles—throughout their lives. Leaf and internode shapes of climbers at the early stage resemble those of nonclimbers, but internodes later become longer (internode length/diameter ratio ≥10) and petioles become shorter and developed one of two kinds of climbing organs at the adult stage. These developmental changes reduce self-shading within the crown. Some climbers have dwarf blades at the early stage and skip the rosette form. Principal component analysis of biomass allocation indicated that growth strategies to attain the adult stage are diverse. These results suggest that rattans reach maturity at different phases along a series of ontogenetic development stages and generate diverse growth forms. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Suppressive effect of microRNA319 expression on rice plant height

Key message

miR319 was identified as a dwarf-inducing gene from Shiokari and its dwarf near isogenic line, and its transgenic rice showed a reduced plant height. This finding reveals the potential application of miR319 in future molecular breeding.

Abstract

It is well known that microRNAs (miRNAs) play important roles in plant physiology, especially in development and stress responses. However, little is known about the role of miRNAs in plant height. In this study, the rice cultivar Shiokari and its dwarf near isogenic line Shiokari-d6 were analysed to identify and characterize plant height-associated miRNAs. This anatomic and morphological investigation revealed that the major cause of the shorter height of Shiokari-d6 is the significantly dis-elongated internodes, particularly the second internode and those underneath it. The results of miRNA microarray profiling and real-time RT-PCR indicated that miR319 is expressed at a significantly higher level in Shiokari-d6 than in Shiokari. Transgenic rice overexpressing miR319 in Oryza sativa L. cv. Tainung 67 generated through Agrobacterium-mediated transformation had a stable dwarf phenotype regardless of whether the plants were from the T1 or T2 generation. We also found that the internodes of miR319-overexpressing rice are shortened, particularly the third internode and those underneath it. Furthermore, we identified three putative miR319 target genes that were previously uncharacterized with expression levels that were negatively correlated with the expression of miR319. In conclusion, miR319 is the first miRNA proposed to be involved in plant height regulation, and its function may influence the elongation of internodes, which leads to decreased plant height.

     

Characterization and genetic mapping of a Photoperiod-sensitive dwarf 1 locus in rice (Oryza sativa L.)

Plant height is an important agronomic trait for crop architecture and yield. Most known factors determining plant height function in gibberellin or brassinosteroid biosynthesis or signal transduction. Here, we report a japonica rice (Oryza sativa ssp. japonica) dominant dwarf mutant, Photoperiod-sensitive dwarf 1 (Psd1). The Psd1 mutant showed impaired cell division and elongation, and a severe dwarf phenotype under long-day conditions, but nearly normal growth in short-day. The plant height of Psd1 mutant could not be rescued by gibberellin or brassinosteroid treatment. Genetic analysis with R₁ and F₂ populations determined that Psd1 phenotype was controlled by a single dominant locus. Linkage analysis with 101 tall F₂ plants grown in a long-day season, which were derived from a cross between Psd1 and an indica cultivar, located Psd1 locus on chromosome 1. Further fine-mapping with 1017 tall F₂ plants determined this locus on an 11.5-kb region. Sequencing analysis of this region detected a mutation site in a gene encoding a putative lipid transfer protein; the mutation produces a truncated C-terminus of the protein. This study establishes the genetic foundation for understanding the molecular mechanisms regulating plant cell division and elongation mediated by interaction between genetic and environmental factors.     

Changes in total nitrogen, soluble protein, and peroxidases in the expanding leaf zone of eastern cottonwood

Nitrogen content and soluble protein and anodal peroxidase banding in acrylamide gel changed with leaf and internode development in the expanding leaf zone of eastern cottonwood (Populus deltoides Bartr.). Nitrogen per unit leaf area was high near the apex and decreased to a constant value at the sixth node below it. Soluble protein banding was qualitatively similar for leaves and internodes in this zone, but anodal peroxidases differed between leaves and internodes. The major leaf peroxidase band was absent from the second leaf below the apex but present in the fourth and sixth leaves; its appearance and intensification seemed to parallel the development of photosynthetic activity. The major internode peroxidase band was present in the apex, second, fourth, and sixth internodes, and intensified during internodal development. It is suggested that these two “isoenzymes” may have different functions in vivo.     

Growth Pattern of Tall and Short Lines of Rice and their Response to Gibberellin

The growth pattern of two types of dwarfness in rice was studied.The difference in height of the tall and short Peta plants wasapparent even at the seedling stage. In the dwarf and in thenormal Century Patna 231 the difference in height was apparentonly after panicle initiation. The four lines had the same numberof elongated internodes, with the taller lines having longerinternodes. The application of gibberellin resulted in increased plant height,longer internodes, blades and leaf sheaths, and decreased tillernumber. Century Patna 231-dwarf showed the least, and shortPeta the greatest, response to gibberellin application.