Contrasting Strategies of Alfalfa Stem Elongation in Response to Fall Dormancy in Early Growth Stage: The Tradeoff between Internode Length and Internode Number

Fall dormancy (FD) in alfalfa (Medicago sativa L.) can be described using 11 FD ratings, is widely used as an important indicator of stress resistance, productive performance and spring growth. However, the contrasting growth strategies in internode length and internode number in alfalfa cultivars with different FD rating are poorly understood. Here, a growth chamber study was conducted to investigate the effect of FD on plant height, aboveground biomass, internode length, and internode number in alfalfa individuals in the early growth stages. In order to simulate the alfalfa growth environment in the early stage, 11 alfalfa cultivars with FD ratings from one to 11 were chosen and seeded at the greenhouse, and then were transplanted into an artificial growth chamber. The experimental design was a randomized complete block in a split-plot arrangement with three replicates. Plant height, above-ground biomass, internode length, and internode number were measured in early growth stage in all individuals. Our findings showed that plant height and the aboveground biomass of alfalfa did not significantly differ among 11 different FD rated cultivars. Also, internode length and internode number positively affected plant height and the aboveground biomass of alfalfa individuals and the average internode length significantly increased with increasing FD rating. However, internode number tended to sharply decline when the FD rating increased. Moreover, there were no correlations, slightly negative correlations, and strongly negative correlations between internode length and internode number in alfalfa individuals among the three scales, including within-FD ratings, within-FD categories and inter-FD ratings, respectively. Therefore, our results highlighted that contrasting growth strategies in stem elongation were adopted by alfalfa with different FD ratings in the early growth stage. Alfalfa cultivars with a high FD rating have longer internodes, whereas more dormant alfalfa cultivars have a larger number of internodes. There were tradeoffs between internode length and internode number in response to FD in alfalfa, which reflected certain scale-dependence.     

Internode length in Pisum

The influence of the Na and Le genes in peas on gibberellin (GA) levels and metabolism were examined by gas chromatographic-mass spectrometric analysis of extracts from a range of stem-length genotypes fed with [¹³C, ³H]GA₂₀. The substrate was metabolised to [¹³C, ³H]GA₁, [¹³C, ³H]GA₈ and [¹³C, ³H]GA₂₉ in the immature, expanding apical tissue of all genotypes carrying Le. In contrast, [¹³C, ³H]GA₂₉ and, in one line, [¹³C, ³H]GA₂₉-catabolite, were the only products detected in plants homozygous for the le gene. These results confirm that the Le gene in peas controls the 3-hydroxylation of GA₂₀ to GA₁. Qualitatively the same results were obtained irrespective of the genotype at the Na locus. In all Na lines the [¹³C, ³H]GA₂₀ metabolites were considerably diluted by endogenous [¹²C]GAs, implying that the metabolism of [¹³C, ³H]GA₂₀ mirrored that of endogenous [¹²C]GA₂₀. In contrast, the [¹³C, ³H]GA₂₀ metabolites in na lines showed no dilution with [¹²C]GAs, confirming that the na mutation prevents the production of C₁₉-GAs. Estimates of the levels of endogenous GAs in the apical tissues of Na lines, made from the ¹²C:¹³C isotope ratios and the radioactivity recovered in respective metabolites, varied between 7 and 40 ng of each GA per plant in the tissue expanded during the 5 d between treatment with [¹³C, ³H]GA₂₀ and extraction. No [¹²C]GA₁ and only traces of [¹²C]GA₈ (in one line) were detected in the two Na le lines examined. These results are discussed in relation to recent observations on dwarfism in rice and maize.Abbreviations GA_n gibberellin A_n - GC-MS gas chromatography-mass spectrometry - HPLC high-pressure liquid chromatography     

Internode Length in Pisum. A New Allele at the le Locus

In Pisum sativum L. a third, more severe, allele at the internodelength locus le is identified and named le^d. Plants homozygousfor le^d possess shorter internodes and appear relatively lessresponsive to GA₂₀ than comparable le (dwarf) plants. Gene le^dmay act by reducing the 3ß-hydroxylation of GA₂₀ tothe highly active GA₁ more effectively than does gene le. Theresults indicate that le is a leaky mutant and therefore thatendogenous GA₁ influences internode elongation in dwarf (le)plants. Pisum sativum, peas, internode length, genetics, gibberellin, dwarf elongation     

The Relationship of Endogenous Gibberellins to Light-regulated Stem Elongation Rates in Dwarf and Normal Cultivars of Pisum sativum L.

Dwarf cultivar Progress No. 9 and normal cultivar Alaska ofPisum sativum L. were grown under conditions of darkness orred light. Red light decreased the stem elongation rate of bothcultivars. Gibberellins present during the linear phase of stemelongation were isolated and the two main components were tentativelyidentified by gas chromato-chromatography and mass spectrometryas Gibberellin A₁ and A₅. Both gibberellins varied quantitativelywithin and between cultivar and treatment groups, and the amountspresent were inversely related to stem elongation rate. Althoughthe stem elongation rate of plants grown under red light wasrepressed, endogenous gibberellin was not limiting and was asmuch as two-fold higher in red light-grown plants than in dark-grownplants. The levels of endogenous gibberellin in dawrf plantsindicated that the genetic growth limitation was not due toa gibberellin deficiency. (Received May 26, 1981; Accepted January 28, 1982)     

Internode Length in Pisum: the Response of le na Scions Grafted to Na Stocks

The recessive gene na results in peas with extremely short internodes(phenotype nana). In intact plants, na prevents expression ofthe Le/le gene pair which determine the tall/dwarf difference.na blocks a step early in the gibberellin biosynthetic pathwaywhile le prevents conversion of gibberellin A₂₀ to GA₁. Whengrafted to leafy Na stocks, le na and Le na scions become phenotypicallydwarf and tall, respectively. Hence, Na stocks provide a graft-transmissiblesubstance which promotes elongation of na scions and allowsexpression of the Le/le difference. Pisum, internode length, grafting, genotype     

Internode Length in Pisum: Variation in Response to a Daylength Extension with Incandescent Light

Lines representing a range of internode length and floweringgenotypes in Pisum sativum L. were grown in 8 h of daylightfollowed by either 16 h of darkness or incandescent light. Thestem elongation response index (RI = length in 24 h ÷length in 8 h) was least in the very short internode nana types,which are grossly deficient in gibberellins (GAs), and the verylong internode slender types, which behave as if saturated withGAs. The common tall (genotype Le) and dwarf (le) types (lepartially blocks conversion of GA₂₀ to the active form, GA₁)were all markedly responsive but the peak RI (based on the mostresponsive internode) was less in tall lines (1.79 to 2.78)than in dwarf lines (2.32 to 5.01) and the peak RI tended tooccur about three to four internodes earlier in tall than indwarf lines. The cry⁸ mutation reduced the RI. (Duplicate lengthloci La and Cry are probably concerned with GA reception.) Amongle dwarf lines, genotype La cry⁸, was generally less responsivethan La Cry, La cry^c and la Cry. Data from crosses showed thaton either an le La or le la background cry⁸ segregates had alower RI than cry⁸ segregates. On an le la background, cry⁸plants were shorter than cry^c plants, cry⁸ was partially dominantto cry⁸ and segregation was clear only in long days. On an lela background, cry^c plants were shorter than cry^c plants, cry⁸was partially dominant to cry⁸ and segregation was clear inlong or short days. The very high peak RI (5.0) of the microcryptodwarfline, L57, appeared to result, in part, from a marked foreshorteningof internodes 4 to 10 in the 8 h regime. In the 24 h regimeL57 (lm) had a fairly similar growth pattern to normal (Lm)cryptodwarf types. The peak RI tended to occur at a lower internode in early thanlate flowering lines, especially among dwarf types, and genotypeswith a day neutral flowering habit (genotype sn or dne) wereless responsive than their photoperiodic counterparts (Sn Dne). White fluorescent light, given as a daylength extension, wasmuch less effective than incandescent light at stimulating stemelongation suggesting control through the phytochrome equilibrium(P_tr/P_total). Pisum sativum, garden pea, daylength extension, flowering, genotype, gibberellin, hormone receptor, incandescent light, internode length, phytochrome, stem elongation     

Gibberellin Substitution for the Requirement of the Cotyledons in Stem Elongation in Pisum sativum Seedlings

The removal of the cotyledons from 8-day-old light-grown Pisum sativum cv. Alaska seedlings caused a reduction in the rate of stem elongation to 50% of the intact control value. Gibberellic acid restored the stem elongation rate of decotylized plants to the level of the intact controls. The effect of decotylization was to lower both the rate of node formation and the rate of internode elongation. The steady state rate of internode elongation was reduced to 50% of the control rate by decotylization. Applied gibberellic acid did not restore the normal rate of node formation nor the lag in internode elongation caused by decotylization, but gibberellic acid did restore the normal steady state rate of internode elongation. Analysis of variance demonstrated an interaction between the cotyledons and applied gibberellic acid. 2-Isopropyl-4-dimethylamino-5-methyl phenyl-1-piperidine carboxylate methyl chloride inhibited internode elongation to the same extent in both intact and decotylized plants. The results indicate that the cotyledons are an effective source of gibberellin for the young pea seedling.     

Identification of Dw1, a Regulator of Sorghum Stem Internode Length

Sorghum is an important C4 grain and grass crop used for food, feed, forage, sugar, and biofuels. In its native Africa, sorghum landraces often grow to approximately 3–4 meters in height. Following introduction into the U.S., shorter, early flowering varieties were identified and used for production of grain. Quinby and Karper identified allelic variation at four loci designated Dw1-Dw4 that regulated plant height by altering the length of stem internodes. The current study used a map-based cloning strategy to identify the gene corresponding to Dw1. Hegari (Dw1dw2Dw3dw4) and 80M (dw1dw2Dw3dw4) were crossed and F₂ and HIF derived populations used for QTL mapping. Genetic analysis identified four QTL for internode length in this population, Dw1 on SBI-09, Dw2 on SBI-06, and QTL located on SBI-01 and SBI-07. The QTL on SBI-07 was ~3 Mbp upstream of Dw3 and interacted with Dw1. Dw1 was also found to contribute to the variation in stem weight in the population. Dw1 was fine mapped to an interval of ~33 kbp using HIFs segregating only for Dw1. A polymorphism in an exon of Sobic.009G229800 created a stop codon that truncated the encoded protein in 80M (dw1). This polymorphism was not present in Hegari (Dw1) and no other polymorphisms in the delimited Dw1 locus altered coding regions. The recessive dw1 allele found in 80M was traced to Dwarf Yellow Milo, the progenitor of grain sorghum genotypes identified as dw1. Dw1 encodes a putative membrane protein of unknown function that is highly conserved in plants.     

Internode length in Pisum. Gene lkd

A new, single gene, recessive internode length mutant in Pisum, lkd, is described. The internodes of lkd plants are ca. 40% shorter than comparable Lkd plants and this difference appears greater in the dark than in the light. The mutant does not appear to be dwarfed due to modified gibberellin (GA) levels, as determined by gas-chromatography-selected ion monitoring (GC-SIM) for GA₁ and GA₂₀. In relative terms, the mutant responds as well as the wild-type to applied GA₁. However due to its initial short stature it does not elongate to the same extent as the wild-type to high doses of GA₁ suggesting that some other factor, unrelated to GA levels or perception is probably limiting growth in this mutant. Author for correspondence     

Control of Internode Length in Pisum sativum (Further Evidence for the Involvement of Indole-3-Acetic Acid)

The effects of altered endogenous indole-3-acetic (IAA) levels on elongation in garden pea (Pisum sativum L.) plants were investigated. The auxin transport inhibitors 2,3,5-triiodobenzoic acid (TIBA) and 9-hydroxyfluorene-9-carboxylic acid (HFCA) were applied to elongating internodes of wild-type and mutant lkb plants. The lkb mutant was included because elongating lkb internodes contained 2- to 3-fold less free IAA than those of the wild type. In the wild type, TIBA reduced both the IAA level and internode elongation below the site of application. Both TIBA and HFCA strongly promoted the elongation of lkb internodes and also raised IAA levels above the application site. The synthetic auxin 2,4-dichlorophenoxyacetic acid (2,4-D) also markedly increased internode elongation in lkb plants and virtually restored petioles and tendrils to their wild-type length. In contrast, treatment of wild-type plants with TIBA, HFCA, or 2,4-D caused little or no increase in elongation above the application site. The ethylene synthesis inhibitor aminoethoxyvinylglycine also increased stem elongation in lkb plants, and combined application of HFCA and aminoethoxy-vinylglycine restored lkb internodes to the wild-type length. It is concluded that the level of IAA in wild-type internodes is necessary for normal elongation, and that the reduced stature of lkb plants is at least partially attributable to a reduction in free IAA level in this mutant.     

Internode length in Pisum. Action of gene lw

Jolly, C. J., Reid, J. B. and Ross, J. J. 1987. Internode length in Pisum. Action of gene lw.
Mutant K29 of Pisum sativum L. is shown to possess a recessive gene at a new locus, lw , which results in reduced internode length, delayed flowering and increased symptoms of water congestion compared with the parental cv. Torsdag. The interaction of gene lw with the internode length genes na, le, la and cry ⁵ is examined. Extracts from the shoots of Iw plants are shown to contain similar levels of gibberellin (GA)-like substances to comparable Lw plants, but Iw plants do not elongate to the same extent as Lw plants when treated with GA₁₉ GA₁₉, or GA₂₀. The effect of gene Iw is not graft-transmissible. Unlike essentially isogenic dwarf lines possessing the GA-synthesis genes le, Ih or Is, lw plants show a relative increase in elongation similar to Torsdag in response to photoperiod extensions from sources rich in far-red light. These results suggest that gene lw probably does not reduce elongation by influencing GA-synthesis and that the response to photoperiod extensions with far-red light may depend on the level of GA.     

Isolation and Characterization of Escherichia Coli Mutants with Altered Rates of Deletion Formation

Using site-specific mutagenesis in vitro we constructed a genetic system to detect mutants with altered rates of deletion formation between short repeated sequences in Escherichia coli. After in vivo mutagenesis with chemical mutagens and transposons, the system allowed the identification of mutants with either increased or decreased deletion frequencies. One mutational locus, termed mutR, that results in an increase in deletion formation, was studied in detail. The mutR gene maps at 38.5 min on the E. coli genetic map. Since the precise excision of many transposable elements is also mediated at short repeated sequences, we investigated the effects of the mutant alleles, as well as recA, on precise excision of the transposon Tn9. Neither mutR nor recA affect precise excision of the transposon Tn9, from three different insertions in lacI, whereas these alleles do affect other spontaneous deletions in the same system. These results indicate that deletion events leading to precise excision occur principally via a different pathway than other random spontaneous deletions. It is suggested that, whereas precise excision occurs predominantly via a pathway involving replication enzymes (for instance template strand slippage), deletions on an F'factor are stimulated by recombination enzymes.     

Altered Intersubunit Communication Is the Molecular Basis for Functional Defects of Pathogenic p97 Mutants

The human AAA ATPase p97 is a molecular chaperone essential in cellular proteostasis. Single amino acid substitutions in p97 have been linked to a clinical multiple-disorder condition known as inclusion body myopathy associated with Paget''s disease of the bone and frontotemporal dementia. How the mutations affect the molecular mechanism that governs the function of p97 remains unclear. Here, we show that within the hexameric ring of a mutant p97, D1 domains fail to regulate their respective nucleotide-binding states, as evidenced by the lower amount of prebound ADP, weaker ADP binding affinity, full occupancy of adenosine-5′-O-(3-thiotriphosphate) binding, and elevated overall ATPase activity, indicating a loss of communication among subunits. Defective communication between subunits is further illustrated by altered conformation in the side chain of residue Phe-360 that probes into the nucleotide-binding pocket from a neighboring subunit. Consequently, conformations of N domains in a hexameric ring of a mutant p97 become uncoordinated, thus impacting its ability to process substrate.     

Structural Basis of a Histone H3 Lysine 4 Demethylase Required for Stem Elongation in Rice

Histone lysine methylation is an important epigenetic modification in regulating chromatin structure and gene expression. Histone H3 lysine 4 methylation (H3K4me), which can be in a mono-, di-, or trimethylated state, has been shown to play an important role in gene expression involved in plant developmental control and stress adaptation. However, the resetting mechanism of this epigenetic modification is not yet fully understood. In this work, we identified a JmjC domain-containing protein, JMJ703, as a histone lysine demethylase that specifically reverses all three forms of H3K4me in rice. Loss-of-function mutation of the gene affected stem elongation and plant growth, which may be related to increased expression of cytokinin oxidase genes in the mutant. Analysis of crystal structure of the catalytic core domain (c-JMJ703) of the protein revealed a general structural similarity with mammalian and yeast JMJD2 proteins that are H3K9 and H3K36 demethylases. However, several specific features were observed in the structure of c-JMJ703. Key residues that interact with cofactors Fe(II) and N-oxalylglycine and the methylated H3K4 substrate peptide were identified and were shown to be essential for the demethylase activity in vivo. Several key residues are specifically conserved in known H3K4 demethylases, suggesting that they may be involved in the specificity for H3K4 demethylation.     

Internode length in Pisum. Gibberellins and the slender phenotype

Pea plants ( Pisum sativum L.) possessing the slender phenotype (conferred by the gene combination la cry^s ) have extremely long, thin internodes and are phenotypically similar to dwarf plants (possessing genes La and/or Cry ) that have been treated with a non-limiting dose of gibberellin (GA₃). In contrast to tall and dwarf plants, slender plants are virtually insensitive to treatment with AMO 1618, PP333 or GA₃ and addition of the "gibberellin-less" mutant gene na does not alter the phenotype of slender plants. Na slender segregates possessed lower levels of gibberellin-like substances than comparable dwarf segregates when extracts from shoots were assayed using the lettuce hypocotyl or rice seedling bioassays. In addition, na slenders possessed little or no gibberellin-like activity even though they possessed a slender phenotype. Thus the gene combination la cry^s causes slender plants to respond as if they are saturated with gibberellins for growth. In addition, the gene combinations la cry^s and le la cry^c (allele cry^c is less extreme in effect than cry^s ) are shown to be almost completely epistatic to the alleles at the na locus. All these results suggest that gibberellin levels are not important in determining the internode length of slender peas (genotype la cry^s ). The possible mechanisms by which this could occur are discussed.     

Blue-Light Regulation of Epicotyl Elongation in Pisum sativum

Blue light is known to induce suppression of stem elongation. To avoid the complication of blue-light-induced transformation of phytochrome we have adapted the procedure of measuring blue-light-induced suppression of stem elongation in Pisum sativum L. var Alaska grown under continuous red light. The resulting fluence-response curve for suppression of epicotyl elongation measured twenty-four hours after a blue-light treatment is bell-shaped, with the peak of suppression between 10⁰ and 10¹ micromoles per square meter, and no suppression at 10⁴ micromoles per square meter. Suppression is first observed 5 and 11 hours after the blue-light treatment for the fourth and third internodes, respectively. No significant differences in elongation rates were noted for the 10⁴ micromoles per square meter treated seedlings throughout the 24 hour period. Reciprocity holds for both third and fourth internodes in response to 10¹ and 10⁴ micromoles per square meter of blue light over the range of irradiation times tested (10⁰ to 10⁴ seconds, 10¹ micromoles per square meter; 10⁰ to 10³ seconds, 10⁴ micromoles per square meter). In contrast to the bell-shaped fluence-response obtained for epicotyl elongation, measurements of chlorophyll and carotenoid accumulation indicate increasing accumulation with increasing fluence.     

IAA和GA3在调控豌豆黄化苗茎切段伸长生长中的相互作用

IAA和GA3均能促进豌豆黄化苗茎切段的伸长。IAA效应可以为GA的合成抑制剂S-3307抑制,GA3效应同样也为IAA的运输抑制剂TIBA所抑制,并且分别再施用GA3和IAA后,抑制效应又能有所解除。观察顶端切半茎切段的结果表明,IAA主要促进茎切段表皮细胞的伸长,而GA3可能主要促进内部组织细胞的伸长。观察切段横纵切片的结果则显示,IAA促进皮层细胞的伸长和增大,而GA3只促进皮层细胞的伸长。这些结果说明两者是通过不同的作用部位和方式共同调节豌豆茎切段伸长生长的。