Auxin

Native auxins

indole-3-acetic acid (IAA) is the most abundant and the basic auxin natively occurring and functioning in plants. It generates the majority of auxin effects in intact plants, and is the most potent native auxin.

There are four more endogenously synthesized auxins in plants.^[1][2]
All auxins are compounds with aromatic ring and a carboxylic acid group:^[2][3]

4-Chloroindole-3-acetic acid (4-CI-IAA)

2-phenylacetic acid (PAA)

Indole-3-butyric acid (IBA)

Indole-3-propionic acid (IPA)

For representatives of synthetic auxins, see: Auxin § Synthetic auxins.

Auxins (plural of auxin /ˈɔːksᵻn/) are a class of plant hormones (or plant growth substances) with some morphogen-like characteristics. Auxins have a cardinal role in coordination of many growth and behavioral processes in the plant's life cycle and are essential for plant body development. Auxins and their role in plant growth were first described by the Dutch scientist Frits Warmolt Went.^[4] Kenneth V. Thimann was the first to isolate one of these phytohormones and determine its chemical structure as indole-3-acetic acid (IAA). Went and Thimann co-authored a book on plant hormones, Phytohormones, in 1937.

Overview

Auxins were the first of the major plant hormones to be discovered. They derive their name from the Greek word αυξειν (auxein – "to grow/increase"). Auxin (namely IAA) is present in all parts of a plant, although in very different concentrations. The concentration in each position is crucial developmental information, so it is subject to tight regulation through both metabolism and transport. The result is the auxin creates "patterns" of auxin concentration maxima and minima in the plant body, which in turn guide further development of respective cells, and ultimately of the plant as a whole.

The (dynamic and environment responsive) pattern of auxin distribution within the plant is a key factor for plant growth, its reaction to its environment, and specifically for development of plant organs^[5][6] (such as leaves or flowers). It is achieved through very complex and well coordinated active transport of auxin molecules from cell to cell throughout the plant body — by the so-called polar auxin transport.^[5] Thus, a plant can (as a whole) react to external conditions and adjust to them, without requiring a nervous system. Auxins typically act in concert with, or in opposition to, other plant hormones. For example, the ratio of auxin to cytokinin in certain plant tissues determines initiation of root versus shoot buds.

On the molecular level, all auxins are compounds with an aromatic ring and a carboxylic acid group.^[3] The most important member of the auxin family is indole-3-acetic acid (IAA),^[1] which generates the majority of auxin effects in intact plants, and is the most potent native auxin. And as native auxin, its stability is controlled in many ways in plants, from synthesis, through possible conjugation to degradation of its molecules, always according to the requirements of the situation. However, molecules of IAA are chemically labile in aqueous solution, so it is not used commercially as a plant growth regulator.

• Five naturally occurring (endogenous) auxins in plants include indole-3-acetic acid, 4-chloroindole-3-acetic acid, phenylacetic acid, indole-3-butyric acid, and indole-3-propionic acid.^[1][2] However, most of the knowledge described so far in auxin biology and as described in the article below, apply basically to IAA; the other three endogenous auxins seems to have rather marginal importance for intact plants in natural environments. Alongside endogenous auxins, scientists and manufacturers have developed many synthetic compounds with auxinic activity.
• Synthetic auxin analogs include 1-naphthaleneacetic acid, 2,4-dichlorophenoxyacetic acid (2,4-D),^[1] and many others.

Some synthetic auxins, such as 2,4-D and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T), are used also as herbicides. Broad-leaf plants (dicots), such as dandelions, are much more susceptible to auxins than narrow-leaf plants (monocots) such as grasses and cereal crops, so these synthetic auxins are valuable as synthetic herbicides.

Discovery of auxin

Charles Darwin

In 1881, Charles Darwin and his son Francis performed experiments on coleoptiles, the sheaths enclosing young leaves in germinating grass seedlings. The experiment exposed the coleoptile to light from a unidirectional source, and observed that they bend towards the light.^[7] By covering various parts of the coleoptiles with a light-impermeable opaque cap, the Darwins discovered that light is detected by the coleoptile tip, but that bending occurs in the hypocotyl. However the seedlings showed no signs of development towards light if the tip was covered with an opaque cap, or if the tip was removed. The Darwins concluded that the tip of the coleoptile was responsible for sensing light, and proposed that a messenger is transmitted in a downward direction from the tip of the coleoptile, causing it to bend.^[8]

Peter Boysen-Jensen

In 1913, Danish scientist Peter Boysen-Jensen demonstrated that the signal was not transfixed but mobile. He separated the tip from the remainder of the coleoptile by a cube of gelatine which prevented cellular contact, but allowed chemicals to pass through. The seedlings responded normally bending towards the light. However, when the tip was separated by an impermeable substance, there was no curvature of the stem.^[9]

Frits Went

In 1928, the Dutch botanist Frits Warmolt Went showed that a chemical messenger diffuses from coleoptile tips. Went's experiment identified how a growth promoting chemical causes a coleoptile to grow towards light. Went cut the tips of the coleoptiles and placed them in the dark, putting a few tips on agar blocks that he predicted would absorb the growth-promoting chemical. On control coleoptiles, he placed a block that lacked the chemical. On others, he placed blocks containing the chemical, either centred on top of the coleoptile to distribute the chemical evenly or offset to increase the concentration on one side.^[8]

When the growth-promoting chemical was distributed evenly the coleoptile grew straight. If the chemical was distributed unevenly, the coleoptile curved away from the side with the cube, as if growing towards light, even though it was grown in the dark. Went later proposed that the messenger substance is a growth-promoting hormone, which he named auxin, that becomes asymmetrically distributed in the bending region. Went concluded that auxin is at a higher concentration on the shaded side, promoting cell elongation, which results in a coleoptiles bending towards the light.^[9]

Hormonal activity

Auxins help development at all levels in plants, from the cellular level, through organs, and ultimately to the whole plant.

Molecular mechanisms

Auxin molecules present in cells may trigger responses directly through stimulation or inhibition of the expression of sets of certain genes^[10] or by means independent of gene expression. Auxin transcriptionally activates four different families of early genes (aka primary response genes), so-called because the components required for the activation are preexisting, leading to a rapid response.^[11] The families are glutathione S-transferases, auxin homeostasis proteins like GH3, SAUR genes of currently unknown function, and the Aux/IAA repressors.^[12]

Aux/IAA, ARF, TIR1, SCF auxin regulatory pathways

The Aux/IAA repressors provide an example of one of the pathways leading to auxin induced changes of gene expression. This pathway involves the protein families TIR1 (transport inhibitor response1), ARF (auxin response factor), Aux/IAA transcriptional repressors, and the ubiquitin ligase complex that is a part of the ubiquitin-proteasome protein degradation pathway.^[13][14] ARF proteins have DNA binding domains and can bind promoter regions of genes and activate or repress gene expression. Aux/IAA proteins can bind ARF proteins sitting on gene promoters and prevent them from doing their job.

TIR1 proteins are F-box proteins that have three different domains giving them the ability to bind to three different ligands: an SCF^TIR1 ubiquitin ligase complex (using the F-box domain), auxin^[15] (so TIR1 proteins are auxin receptors), and Aux/IAA proteins (via a degron domain). Upon binding of auxin, a TIR1 protein's degron domain has increased affinity for Aux/IAA repressor proteins, which when bound to TIR1 and its SCF complex undergo ubiquitination and subsequent degradation by a proteasome. The degradation of Aux/IAA proteins frees ARF proteins to activate or repress genes at whose promoters they are bound.^[16]

Within a plant, elaboration of the Aux/IAA repressor pathway takes place via diversification of the TIR1, ARF, and Aux/IAA protein families. Each family may contain many similar-acting proteins, differing in qualities such as degree of affinity for partner proteins, amount of activation or repression of target gene transcription, or domains of expression (e.g. different plant tissues might express different members of the family, or different environmental stresses might activate expression of different members). Such elaboration permits the plant to use auxin in a variety of ways depending on the needs of the tissue and plant.

Other auxin regulatory pathways

Another protein, auxin-binding protein 1 (ABP1), is a putative receptor for a different signaling pathway, but its role is as yet unclear. Electrophysiological experiments with protoplasts and anti-ABP1 antibodies suggest ABP1 may have a function at the plasma membrane, and cells can possibly use ABP1 proteins to respond to auxin through means faster and independent of gene expression.

On a cellular level

On the cellular level, auxin is essential for cell growth, affecting both cell division and cellular expansion. Auxin concentration level, together with other local factors, contributes to cell differentiation and specification of the cell fate.

Depending on the specific tissue, auxin may promote axial elongation (as in shoots), lateral expansion (as in root swelling), or isodiametric expansion (as in fruit growth). In some cases (coleoptile growth), auxin-promoted cellular expansion occurs in the absence of cell division. In other cases, auxin-promoted cell division and cell expansion may be closely sequenced within the same tissue (root initiation, fruit growth). In a living plant, auxins and other plant hormones nearly always appear to interact to determine patterns of plant development.

Organ patterns

Growth and division of plant cells together result in growth of tissue, and specific tissue growth contributes to the development of plant organs.

Growth of cells contributes to the plant's size, unevenly localized growth produces bending, turning and directionalization of organs- for example, stems turning toward light sources (phototropism), roots growing in response to gravity (gravitropism), and other tropisms originated because cells on one side grow faster than the cells on the other side of the organ. So, precise control of auxin distribution between different cells has paramount importance to the resulting form of plant growth and organization.

Auxin transport and the uneven distribution of auxin

To cause growth in the required domains, auxins must of necessity be active preferentially in them. Local auxin maxima can be formed by active biosynthesis in certain cells of tissues, for example via tryptophan-dependent pathways,^[17] but auxins are not synthesized in all cells (even if cells retain the potential ability to do so, only under specific conditions will auxin synthesis be activated in them). For that purpose, auxins have to be not only translocated toward those sites where they are needed, but also they must have an established mechanism to detect those sites. For that purpose, auxins have to be translocated toward those sites where they are needed. Translocation is driven throughout the plant body, primarily from peaks of shoots to peaks of roots (from up to down).

For long distances, relocation occurs via the stream of fluid in phloem vessels, but, for short-distance transport, a unique system of coordinated polar transport directly from cell to cell is exploited. This short-distance, active transport exhibits some morphogenetic properties.

This process, polar auxin transport, is directional, very strictly regulated, and based in uneven distribution of auxin efflux carriers on the plasma membrane, which send auxins in the proper direction. While PIN-FORMED (PIN) proteins are vital in transporting auxin in a polar manner,^[6][18] the family of AUXIN1/LIKE-AUX1 (AUX/LAX) genes encodes for non-polar auxin influx carriers.^[19]

The regulation of PIN protein localisation in a cell determines the direction of auxin transport from cell, and concentrated effort of many cells creates peaks of auxin, or auxin maxima (regions having cells with higher auxin – a maximum).^[6] Proper and timely auxin maxima within developing roots and shoots are necessary to organise the development of the organ.^[5][20][21] Surrounding auxin maxima are cells with low auxin troughs, or auxin minima. For example, in the Arabidopsis fruit, auxin minima have been shown to be important for its tissue development.^[22]

Organization of the plant

As auxins contribute to organ shaping,^[5][6] they are also fundamentally required for proper development of the plant itself.^[5] Without hormonal regulation and organization, plants would be merely proliferating heaps of similar cells. Auxin employment begins in the embryo of the plant, where directional distribution of auxin ushers in subsequent growth and development of primary growth poles, then forms buds of future organs. Next, it helps to coordinate proper development of the arising organs, such as roots, cotyledons and leaves and mediates long distance signals between them, contributing so to the overall architecture of the plant.^[5] Throughout the plant's life, auxin helps the plant maintain the polarity of growth,^[5] and actually "recognize" where it has its branches (or any organ) connected.

An important principle of plant organization based upon auxin distribution is apical dominance, which means the auxin produced by the apical bud (or growing tip) diffuses (and is transported) downwards and inhibits the development of ulterior lateral bud growth, which would otherwise compete with the apical tip for light and nutrients. Removing the apical tip and its suppressively acting auxin allows the lower dormant lateral buds to develop, and the buds between the leaf stalk and stem produce new shoots which compete to become the lead growth. The process is actually quite complex, because auxin transported downwards from the lead shoot tip has to interact with several other plant hormones (such as strigolactones or cytokinins) in the process on various positions along the growth axis in plant body to achieve this phenomenon. This plant behavior is used in pruning by horticulturists.

Finally, the sum of auxin arriving from stems to roots influences the degree of root growth. If shoot tips are removed, the plant does not react just by outgrowth of lateral buds — which are supposed to replace to original lead. It also follows that smaller amount of auxin arriving to the roots results in slower growth of roots and the nutrients are subsequently in higher degree invested in the upper part of the plant, which hence starts to grow faster.

Effects

A healthy Arabidopsis thaliana plant (left) next to an auxin signal-transduction mutant with a repressed response to auxin.

Crown galls are caused by Agrobacterium tumefaciens bacteria; they produce and excrete auxin and cytokinin, which interfere with normal cell division and cause tumors.

Auxin participates in phototropism, geotropism, hydrotropism and other developmental changes. The uneven distribution of auxin, due to environmental cues, such as unidirectional light or gravity force, results in uneven plant tissue growth, and generally, auxin governs the form and shape of plant body, direction and strength of growth of all organs, and their mutual interaction.^[6]

Auxin stimulates cell elongation by stimulating wall-loosening factors, such as elastins, to loosen cell walls. The effect is stronger if gibberellins are also present. Auxin also stimulates cell division if cytokinins are present. When auxin and cytokinin are applied to callus, rooting can be generated if the auxin concentration is higher than cytokinin concentration. Xylem tissues can be generated when the auxin concentration is equal to the cytokinins.

Auxin also induces sugar and mineral accumulation at the site of application.

Wound response

Auxin induces the formation and organization of phloem and xylem. When the plant is wounded, the auxin may induce the cell differentiation and regeneration of the vascular tissues.^[23]

Root growth and development

Auxins promote root initiation.^[24] Auxin induces both growth of pre-existing roots and adventitious root formation, i.e., branching of the roots. As more native auxin is transported down the stem to the roots, the overall development of the roots is stimulated. If the source of auxin is removed, such as by trimming the tips of stems, the roots are less stimulated accordingly, and growth of stem is supported instead.

In horticulture, auxins, especially NAA and IBA, are commonly applied to stimulate root initiation when rooting cuttings of plants. However, high concentrations of auxin inhibit root elongation and instead enhance adventitious root formation. Removal of the root tip can lead to inhibition of secondary root formation.

Apical dominance

Auxin induces shoot apical dominance; the axillary buds are inhibited by auxin, as a high concentration of auxin directly stimulates ethylene synthesis in axillary buds, causing inhibition of their growth and potentiation of apical dominance. When the apex of the plant is removed, the inhibitory effect is removed and the growth of lateral buds is enhanced. Auxin is sent to the part of the plant facing away from the light, where it promotes cell elongation, thus causing the plant to bend towards the light.^[25]

Fruit growth and development

Auxin is required for fruit growth and development and delays fruit senescence. When seeds are removed from strawberries, fruit growth is stopped; exogenous auxin stimulates the growth in fruits with seeds removed. For fruit with unfertilized seeds, exogenous auxin results in parthenocarpy ("virgin-fruit" growth).

Fruits form abnormal morphologies when auxin transport is disturbed.^[26] In Arabidopsis fruits, auxin controls the release of seeds from the fruit (pod). The valve margins are a specialised tissue in pods that regulates when pod will open (dehiscence). Auxin must be removed from the valve margin cells to allow the valve margins to form. This process requires modification of the auxin transporters (PIN proteins).^[22]

Flowering

Auxin plays also a minor role in the initiation of flowering and development of reproductive organs. In low concentrations, it can delay the senescence of flowers. A number of plant mutants have been described that affect flowering and have deficiencies in either auxin synthesis or transport. In maize, one example is bif2 barren inflorescence2.^[27]

Ethylene biosynthesis

In low concentrations, auxin can inhibit ethylene formation and transport of precursor in plants; however, high concentrations can induce the synthesis of ethylene.^[28] Therefore, the high concentration can induce femaleness of flowers in some species.

Auxin inhibits abscission prior to formation of abscission layer, and thus inhibits senescence of leaves.

Synthetic auxins

In the course of research on auxin biology, many compounds with noticeable auxin activity were synthesized. Many of them had been found to have economical potential for man-controlled growth and development of plants in agronomy. Synthetic auxins include the following compounds:

Auxins are toxic to plants in large concentrations; they are most toxic to dicots and less so to monocots. Because of this property, synthetic auxin herbicides, including 2,4-D and 2,4,5-T, have been developed and used for weed control.

However, some exogenously synthesized auxins, especially 1-naphthaleneacetic acid (NAA) and indole-3-butyric acid (IBA), are also commonly applied to stimulate root growth when taking cuttings of plants or for different agricultural purposes such as the prevention of fruit drop in orchards.

Used in high doses, auxin stimulates the production of ethylene. Excess ethylene (also native plant hormone) can inhibit elongation growth, cause leaves to fall (abscission), and even kill the plant. Some synthetic auxins, such as 2,4-D and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) were marketed also as herbicides. Dicots, such as dandelions, are much more susceptible to auxins than monocots, such as grasses and cereal crops. So, these synthetic auxins are valuable as synthetic herbicides. 2,4-D was the first widely used herbicide, and it is still so.^[29] 2,4-D was first commercialized by the Sherwin-Williams company, and saw use in the late 1940s. It is easy and inexpensive to manufacture.

Herbicide manufacture

The defoliant Agent Orange, used extensively by British forces in the Malayan Emergency and American forces in the Vietnam War, was a mix of 2,4-D and 2,4,5-T. The compound 2,4-D is still in use and is thought to be safe, but 2,4,5-T was more or less banned by the U.S. Environmental Protection Agency in 1979. The dioxin TCDD is an unavoidable contaminant produced in the manufacture of 2,4,5-T. As a result of the integral dioxin contamination, 2,4,5-T has been implicated in leukemia, miscarriages, birth defects, liver damage, and other diseases.

See also

• Auxin binding protein
• Fusicoccin
• Herbicide; specifically, see the chapter: ^#History
  • Phenoxy herbicide
• Pruning fruit trees
• Tropism
• Witch's broom
• Toshio Murashige
• Folke K. Skoog
• Kenneth V. Thimann

References