A Rapid and Simple Procedure for Purification of Indole-3-Acetic Acid Prior to GC-SIM-MS Analysis

Chen, Kai-Hsien; Miller, Anita N.; Patterson, Glenn W.; Cohen, Jerry D.

doi:10.1104/pp.86.3.822

Cited by 177 publications

(137 citation statements)

References 15 publications

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“…In this process, as previously proposed by Chory (1993), light seems to play an important role by stimulating the production of the shoot, which was the main sink to the products of xyloglucan. The reductions of the rate of xyloglucan mobilization by excision of the shoot or by isolation of cotyledons appear to be related to the observed re- Peres et al, 1997) and by GC-SIM-MS (B; Chen et al, 1988). In A, the IAA was measured in the attached cotyledons from intact seedlings of H. courbaril growing in light, darkness, or with shoot excised above the cotyledons insertions (shoot excised) at 34 d. The only isolated cotyledons used as a reference in this case were the ones detached at 34 d and subsequently kept in the darkness.…”

Section: Discussionmentioning

confidence: 93%

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The Control of Storage Xyloglucan Mobilization in Cotyledons of Hymenaea courbaril

et al. 2004

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Hymenaea courbaril is a leguminous tree species from the neotropical rain forests. Its cotyledons are largely enriched with a storage cell wall polysaccharide (xyloglucan). Studies of cell wall storage polymers have been focused mostly on the mechanisms of their disassembly, whereas the control of their mobilization and the relationship between their metabolism and seedling development is not well understood. Here, we show that xyloglucan mobilization is strictly controlled by the development of first leaves of the seedling, with the start of its degradation occurring after the beginning of eophyll (first leaves) expansion. During the period of storage mobilization, an increase in the levels of xyloglucan hydrolases, starch, and free sugars were observed in the cotyledons. Xyloglucan mobilization was inhibited by shoot excision, darkness, and by treatment with the auxin-transport inhibitor N-1-naphthylphthalamic acid. Analyses of endogenous indole-3-acetic acid in the cotyledons revealed that its increase in concentration is followed by the rise in xyloglucan hydrolase activities, indicating that auxin is directly related to xyloglucan mobilization. Cotyledons detached during xyloglucan mobilization and treated with 2,4-dichlorophenoxyacetic acid showed a similar mobilization rate as in attached cotyledons. This hormonal control is probably essential for the ecophysiological performance of this species in their natural environment since it is the main factor responsible for promoting synchronism between shoot growth and reserve degradation. This is likely to increase the efficiency of carbon reserves utilization by the growing seedling in the understorey light conditions of the rain forest.

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Section: Discussionmentioning

confidence: 93%

“…The GC-SIM-MS analyses of free endogenous IAA were performed according to Chen et al (1988). Five fresh cotyledons (per date and treatment) were chopped into small pieces, weighed, and divided into three subsamples (0.5 g).…”

Section: Measurements Of Endogenous Iaamentioning

confidence: 99%

The Control of Storage Xyloglucan Mobilization in Cotyledons of Hymenaea courbaril

et al. 2004

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“…Total IAA was determined essentially by the method of Chen et al (1988). The extract was hydrolyzed with 7 N NaOH at 100°C for 3 h using oxygen-scrubbed N, (Bialek and Cohen, 1989).…”

Section: Iaa Analysismentioning

confidence: 99%

Selection and Characterization of [alpha]-Methyltryptophan-Resistant Lines of Lemna gibba Showing a Rapid Rate of Indole-3-Acetic Acid Turnover

1995

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Turnover rate is an important aspect of the regulation of plant processes by plant growth substances. To study turnover of indole-3-acetic acid (IAA), two a-methyltryptophan-resistant lines (MTR1 and MTR2) of Lemna gibba were generated by nitrosomethyl urea treatment of an inbred line derived from L. gibba G-3. In this report we describe: (a) the development of a selection system using this near isogenic line of L. gibba; (b) techniques for chemical mutation of the lines and selection for a-methyltryptophan resistance; and (c) the partia1 characterization of the selected lines. MTR lines contained 3-fold higher levels of anthranilate synthase activity. The enzyme in the MTR lines required higher levels of tryptophan for feedback inhibition. MTR lines also contained 8-fold higher levels of tryptophan, 3-fold higher levels of free IAA, and similar levels of total IAA compared to the inbred line. Turnover rates in the inbred and selected lines were calculated, using the first-order rate equation, based on the decrease over time in isotopic enrichment of 13C,-IAA introduced into L. gibba during a 1-h pulse period. lsotope enrichment in IAA was determined by using gas chromatography-mass spectrometry. Both MTR lines had an approximately 10-fold higher rate of IAA turnover than the parent inbred lhe.IAA plays an important role in plant growth, development, and responses to the environment, yet after more than a century of study we still have large gaps in our knowledge of fundamental aspects of how the hormone acts and what controls its metabolism. One aspect of plant hormone metabolism that has attracted attention over the last 15 years is the rate of hormone turnover (Epstein et al., 1980;Nonhebel and Cooney, 1990). Hormonal homeostasis requires that increased rates of synthesis of IAA be offset by increasing rates of degradation, resulting in more rapid turnover. Studies of hormone turnover have been limited in number in part by lack of plant systems that overcome the difficulties of labeling interna1 pools. In many cases, IAA levels in plant tissues remain relatively constant, but changes in the rates of IAA biosynthesis and degradation are seen only as changes in turnover rate. To date we have information on turnover in only a few plants and no knowledge of how turnover might change relative to genetic, developmental, and environmental variation. Turnover can be expected to be an important metabolic parameter for hormone action, since rates of turnover must be rapid relative to the processes that are controlled by the particular hormone (see Cohen, 1983). A mechanism linking turnover and hormone action may involve the co-oxidation of plant fatty acids (Reineke, 1990).Studies of IAA biosynthesis have shown the utility of using mutants to study IAA and Trp metabolism (Wright et al., 1991;Normanly et al., 1993). Unlike mutants used for studies of metabolic pathways, those most useful for understanding the function of IAA turnover would not be auxotrophs, but rather would be modified in metabolism so as to result in...

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“…Specifically, CM was collected from the following: three separate 4-d-old xylogenic cultures (112 mL total volume), six separate 9-d-old xylogenic cultures (265 mL total volume), and five separate 9-d-old nonxylogenic cultures (158 mL total volume). Total IAA (free IAA plus conjugated IAA) was quantitated using [ 3 H]IAA as a radioactive tracer and [ 13 C 61 IAA as an internal standard, as previously described (Chen et al, 1988;Cohen et al, 1992). Briefly, after evaporating in vacuo the medium was subjected to alkaline hydrolysis (3 h, 7 N NaOH, 100°C) then purified on a Prep Sep C ]8 disposable column (Fisher Scientific) and by HPLC on a C 18 reverse-phase column.…”

Section: Analysis Of Total Iaamentioning

confidence: 99%

“…Briefly, after evaporating in vacuo the medium was subjected to alkaline hydrolysis (3 h, 7 N NaOH, 100°C) then purified on a Prep Sep C ]8 disposable column (Fisher Scientific) and by HPLC on a C 18 reverse-phase column. The IAA fraction was methylated and analyzed by GC-selected ion-monitoring MS (5890 series II GC/5971A MS, Hewlett Packard), according to published methods (Chen et al, 1988;Cohen et al, 1992).…”

Section: Analysis Of Total Iaamentioning

confidence: 99%

A Secreted Factor Inducs Cell Expansion and Formation of Metaxylem-Like Tracheary Elements in Xylogenic Suspension Cultures of Zinnia

1997

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Conditioned medium from mesophyll cell-suspension cultures of Zinnia eregans 1. has striking effects on cell expansion and tracheary element differentiation when applied to cultures of freshly isolated mesophyll cells. These effects include (a) induction of early cell expansion, (b) delay in differentiation by 48 h or more, (c) reduction in the synchrony of differentiation, and (d) early formation of very large, metaxylem-like tracheary elements. Like reduced osmotic potential and buffering at pH 5.5, conditioned medium appears to have its primary effect on cell expansion. Partia1 characterization of the expansion-inducing factor indicates that it is heat stable, of low molecular mass, and is resistant to protease. It also binds reversibly to concanavalin A but is not adsorbed by charcoal. We suggest that the secreted factor may be an oligosaccharide involved in the coordination of cell expansion and differentiation and the regulation of the protoxylem-like to metaxylem-like transition in xylogenic suspension cultures.In apical meristems cell expansion and cell differentiation are coordinated to give rise to highly organized tissues. For example, protoxylem TEs, which develop while surrounding cortical cells are expanding, deposit secondary cell walls in annular or helical patterns that allow the TEs to expand longitudinally in spite of the inelasticity of the secondary wall. As cell expansion ceases, developing metaxylem TEs deposit reticulate or pitted secondary cell walls that resist stretching and are also better able to withstand the pressure differential that develops between TEs and surrounding tissues during transpiration.Correlations between patterns of cell expansion and cell and tissue differentiation have been noted in differentiating culture systems, including embryogenic carrot (Daucus carota L.) cell-suspension cultures (van Engelen and de Vries, 1992), tobacco (Nicotiana tabacum L.) thin-layer cultures (Tran Thanh Van and Mutaftschiev, 1990), and xylogenic suspension cultures (Roberts and Haigler, 1994). In developing xylem the abundance of TEs (Torrey et al., 1971) and pattern of secondary-cell-wall deposition

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A Rapid and Simple Procedure for Purification of Indole-3-Acetic Acid Prior to GC-SIM-MS Analysis

Cited by 177 publications

References 15 publications

The Control of Storage Xyloglucan Mobilization in Cotyledons of Hymenaea courbaril

The Control of Storage Xyloglucan Mobilization in Cotyledons of Hymenaea courbaril

Selection and Characterization of [alpha]-Methyltryptophan-Resistant Lines of Lemna gibba Showing a Rapid Rate of Indole-3-Acetic Acid Turnover

A Secreted Factor Inducs Cell Expansion and Formation of Metaxylem-Like Tracheary Elements in Xylogenic Suspension Cultures of Zinnia

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