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Nutrient and hormone levels in cotton ovules during embryony

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Abstract

In vitro zygotic and somatic embryogenesis protocols rely on nutrient and hormone levels from media to satisfy the physiological and developmental requirements of embryony. To better understand these requirements for cotton, we quantified levels of major and minor elements, carbohydrates, NH4 +, free amino acids and six hormones in whole cotton ovules (with fibers removed), nucelli (ovules with integuments removed), or ovule fluid (extracted from the endosperm region). Samples were collected from field-grown cotton at 1–18 days-past-anthesis (DPA) during each of three growing seasons. Replication across 2 years was obtained for carbohydrates, NH4 +, free amino acids and hormones from nucellus samples. The year effect was large primarily for hormones only. The most abundant minerals across tissue types and years were K, P, Mg and S. Potassium was the most abundant at 260, 600 and 1,660 mmol kg−1 dry mass (DM) in nucelli, whole ovules and ovule fluid, respectively. Magnesium, Ca, Zn and Mn levels were 2–8-fold higher in ovule fluid compared to whole ovules or nucelli. In the free amino acid plus NH4 + category, NH4 +, alanine, serine, glycine, asparagine (plus aspartic acid), glutamine (plus glutamic acid), leucine, threonine and arginine predominated in nucelli and ovule fluid, and levels tended to be higher in the older samples across years and tissue types. Fructose and glucose levels also increased with age with very high levels being found in late DPA ovule fluid. Arabinose, inositol and melibiose were also prominent sugars. Indole-3-acetic acid levels were similar between nucelli and ovule fluid and ranged from 10 to 80 μmol kg−1 DM. An abscisic acid spike, from 15 to 400 μmol kg−1 DM, occurred in nucelli and whole ovules from 2 to 8 DPA. Thereafter, abscisic acid levels remained between 5 and 10 μmol kg−1 DM. Zeatin and zeatin riboside were the most abundant cytokinins, and levels of these hormones fluctuated between 1 and 4 μmol kg−1 DM in both nucelli and ovule fluid.

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Abbreviations

Ala:

Alanine

Ara:

Arabinose

Arg:

Arginine

Asn:

Asparagine

Asp:

Aspartic acid

Asx:

Asparagine and aspartic acid

Cys:

Cystine

DHZ:

Dihydrozeatin

DHZR:

Dihydrozeatin riboside

DPA:

Days past anthesis

DM:

Dry mass

ELISA:

Enzyme linked immunosorbent assay

FM:

Fresh mass

Fru:

Fructose

Fu:

Fucose

GA:

Gibberellic acid

Glu:

Glutamic acid

Gluc:

Glucose

Gln:

Glutamine

Glx:

Glutamine and glutamic acid

Gly:

Glycine

His:

Histidine

HPLC:

High pressure liquid chromatography

IAA:

Indole-3-acetic acid

Ino:

Inositol

IP:

Isopentenyl adenine

IPA:

Isopentenyl adenosine

Ise:

Isoleucine

Leu:

Leucine

Lys:

Lysine

Mel:

Melibiose

Met:

Methionine

Phe:

Phenylalanine

Pro:

Proline

Ser:

Serine

Suc:

Sucrose

Thr:

Threonine

Tyr:

Tyrosine

Val:

Valine

Z:

Zeatin

ZR:

Zeatin riboside

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Acknowledgments

We thank Landon Farmer, Becky Kowallis, Laurie Gilbert, Chester Ogborn and Gordon Reese for technical assistance; Norma Trolinder and Linda Koonce for growing, harvesting and sending cotton bolls to Utah, and for technical discussions concerning cotton tissue culture; Dr. Philip Harrison for assistance with carbohydrate analyses; and Dr. Jan Kotuby-Amacher for mineral analyses. This research was supported by an U.S. Department of Agriculture, National Research Initiative, Competitive Grants Program award, No. 91-37300-6457; a Centers of Excellence grant (CVAST) from the State of Utah; a Department of Energy, Idaho Field Office Contract (DE-AC07-94ID13223); an Associated Western Universities, Inc., graduate student fellowship to RJF; and the Utah Agricultural Experiment Station, Utah State University, Logan, UT 84322-4810, USA. This paper is approved as Utah Agricultural Experiment Station journal paper number 8073.

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Authors and Affiliations

  1. Fennemore Craig, 3003 North Central Avenue Suite 2600, Phoenix, AZ, 85012-2913, USA

    Rodney J. Fuller

  2. Plants, Soils, and Climate Department, Utah State University, Logan, UT, 84322-4820, USA

    John G. Carman

  3. Idaho National Laboratory, P.O. Box 1625, Idaho Falls, ID, 83415-3710, USA

    J. Richard Hess

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  1. Rodney J. Fuller
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  2. John G. Carman
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  3. J. Richard Hess
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Correspondence to John G. Carman.

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Fuller, R.J., Carman, J.G. & Hess, J.R. Nutrient and hormone levels in cotton ovules during embryony. Plant Cell Tiss Organ Cult 99, 183–192 (2009). https://doi.org/10.1007/s11240-009-9591-1

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