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Understanding “Stress” and How To Measure It

Understanding what stress in seeds is, and developing analytical methods in the search for potential markers of seed viability.

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High performance liquid chromatography is used to measure antioxidants and pigments in seeds
Wolfgang Stuppy

High-Performance Liquid Chromatography (HPLC) is one of the analytical methods used to measure chemical compounds involved in the seed stress response.

Conventional germination tests are convenient to assess seed viability in species of which certain seed traits are known. However, germination tests have been developed for crops, but not for the majority of the species stored in the Millennium Seed Bank, where we are challenged with a biodiversity of thousands of wild species.

Seed stress arises following exposure to unfavourable conditions both during development on the mother plant and following dispersal. Environmental factors such as temperature and humidity can affect seed development and also have consequences for seed longevity. In addition, exposure to toxic pollutants e.g. heavy metals can result in stress leading to reduction in seed viability.

We endeavour to use methods of biochemistry and molecular biology to find markers of viability and germination to diagnose and quantify ‘stress’, and to understand the regulatory switchboards that induce stress responses leading to acclimation and adaption. This project is one of a series in the research theme ‘Stress and Survival”.

Prior to stress marker development it must be understood what “stress” in seeds is, and if and how we can “put a number on it” as argued by Lord Kelvin in his famous quote:

“When you can measure what you are speaking about and express it in numbers you know something about it; but when you cannot measure it, when you cannot express it in numbers, your knowledge is of a meagre and unsatisfactory kind” (Kelvin, 1883).

We have recently developed a new stress model for seeds (see Annex 1), based on the General Adaptation Syndrome (Selye, 1936), which is the perhaps most accepted biomedical stress model since 1936. In the ‘alarm phase’ of this triphasic seed stress model, a stress factor is perceived by a signalling network of plant hormones and reactive oxygen species (ROS), and signals are transduced to activate the stress response systems. Under continuing stress, the ‘resistance phase’ is reached when the stress response is sufficient to protect cells from damage and to repair damage suffered, maintaining plant viability. The ‘exhaustion phase’ is reached when the protection and repair mechanisms fail, leading to the breakdown of essential molecules, including DNA, proteins and lipids, ultimately resulting in seed death.

The seed stress response is being investigated using a range of analytical approaches including omics-based methodology to gain a detailed picture of the processes initiated following exposure to stressful conditions. The analysis of certain compounds that respond to stressful environmental conditions is often a major challenge in itself. The chemical composition of plant seeds is dominated by their reserves, carbohydrates, lipids and proteins. When co-extracted with, for example nucleic acids, these compounds can interfere with conventional assays. In addition, many seeds are rich in (poly)phenols, which can be oxidized to quinones during extraction and then covalently bind to the analyte, for example, antioxidants, leading to erroneous results. Modifying existing methods, finding adequate combinations of existing methods to deal with several interferents simultaneously, or developing new assays for reproducible and reliable measurement of a certain group of chemical compounds is a challenging and often very time-consuming task for the analytical chemist, and we put significant effort into developing appropriate assays for our research.

 

 

Project partners and collaborators

China

 Kunming Institute of Botany, Chinese Academy of Sciences, Beijing
Graduate University of the Chinese Academy of Sciences, Beijing

France

University of Paris 6
INRA - Centre de Versailles

Germany

Albert-Ludwigs-University Freiburg
Center for Biotechnology, University of Bielefeld

Russia

Russian Academy of Sciences

South Africa

University of Kwa-Zulu Natal

Spain

Universidad de Salamanca

Project funders

UKMSBP

Annex material

Annex 1: The seed stress concept model. Read the definitive version of the New Phytologist article online

Key papers published since 2006

Kranner I and Colville L. 2011. Metals and seeds: Biochemical and molecular implications and their significance for seed germination. Environmental and Experimental Botany 72: 93-105.

Bailly C and Kranner I. 2011. Analyses of reactive oxygen species and antioxidants in relation to seed longevity and germination. In Kermode A (ed) Seed Dormancy: Methods and Protocols, Springer Science+Business Media, LLC, pp 343-367.

Kranner I, Beckett RP, Minibayeva FV and Seal CE. 2010. Tansley review: What is stress? Concepts, definitions and applications in seed science. New Phytologist 188: 655-673.

Seal CE, Pritchard HW and Kranner I. 2008. Quantification of seed oil from species with varying oil content using supercritical fluid extraction. Phytochemical Analysis 19, 493-498.

Birtic, S., & Kranner, I. (2006) Isolation of high-quality RNA from polyphenol-, polysaccharide- and lipid-rich seeds. Phytochemical Analysis 17: 144-148. 

Conferences and workshops

Since 2006 7 oral and poster contributions, 5 of which were plenary or keynote lectures, have been presented at 7 international conferences, including:

Meeting of the Italian Botanical Society, Plant Cell Biology Group (Rome, Italy 2011)

9th Conference of the International Society for Seed Science (in Olsztyn, Poland 2008)

3rd International Congress on Stress Responses in Biology and Medicine and 2nd World Conference of Stress (in Budapest, Hungary 2007)

 

 

 

 

Project team

Seed Conservation Department

Louise Colville, Charlotte Seal, Ilse Kranner

 

Project Leader: