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The “Protocol for Comparative Seed Longevity Testi The “Protocol for Comparative Seed Longevity Testing” is an official technical information sheet from the Millennium Seed Bank (MSB) that describes a standardized method used to compare the seed longevity of different plant species stored in conservation collections. The goal of the protocol is to generate a seed survival curve that reveals how quickly seed viability declines under controlled ageing conditions, allowing species to be ranked into longevity categories.
The method uses controlled rehydration followed by accelerated ageing. Seeds are first equilibrated at 47% relative humidity (RH) and 20°C to stabilize moisture content. They are then transferred to an ageing environment of 60% RH and 45°C, created using non-saturated lithium chloride (LiCl) solutions inside airtight containers. These uniform conditions ensure that all seed samples experience identical ageing stress.
During the ageing process, samples of 50 seeds are removed on a scheduled series of days (1, 2, 5, 9, 20, 30, 50, 75, 100, and 125). Each sample undergoes germination testing for at least 42 days, followed by a “cut test” to assess seed viability and identify empty, infested, or abnormal seeds. The resulting data are used to plot viability decline curves, typically analyzed using probit analysis and the Ellis & Roberts viability equation. A key output is p50, the time it takes for seed viability to drop to 50%, which enables clear comparisons across species and against two known “marker species” used by MSB.
The document also includes detailed preparation steps, practical guidance for ensuring accurate humidity control, tips for handling different seed types, and recommended equipment (such as hygrometers, fan-assisted ovens, airtight containers, and statistical software). It emphasizes that although the method does not predict exact natural longevity, it reliably ranks species and helps identify factors—such as seed maturity or post-harvest handling—that influence long-term seed survival.
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