Seed testing

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Seed germination testing Seed germination test. State Forest.jpg
Seed germination testing

Seed testing is an analysis of seeds for viability.[ citation needed ] Seed testing performed for a number of reasons, including research purposes or to determine if seed storage techniques are functioning. Common seed tests include Germination tests, Viability tests, purity tests and weed tests. The first two are common for scientific research. For commercially sold seed, all four of these tests are done in dedicated laboratories by trained and usually certified analysts. The tests are designed to evaluate the quality of the seed lot being sold. [1]

Contents

Germination tests

A germination test reports the percentage of seed that germinated. In commercial settings, tests are usually made in either 200 or 400 seed samples.

Viability test

The Tetrazolium Chloride (TZ) test, often called the quick germination test, is a chemical test used to determine seed viability, and results are usually available within 24 to 48 hours The TZ test differs from a germination test in that the TZ test can give you an early and quick snapshot of seed viability but is not a replacement for the more comprehensive seed germination test. In Canada, the TZ test is not officially recognized by the CFIA (except for western wheatgrass where the TZ result may be added to the germination for a final germination total). In the United States, the TZ test can be used as a replacement for a germination test, although a follow-up germination test is usually recommended.

Advantages of the TZ test

Disadvantages of the TZ test

Procedure

A viability test (TZ test) is a test for viability that involves the steps: preconditioning (imbibition) and then preparation and staining (sometimes cutting the seed and then soaking the seed in a 2,3,5 triphenyl tetrazolium chloride solution).

Seeds are soaked in water overnight. They may be pre-moistened, in which case the seeds are allowed to imbibe water between a moistened germination paper blotter. Seeds are then dissected, either longitudinally or transversely, with a scalpel so that the embryo is exposed to the tetrazolium chloride solution. One half of this seed is used for the test and the other half is discarded. Staining is done with a solution of 2, 3, 5 triphyenyl tetrazolium chloride (a salt) is added to water to form a colorless solution. The seeds are placed in a 1% solution (for legumes and grasses that are not bisected), or a dilute 0.1% solution for bisected grasses and cereals. Seed coats of legumes must usually be removed or peeled back before examination. Care must be taken to prevent breaking of radicles and other damage to the seeds.

Evaluation

Dehydrogenase enzymes present in living tissue reduce the tetrazolium chloride to formazan, a reddish, water-insoluble compound. This reaction occurs in or near living cells, which are releasing hydrogen in respiration processes. Depending on size, all seeds are examined under a microscope at 10-30 power. Larger seeds, such as peas, may be examined without a microscope. Analysts look for three things: Sound tissues, weak living tissues or dead tissues. Sound tissues produce a normal red color and resist the penetration of tetrazolium. The rate of hydrogen released in sound tissue is slow in comparison to that in partially weakened tissues. Weak living tissues produce an abnormal color. These tissues have lost some of their initial resistance to the penetration of tetrazolium. Respiration is accelerated and formazan is produced rapidly. During the early stages of deterioration, these tissues become darker red (bruised) quicker than sound, healthy tissues. Dead tissues do not stain, remaining usually white (aged tissue) because the lack of respiration prevents the production of formazan.

TZ test results are recorded as a percentage and are usually reported in the remarks section of the Certificate of Seed Analysis issued by the seed laboratory. Hard seeds are to be reported (if applicable) as a percentage and are included in the total percent viable seed. Regardless of how they are reported, TZ test results for viability give you an estimate of the maximum percentage of seeds that have the potential to produce normal seedlings.

Purity test

A purity test reports the percentage of seed described on the label that is actually found in the quantity of seed.

Weed test

A weed test examines a sample of seed and identifies every seed that is different from the labeled seed kind.

Related Research Articles

<span class="mw-page-title-main">Seed</span> Embryonic plant enclosed in a protective outer covering

In botany, a seed is a plant embryo and food reserve enclosed in a protective outer covering called a seed coat (testa). More generally, the term "seed" means anything that can be sown, which may include seed and husk or tuber. Seeds are the product of the ripened ovule, after the embryo sac is fertilized by sperm from pollen, forming a zygote. The embryo within a seed develops from the zygote and grows within the mother plant to a certain size before growth is halted.

Hygroscopy is the phenomenon of attracting and holding water molecules via either absorption or adsorption from the surrounding environment, which is usually at normal or room temperature. If water molecules become suspended among the substance's molecules, adsorbing substances can become physically changed, e.g. changing in volume, boiling point, viscosity or some other physical characteristic or property of the substance. For example, a finely dispersed hygroscopic powder, such as a salt, may become clumpy over time due to collection of moisture from the surrounding environment.

<span class="mw-page-title-main">Seed bank</span> Backup seed storage

A seed bank stores seeds to preserve genetic diversity; hence it is a type of gene bank. There are many reasons to store seeds. One is to preserve the genes that plant breeders need to increase yield, disease resistance, drought tolerance, nutritional quality, taste, etc. of crops. Another is to forestall loss of genetic diversity in rare or imperiled plant species in an effort to conserve biodiversity ex situ. Many plants that were used centuries ago by humans are used less frequently now; seed banks offer a way to preserve that historical and cultural value. Collections of seeds stored at constant low temperature and low moisture are guarded against loss of genetic resources that are otherwise maintained in situ or in field collections. These alternative "living" collections can be damaged by natural disasters, outbreaks of disease, or war. Seed banks are considered seed libraries, containing valuable information about evolved strategies to combat plant stress, and can be used to create genetically modified versions of existing seeds. The work of seed banks often span decades and even centuries. Most seed banks are publicly funded and seeds are usually available for research that benefits the public.

<span class="mw-page-title-main">Germination</span> Process by which an organism grows from a spore or seed

Germination is the process by which an organism grows from a seed or spore. The term is applied to the sprouting of a seedling from a seed of an angiosperm or gymnosperm, the growth of a sporeling from a spore, such as the spores of fungi, ferns, bacteria, and the growth of the pollen tube from the pollen grain of a seed plant.

<span class="mw-page-title-main">Sprouting</span> Practice of germinating seeds to be eaten raw or cooked

Sprouting is the natural process by which seeds or spores germinate and put out shoots, and already established plants produce new leaves or buds, or other structures experience further growth.

Cytotoxicity is the quality of being toxic to cells. Examples of toxic agents are toxic metals, toxic chemicals, microbe neurotoxins, radiation particles and even specific neurotransmitters when the system is out of balance. Also some types of venom, e.g. from the puff adder or brown recluse spider are toxic to cells.

<span class="mw-page-title-main">Microscope slide</span> Thin, flat piece of glass onto which a sample is placed to be examined under a microscope

A microscope slide is a thin flat piece of glass, typically 75 by 26 mm and about 1 mm thick, used to hold objects for examination under a microscope. Typically the object is mounted (secured) on the slide, and then both are inserted together in the microscope for viewing. This arrangement allows several slide-mounted objects to be quickly inserted and removed from the microscope, labeled, transported, and stored in appropriate slide cases or folders etc.

<span class="mw-page-title-main">Staining</span> Technique used to enhance visual contrast of specimens observed under a microscope

Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Stains and dyes are frequently used in histology, in cytology, and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses of diseases at the microscopic level. Stains may be used to define biological tissues, cell populations, or organelles within individual cells.

<span class="mw-page-title-main">Histopathology</span> Microscopic examination of tissue in order to study and diagnose disease

Histopathology is the microscopic examination of tissue in order to study the manifestations of disease. Specifically, in clinical medicine, histopathology refers to the examination of a biopsy or surgical specimen by a pathologist, after the specimen has been processed and histological sections have been placed onto glass slides. In contrast, cytopathology examines free cells or tissue micro-fragments.

<span class="mw-page-title-main">MTT assay</span> Colorimetric analysis for measuring activity of cellular enzymes that reduce a tetrazolium dye

The MTT assay is a colorimetric assay for assessing cell metabolic activity. NAD(P)H-dependent cellular oxidoreductase enzymes may, under defined conditions, reflect the number of viable cells present. These enzymes are capable of reducing the tetrazolium dye MTT, which is chemically 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, to its insoluble formazan, which has a purple color. Other closely related tetrazolium dyes including XTT, MTS and the WSTs, are used in conjunction with the intermediate electron acceptor, 1-methoxy phenazine methosulfate (PMS). With WST-1, which is cell-impermeable, reduction occurs outside the cell via plasma membrane electron transport. However, this traditionally assumed explanation is currently contended as proof has also been found of MTT reduction to formazan in lipidic cellular structures without apparent involvement of oxidoreductases.

Zenker's fixative is a rapid-acting fixative for animal tissues. It is employed to prepare specimens of animal or vegetable tissues for microscopic study. It provides excellent fixation of nuclear chromatin, connective tissue fibers and some cytoplasmic features, but does not preserve delicate cytoplasmic organelles such as mitochondria. Helly's fixative is preferable for traditional dye staining of mitochondria. Zenker's fixative permeabilises the plasma, but not the nuclear membrane. It can therefore be used to selectively stain mitotic cells with antibodies against chromatin

A vital stain in a casual usage may mean a stain that can be applied on living cells without killing them. Vital stains have been useful for diagnostic and surgical techniques in a variety of medical specialties. In supravital staining, living cells have been removed from an organism, whereas intravital staining is done by injecting or otherwise introducing the stain into the body. The term vital stain is used by some authors to refer to an intravital stain, and by others interchangeably with a supravital stain, the core concept being that the cell being examined is still alive. In a more strict sense, the term vital staining has a meaning contrasting with supravital staining. While in supravital staining the living cells take up the stain, in "vital staining" – the most accepted but apparently paradoxical meaning of this term, the living cells exclude the stain i.e. stain negatively and only the dead cells stain positively and thus viability can be assessed by counting the percentage of total cells that stain negatively. Very bulky or highly charged stains that don't cross live plasma membrane are used as vital stains and supravital stains are those that are either small or are pumped actively into live cells. Since supravital and intravital nature of the staining depends on the dye, a combination of supravital and vital dyes can also be used in a sophisticated way to better classify cells into distinct subsets.

Imbibition is a special type of diffusion that takes place when liquid is absorbed by solids-colloids causing an increase in volume. Water surface potential movement takes place along a concentration gradient; some dry materials absorb water. A gradient between the absorbent and the liquid is essential for imbibition. For a substance to imbibe a liquid, there must first be some attraction between them. Imbibition occurs when a wetting fluid displaces a non-wetting fluid, the opposite of drainage in which a non-wetting phase displaces the wetting fluid. The two processes are governed by different mechanisms. Imbibition is also a type of diffusion since water movement is along the concentration gradient. Seeds and other such materials have almost no water hence they absorb water easily. Water potential gradient between the absorbent and liquid imbibed is essential for imbibition.

<span class="mw-page-title-main">Neutral red</span> Chemical compound

Neutral red is a eurhodin dye used for staining in histology. It stains lysosomes red. It is used as a general stain in histology, as a counterstain in combination with other dyes, and for many staining methods. Together with Janus Green B, it is used to stain embryonal tissues and supravital staining of blood. Can be used for staining Golgi apparatus in cells and Nissl granules in neurons.

<span class="mw-page-title-main">Uranyl acetate</span> Chemical compound

Uranyl acetate is the acetate salt of uranium oxide, a toxic yellow-green powder useful in certain laboratory tests. Structurally, it is a coordination polymer with formula UO2(CH3CO2)2(H2O)·H2O.

<span class="mw-page-title-main">Silver chromate</span> Chemical compound

Silver chromate is an inorganic compound with formula Ag2CrO4 which appears as distinctively coloured brown-red crystals. The compound is insoluble and its precipitation is indicative of the reaction between soluble chromate and silver precursor salts (commonly potassium/sodium chromate with silver nitrate). This reaction is important for two uses in the laboratory: in analytical chemistry it constitutes the basis for the Mohr method of argentometry, whereas in neuroscience it is used in the Golgi method of staining neurons for microscopy.

<span class="mw-page-title-main">Tetrazolium chloride</span> Redox indicator

Triphenyl tetrazolium chloride (TTC), or simply tetrazolium chloride is a redox indicator commonly used in biochemical experiments especially to indicate cellular respiration. It is a white crystalline powder, soluble in water, ethanol and acetone but insoluble in ether.

The formazans are compounds of the general formula [R-N=N-C(R')=N-NH-R"], formally derivatives of formazan [H2NN=CHN=NH], unknown in free form.

<i>Tylosema esculentum</i> Species of flowering plant

Tylosema esculentum, with common names gemsbok bean and marama bean or morama bean, is a long-lived perennial legume native to arid areas of southern Africa. Stems grow at least 3 metres (9.8 ft), in a prostrate or trailing form, with forked tendrils that facilitate climbing. A raceme up to 25 millimetres (1 in) long, containing many yellow-orange flowers, ultimately produces an ovate to circular pod, with large brownish-black seeds.

<span class="mw-page-title-main">Oldest viable seed</span> Oldest seed known to have grown into a full plant

There have been several seeds known at different times as the oldest viable seed.

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