Name: reliable method for assessing seed germination, dormancy, and mortality under field conditions
Uploaded: 2016-11-06T13:00:00
Description: Watch this Scientific Journal Video about Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions at JoVE.com

This work was supported by Biotech Risk Assessment Grant Program competitive grand no. 2006-39454-17438 to A. Snow, K. Mercer, and H. Alexander from the United States Department of Agriculture, National Institute of Food and Agriculture. Experiments using this method were conducted at and supported by the University of Kansas Field Station, a research unit of the Kansas Biological Survey and the University of Kansas. The authors would like to thank P. Jourdan and E. Regnier for helpful reviews on earlier versions of this manuscript. Additionally, this work was aided by the contributions of the staff at the University of Kansas Field Station, Waterman Farm at the Ohio State University (OSU), the USDA Ornamental Plant Germplasm Center at OSU, and the Seed Biology Lab in the Department of Horticulture and Crop Science at OSU, especially E. Renze, S. Stieve, A. Evans, and E. Grassbaugh, for technical support.

1. Saml Frø fra flere Dyreart eller kontrollerede krydsninger af en enkelt art Bemærk: Dette eksempel brugt frø fra 15 cross typer inden arten Helianthus annuus (solsikke) ved hjælp af naturen, hybrid, og afgrødetyper som moderens (seed producerende) forælder. <ol><li> Ved slutningen af ​​vækstsæsonen, indsamle modne frø hoveder i mærkede poser. Clean frø fra bukkene og sted frø i kuverter mærket med forældrenes krydstype i standardformat (dvs. moderens x fædrene). </li><li...

<ol>
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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+classification+system+for+seed+dormancy.">A classification system for seed dormancy.</a> Seed Sci Res. 14 (1), 1-16 (2004).</li><li>Donohue, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Environmental+and+genetic+influences+on+the+germination+of+Arabidopsis+thaliana+in+the+field.">Environmental and genetic influences on the germination of Arabidopsis thaliana in the field.</a> Evolution. 54 (4), 740-757 (2005).</li><li>Roach, D. A., Wulff, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Maternal+effects+in+plants.">Maternal effects in plants.</a> Ann Rev Ecol Syst. 18, 209-235 (1987).</li><li>Pace, B. A., Alexander, H. M., Emry, J. D., Mercer, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Seed+fates+in+crop-wild+hybrid+sunflower:+crop+allele+and+maternal+effects.">Seed fates in crop-wild hybrid sunflower: crop allele and maternal effects.</a> Evol Appl. 8 (2), 121-132 (2015).</li><li>Alexander, H. M., Emry, D. J., Pace, B. A., Kost, M. A., Sparks, K. A., Mercer, K. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Roles+of+maternal+effects+and+nuclear+genetic+composition+change+across+the+life+cycle+of+crop-wild+hybrids.">Roles of maternal effects and nuclear genetic composition change across the life cycle of crop-wild hybrids.</a> Am J Bot. 101 (7), 1176-1188 (2014).</li><li>Stewart, N. C., Matthew, J., Halfhill, D., Warwick, S. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgene+introgression+from+genetically+modified+crops+into+their+wild+relatives.">Transgene introgression from genetically modified crops into their wild relatives.</a> Genetics. 4, 806-817 (2003).</li><li>Mercer, K. L., Shaw, R. G., Wyse, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+germination+of+diverse+crop-wild+hybrid+sunflower+seeds.">Increased germination of diverse crop-wild hybrid sunflower seeds.</a> Ecol Appl. 16, 845-854 (2006).</li><li>Delouche, J. C., Still, T. W., Rapset, M., Lienhard, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+tetrazolium+test+for+seed+viability.">The tetrazolium test for seed viability.</a> Mississippi Sta Uni Ag Exp Sta Techn Bull. 51, 1-63 (1962).</li><li>Association of Official Seed Analysts. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Tetrazolium Testing Handbook. , (2010).</li><li>Alexander, H. M., Emry, D. J., Pace, B. A., Kost, M. A., Sparks, K. A., Mercer, K. 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Her præsenterer vi metoder til at bruge frø nedgravning strips til at observere frøspiring, hviletilstand og dødelighed af forskellige frø bestande på forud valgte tidsperioder på området. Fordelene ved at anvende strimler snarere end individuelle pakker ligger i (1) hastigheden af ​​strimlen og rum konstruktion løbet oprettelse af individuelle pakker; og (2) den lethed og hastighed af fjernelse flere rum i en bevægelse uden fare for at udelade en pakke eller fjerne en utilsigtet. Som to af fjernelse datoe...

Det overordnede mål med denne metode er at pålidelig vurdering frø overlevelse over tid under markforhold. Soil frøbanker er en reserve af spredte, levedygtige endnu uspirede frø fordelt enten på jordoverfladen, inden overfladen kuld, eller i jorden profil, der vil kunne overleve forbigående eller i mange år 1,2. Når frø nedgravning metoder svarende til dem, der præsenteres her blev anvendt på en 17-årig undersøgelse ved hjælp af flere dusin arter blev levedygtige frø findes i mange af de testede tre arter. Seed vækstdvale er en blok til frø spiring indtil passende kombination af betingelser for sætteplante overlevelse opstår fire. Resterende hvilende kan tillade frø at overleve barske forhold, såsom lave vintertemperaturer, næringsstof begrænsning eller sæsonbestemt tørke, indtil en ekstern udløser hviletilstand-release giver mulighed for spiring. Udløser til hviletilstand-release kan variere fra udsættelse for udvidede kolde, forbindelser efterladt af fire, eller fysisk angreb på frøskallen gennem slid eller kontakt med mave dyr syrer 5. Som spireevne stikord kan være slægter eller arter specifikke og ofte skyldes fortiden naturlig udvælgelse, utilpasset frøspiring er, at der opstår på et upassende tidspunkt, og kan resultere i frø eller sætteplante dødelighed eller dårlig sætteplante vækst. Mens vækstdvale er blevet klassificeret i en række typer baseret på mekanismerne i hviletilstand frigivelse (fx fysisk vækstdvale, fysiologisk vækstdvale), 6 frø hviletilstand er fortsat en af de mindst forståede emner i plantebiologi. Således feltstudier, der tillader vurdering af status for de enkelte frø eller grupper af frø i henhold til relevante økologiske forhold har højere forklaringsgrad end dem, der blot stole på standard spireevne test i laboratoriet. Udnyttelse af kendte frø egenskaber kan give indsigt i mekanismerne i dvale. Kontrol af frø vækstdvale er complex, herunder genetisk kontrol af fysiologiske og morfologiske faktorer. Mens en fuld forståelse af bredden i hviletilstand mekanismer er endnu ikke klarlagt, har en generel model opstået, indebærer en feedback-forhold mellem de to plantehormoner gibberellinsyre (GA) og abscisinsyre (ABA) 7. I denne generaliseret model for frø med en fysiologisk komponent til deres hviletilstand, GA tjener som signal for hviletilstand frigivelse, mens ABA tjener til at opretholde den sovende tilstand. Maternal genetiske virkninger samt den maternelle vækstmiljø kan påvirke vækstdvale og andre frø træk, såsom størrelse, gennem moderen genererede væv og udviklingsmæssige signaler 8. Maternelt genererede eksterne strukturer (eller frø belægninger) kan opretholde hviletilstand, til tider i kombination med fysiologiske signaler. Da maternelle frø belægninger styres af moderen plantens gener, kan de ikke afspejler frøet faktiske nukleare genetiske make-up. Vi har osed de Helianthus annuus nøddefrugt fra en vifte af afgrøde-vilde hybrid krydser at drille disse mødres vs. embryo genetiske effekter på frø egenskaber 9,10. Således studiedesign, der omfatter forskellige arter, cross typer eller genotyper kan indsamle oplysninger om økologi og genetik frø hviletilstand, spiring og overlevelse. Et vigtigt eksempel på, hvordan frøspiring og overlevelse fænotyper kan påvirke populationsdynamik kan ses i afgrødespecifikke vilde hybrid zoner. Udvælgelse under domesticering af dyrkede planter fjerner det meste vækstdvale og reducerer et frø overlevelsesevne uden for vækstsæsonen. Alligevel gen flow, eller hybridisering mellem dyrkede og vilde typer i afgrødespecifikke vilde hybrid zoner kan genindføre afgrøde alleler (eller genetiske varianter) i en vild population, der kan påvirke de frø bank dynamik. Hybrider mellem dyrkede og vilde slægtninge potentielt findes i afgrødespecifikke vilde hybrid zoner kan haveen række mellemliggende hviletilstand fænotyper, med kun et par fænotyper forventes at overleve betingelserne uden for dyrkning (f.eks vintermånederne) 11. Formålet med dette manuskript er at vise, hvordan, ved hjælp af frø begravelse strimmel metode, kan vi vurdere spiring, vækstdvale, og overlevelse af en række frø typer på forskellige tidsperioder for at undersøge deres naturlige variation under markforhold. I vores eksempel, vi ansat solsikkefrø fra 15 afgrødespecifikke vilde typer hybrid cross da vi er interesseret i maternelle og embryonale genetiske effekter på frø egenskaber.

<table border="0" cellpadding="0" cellspacing="0" width="440">
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		<col />
		<col span="2" />
	</colgroup>
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:240px;">Small coin envelopes</td>
			<td style="width:72px;">Any</td>
			<td style="width:64px;"></td>
			<td style="width:64px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Large coin envelopes</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">fine meshed polyester mosquito netting</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">high-temperature glue gun</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">high-temperature glue stick refills</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Industrial permenant markers</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">plastic garden labels</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">scissors</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Shovel</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Metal mesh hardward cloth</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Surveyor&#39;s flags, multiple colors</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Wet newspaper</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">cooler</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">blotter paper</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">petri dishes</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Temp. controlled growth chamber</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">razor blades</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">petri dishes</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tetrazolium chloride</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">water</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">heat incubator</td>
			<td>Any</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

reliable method for assessing seed germination, dormancy, and mortality under field conditions

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester fabric and glue can be folded and glued to construct a strip of compartments that house seeds and identifying information, while allowing contact with soil leachate, water, microorganisms, and ambient temperature. Strips may be constructed with a wide range of compartment numbers and sizes and allow the researcher to house a variety of genotypes within a single species, different species, or seeds that have experienced different treatments. As opposed to individual seed packets, strips are more easily retrieved as a unit. While replicate packets can be included within a strip, different strips can act as blocks or can be retrieved at different times for observation of seed behavior over time. We used a high temperature glue gun to delineate compartments and sealed the strips once the seed and tags identifying block and removal times were inserted. The seed strips were then buried in the field at the desired depth, with the location marked for later removal. Burrowing animal predators were effectively excluded by use of a covering of metal mesh hardware cloth on the soil surface. After the selected time interval for burial, strips were dug up and seeds were assessed for germination, dormancy and mortality. While clearly dead seeds can often be distinguished from ungerminated living ones by eye, dormant seeds were conclusively identified using a standard Tetrazolium chloride colorimetric test for seed viability.

Cross typer med varieret moderens herkomst og afgrøde allel procent (tabel 1) afveg tværs fjernelse datoer i procent spiret, uspiret, og døde frø (fig. 2 og 3). Brug af TZ afprøvning af uspirede frø, fandt vi nogle virkelig hvilende frø ved den anden fjernelse (tidligt forår) (tabel 2), mens alle frø uspirede af den tredje fjernelse (forår) sig at være virkelig hvilende. <p class="jove_content" fo:keep-tog...

Watch this Scientific Journal Video about Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions at JoVE.com

Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions

Her præsenterer vi en protokol til vurdering af frø efterladte, spiring og vækstdvale under markforhold ved hjælp begravet, mærket frø strips og tetrazoliumchlorid (TZ) levedygtighed test.

Pålidelig metode til at vurdere Seed Spiring, dvaletilstand, og Dødelighed under markforhold

We describe techniques for approximating seed bank dynamics over time using Helianthus annuus as an example study species. Strips of permeable polyester ...

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