Periodic Reporting for period 1 - POP1Seed (Population-wide analysis of molecular and metabolic features intrinsic to dormancy variability in single seeds)
Okres sprawozdawczy: 2021-12-01 do 2023-11-30
Seed physiological quality is pivotal to ensure rapid and homogeneous seedling establishment required for stable yield and is a major economic challenge for the seed industry. Besides germination, vigor and storability or longevity, seed physiological quality also includes seed dormancy, i.e. the seed physiological stage preventing germination even under favorable conditions. While seed dormancy has been widely studied at population level, little has been done at single seed level. However, homogeneous seed lot production is crucial in plant breeding in the context of climate change.
In this project, POP1SEED, we aim to decipher novel regulators of single seed dormancy by GWAS approach. There are two sub-objectives:
(i) Characterize single seed variability within A. thaliana accessions at both physiological and metabolic levels and identify new putative genomic regions involved in single seeds dormancy depth by Genome Wide Analysis (GWAS)
(ii) Identify whether dormancy variability is cis or trans regulated and to identify
the putative candidates at gene level using single seed RNA sequencing
In this project, POP1SEED, we aim to decipher novel regulators of single seed dormancy by GWAS approach. There are two sub-objectives:
(i) Characterize single seed variability within A. thaliana accessions at both physiological and metabolic levels and identify new putative genomic regions involved in single seeds dormancy depth by Genome Wide Analysis (GWAS)
(ii) Identify whether dormancy variability is cis or trans regulated and to identify
the putative candidates at gene level using single seed RNA sequencing
From 01/12/2021 to 31/08/2022, focus has been made on the first objective of the project. This includes:
(i) Accessions have been selected based on their diversity and their availability of sequences in www.1001genomes.com then ordered or NASC. Genotypes have been grown accordingly to the data provided by NASC on flowering time and vernalization needs in order to grow all the genotypes in triplicates randomized within the same greenhouse to obtain comparable levels of seed production. Out of 193 genotypes selected, 186 genotypes reached seed production.
Next, we analysed dormancy at single seed level. We established a protocol to quantify single seed dormancy using parameters of U1684 and U1090 (Uniformity 16/10 to 84/90, time of imbibition to go from 16/10% of total germination to 84/90%) as calculated automatically using Germinator (Joosen et al., 2010). Experimental time has been done per genotype depending on their dormancy level as checked on a weekly basis on total seed pool.
Other physiological parameters have also been recorded: number of seeds per silique (186 genotypes), individual seed size using (BoxSeed®), plant height and total number of seeds per plant (yield). Those data can further be used by the laboratory to perform GWAS on individual seed size variation.
Out of our preliminary data, we have been able to characterize genotypes with variation in single seed dormancy levels. First, we characterized candidates displaying opposite single seed dormancy phenotypes, i.e. high and low. Those genotypes can be then later analysed for single seed RNAseq and help with the identification of new putative regions involved in single seed dormancy.
Within the timeframe of the project (01/12/2021 to 31/08/2022), we have been able to produce the plant material and start the physiological characterization of the accessions. Low and high varaibel accessions has been selected and interesting candidates have been found in order to analyse further the genetic regulation of single seed dormancy.
Those preliminary data have been presented and discussed as a poster at two scientific meetings:
As a poster and oral at “IHC2022” (International Horticultural Congress 2022).
Poster: “A floral induction system to analyze seed lot intra variability in Arabidopsis and perspectives for horticultural species seed production”
Oral: “Role of seeds and seed sector in human health : « One Health » approach”
and as a poster at "10th conference of the Polish Society of Plant Experimental Biology".
“Single seed dormancy in Arabidopsis thaliana: identification of novel regulators and perspectives of application”
(i) Accessions have been selected based on their diversity and their availability of sequences in www.1001genomes.com then ordered or NASC. Genotypes have been grown accordingly to the data provided by NASC on flowering time and vernalization needs in order to grow all the genotypes in triplicates randomized within the same greenhouse to obtain comparable levels of seed production. Out of 193 genotypes selected, 186 genotypes reached seed production.
Next, we analysed dormancy at single seed level. We established a protocol to quantify single seed dormancy using parameters of U1684 and U1090 (Uniformity 16/10 to 84/90, time of imbibition to go from 16/10% of total germination to 84/90%) as calculated automatically using Germinator (Joosen et al., 2010). Experimental time has been done per genotype depending on their dormancy level as checked on a weekly basis on total seed pool.
Other physiological parameters have also been recorded: number of seeds per silique (186 genotypes), individual seed size using (BoxSeed®), plant height and total number of seeds per plant (yield). Those data can further be used by the laboratory to perform GWAS on individual seed size variation.
Out of our preliminary data, we have been able to characterize genotypes with variation in single seed dormancy levels. First, we characterized candidates displaying opposite single seed dormancy phenotypes, i.e. high and low. Those genotypes can be then later analysed for single seed RNAseq and help with the identification of new putative regions involved in single seed dormancy.
Within the timeframe of the project (01/12/2021 to 31/08/2022), we have been able to produce the plant material and start the physiological characterization of the accessions. Low and high varaibel accessions has been selected and interesting candidates have been found in order to analyse further the genetic regulation of single seed dormancy.
Those preliminary data have been presented and discussed as a poster at two scientific meetings:
As a poster and oral at “IHC2022” (International Horticultural Congress 2022).
Poster: “A floral induction system to analyze seed lot intra variability in Arabidopsis and perspectives for horticultural species seed production”
Oral: “Role of seeds and seed sector in human health : « One Health » approach”
and as a poster at "10th conference of the Polish Society of Plant Experimental Biology".
“Single seed dormancy in Arabidopsis thaliana: identification of novel regulators and perspectives of application”
The project has been conducted for 8 months.
The work acomplished so far led to the screening of single seed dormancy variation among Arabidopsis thaliana accessions. It also led to the identification of contrasted genotypes, displaying high and low single seed dormancy variation.
Those data will be further used within the laboratory to identify novel regulators of single seed dormancy.
The work acomplished so far led to the screening of single seed dormancy variation among Arabidopsis thaliana accessions. It also led to the identification of contrasted genotypes, displaying high and low single seed dormancy variation.
Those data will be further used within the laboratory to identify novel regulators of single seed dormancy.