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#### Chlorophyll fluorescence # Chlorophyll fluorescence
## Assay Description and Instrumentation
Fluorescence data was collected using a MINI-PAM-II fluorometer (Walz Heinz GmbH, Effeltrich, Fluorescence data was collected using a MINI-PAM-II fluorometer (Walz Heinz GmbH, Effeltrich,
Germany). Germany).
...@@ -8,7 +10,7 @@ Germany). ...@@ -8,7 +10,7 @@ Germany).
<img src="Quinoaimage4.jpeg" width="40%" /> <img src="Quinoaimage4.jpeg" width="40%" />
</p> </p>
**To conduct measurements:** ### Protocol
- The leaf clip was attached to a leaf and measurements of - The leaf clip was attached to a leaf and measurements of
photosynthetically active radiation (PAR) and the photosynthetic efficiency of photosystem II (PhiPSII) were photosynthetically active radiation (PAR) and the photosynthetic efficiency of photosystem II (PhiPSII) were
...@@ -22,8 +24,11 @@ recorded. ...@@ -22,8 +24,11 @@ recorded.
measured 48 days after sowing between 15:00 and 17:00. All blocks were measured 50 days after sowing measured 48 days after sowing between 15:00 and 17:00. All blocks were measured 50 days after sowing
between 10:24 and 12:49.** between 10:24 and 12:49.**
### Calibration and Quality Control
The fluorescence kinetics of saturation pulse analyses for measurements were inspected using WinControl-3 Software (version 3.29-rev.1112) (Walz Heinz GmbH, Effeltrich, Germany). The fluorescence kinetics of saturation pulse analyses for measurements were inspected using WinControl-3 Software (version 3.29-rev.1112) (Walz Heinz GmbH, Effeltrich, Germany).
Measurements that did not pass inspection were discarded from subsequent analyses. Measurements that did not pass inspection were discarded from subsequent analyses.
### Data Processing
PhiPSII values were clustered according to genotype, treatment and PAR level using custom R scripts. PhiPSII values were clustered according to genotype, treatment and PAR level using custom R scripts.
**PAR was divided into four levels:** **PAR was divided into four levels:**
...@@ -34,6 +39,15 @@ PhiPSII values were clustered according to genotype, treatment and PAR level usi ...@@ -34,6 +39,15 @@ PhiPSII values were clustered according to genotype, treatment and PAR level usi
Agglomerative hierarchical clustering was applied on the scaled PhiPSII means for all plots using the ‘agnes’ function of the R package ‘cluster’ [1] with a cut-off of five clusters. Agglomerative hierarchical clustering was applied on the scaled PhiPSII means for all plots using the ‘agnes’ function of the R package ‘cluster’ [1] with a cut-off of five clusters.
### Software and tools
- WinControl-3 Software (version 3.29-rev.1112) (Walz Heinz GmbH, Effeltrich, Germany)
- R (with package 'cluster')
### Assay Date
This assay was performed at the visible inflorescence stage of development (BBCH 59); 46-50 days after sowing
Reference: Reference:
[1] Maechler, M.; Rousseeuw, P.; Struyf, A.; Hubert, M.; Hornik, K.; Studer, M.; Roudier, P.; Gonzalez, J.; Kozlowski, K.; Schubert, E.; et al. “Finding Groups in Data”: Cluster Analysis Extended Rousseeuw et al. Available online: https://cran.r-project.org/web/packages/cluster/cluster.pdf (accessed on 26 February 2021). [1] Maechler, M.; Rousseeuw, P.; Struyf, A.; Hubert, M.; Hornik, K.; Studer, M.; Roudier, P.; Gonzalez, J.; Kozlowski, K.; Schubert, E.; et al. “Finding Groups in Data”: Cluster Analysis Extended Rousseeuw et al. Available online: https://cran.r-project.org/web/packages/cluster/cluster.pdf (accessed on 26 February 2021).
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