mRNASeq DE analysis: calculating summary effect sizes for leaf, BT642Year1
This document has last been compiled on 2021-12-14 21:28:35.
Bringing in the effect sizes
We bring in the per-timepoint estimates of effect size from results/BT642Year1/DE_timepoint.
Time Interval Effect Size Summaries
Using the estimates of log-fold change at different time points, we will also calculate cumulative measures of log-fold change over wider periods of time.
The basic measure of log fold-change is the cumulative mean of the absolute log-fold change, with a sign (postive/negative) being imposed based on the sign of the mean of the log-fold change (i.e. without the absolute values):
\[sign(\frac{1}{N}\sum_{i=1}^{N}lfc_i)\times\frac{1}{N}\sum_{i=1}^{N}|lfc_i|.\] \(N\) is the number of the time points of our time period of interest (e.g. drought time points, water recovery time points, etc).
The column above (_lfc) is used for the log-fold change value between the drought condition and the control for a particular timepoint.
Note that the code also calculates this same summary value using the test statistic (_stat), basically a t-statistic.
In addition to all timepoints, the time periods that are considered are summarized over intervals (defined in sharedDataCommands.R). Below is a summary of the time periods for which effects are calculated. This summary is the maximum possible timepoints that could be included – if a particular timepoint is missing (e.g. missing due to not being sampled that year or samples failing ) that is not necessarily indicated below.
## Description.Preflowering.DroughtDiff.AllTps
## 1 All timepoints from Preflowering design that different from control
## 2 All timepoints from Preflowering design that different from control
## 3 All timepoints from Preflowering design that different from control
## 4 All timepoints from Preflowering design that different from control
## 5 All timepoints from Preflowering design that different from control
## 6 All timepoints from Preflowering design that different from control
## 7 All timepoints from Preflowering design that different from control
## 8 All timepoints from Preflowering design that different from control
## 9 All timepoints from Preflowering design that different from control
## 10 All timepoints from Preflowering design that different from control
## 11 All timepoints from Preflowering design that different from control
## Description.Preflowering.DroughtDiff.DroughtTps
## 1 Preflowering design, under drought
## 2 Preflowering design, under drought
## 3 Preflowering design, under drought
## 4 Preflowering design, under drought
## 5 Preflowering design, under drought
## 6 Preflowering design, under drought
## 7 Preflowering design, under drought
## 8 Preflowering design, under drought
## 9 Preflowering design, under drought
## 10 Preflowering design, under drought
## 11 Preflowering design, under drought
## Description.Preflowering.DroughtDiff.RecoveryTps
## 1 Preflowering design, recovery
## 2 Preflowering design, recovery
## 3 Preflowering design, recovery
## 4 Preflowering design, recovery
## 5 Preflowering design, recovery
## 6 Preflowering design, recovery
## 7 Preflowering design, recovery
## 8 Preflowering design, recovery
## 9 Preflowering design, recovery
## 10 Preflowering design, recovery
## 11 Preflowering design, recovery
## Description.Preflowering.DroughtDiff.EarlyDroughtTps
## 1 Preflowering design, under early drought
## 2 Preflowering design, under early drought
## 3 Preflowering design, under early drought
## 4 Preflowering design, under early drought
## 5 Preflowering design, under early drought
## 6 Preflowering design, under early drought
## 7 Preflowering design, under early drought
## 8 Preflowering design, under early drought
## 9 Preflowering design, under early drought
## 10 Preflowering design, under early drought
## 11 Preflowering design, under early drought
## Description.Preflowering.DroughtDiff.LateDroughtTps
## 1 Preflowering design, under late drought
## 2 Preflowering design, under late drought
## 3 Preflowering design, under late drought
## 4 Preflowering design, under late drought
## 5 Preflowering design, under late drought
## 6 Preflowering design, under late drought
## 7 Preflowering design, under late drought
## 8 Preflowering design, under late drought
## 9 Preflowering design, under late drought
## 10 Preflowering design, under late drought
## 11 Preflowering design, under late drought
## Description.Preflowering.DroughtDiff.EarlyRecoveryTps
## 1 Preflowering design, under early recovery
## 2 Preflowering design, under early recovery
## 3 Preflowering design, under early recovery
## 4 Preflowering design, under early recovery
## 5 Preflowering design, under early recovery
## 6 Preflowering design, under early recovery
## 7 Preflowering design, under early recovery
## 8 Preflowering design, under early recovery
## 9 Preflowering design, under early recovery
## 10 Preflowering design, under early recovery
## 11 Preflowering design, under early recovery
## Description.Preflowering.DroughtDiff.LateRecoveryTps
## 1 Preflowering design, under late recovery
## 2 Preflowering design, under late recovery
## 3 Preflowering design, under late recovery
## 4 Preflowering design, under late recovery
## 5 Preflowering design, under late recovery
## 6 Preflowering design, under late recovery
## 7 Preflowering design, under late recovery
## 8 Preflowering design, under late recovery
## 9 Preflowering design, under late recovery
## 10 Preflowering design, under late recovery
## 11 Preflowering design, under late recovery
## Description.Postflowering.DroughtDiff.AllTps
## 1 All timepoints from Postflowering design that different from control
## 2 All timepoints from Postflowering design that different from control
## 3 All timepoints from Postflowering design that different from control
## 4 All timepoints from Postflowering design that different from control
## 5 All timepoints from Postflowering design that different from control
## 6 All timepoints from Postflowering design that different from control
## 7 All timepoints from Postflowering design that different from control
## 8 All timepoints from Postflowering design that different from control
## 9 All timepoints from Postflowering design that different from control
## 10 All timepoints from Postflowering design that different from control
## 11 All timepoints from Postflowering design that different from control
## Description.Postflowering.DroughtDiff.DroughtTps
## 1 Postflowering design, under drought
## 2 Postflowering design, under drought
## 3 Postflowering design, under drought
## 4 Postflowering design, under drought
## 5 Postflowering design, under drought
## 6 Postflowering design, under drought
## 7 Postflowering design, under drought
## 8 Postflowering design, under drought
## 9 Postflowering design, under drought
## 10 Postflowering design, under drought
## 11 Postflowering design, under drought
## Description.Postflowering.DroughtDiff.EarlyDroughtTps
## 1 Postflowering design, early drought
## 2 Postflowering design, early drought
## 3 Postflowering design, early drought
## 4 Postflowering design, early drought
## 5 Postflowering design, early drought
## 6 Postflowering design, early drought
## 7 Postflowering design, early drought
## 8 Postflowering design, early drought
## 9 Postflowering design, early drought
## 10 Postflowering design, early drought
## 11 Postflowering design, early drought
## Description.Postflowering.DroughtDiff.LateDroughtTps
## 1 Postflowering design, late drought
## 2 Postflowering design, late drought
## 3 Postflowering design, late drought
## 4 Postflowering design, late drought
## 5 Postflowering design, late drought
## 6 Postflowering design, late drought
## 7 Postflowering design, late drought
## 8 Postflowering design, late drought
## 9 Postflowering design, late drought
## 10 Postflowering design, late drought
## 11 Postflowering design, late drought
## TimePoints
## 1 21,28,35,42,49,56,63,70,77,84,91,98,105,112,119
## 2 21,28,35,42,49,56
## 3 63,70,77,84,91,98,105,112,119
## 4 21,28,35
## 5 42,49,56
## 6 63
## 7 70,77,84,91,98,105,112,119
## 8 70,77,84,91,98,105,112,119
## 9 70,77,84,91,98,105,112,119
## 10 70,77
## 11 84,91,98,105,112,119
## columnName
## 1 Preflowering.DroughtDiff.AllTps.X
## 2 Preflowering.DroughtDiff.DroughtTps.X
## 3 Preflowering.DroughtDiff.RecoveryTps.X
## 4 Preflowering.DroughtDiff.EarlyDroughtTps.X
## 5 Preflowering.DroughtDiff.LateDroughtTps.X
## 6 Preflowering.DroughtDiff.EarlyRecoveryTps.X
## 7 Preflowering.DroughtDiff.LateRecoveryTps.X
## 8 Postflowering.DroughtDiff.AllTps.X
## 9 Postflowering.DroughtDiff.DroughtTps.X
## 10 Postflowering.DroughtDiff.EarlyDroughtTps.X
## 11 Postflowering.DroughtDiff.LateDroughtTps.XNote that Postflowering.DroughtDiff.AllTps and Postflowering.DroughtDiff.DroughtTps are equivalent, by definition. We include both so that they mirror the Preflowering results
These effects will be calculated and added to the effects per timepoint (i.e. results of moanin calculations minus p-value) and saved to a separate file from the moanin results. The column names of these effects are as in the above table, only with X replaced with information that indicates the genotype and whether calculated on stat or lfc. (Note that we calculate the stat summary because it is used in extracting genotype differences.)
Calculating Effect size summaries
## Calculating Preflowering.DroughtDiff.AllTps with timepoints:
## [1] 21 28 35 42 49 56 63 70 77 84 91 98 105 112 119
##
## Calculating Preflowering.DroughtDiff.DroughtTps with timepoints:
## [1] 21 28 35 42 49 56
##
## Calculating Preflowering.DroughtDiff.RecoveryTps with timepoints:
## [1] 63 70 77 84 91 98 105 112 119
##
## Calculating Preflowering.DroughtDiff.EarlyDroughtTps with timepoints:
## [1] 21 28 35
##
## Calculating Preflowering.DroughtDiff.LateDroughtTps with timepoints:
## [1] 42 49 56
##
## Calculating Preflowering.DroughtDiff.EarlyRecoveryTps with timepoints:
## [1] 63
##
## Calculating Preflowering.DroughtDiff.LateRecoveryTps with timepoints:
## [1] 70 77 84 91 98 105 112 119
##
## Calculating Postflowering.DroughtDiff.AllTps with timepoints:
## [1] 70 77 84 91 98 105 112 119
##
## Calculating Postflowering.DroughtDiff.DroughtTps with timepoints:
## [1] 70 77 84 91 98 105 112 119
##
## Calculating Postflowering.DroughtDiff.EarlyDroughtTps with timepoints:
## [1] 70 77
##
## Calculating Postflowering.DroughtDiff.LateDroughtTps with timepoints:
## [1] 84 91 98 105 112 119
##
## Calculating Preflowering.DroughtDiff.AllTps with timepoints:
## [1] 21 28 35 42 49 56 63 70 77 84 91 98 105 112 119
##
## Calculating Preflowering.DroughtDiff.DroughtTps with timepoints:
## [1] 21 28 35 42 49 56
##
## Calculating Preflowering.DroughtDiff.RecoveryTps with timepoints:
## [1] 63 70 77 84 91 98 105 112 119
##
## Calculating Preflowering.DroughtDiff.EarlyDroughtTps with timepoints:
## [1] 21 28 35
##
## Calculating Preflowering.DroughtDiff.LateDroughtTps with timepoints:
## [1] 42 49 56
##
## Calculating Preflowering.DroughtDiff.EarlyRecoveryTps with timepoints:
## [1] 63
##
## Calculating Preflowering.DroughtDiff.LateRecoveryTps with timepoints:
## [1] 70 77 84 91 98 105 112 119
##
## Calculating Postflowering.DroughtDiff.AllTps with timepoints:
## [1] 70 77 84 91 98 105 112 119
##
## Calculating Postflowering.DroughtDiff.DroughtTps with timepoints:
## [1] 70 77 84 91 98 105 112 119
##
## Calculating Postflowering.DroughtDiff.EarlyDroughtTps with timepoints:
## [1] 70 77
##
## Calculating Postflowering.DroughtDiff.LateDroughtTps with timepoints:
## [1] 84 91 98 105 112 119We give a quick summary of the log-foldchange of these results:
## BT642:## Preflowering.AllTps Preflowering.DroughtTps Preflowering.RecoveryTps
## Min. :-2.54031 Min. :-3.19886 Min. :-2.81958
## 1st Qu.:-0.24130 1st Qu.:-0.30025 1st Qu.:-0.20793
## Median : 0.16114 Median : 0.12223 Median : 0.11680
## Mean : 0.04613 Mean : 0.01992 Mean : 0.03259
## 3rd Qu.: 0.31122 3rd Qu.: 0.32826 3rd Qu.: 0.27108
## Max. : 4.04012 Max. : 7.43195 Max. : 3.37144
## Preflowering.EarlyDroughtTps Preflowering.LateDroughtTps
## Min. :-2.46043 Min. :-4.53766
## 1st Qu.:-0.27012 1st Qu.:-0.29109
## Median : 0.06254 Median : 0.12817
## Mean : 0.01837 Mean : 0.03395
## 3rd Qu.: 0.29364 3rd Qu.: 0.36360
## Max. : 5.58198 Max. : 9.28192
## Preflowering.EarlyRecoveryTps Preflowering.LateRecoveryTps
## Min. :-4.47642 Min. :-3.09232
## 1st Qu.:-0.25371 1st Qu.:-0.19632
## Median :-0.01313 Median : 0.11886
## Mean : 0.04013 Mean : 0.03184
## 3rd Qu.: 0.26070 3rd Qu.: 0.25706
## Max. : 8.17318 Max. : 2.86386
## Postflowering.AllTps Postflowering.DroughtTps Postflowering.EarlyDroughtTps
## Min. :-3.6662 Min. :-3.6662 Min. :-3.51894
## 1st Qu.:-0.2563 1st Qu.:-0.2563 1st Qu.:-0.19977
## Median : 0.1526 Median : 0.1526 Median : 0.06656
## Mean : 0.0415 Mean : 0.0415 Mean : 0.02740
## 3rd Qu.: 0.3307 3rd Qu.: 0.3307 3rd Qu.: 0.24119
## Max. : 4.6112 Max. : 4.6112 Max. : 5.38534
## Postflowering.LateDroughtTps
## Min. :-3.96373
## 1st Qu.:-0.26681
## Median : 0.16356
## Mean : 0.04686
## 3rd Qu.: 0.36119
## Max. : 4.42581And the percentage of genes past certain quantiles:
Proportion absolute lfc greater in BT642:
| % abs(lfc)> 0.8 | % abs(lfc)> 1 | % abs(lfc)> 1.5 | % abs(lfc)> 2 | |
|---|---|---|---|---|
| Preflowering.AllTps | 0.065 | 0.032 | 0.006 | 0.002 |
| Preflowering.DroughtTps | 0.116 | 0.070 | 0.025 | 0.009 |
| Preflowering.RecoveryTps | 0.042 | 0.021 | 0.004 | 0.001 |
| Preflowering.EarlyDroughtTps | 0.079 | 0.043 | 0.011 | 0.004 |
| Preflowering.LateDroughtTps | 0.159 | 0.109 | 0.048 | 0.023 |
| Preflowering.EarlyRecoveryTps | 0.135 | 0.092 | 0.040 | 0.019 |
| Preflowering.LateRecoveryTps | 0.035 | 0.018 | 0.004 | 0.001 |
| Postflowering.AllTps | 0.093 | 0.055 | 0.018 | 0.007 |
| Postflowering.DroughtTps | 0.093 | 0.055 | 0.018 | 0.007 |
| Postflowering.EarlyDroughtTps | 0.086 | 0.054 | 0.019 | 0.008 |
| Postflowering.LateDroughtTps | 0.104 | 0.061 | 0.021 | 0.007 |
Max Summaries
We will create another global summary that is the max of log-fold changes either across
- All timepoints
- The summary of the timepoints
We consider both of these when we use results from only RT430 or BT642 or when we take the max across both genotypes.
We give a quick summary of the log-foldchange of these results:
## BT642:## Preflowering.MaxOfIntervals Preflowering.MaxOfTps Postflowering.MaxOfIntervals
## Min. :-2.8505 Min. :-2.54031 Min. :-1.3072
## 1st Qu.:-0.1692 1st Qu.:-0.24130 1st Qu.: 0.1549
## Median : 0.2441 Median : 0.16114 Median : 0.2859
## Mean : 0.2894 Mean : 0.04613 Mean : 0.3579
## 3rd Qu.: 0.5376 3rd Qu.: 0.31122 3rd Qu.: 0.4845
## Max. : 9.2819 Max. : 4.04012 Max. : 5.3853
## Postflowering.MaxOfTps
## Min. :-3.21561
## 1st Qu.:-0.25495
## Median : 0.14519
## Mean : 0.03612
## 3rd Qu.: 0.30985
## Max. : 4.15867And the percentage of genes past certain quantiles:
## Proportion absolute lfc greater in BT642:
## % abs(lfc)> 0.8 % abs(lfc)> 1 % abs(lfc)> 1.5
## Preflowering.MaxOfIntervals 0.15422745 0.10432894 0.047490913
## Preflowering.MaxOfTps 0.06500496 0.03214842 0.006136997
## Postflowering.MaxOfIntervals 0.09351839 0.05480810 0.019166313
## Postflowering.MaxOfTps 0.07694850 0.04357268 0.012321201
## % abs(lfc)> 2
## Preflowering.MaxOfIntervals 0.025256102
## Preflowering.MaxOfTps 0.001746684
## Postflowering.MaxOfIntervals 0.007600434
## Postflowering.MaxOfTps 0.004248690We do a quick pairs plot to compare these values
Merge and Add Global Splines Results
We also will merge in the global splines summary of effect. This should be equal to the DroughtDiff.AllTps in the above, but it is somewhat different. It’s not clear the source of the differences, but these are calculated from the function in the package moanin.
We draw some basic pairs plots to compare them:
Save individual timepoints
The final effects file also contains the individual timepoints results (of stat and lfc). We are going to clean up the column names and add that to our results. The columns will now be of the form Preflowering.DroughtDiff.Tp.14_lfc where they were previously of the form Group_BT642_Preflowering.14-Group_BT642_Control.14_stat
## Old Results:## Gene
## 1 SbiBTX642.01G000200.v1.1
## 2 SbiBTX642.01G000400.v1.1
## 3 SbiBTX642.01G000500.v1.1
## 4 SbiBTX642.01G000600.v1.1
## 5 SbiBTX642.01G000900.v1.1
## 6 SbiBTX642.01G001000.v1.1
## Group_BT642_Preflowering.21-Group_BT642_Control.21_lfc
## 1 -0.29252544
## 2 -0.38953399
## 3 -0.33991469
## 4 -0.71076026
## 5 -0.03202144
## 6 -0.10404199
## Group_BT642_Preflowering.28-Group_BT642_Control.28_lfc
## 1 -0.0008505584
## 2 -0.1332127352
## 3 0.2326486856
## 4 -0.0703141819
## 5 0.5748316901
## 6 0.4443493292
## Group_BT642_Preflowering.35-Group_BT642_Control.35_lfc
## 1 -0.1712599
## 2 -0.2902600
## 3 -0.1820348
## 4 -0.2352668
## 5 0.5828230
## 6 0.1755099
## Group_BT642_Preflowering.42-Group_BT642_Control.42_lfc
## 1 -0.05493845
## 2 0.02436610
## 3 -0.03868713
## 4 -0.45653764
## 5 0.70658462
## 6 0.42970430## New Results:## Gene Preflowering.DroughtDiff.Day021.BT642_lfc
## 1 SbiBTX642.01G000200.v1.1 -0.29252544
## 2 SbiBTX642.01G000400.v1.1 -0.38953399
## 3 SbiBTX642.01G000500.v1.1 -0.33991469
## 4 SbiBTX642.01G000600.v1.1 -0.71076026
## 5 SbiBTX642.01G000900.v1.1 -0.03202144
## 6 SbiBTX642.01G001000.v1.1 -0.10404199
## Preflowering.DroughtDiff.Day028.BT642_lfc
## 1 -0.0008505584
## 2 -0.1332127352
## 3 0.2326486856
## 4 -0.0703141819
## 5 0.5748316901
## 6 0.4443493292
## Preflowering.DroughtDiff.Day035.BT642_lfc
## 1 -0.1712599
## 2 -0.2902600
## 3 -0.1820348
## 4 -0.2352668
## 5 0.5828230
## 6 0.1755099
## Preflowering.DroughtDiff.Day042.BT642_lfc
## 1 -0.05493845
## 2 0.02436610
## 3 -0.03868713
## 4 -0.45653764
## 5 0.70658462
## 6 0.42970430Saving Results
## Saving effect size of leaf to results/BT642Year1/data