50 years of bioscience statistics

Earlier this year I had an enquiry from Carey Langley of VSNi as to why I had not renewed my Genstat licence. The truth was simple – I have decided to fully retire after 50 years as an agricultural entomologist / applied biologist / consultant. This prompted some reflections about the evolution of bioscience data analysis that I have experienced over that half century, a period during which most of my focus was the interaction between insects and their plant hosts; both how insect feeding impacts on plant growth and crop yield, and how plants impact on the development of the insects that feed on them and on their natural enemies.

Where it began – paper and post

My journey into bioscience data analysis started with undergraduate courses in biometry – yes, it was an agriculture faculty, so it was biometry not statistics. We started doing statistical analyses using full keyboard Monroe calculators (for those of you who don’t know what I am talking about, you can find them here).  It was a simpler time and as undergraduates we thought it was hugely funny to divide 1 by 0 until the blue smoke came out…

After leaving university in the early 1970s, I started working for the Agriculture Department of an Australian state government, at a small country research station. Statistical analysis was rudimentary to say the least. If you were motivated, there was always the option of running analyses yourself by hand, given the appearance of the first scientific calculators in the early 1970s. If you wanted a formal statistical analysis of your data, you would mail off a paper copy of the raw data to Biometry Branch… and wait.  Some months later, you would get back your ANOVA, regression, or whatever the biometrician thought appropriate to do, on paper with some indication of what treatments were different from what other treatments.  Dose-mortality data was dealt with by manually plotting data onto probit paper. 

Enter the mainframe

In-house ANOVA programs running on central mainframes were a step forward some years later as it at least enabled us to run our own analyses, as long as you wanted to do an ANOVA…. However, it also required a 2 hours’ drive to the nearest card reader, with the actual computer a further 1000 kilometres away.… The first desktop computer I used for statistical analysis was in the early 1980s and was a CP/M machine with two 8-inch floppy discs with, I think, 256k of memory, and booting it required turning a key and pressing the blue button - yes, really! And about the same time, the local agricultural economist drove us crazy extolling the virtues of a program called Lotus 1-2-3!

Having been brought up on a solid diet of the classic texts such as Steele and Torrie, Cochran and Cox and Sokal and Rohlf, the primary frustration during this period was not having ready access to the statistical analyses you knew were appropriate for your data. Typical modes of operating for agricultural scientists in that era were randomised blocks of various degrees of complexity, thus the emphasis on ANOVA in the software that was available in-house. Those of us who also had less-structured ecological data were less well catered for.

My first access to a comprehensive statistics package was during the early to mid-1980s at one of the American Land Grant universities. It was a revelation to be able to run virtually whatever statistical test deemed necessary. Access to non-linear regression was a definite plus, given the non-linear nature of many biological responses. As well, being able to run a series of models to test specific hypotheses opened up new options for more elegant and insightful analyses. Looking back from 2021, such things look very trivial, but compared to where we came from in the 1970s, they were significant steps forward.

Enter Genstat

My first exposure to Genstat, VSNi’s stalwart statistical software package, was Genstat for Windows, Third Edition (1997). Simple things like the availability of residual plots made a difference for us entomologists, given that much of our data had non-normal errors; it took the guesswork out of whether and what transformations to use. The availability of regressions with grouped data also opened some previously closed doors. 

After a deviation away from hands-on research, I came back to biological-data analysis in the mid-2000s and found myself working with repeated-measures and survival / mortality data, so ventured into repeated-measures restricted maximum likelihood analyses and generalised linear mixed models for the first time (with assistance from a couple of Roger Payne’s training courses in Hobart and Queenstown). Looking back, it is interesting how quickly I became blasé about such computationally intensive analyses that would run in seconds on my laptop or desktop, forgetting that I was doing ANOVAs by hand 40 years earlier when John Nelder was developing generalised linear models. How the world has changed!

Partnership and support

Of importance to my Genstat experience was the level of support that was available to me as a Genstat licensee. Over the last 15 years or so, as I attempted some of these more complex analyses, my aspirations were somewhat ahead of my abilities, and it was always reassuring to know that Genstat Support was only ever an email away. A couple of examples will flesh this out. 

Back in 2008, I was working on the relationship between insect-pest density and crop yield using R2LINES, but had extra linear X’s related to plant vigour in addition to the measure of pest infestation. A support-enquiry email produced an overnight response from Roger Payne that basically said, “Try this”. While I slept, Roger had written an extension to R2LINES to incorporate extra linear X’s. This was later incorporated into the regular releases of Genstat. This work led to the clearer specification of the pest densities that warranted chemical control in soybeans and dry beans (https://doi.org/10.1016/j.cropro.2009.08.016 and https://doi.org/10.1016/j.cropro.2009.08.015).

More recently, I was attempting to disentangle the effects on caterpillar mortality of the two Cry insecticidal proteins in transgenic cotton and, while I got close, I would not have got the analysis to run properly without Roger’s support. The data was scant in the bottom half of the overall dose-response curves for both Cry proteins, but it was possible to fit asymptotic exponentials that modelled the upper half of each curve. The final double-exponential response surface I fitted with Roger’s assistance showed clearly that the dose-mortality response was stronger for one of the Cry proteins than the other, and that there was no synergistic action between the two proteins (https://doi.org/10.1016/j.cropro.2015.10.013) 

The value of a comprehensive statistics package

One thing that I especially appreciate about having access to a comprehensive statistics package such as Genstat is having the capacity to tease apart biological data to get at the underlying relationships. About 10 years ago, I was asked to look at some data on the impact of cold stress on the expression of the Cry2Ab insecticidal protein in transgenic cotton. The data set was seemingly simple - two years of pot-trial data where groups of pots were either left out overnight or protected from low overnight temperatures by being moved into a glasshouse, plus temperature data and Cry2Ab protein levels. A REML analysis, and some correlations and regressions enabled me to show that cold overnight temperatures did reduce Cry2Ab protein levels, that the effects occurred for up to 6 days after the cold period and that the threshold for these effects was approximately 14 Cº (https://doi.org/10.1603/EC09369). What I took from this piece of work is how powerful a comprehensive statistics package can be in teasing apart important biological insights from what was seemingly very simple data. Note that I did not use any statistics that were cutting edge, just a combination of REML, correlation and regression analyses, but used these techniques to guide the dissection of the relationships in the data to end up with an elegant and insightful outcome.

Final reflections

Looking back over 50 years of work, one thing stands out for me: the huge advances that have occurred in the statistical analysis of biological data has allowed much more insightful statistical analyses that has, in turn, allowed biological scientists to more elegantly pull apart the interactions between insects and their plant hosts. 

For me, Genstat has played a pivotal role in that process. I shall miss it.

Dr John Rogers

Research Connections and Consulting

St Lucia, Queensland 4067, Australia

Phone/Fax: +61 (0)7 3720 9065

Mobile: 0409 200 701

Email: john.rogers@rcac.net.au

Alternate email: D.John.Rogers@gmail.com