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Background  There are a number of unresolved issues in the design of experiments in greenhouses. They include whether statistical designs should be used and, if so, which designs should be used. Also, are there thigmomorphogenic or other effects arising from the movement of plants on conveyor belts within a greenhouse? A two-phase, single-line wheat experiment involving four tactics was conducted in a conventional greenhouse and a fully-automated phenotyping greenhouse (Smarthouse) to investigate these issues.    Results and discussion  Analyses of our experiment show that there was a small east–west trend in total area of the plants in the Smarthouse. Analyses of the data from three multiline experiments reveal a large north–south trend. In the single-line experiment, there was no evidence of differences between trios of lanes, nor of movement effects. Swapping plant positions during the trial was found to decrease the east–west trend, but at the cost of increased error variance. The movement of plants in a north–south direction, through a shaded area for an equal amount of time, nullified the north–south trend. An investigation of alternative experimental designs for equally-replicated experiments revealed that generally designs with smaller blocks performed best, but that (nearly) trend-free designs can be effective when blocks are larger.    Conclusions  To account for variation in microclimate in a greenhouse, using statistical design and analysis is better than rearranging the position of plants during the experiment. For the relocation of plants to be successful requires that plants spend an equal amount of time in each microclimate, preferably during comparable growth stages. Even then, there is no evidence that this will be any more precise than statistical design and analysis of the experiment, and the risk is that it will not be successful at all. As for statistical design and analysis, it is best to use either (i) smaller blocks, (ii) (nearly) trend-free arrangement of treatments with a linear trend term included in the analysis, or, as a last resort, (iii) blocks of several complete rows with trend terms in the analysis. Also, we recommend that the greenhouse arrangement parallel that in the Smarthouse, but with randomization where appropriate.

Accounting for variation in designing greenhouse experiments with special reference to greenhouses containing plants on conveyor systems