You are on page 1of 32

Designing Experiments with Orchids

Presented by Ian Parsons.

Designing Experiments with Orchids


Why experiment with orchids?

Designing Experiments with Orchids


Why experiment with orchids?

We may need to carry out experiments with orchids to estabish facts about optimum growing conditions.

Designing Experiments with Orchids


Why experiment with orchids? We may need to carry out experiments with orchids to estabish facts about optimum growing conditions.

Nutrients Composts Growth Supplements Pots Other factors

Designing Experiments with Orchids


Having established the reasons why experiments with orchids may be carried out, the factors affecting orchids in particular must now be considered.

Designing Experiments with Orchids


Having established the reasons why experiments with orchids may be carried out, the factors affecting orchids in particular must now be considered.

Orchids are relatively expensive, so small designs are appropriate.

Designing Experiments with Orchids


Having established the reasons why experiments with orchids may be carried out, the factors affecting orchids in particular must now be considered.

Orchids are relatively expensive, so small designs are appropriate.


Therefore use Non-Destructive testing. Quantitative Sampling. Relative Measurements.

Designing Experiments with Orchids

Non Destructive Testing

Dry Weight and other mass measurements are the usual method employed in plant experiments, but not with orchids.
Quantitative Sampling is best for orchids.

Counting the number of shoots, number of leaves, number of flower spikes. Measuring the lengths of leaves and shoots. Relatives measures, where initial plant size is measured, and subsequent sampling provides relative increases (or otherwise!).

Designing Experiments with Orchids


Statistics

Initial steps Design of experimental layout

Analysis of results
Repeatability (tell everything, hide nothing!) Defined limits of experiments (know the boundaries)

Designing Experiments with Orchids


Statistics

Initial steps Design of experimental layout

Analysis of results
Repeatability (tell everything, hide nothing!) Defined limits of experiments (know the boundaries)

Designing Experiments with Orchids


Statistics

Initial steps

A Null Hypothesis, for example, that treatments have no measurable effects on the plants. A Control treatment, an example is that of no treatment at all. Replication, using as many different samples (plants) in an experiment as possible. Randomization, to reduce the effects of external variables such as light, heat, humidity, draughts.

Designing Experiments with Orchids


Statistics

Design of experimental layout

Latin Squares.

A 4 x 4 Latin Square :A B C D B C D A C D A B D A B C A regular square, no letters repeated in either rows or columns.

A
D C B

D
C B A

C
B A D

B
A D C A randomized square, no letters repeated in either rows or columns.

Designing Experiments with Orchids


Statistics

Design of experimental layout

Graeco-Latin Squares.

A 5 x 5 Greaco-Latin Square :A C D E C E A B B D E A A randomised square, no Greek or Latin Letters repeated in either rows or columns, and no combinations are repeated or absent. Each Latin letter can represent a level of one nutient. Each Greek letter can represent a level of one growth supplement

D A B C

E B C D

Designing Experiments with Orchids


Collecting Data
Measuring Shoot lengths. One plant. Initial Measurements. (A) Subsequent Measurements. (B) Relative increases. (C=B-A) Example : Plant A (3 Shoots lengths in mm)

B Shoot 1 Shoot 2 168

A 117 60 52 229

C (result) 51 6 0 57

66
52

Shoot 3 Total

286

Designing Experiments with Orchids


Analysis of results

Designing Experiments with Orchids

Designing Experiments with Orchids

Designing Experiments with Orchids


Missing Values
Values in the data may be lost for several reasons. For example, a plant may die, or may be eaten by a pest. In a case of loss, some value should be inserted in to the data table to replace the missing values. It may not be wise to try to replace more that 2 values in a 5 X 5 design because for each missing value the total degrees of freedom must be reduced by one. This may leave only a few degrees of freedom for the Error Mean Square, and 6 is the minimum acceptable. If there are more than 2 losses in a 5 X 5 design then the experiment needs to be repeated. There are two ways to estimate a value to replace one that has been lost.

The first is to use the overall mean from the remaining good values, and the second is to estimate a missing value from its component contributing parts.

Designing Experiments with Orchids


Missing Values (continued...)
Using the overall mean may be sufficient as the Error Mean Square is computed from the square of deviations from the mean, by subtraction of the component Mean Squares from the Total Mean Squares. This gives rise to a deviation of zero for the missing value(s) from the overall mean, hence its acceptability.

The second method is slightly more accurate in that it takes in to consideration the variability of each of the component contributions to the specific cell in the design for which the data is missing.
The next slide shows the formula.

Designing Experiments with Orchids


Missing Values (continued...)
The general form for this is :-

yijkl = + k + l + i + j + ijkl
Where i,j,k,l are each 1 to 5 in this example.

y ijkl

= the observation in the i th row, the j th column, and receiving the k th Latin treatment and the l th Greek treatment.

= The overall mean.


th k = the effect of the k Latin treatment, th

i = the effect of the i ijkl = random error.

row ,

j = the effect of the j

= the effect of the l th Greek treatment.


th

column.

Designing Experiments with Orchids


Missing Values (continued...)
The Missing Values can be computed from adding to the overall mean the means of the remaining values in each row, column, Latin treatment, and Greek treatment corresponding to the position of the missing cell.
These result is then averaged to find the best estimate of the missing value.

In the final analysis, the Error Degrees of Freedom have to be reduced by one for each missing value.
This method is more accurate than just using the overall mean, but for large variance, a Bias may have to be subtracted from the Error Sum of Squares. This is beyond the scope of this presentation.

Designing Experiments with Orchids


Basic Compost Mix :Equal parts (1) Fine Bark (2) Perlite (3) Sphagnum Moss (chopped)

Designing Experiments with Orchids


25 small plants of Dendrobium kingianum.

Designing Experiments with Orchids

A potted and labeled plant.

Designing Experiments with Orchids

Plants ready for the bench.

Designing Experiments with Orchids

Plants in the greenhouse during an experiment.

Designing Experiments with Orchids


Treatments applied to the Greek and Latin components.

The Latin component of the design was 5 levels of nutrients where each level was as follows :A B C D E No nutrient. 1.25 ml/litre, corresponding to half recommended strength. 2.5 ml/litre, corresponding to recommended normal strength. 5 ml/litre, double strength. 10 ml/litre, quadruple strength.

The formulation of the nutrient mix is 1.8:1.2:2.1, with additional micro-elements :% w/v Nitrogen, in the form of soluble Nitrates 1.83 Phosphorous pentoxide, soluble 1.19 Potassium oxide 2.15 Calcium oxide 1.72 Copper chelated by EDTA 0.002 Iron chelated by EDTA 0.040 Manganese chelated by EDTA 0.010 Zinc chelated by EDTA 0.0025
(No Urea or Ammonia in this formulation)

This is similar to Growth Technology Orchid Focus Grow, but without supplements.

Designing Experiments with Orchids


Treatments applied to the Greek and Latin components.

The Greek component of the design was 5 levels of growth supplements where each level was as follows : No supplement. 3.75 ml/litre, corresponding to half recommended strength. 7.5 ml/litre, corresponding to recommended normal strength. 15 ml/litre, double strength. 30 ml/litre, quadruple strength.

The formulation of the supplement mix is :Humic Acid, in the form of soluble salts Fulvic Acid, in the form of soluble salts 53% 17%

This is diluted in steps to form a solution of 7.5 ml/litre For the recommended normal strength. This represents a solution of 0.0075% supplements per litre.
These supplements are part of Growth Technology Orchid Focus Grow and Bloom.

Designing Experiments with Orchids


Advantages and Disadvantages of Graeco-Latin Square Designs.

Disadvantages

Requires n2 combinations of treatments, in these examples, where n=5, Each plant requires a different mix of nutrients and growth supplements.
Finding n2 similar plants may be a problem! The design provides no analysis of interactions. As all combinations are represented in each in only one plant, there is no room for replication, and interactions must be analyzed Using a Randomized Block design. A small design as described here does not allow for a large number of missing values. Enlarging a Graeco-Latin square requires stepping up to a 7 X 7 design, as Euler's Conjecture suggested that 4n+2 squares do not exist, so a 6 X 6 is not possible.

Designing Experiments with Orchids


Advantages and Disadvantages of Graeco-Latin Square Designs.

Advantages

The design provides a good range of options for any form of treatments that will be applied to the plants.
Recording results is not too onerous. A simple analysis is provided for the design in the form of an Excel spreadsheet, with graphical displays of results, and this can be copied and reused. Computation of up to 2 missing values is possible without the need to redesign. Space required to carry out an experiment is not too large, even for an Amateur or hobby grower. The costs of treatments is not too large for this design, and yet it copes with variable conditions in a compact space. The design is repeatable, so that another independent researcher can test the results.

Designing Experiments with Orchids


Further Experiments
Several types of experiments offer themselves for examination. Some that need investigation are listed as follows :

Further nutrient experiments e.g. the composition of the most successful nutrients. Composts, and their effects on quality of orchids. Types of pots, plastic, terra-cotta, clear, opaque, open mesh etc. Growth supplements, which to use for best results? Plant hormones, for growth, and for flower. Repeat experiments for different orchids.

More difficult, but possible in controlled conditions :

Effects of light, temperature and humidity with combinations of the above.

Designing Experiments with Orchids


Notes and references
Graeco-Latin squares have been used in experimental design for over a century, and are experiencing a renaissance in communications theory and practice at at the present time. The Swiss mathematician Euler (1707-1783) was fascinated by them, and did much work describing methods for their construction, and put forward the conjecture (1750) that 4n+2 (n=1 to infinity) Graeco-Latin squares did not exist. It was not until 1959 that Parker, Bose, and Shrikhande presented their paper showing Euler's conjecture to be false for all n 10. Thus, Graeco-Latin squares exist for all orders n 3 except n = 6. Any number of papers can be found on the internet describing Graeco-Latin Squares, and their use in experiments design, along with the statistical analyses associated with them. This presentation, and the spread sheet for the statistical analysis can be found on my website as follows :Statistical Analysis - http://www.iantparsons.creators.co.uk/Experiments/Analysis.xls Metric Rulers - http://www.iantparsons.creators.co.uk/Experiments/rulers_metric.pdf Presentation - http://www.iantparsons.creators.co.uk/Experiments/Presentation.odp PowerPoint - http://www.iantparsons.creators.co.uk/Experiments/Presentation.ppt

You might also like