EXAM QUESTION FEEDBACK EXAMPLE MODULE & ASSESSMENT INSTRUMENT COMMENTS Exam Question: 1. Explain how expression of Ti plasmid T-DNA genes can lead to overproduction of plant growth regulators by genetically modified plant cells. Specimen Answer: T-DNA Gene 1 Tryptophan-2- Monooxygenase converts Tryptophan to Indole-3- Acetamide, in turn converted by Gene 2 Indole-3-Acetamide Hydrolase into highly active auxin Indole-3-Acetic Acid. Nett effect of expressing these two T-DNA genes (Gene 1 and Gene 2) is to increase active auxin concentrations in the plant cells by up to 1,500x. Causes excessive rootiness and/or the formation of fluffy white roots at the site of expression. T-DNA Gene 4 involved in complex cascade leading to overproduction of Zeatin-type cytokinins, by up to 1,200x in genetically modified plant cells. Causes excessive shootiness and cell division. Overproduction of the two together causes massive uncontrolled cell proliferation, leading to growth of characteristic crown gall tumour in dicotyledonous plant hosts. Narrative Commentary: Students answered this question generally very well, with the vast majority being able to explain the differing effects of expressing Genes 1 & 2 (for Auxin production) and Gene 4 (for Cytokinin production) at high levels in plant cells. A minority of students appeared to get Auxins and Cytokinins mixed up in their answers and therefore were suggesting that the wrong type of plant growth regulator promoted rooty or shooty phenotypes. A very small number of students were unable to provide any meaningful information on this topic. For Future Learning: Students may wish to consider enhancing their knowledge of T-DNA structure and function, for example by using the hand-outs provided on GCU Learn for Lecture 1 of this segment, or by reading one of the recommended references, such as Agrobacterium Protocols, Chapter 7. Students are reminded that it is the balance (or ratio) of Auxins: Cytokinins that will determine whether a dicot plant cell is programmed towards trying to divide and form root-, or divide and form shoot- initiating cells. Cells expressing high concentrations of both types of plant growth regulators will enter a phase of uncontrolled cell division, as known as a crown gall tumour, from which the term ‘crown gall disease’ of dicot plants is derived. Use of an appropriate diagram may also be helpful, to prompt recall of thoughts about these genes and the effects of their expression upon plant cells.