Rebel xsi

Trees were monitored for flower formation each spring. Flowers were imaged using a Canon Rebel XSI digital camera. The average number of stigmas per flower was determined by counting the number of stigmas present in 30 total flowers from three trees (ten flowers per tree, from two clusters of 5 flowers chosen for similar amounts of floral opening) from each event, and from three control trees. A t-test was used to determine if the number of stigmas per flower were significantly different. Representative flowers were dissected and the floral organs imaged using a Keyence VHX-1000 digital microscope. Anthers and petaloid anthers were cross sectioned for imaging of pollen grains. Organs of flowers used for floral montages were removed and placed in order of phylotaxy, beginning with the sepals and ending with the gynoecium.

Non-transgenic apple pollen (variety Idared) was used to hand-pollinate selected flowers, pollinated clusters were labeled and monitored for fruit formation. All apples retained on trees were collected, sorted by tree and by category (apples from non-pollinated flowers, apples from cross-pollinated flowers), weighed, and photographed using a Canon Rebel XSI digital camera. Fruits were hand-sectioned with a sharp knife to examine internal fruit morphology and to count the seeds present in each fruit. Seeds were classified as developed (large, plump) or undeveloped (small, shriveled) based on visual inspection. Cross-sectioned fruits were used to count the numbers of locules present in each apple. Sectioned fruit pieces were dipped in a 3% citric acid solution to delay browning prior to photography. Once fruits were photographed, seeds were removed and retained for germination testing. T-tests were used to determine if the average fruit weights, seed numbers, seeds per locule, and locules per fruit were different between controls and transgenic events.