Hi, Good Afternoon,
Kindly follow the methodology,
Seeds of a single species were sown into 24 L pots (40*30*20 cm 3; length:width: height) and seedlings thinned after emergence leaving a total number of 12 plants per pot. Three replicates were established for each species. The potting mix used was Pindstrup Substrate No. 1, 0-20 mm sieved and with a pH 6 (Pindstrup Mosebrug A/S Denmark). Pots were watered for four six minutes per day with a nutrient solution (Pioner NPK MAKRO 14323+Mg) which contained the following nutrients: 14.5% Ntotal, 10.7% NO 3N, 3.8% NH4N, 2.9% water soluble P, 23.1% water soluble K, 3.0% water soluble Mg and 3.9% water soluble S. The nutrient solution was mixed with water to a EC=1.3 mS and regulated with 62% HNO 3 to a pH of 5.5. Temperature in the greenhouse was kept at 15ºC during the day and 12ºC during the night with a photoperiod of 13-11 hours (day-night).
Then, you can use the statistical analysis like as follows:
Samples were taken at four different key stages of plant development: leaf, bud, flower and seed. The stages chosen were, according to Berkenkamp (1973): 1.4 (four leaves), 2.3 (pedicels elongating), 3.1 (many flowers open) and 4.0 (seeds in lower pods full size). A total of three plants per species (one plant per replicate) were sampled at each growth stage. Plants were removed intact from the pots, the roots carefully washed with tap water and the whole intact plants were then freeze-dried.
Once dried, the plants were separated into roots, stems, leaves and reproductive tissues and their weight determined. The material was then thoroughly milled and kept dried until extraction. Glucosinolates were extracted, desulphoglucosinolates produced and the individual compounds were determined by MECC according to standard procedures with use of two internal standards as described elsewhere.
Kindly refer: Bjerg and Sorensen et al - 1987; Michaelsen et al -1992; Sorensen et al - 1999
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