For our experiments we modified two media; agar was reduced from 10 to 6 g l−1 and sucrose from 30 to 20 g l−1 in
MS medium (hereafter referred to as modified MS) and agar was reduced from 16 to 6 g l−1 and fructose was replaced with sucrose in Robert Ernst medium (hereafter referred to as modified RE; cf. Arditti, 1982). To assess the effect of reduced salt and addition of coconut water (CW) as a supplement, the harvested seeds were tested in six compositions: (1) 1/2 strength modified MS;
(2) 1/2 strength modified MS supplemented with 10% CW;
(3) 1/4 strength modified MS;
(4) 1/4 strength
modified MS supplemented with 10% CW;
(5) modified RE with 1.0 g l−1 activated charcoal;
(6) modified RE with 1.0 g l−1
activated charcoal and 10% CW. In all media pHwas adjusted to 5.8 using 1 mol l−1 NaOHor 1 mol l−1 HCl. Each treatment was replicated 15 times in 200 ml sterile clear glass jars with 30 ml of media solution. Approximately 100 seeds, each sterilized for 5 min in 1% NaOCl, were placed in each jar under sterile conditions. Seed placement was done by suspending seeds in 0.1% autoclaved Agar solution using a sterile syringe and pipetting out 1 ml for each replicate. After seed placement, the exact number of seeds in each jar was counted. Seeds were then allowed to germinate in tissue culture rooms under a 12 h dark and 12 h light cycle. We used cool white fluorescent lights with approximately 200 μ molm−2 s−1, for the photoperiod cycle condition. In orchids germination and seedling development are divided into six stages including stage 0 the ungerminated seed and were scored as described by Arditti (1967). Germination (stage 1), defined as imbibed seeds still covered by testa and with a white and green color visual to the eye, was assessed three months after seed sowing
For our experiments we modified two media; agar was reduced from 10 to 6 g l−1 and sucrose from 30 to 20 g l−1 inMS medium (hereafter referred to as modified MS) and agar was reduced from 16 to 6 g l−1 and fructose was replaced with sucrose in Robert Ernst medium (hereafter referred to as modified RE; cf. Arditti, 1982). To assess the effect of reduced salt and addition of coconut water (CW) as a supplement, the harvested seeds were tested in six compositions: (1) 1/2 strength modified MS; (2) 1/2 strength modified MS supplemented with 10% CW; (3) 1/4 strength modified MS; (4) 1/4 strengthmodified MS supplemented with 10% CW; (5) modified RE with 1.0 g l−1 activated charcoal; (6) modified RE with 1.0 g l−1activated charcoal and 10% CW. In all media pHwas adjusted to 5.8 using 1 mol l−1 NaOHor 1 mol l−1 HCl. Each treatment was replicated 15 times in 200 ml sterile clear glass jars with 30 ml of media solution. Approximately 100 seeds, each sterilized for 5 min in 1% NaOCl, were placed in each jar under sterile conditions. Seed placement was done by suspending seeds in 0.1% autoclaved Agar solution using a sterile syringe and pipetting out 1 ml for each replicate. After seed placement, the exact number of seeds in each jar was counted. Seeds were then allowed to germinate in tissue culture rooms under a 12 h dark and 12 h light cycle. We used cool white fluorescent lights with approximately 200 μ molm−2 s−1, for the photoperiod cycle condition. In orchids germination and seedling development are divided into six stages including stage 0 the ungerminated seed and were scored as described by Arditti (1967). Germination (stage 1), defined as imbibed seeds still covered by testa and with a white and green color visual to the eye, was assessed three months after seed sowing
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