The Influence Of Priming Two Cucumber Cultivar Seeds

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J. Duhok Univ. Vol.13, No.1, (Agri. And Vet. Sciences) Pp 112-121, 2010 RESULTS AND DISCUSSION Effect of Auxin and Cytokinine in callus induction: The most important factors contributing the induction of somatic embryo from the callus, plant growth regulators and medium formulation (Buyukalaca and Mairtuna, 1996). Among the plant growth regulators, generally auxin is known to be essential for the induction of somatic embryogenesis and 2,4-D is the most commonly used auxin (Ammirato, 1983). Morever, a combinations of 2,4-D or NAA with cytokinine was reported to be essential for induction of callus and somatic embryos (Jarsrai et al, 2003). Certain cells may need simple MS medium for the induction of somatic embryos and further development (Jarsrai et al, 1999). To determine the best callus induction response from leaves explant of different types, various concentrations and combinations of plant growth regulators were tried (Table 1). The best results of callus induction response (100%) in whole area of leaf explant was observed on MS medium supplemented with 2 mgl -1 BA combined with (0.2 or 0.4 mgl -1 ) 2,4-D (Fig 2. a) under dark condition followed by MS medium supplemented with (1, 2) mgl -1 BA + (0.2, 0.4) mgl -1 2,4-D (80%) callus was formed on the three fourths of explant under light condition (Fig 2. b). The lowest callus induction response (30%) was observed on MS medium supplemented with 1 mgl -1 BA + 0.4 mgl -1 NAA formed on the two fourths of explant under dark condition (Fig 2. c) followed by MS medium supplemented with 1 mgl -1 BA + (0.2, 0.4 mgl -1 ) NAA (20%) callus was formed on the one fourth of explant under light condition (Fig 2. d) however callus was not formed at all in MS medium free-hormone (control) under both conditions (Fig 2. e). Presad et al (2006) 111 A B C reported callus development from Ipomoea aqatica leaves segment cultured on MS medium with different concentrations of BA combine with 2,4-D. Nurazan et al (2009) and Lima et al (2008) obtained callus in Cananga odorata and Croton uruerana using different levels of 3 mgl- 1 NAA or 3 mgl -1 2,4-D from petal explant and leave segments respectively. In the same table, the average callus fresh weight was obtained on MS medium supplemented with 2 mgl -1 BA + 0.2 mgl -1 2,4-D with an average of (5.39) mg/ culture. Medium containing less than 2 mgl -1 BA + 0.2 mgl -1 2,4-D and higher than produced fewer fresh weight of callus / culture under dark condition and the highest percentage of embryo induction was (98%) in the same condition and medium while the average callus fresh weight (3.91) mg/ culture was obtained on MS medium supplemented with 2 mgl -1 BA + 0.2 mgl -1 2,4-D and the highest percentage of embryo induction (89%) was obtained in MS medium supplemented with 2 mgl -1 BA + 0.4 mgl -1 2,4-D under light condition . In vitro grown leaf disk did not show any callus production on MS medium free-hormone treatment (control) (Fig 2.e). Marconi and Radice (1997) reported somatic embryo from Codiaeum variegatum L. leaves explant cultured on MS medium containing 1mgl -1 2,4-D + 1mgl -1 BA. Conversely, Manohari et al (2008) concluded that in leaf explant of Elettaria cardamonum Malon, the callus induction was achieved on MS medium with combinations of NAA and BAP. The results of this work support those found by other authors. For instance, Machii (1999) did not observed callus formation in leaf explant of Morus alba L. in the absence of 2,4-D and Vietez and san-Jose (1996) did not obtained callus in leaf explant of Fagus silvatica L. in the presence of BA. Fig. (2): Callus induction from leaf of (Codiaeum variegatum). A: callus was formed on whole area of explant. B: callus was formed on the three fourths of explant. C: callus was formed on the two fourths of explant. D: callus was formed on the one fourth of explant. E: callus was not formed at all. D E
J. Duhok Univ. Vol.13, No.1, (Agri. And Vet. Sciences) Pp 112-121, 2010 Table (1): Effects of plant growth regulators on callus and somaic embryogenesis from leaf explant of Codiaeum variegatum after 6-8 weeks of culture in MS medium. Growth regulator mg l -1 Callus induction % * Means followed by the same letter within each character (column) do not differ significantly (P≤0.05) according to Duncan's Multiple Range Test (Duncan ,1955). ++++: callus was formed on whole area of explant. +++: callus was formed on the three fourths of explant. ++: callus was formed on the two fourths of explant. +: callus was formed on the one fourths of explant. -: callus was not formed at all. Effect of different concentrations of BA or Kin alone on shoot formation from callus induction: Data on shoot formation revealed that when the concentration of BA was increased from 2.5 mgl -1 to 5 mgl -1 , the rate of shoot formation was also increased. Maximum number of somatic embryos induced from callus, maximum shoots formation and highest number of leaves per culture were obtained at 2.5 mgl -1 BA which was 7.95 embryo/ culture, 4.58 shoots/ culture and 3.13 leaves/ culture respectively after 6-8 weeks of culture inoculation (Table 2, Fig. 3 a-c). With further increase in concentration of BA, 7.5 mgl -1 BA produced 2.25 shoots/ culture while at MS medium free-hormone, the medium did not formed any shoots per culture, and the highest length of shoots (2.89 cm )was found on MS medium supplemented 7.5 mgl -1 compared with 10 mgl -1 BA and control treatment which was (1.50 cm). Many researchers have also reported positive effect of BA on shoot multiplication and proliferation. Rahaman et al (2004) cultured leaf segments of Kaempferia galangal L. on different media and reported a number of somatic Dark condition Light condition Average callus fresh weight (mg) Embryo induction % Callus formation on mature leaf culture Callus induction % Average callus fresh weight (mg) Embryo induction embryonic formation on MS medium with 2 mgl -1 BA plus 0.05 mgl -1 NAA. Similarity, Nakaura (1991) found mass prolific growth of Tea plants using 1 mgl -1 BA. During this investigation, it was also observed that 2.5 mgl -1 of BA was the best concentration for shoot development whereas higher concentrations inhibited it. This was in accordance with the study conducted by Kumar ad Kanwar (2004) in which they reported that lower concentration of BA (1 mgl -1 ) stimulated shoots growth in Gerbera jamesoni, but the higher concentration of BA (5 mgl -1 ) inhibited shoot proliferation. For in vitro shoot formation from callus, MS medium was supplemented with different concentrations of Kin ranging from 2.5 mgl -1 to 10 mgl -1 . It was noticed that when the concentration of Kin was increased to 7.5 mgl -1 , average number of embryo per culture formed was also increased. At this concentration, maximum number (3.70 embryo/ culture) was formed. Maximum shoots formation per culture and highest number of leaves per culture was observed at 7.5 mgl -1 Kin which was 3.45 % Callus formation on mature leaf culture control 0f 0f 0g - 0f 0f 0g - BA+NAA 1+0.2 40c-f 1.23e 66.6bc +++ 20ef 0.92e 0.0g + 1+0.4 30d-f 2.28cd 73.5b +++ 20ef 1.47e 0.0g ++ 2+0.2 70a-d 2.45cd 93.4a +++ 40c-f 2.25cd 27.0e ++ 2+0.4 70a-d 2.57cd 56.6cd +++ 30d-f 2.09d 13.3f ++ BA+2,4-D 1+0.2 90ab 2.84c 46.7d +++ 50b-e 2.46cd 0.0g ++ 1+0.4 80a-c 2.89c 90.0a ++++ 80a-c 2.79cd 13.3f +++ 2+0.2 100a 5.39a 96.6a ++++ 80a-c 3.91b 33.0e +++ 2+0.4 100a 4.19a 86.6a ++++ 70a-d 3.36b 23.3e +++ 111
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The Influence Of Priming Two Cucumber Cultivar Seeds

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