Establishment of Tissue Culture and Genetic Transformation Systems for Thick-peeled Melon'SN-1'
We used the self-pollinated line'SN-1'(Cucumis melo ssp.melo)of muskmelon as the experimental material.By screening conditions for seed and explant treatment,culture medium components,hormone ratios,and co-culture conditions,a tissue culture regeneration and genetic transformation system for the self-pollinated line'SN-1'of muskmelon was established.The results indicated that the germination time of seeds on Agar medium was significantly better than on 1/2MS+Agar and MS+Agar;analyzing the effects of different concentrations(2%,3%,and 4%,v/v)of NaClO disinfectant and treatment times(10,15,20,and 25 min)on seed germination revealed that treatment with 3%NaClO for 15 minutes resulted in significantly higher germination rates and uniformity compared to other treatments;compared to the"binary method,"the"quaternary method"can obtain more cotyledonary nodes without affecting differentiation,significantly improving seed utilization;further analysis found that collecting cotyledonary nodes from seeds with a radicle length of about 5 mm at 48 hours after sowing resulted in significantly better adventitious bud differentiation than other periods;research on the effects of different hormone ratios and concentrations on cotyledonary nodes revealed that the best adventitious bud differentiation was achieved with concentrations of 0.5 mg·L-1 6-BA,0.05 mg·L-1 IAA,and 1 mg·L-1 ABA in the culture medium;when adding 0.4 mg·L-1 IAA to the rooting medium,the induction effect of adventitious roots in the regenerated seedlings was significantly better than other treatments;analysis of the co-culture period and medium hormone ratios after infection revealed that the best cotyledonary node development occurred with a 3-day co-culture period and the addition of 0.5 mg·L-1 6-BA+0.1 mg·L-1 IAA to the medium.
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