Shoot regeneration experiments from growth-point-derived callus were conducted to improve apple genome editing techniques. The plant material studied was in vitro maintained shoots of ‘Fuji’. In the first step, a procedure for shoot formation from apple callus was established. After using the two earlier reported cases’ experimental procedures and media compositions, we investigated the effect of media variations in callus induction, callus multiplication, and shoot induction from the meristem. The procedure that yielded a higher shoot regeneration rate with meristem-derived callus involved shoot multiplication with 1001 medium followed by the Caboni’s callus induction medium, then without callus propagation on liquid medium, and then either Caboni’s or Saito-Suzuki’s shoot induction medium. In this experiment, axillary buds may remain as the shoot apexes were excised at 1 mm. In the next experiment, shoot apexes were excised at 0.5 mm to completely eliminate the axillary bud. Then treatment in the dark was added to the procedure to further improve shoot regeneration rates for the meristem-derived callus. Shoot multiplication medium and shoot induction medium procedures were conducted under dark conditions. This yielded the following optimal procedure: excising meristems from chlorosis shoots after 2–3 months of dark treatment in shoot multiplication medium on 1001; placing the excised meristems in Carboni’s callus induction medium in the dark for 20 days; then transferring the formed callus Saito-Suzuki’s shoot induction medium with incubation in the dark for at least 2 weeks. The shoot regeneration rates of calli treated in the dark for 6 weeks with shoot induction medium reached 68%. Relative to previous reports, this value is considered high for shoot regeneration from calli in apple cultivars.

进行了生长点愈伤组织的芽再生实验，以改进苹果基因组编辑技术。研究的植物材料是在体外保留了“富士”的拍摄。第一步，建立了苹果愈伤组织芽形成的程序。在使用两个早期报道的案例的实验程序和培养基成分后，我们研究了培养基变化对愈伤组织诱导、愈伤组织增殖和分生组织芽诱导的影响。使用分生组织衍生的愈伤组织产生较高芽再生率的过程涉及用1001培养基进行芽增殖，然后用卡博尼愈伤组织诱导培养基进行芽增殖，然后在液体培养基上不进行愈伤组织繁殖，然后用卡博尼或斋藤铃木芽诱导培养基进行芽增殖。在此实验中，当芽尖被切除 1 毫米时，腋芽可能会保留。在接下来的实验中，将芽尖切除0.5毫米以完全消除腋芽。然后在该过程中添加黑暗处理，以进一步提高分生组织衍生愈伤组织的芽再生率。芽增殖培养基和芽诱导培养基程序在黑暗条件下进行。由此产生了以下最佳程序：在1001上的芽增殖培养基中进行2-3个月的黑暗处理后，从失绿芽中切除分生组织；将切下的分生组织置于Carboni's愈伤组织诱导培养基中，在黑暗中放置20天；然后将形成的愈伤组织转移到Saito-Suzuki的芽诱导培养基中，并在黑暗中孵育至少2周。用芽诱导培养基在黑暗中处理6周的愈伤组织芽再生率达到68％。相对于之前的报道，对于苹果品种中愈伤组织的芽再生来说，该值被认为是较高的。