Ricci Angela, Capriotti Luca, Mezzetti Bruno, Navacchi Oriano, Sabbadini Silvia
Department of Agricultural, Food and Environmental Sciences, Marche Polytechnic University, 60131 Ancona, Italy.
Vitroplant Italia, 47521 Cesena, Italy.
Plants (Basel). 2020 Jun 16;9(6):755. doi: 10.3390/plants9060755.
In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 μM N-benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 μM STS or carbenicillin (238 μM) combined with cefotaxime (210 μM) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars.
在本研究中，已建立了一种从桃砧木汉森536叶片体外再生不定芽的高效系统。测试了20种以McCown木本植物培养基（WPM）为基础盐并添加不同浓度和组合植物生长调节剂（PGR）的再生培养基。将3周龄伸长的体外芽培养物中的展开叶及其叶柄用作起始外植体。在富含15.5 μM N-苄基腺嘌呤（BAP）的WPM基础培养基上获得了最高再生率（高达53%）。还研究了乙烯抑制剂硫代硫酸银（STS）和添加到优化再生培养基中的不同抗生素组合对叶片再生的影响。与对照相比，使用10 μM STS或羧苄青霉素（238 μM）与头孢噻肟（210 μM）组合可显著增加每片叶再生芽的平均数量。体外芽最终伸长、生根并在温室中成功驯化。本研究取得的结果增进了对影响 spp.叶片器官发生因素的了解，所描述的再生方案对于优化汉森536及其他桃砧木和品种的新遗传转化程序看起来很有前景。
