Vaccinium species such as blueberry, cranberry and bilberry are widely planted in the world and have high economic values. Current varieties are mostly generated by conventional breeding methods, and application of biotechnology especially genetic modification in the breeding process is seldom. Moreover, mechanistic studies of orphan genes in Vaccinium species are more and more important to facilitate the improvement of yield, quality, resistance to abiotic and biotic stresses in breeding, which requires efficient genetic transformation systems. In this study, we select 19 Vaccinium varieties to investigate the effects of variety specificity, explant types and culture schemes on callus development, adventitious bud regeneration and transformation efficiency. The wood plant medium (WPM) contains sufficient nutrients for the effective callus induction from mature leaves of plants grown in the field. Both zeatin and indole-3-butyric acid with the concentration of 2.0 mg L⁻¹ and 1.0 mg L⁻¹, respectively, are critical for adventitious bud regeneration from a variety of Vaccinium leaves. Different Vaccinium varieties have distinct regeneration efficiencies from leaf discs, and the diploid wild Vaccinium reticulatum ‘Red Button’ has the highest efficiency of callus formation and adventitious bud regeneration. Furthermore, we establish an efficient Agrobacterium-mediated genetic transformation system using ‘Red Button’, in which the transformation efficiency of green fluorescent protein (GFP) encoding gene is about 12.82%. Taken together, our study establishes an efficient tissue culture and regeneration system from leaves of Vaccinium varieties, and identify the diploid material ‘Red Button’ as an ideal variety for genetic transformation, which would greatly accelerate the mechanistic studies and breeding program of Vaccinium genus using emerging plant biotechnologies in the future.

蓝莓、蔓越莓、越橘等越橘品种在世界范围内广泛种植，具有很高的经济价值。目前的品种大多是通过常规育种方法产生的，很少在育种过程中应用生物技术特别是转基因技术。此外，越桔物种中孤儿基因的机理研究对于促进育种中产量、质量、对非生物和生物胁迫的抵抗力的提高越来越重要，这需要有效的遗传转化系统。在本研究中，我们选择了 19研究品种特异性、外植体类型和培养方案对愈伤组织发育、不定芽再生和转化效率的影响。木质植物培养基 (WPM) 含有足够的营养物质，可以有效地从田间种植的植物的成熟叶片中诱导愈伤组织。玉米素和吲哚-3-丁酸分别具有2.0 mg L ^-1和1.0 mg L ^-1的浓度，对于从各种越橘叶再生不定芽至关重要。不同的越橘品种具有不同的叶盘再生效率，二倍体野生越桔'Red Button'具有最高的愈伤组织形成和不定芽再生效率。此外，我们利用'Red Button'建立了一个高效的农杆菌介导的遗传转化系统，其中绿色荧光蛋白（ GFP）编码基因的转化效率约为12.82％。综上所述，我们的研究建立了一个高效的越橘叶片组织培养和再生体系，并确定了二倍体材料“红钮扣”作为遗传转化的理想品种，这将大大加快越橘属的机理研究和育种计划。未来新兴的植物生物技术。