Either embryogenesis or indirect organogenesis in sugarcane: Are we missing the key points?

Manoel P L Neto, L. R. Vieira, Pedro Vitor Schumache, Mariele Rossato, Luciano Coutinho Silva, F. J. Pereira, R. Paiva, A. C. Junior
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Abstract

Both in vitro establishment and callogenesis of sugarcane allow a production of quality regenerative material, which is necessary for in vitro clonal propagation and for genetic transformation. In this study, we establish the efficient production of calli from the RB855156, RB92579 and RB867515 cultivars and characterize their regenerative potential in relation to either an embryogenic or an organogenic origin both by morphology and by anatomy. Callogenesis was induced in MS medium with 3.0 mg L-1 2.4-D. Three antioxidants were tested: polyvinylpyrrolidone (150; 300; 600 mg L-1), citric acid (7.5; 15; 30; 60 mg L-1), and ascorbic acid (7.5; 15; 30; 60 mg L-1). The morphological characterization of the calli was performed by visual classification, and the anatomical analyses by light microscopy. The experimental design was completely randomized, containing 150 explants by cultivar to antioxidant evaluations and potential regenerative evaluation within three times of subcultures (84; 112; 140 days). We have attained the key points of our in vitro research. Calli regeneration depended on the oxidation level and genotype. Antioxidants only in the culture medium were not enough to prevent oxidation. However, citric acid (7.5 mg L-1) as a pretreatment of the explant minimized this problem. Bacterial contamination was developed for the three cultivars, inhibiting the establishment to RB867515. The disinfestation protocol was efficient for RB855156 and for RB92579 cultivars. Three varieties of calli differed in the regeneration potential. In addition, histological analysis of the calli unfolded not only that there were structural differences but also that their buds had an organogenic origin
甘蔗的胚胎发生或间接器官发生:我们是否错过了关键点?
甘蔗的离体培养和愈伤组织形成都可以产生优质的再生材料,这是甘蔗离体无性系繁殖和遗传转化所必需的。在本研究中,我们建立了RB855156、RB92579和RB867515愈伤组织的高效生产,并通过形态学和解剖学表征了它们的胚性和器官性再生潜力。在MS培养基中添加3.0 mg L-1 2.4-D诱导形成。测试了三种抗氧化剂:聚乙烯吡咯烷酮(150;300;600 mg L-1),柠檬酸(7.5;15;30;60 mg L-1),抗坏血酸(7.5;15;30;60 mg L-1)。通过视觉分类和光镜解剖对愈伤组织进行形态学表征。试验设计为完全随机化设计,按不同品种共150个外植体,在3次传代培养中进行抗氧化评价和再生潜力评价(84;112;140天)。我们已经达到了体外研究的关键点。愈伤组织再生取决于氧化水平和基因型。仅在培养基中添加抗氧化剂不足以防止氧化。然而,柠檬酸(7.5 mg L-1)作为外植体的预处理最小化了这个问题。3个品种均发现细菌污染,抑制了对RB867515的建立。除虫方案对RB855156和RB92579均有效。3个愈伤组织的再生能力存在差异。此外,对愈伤组织的组织学分析表明,愈伤组织不仅在结构上存在差异,而且其芽具有器官起源
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