Genetic transformation of Eucalyptus camaldulensis by agrobalistic method

Abstract

-Eucalyptus stands in the setting of worldwide forestry due to its adaptability, rapid growth, production of high-quality and low cost of wood pulp fibers. The eucalyptus convetional breeding is impaired mainlly by the long life cycle making the genetic transformation systems an important tool for this purpose. However, this system requires in vitro eficient protocols for plant induction, regeneration and seletion, that allow to obtain transgenic plants from the transformed cell groups. The aim of this work was to evaluate the callus formation and to optimize the leaves and callus genetic transformation protocol by using the Agrobacterium tumefaciens system. Concerning callus formation, two different culture media were evaluated: MS medium supplemented with auxin, cytokinin (M1) and the MS medium with reduced nitrogen concentration and supplemented with auxin, cytokinin coconut water (M2). To establish the leave genetic transformation, those were exposed to agrobiolistics technique (gene gun), to tissue injury, and A. tumesfasciens EHA 105 contening the vetor pCambia 3301 (35S::GUS::NOS), for gene transference and to establish the callus transformation thoses were exposed only to A. tumefasciens. For both experiments, the influence of different infection periods was evaluated. The M2 medium provided the best values for callus sizea and fresh and dry weight. The leaves genetic transformation using the agrobiolistics technique was effective, the gus gene transient expression could be observed. No significant differences were obtained in the infection periods (4, 6 and 8 minutes). The callus genetic transformation with A. tumefaciens also promotend the gus gene transient expression on the callus co-cultiveted for 15 e 30 minutes. The transformed callus was transfered to a regeneration and selection medium and transformed plants were obtained.

Indução de calos em explantes foliares e caulinares de Cumari-do-Pará (Capsicum chinense Jacquin)

“…A cada 5 dias, durante 30 dias, foram feitas observações acerca da indução de calos (IC) e área do explante coberta por células de calos (ACCC). A ACCC foi contabilizada por meio de uma adaptação do método descrito por Mendonça et al (2013), em que a ACCC foi dada pelas porcentagens 0, 10, 25, 50, 75, 90 ou 100% de área do explante coberta por células de calos.…”

Effect of 2,4-D and BA on the establishment of cell suspension from nodes and internodes of Capsicum annuum cv. Etna

“…Treatments were arranged in a completely randomized design. After 49 days, evaluations were done by assessing the percentage of explants where callus induction occurred (%CI); the explant area covered by callus cells (ACCC), according to Mendonça et al (2013), who established the following scores: 0 = 0%, 1 = 25%, 2 = 50%, 3 = 75% and 4 = 100% of leaf area covered by callus; and the fresh weight of the explants, by using a precision scale. Variance analyses and Tukey tests (P<0.05) were performed by using the Assistat 7.5 statistical program.…”

Syzygium cumini (L.) Skeels: Cardiometabolic Properties and Potential Tissue Culture-Based Improvement of Secondary Metabolites Production

“…In another report by Aggarwal et al [75], a procedure for A. tumefaciens mediated T-DNA delivery into the tissues taken from selected E. tereticornis elite clones, and subsequent regeneration of transformed shoots has been developed, with the aim to introduce the useful properties in this commercially important tree species. Similarly agrobalistic method has been used for genetic transformation in E. camaldulensis Dehnh [78]. Stummer et al [77] described a system for transformation and regeneration of transgenic V. grandis J.L.…”