João Paulo Rodrigues Martins

Physiological responses by Billbergia zebrina (Bromeliaceae) when grown under controlled microenvironmental conditions

Sucrose, the most commonly used carbon source in conventional in vitro culture, and limited air exchange in the culture containers are factors that affect the growth of in vitro-cultured plants. They may induce physiological disorders and decrease the survival rate of plants after transfer to ex vitro conditions. The aim of the present study was to analyze the effects of gas exchange and sucrose concentration on Billbergia zebrina plantlets during in vitro propagation. In vitro-established B. zebrina plantlets were transferred to culture media containing 0, 15, 30, 45, or 60 g L-1 sucrose. Two different culture-container sealing systems were compared: lids with a filter (permitting gas exchange) and lids with no filter (blocking fluent gas exchange). Carbohydrate and chlorophyll (Chl a+b) concentrations were analyzed in plantlets at 45-days of culture. The addition of sucrose to the medium reduced the Chl a+b concentration in the plantlets. On the other hand, additional sucrose had a positive effect on the carbohydrate stock formation of the plantlets. The results showed that a photoautotrophic system (air exchange and a sugar-free medium) improves the in vitro propagation of B. zebrina without creating physiological disorders. Key words: Bromeliad, In vitro plant, photoautotrophic growth, physiological disorders, sucrose.

CRESCIMENTO E ASPECTOS SINTOMATOLÓGICOS NA ACLIMATIZAÇÃO DE IPÊ-ROXO

This work evaluated the influence of seal type and substrate type on pre-acclimatization, and symptomatological aspects relating to acclimatization of ipe-roxo. For the pre-acclimatization, ipe-roxo plants obtained through an in vitro zygotic embryo culture were aseptically inoculated in test tubes with different types of seal (cotton plug, plastic cap + parafilm and plastic lid) and different substrates (agar, vermiculite and Plantmax ® ) supplemented with a WPM culture medium with 1 gL -1 of activated charcoal, 10 mgL -1 of citric acid and 30 gL -1 of sucrose added. For the acclimatization, in vitro ipe-roxo plants were transplanted into 56-cm o C and 1 atm for 20 minutes. Once transplanted, the plants were irrigated with different concentrations of the MS medium (25%, 50%, 75%, 100% and 150%), using distilled water as control. After 30 days, the effects of seal type and substrate type on pre-acclimatization were examined. Cotton can be recommended where Plantmax ® or vermiculite is used as substrate. In the acclimatization it was noted that where plants were irrigated with distilled water and concentrations < 50% they showed symptoms of nutritional deficiency. Similar symptoms were observed when using MS medium at a concentration equal to or greater than 100%. The 75% MS concentration was found to be the most effective at maintaining the nutritional vigor of ipe-roxo seedlings in the process of acclimatization .

Effects of 6-benzylaminopurine on photosystem II functionality and leaf anatomy of in vitro cultivated Aechmea blanchetiana

Cytokinins (CKs) are often used during the in vitro cultivation of plant species. However, it is not clear how CKs, such as 6-benzylaminopurine (BAP), affect photosystem PS) II functionality and leaf anatomy over a long period of in vitro plant propagation. The aim of this study was to analyze the residual effects of BAP on the photosynthetic performance and leaf anatomy of Aechmea blanchetiana after 120 d without exposure to CKs. Aechmea blanchetiana plants previously grown in vitro were transferred to Murashige and Skoog (MS) culture media containing 0, 5, 10, 15, or 20 μM BAP. After 60 d on the MS medium with BAP, explants were subcultivated twice on the MS medium without growth regulators, first in a stationary liquid medium for 60 d and then in a solidified medium with 6 g dm-3 agar for 60 d. Leaf anatomy, pigment content, and chlorophyll a fluorescence were assessed for plants from each treatment after 120 d on the CK-free medium. Stomatal density presented a negative linear correlation with BAP concentration. Pigment content decreased in plants subjected to previous BAP exposure. An increase in absorbed energy flux per reaction center (ABS/RC) and a sharp decrease in energy transport flux (ETo/RC) followed by an increase in energy dissipation flux (DIo/RC) also occurred. Furthermore, maximum quantum yield (FV/FM) decreased as a function of BAP concentration. Thus, the use of BAP during in vitro propagation of A. blanchetiana induced long-term physiological defects.

Anatomy and photosystem II activity of in vitro grown Aechmea blanchetiana as affected by 1-naphthaleneacetic acid

Auxins are one of the main regulators of in vitro plant growth and development. However, the mechanisms, by which auxins, such as 1-naphthaleneacetic acid (NAA), affect in vitro root and leaf anatomy and photosystem function, remain unclear. Accordingly, the aim of the present study was to analyze the effect of different NAA concentrations on the anatomy and photosynthetic performance of in vitro-propagated Aechmea blanchetiana and to determine whether such a treatment affects micropropagated plants after acclimatization. In vitro-established A. blanchetiana plants were transferred to culture media that contained 0, 2, 4, or 6 μM NAA, and after 50 d, they were transplanted into plastic seedling trays with a commercial substrate and cultivated for 60 d in a greenhouse. The plants were evaluated after a 50-d in vitro NAA exposure (growth traits, chlorophyll α fluorescence, and root and leaf anatomy) and after 60 d of acclimatization in the greenhouse (root and leaf growth). Changes induced by NAA in root anatomy might improve uptake of minerals and sugars from the medium, thereby increasing the in vitro growth. In the leaves, the lowest chlorenchyma thickness and sclerenchyma area were observed in plants grown without NAA, and NAA exposure also improved photosystem II activity. The highest ex vitro growth rate was observed for plants that were propagated with 4 μM NAA. Therefore, the use of NAA during in vitro propagation can improve the anatomical and physiological quality of A. blanchetiana plants, as well as to improve ex vitro transfer.

Morpho-physiological changes in Billbergia zebrina due to the use of silicates in vitro

The use of silicon in Billbergia zebrina cultivation in vitro is an alternative for optimizing micropropagation of this important ornamental plant species. The objective of the present study was to evaluate the growth and anatomical and physiological alterations in Billbergia zebrina (Herbert) Lindley plants as a function of different sources and concentrations of silicon during in vitro cultivation and acclimatization. The experimental design was completely randomized, with a double factorial arrangement and an additional control treatment (2 x 3 + 1). The first factor was relative to calcium silicate and sodium silicate added to the Murashige & Skoog culture medium; the second factor was related to its concentrations, 0.5, 1.0, and 2.0 mg L-1. After 100 days, their growth, anatomical characteristics, level of silicon and chlorophyll content were evaluated. Growth characteristics were assessed after 60 days of acclimatization period. Plants absorbed more sodium silicate than calcium silicate. This source also stressed the plants impairing their growth, but the highest silicon absorption at 1 mg L-1 attenuated the stressful conditions. The supplementation of the culture medium with calcium silicate led to improved growth, anatomical, and physiological characteristics, which benefited the development of more resistant seedlings with better performance during acclimatization.