Somatic embryogenesis in Agave tequilanaWeber var. Azul was affected by the interaction between the light regimes applied during the induction phase and the expression phase. When embryogenic calli was exposed to white or red light during the expression phase, an average of two germinated embryos per explant was obtained regardless of the light treatment used for callus induction. Conversely, the highest number of germinated embryos, an average of 18 per explant, was obtained when applying either white or red light during the induction phase and then wide-spectrum light during the expression phase. Culture medium had also a great influence in this process, with embryo germination being reduced by up to 70%, depending on the light treatment, when using Schenk and Hildebrandt (SH) medium instead of Murashige and Skoog (MS) medium.
The genus Agave originates from the American continent and grows in arid and semiarid places, being México the center of origin. Many species of the genus are a source of diverse products for human needs, such as food, medicines, fibers, and beverages, and a good source of biomass for the production of biofuels, among many others. These plants are gaining importance as climate change becomes more evident as heat is reaching temperatures above 40 °C worldwide and rains are scarce. Many species of the genus grow in places where other plant species do not survive under severe field conditions, due to their CAM pathway for fixing CO where gas exchange occurs at night when stomata are open, allowing them to avoid excess loss of water. Most of the important species and varieties are usually propagated by offshoots that develop from rhizomes around the mother plant and by bulbils that develop up in the inflorescence, which are produced by the plant mostly when there is a failure in the production of seeds.Areas for commercial plantations are growing worldwide and therefore in the need of big amounts of healthy and good quality plantlets. Although many Agave species produce seeds, it takes longer for the plants to reach appropriate maturity and size for diverse purposes. Micropropagation techniques for the genus Agave offer the opportunity to produce relatively high amounts of plants year around in relatively small spaces in a laboratory. Here, a protocol for micropropagation that has proven good for several Agave species (including species from both subgenera) is presented in detail with two different kinds of explants to initiate the process: rescued zygotic embryos and small offshoots that grow around a mother plant.
We demonstrate the effectiveness of laser-induced fluorescence (LIF) for monitoring the development and stress detection of in vitro tissue cultures in a nondestructive and noninvasive way. The changes in LIF spectra caused by the induction of organogenesis, the increase of the F690/F740 ratio as a result of the stress originated in the organogenic explants due to shoot emergence, and the relationship between fluorescence spectra and shoot development were detected by LIF through closed containers of Saintpaulia ionantha.
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