A brief temperature pulse enhances the competency of microspores for androgenesis in Datura metel

Abstract

Androgenesis may be induced in plants by a stress application on microspores or anthers. Temperature stress treatments have generally been confined to a single temperature regime (above or below ambient) lasting from a few hours to days. We introduced a gradient with two temperature pulses (30 s each) in the stress application on anthers of Datura metel L. by stepping the temperature up and down for a total period of 60 s. Anthers were immersed in sterile water preheated and cooled to the desired temperature and cultured on Nitsch medium. The temperature pulse gradient significantly improved androgenesis compared to single temperature treatments, resulting in increased mean embryogenesis of 128% over control for 45°/15°C, 110% for 45°/10°C, 113% for 40°/10°C and 96% for 45°/5°C. The 45°/10°C gradient also significantly increased the number of dividing microspores observed, after 14 days of anther culture. Besides the differential of the gradient, the temperature limit was important, with anthers not tolerating temperatures beyond 45°C. The temperature pulse gradient applied at an early stage of culture may increase the window of competency of microspores for androgenesis.