Identification of proteins related to microspore embryogenesis responsiveness in anther cultures of winter triticale (\xd7Triticosecale Wittm.)

Monika Krzewska,Ewa Dubas,Iwona Żur,Marta Gawin,Gabriela Gołębiowska-Pikania

doi:10.1007/s10681-017-1978-1

Abstract

For a better understanding of the physiological background of microspore embryogenesis (ME), the protein profile was analyzed in four winter triticale DH lines, which show extremely different embryogenic potential. The analysis were conducted with anthers at the phase of development optimal for ME induction and then after low temperature (LT, 3 weeks at 4 °C) ME-inducing tillers treatment. The sub-proteome of anthers was mapped by two-dimensional gel electrophoresis (2-DE). The protein species significantly more abundant (at least 2-fold) in responsive DH lines after LT treatment were chosen for identification by MALDI-TOF/TOF analysis. In total, 31 protein species were successfully identified as involved in the determination of microspore competence, stress response and in the regulation of ME induction. Microspore competence required sufficient energy supply and efficient system of cell protection that determine survival under prolonged LT stress treatment. LT stress was associated with increased accumulation of proteins typical for cell defence against oxidative stress (e.g., l-ascorbate peroxidase), chaperons (e.g., HSP70) and other enzymes/factors ensuring protein biosynthesis, stability and active cell divisions. Also here, effective cell defence required undisturbed energy supply. Among proteins that accumulated differentially in accordance with microspore embryogenic potential again the most important role seems to be played by the enzymes ensuring energy production and determining ability of plant stress adaptation. Two protein species (enolase, 12S storage protein), proposed earlier as candidates for markers of embryogenesis in other in vitro plant culture systems confirmed their utility for triticale anther cultures.

Highlights

Every living cell possess the whole genetic information that determines the characteristics of a species as a whole and of the particular individual
The term ‘Microspore embryogenesis (ME)’ is often used synonymously with ‘androgenesis’ or ‘pollen embryogenesis’ and in this paper it refers to the process taking place in in vitro cultured anthers in which the cells of male gametophyte were reprogrammed towards embryo-like structures (ELS) production
Incorporation of doubled haploid plants (DHs) technology in breeding programmes increases the efficiency of new cultivar production and accelerates the breeding progress in comparison with traditional methods

Summary

Introduction

Every living cell possess the whole genetic information that determines the characteristics of a species as a whole and of the particular individual. The term ‘ME’ is often used synonymously with ‘androgenesis’ or ‘pollen embryogenesis’ and in this paper it refers to the process taking place in in vitro cultured anthers in which the cells of male gametophyte were reprogrammed towards embryo-like structures (ELS) production. Such more common use of the term ‘microspore embryogenesis’ was proposed by Soriano et al (2013) and Wedzony et al (2015). The progressively growing economic significance of this cereal, resulting from the combination of valuable features such as high yield potential and good grain quality with high tolerance to unfavourable environmental conditions has focused much interest on the identification of genetic factors controlling its embryogenic potential and effective production of DH lines (Gonzalez et al 2005; Krzewska et al 2012; Z_ ur et al 2012, 2013)

Methods

Results

Conclusion