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

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) MEinducing 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.

Glutathione provides antioxidative defence and promotes microspore-derived embryo development in isolated microspore cultures of triticale (× Triticosecale Wittm.)

“…In parallel, in several plant species gene-encoding enzymes involved in antioxidative defence [glutathione- S -transferase (GST), oxalate oxidase] were up-regulated in response to the treatment initiating ME (Vrinten et al 1999; Jacquard et al 2009; Sánchez-Díaz et al 2013; Żur et al 2014b). Increased accumulation of proteins typical for cell defence against oxidative stress (e.g., GST, l -ascorbate peroxidase) during ME induction was also reported (Cordewener et al 2009; Uváčková et al 2012; Krzewska et al 2017). …”

“…The studied DH lines of winter triticale were selected from the mapping population ‘Saka 3006’ × ‘Modus’ derived using the method of distant crosses with maize (Wędzony 2003 ; Tyrka et al 2011 ). The population composed of 90 DH was carefully examined in respect of its embryogenic potential by anther culture method and used as the model in our earlier studies focused on physiological, metabolomic and molecular aspects of ME (Żur et al 2012 , 2014a , b , 2015 ; Krzewska et al 2012 , 2015 , 2017 ). Seven DH lines selected for the first experiment were significantly different in respect of their embryogenic potential (Żur et al 2014a , b , 2015 ).…”

Impact of Ascorbate—Glutathione Cycle Components on the Effectiveness of Embryogenesis Induction in Isolated Microspore Cultures of Barley and Triticale

“…Such a cross-tolerance phenomenon is well known in plants, and in many cases, it is associated with increased ROS generation, which may also serve as signalling molecules activating cellular defence responses [ 43 ]. Several studies have confirmed that ME initiation is associated with the upregulation of genes encoding enzymes involved in antioxidative defence, as well as increased activity and/or accumulation of LMW antioxidants (reviewed in [ 20 , 44 , 45 ] and references therein). Therefore, different types of antioxidants were added to media used for in vitro cultures in order to achieve positive effects on ELS formation and plant regeneration ([ 5 ] and references therein).…”

“…‘Igri’ and spring cv. ‘Golden Promise’, highly differentiated in terms of ME responsiveness [ 16 , 17 , 18 , 19 , 20 ], were used in the study. The seeds of DH lines of triticale were obtained from the State Breeding Institute at the University of Hohenheim (Germany), whereas the seeds of barley cultivars were obtained from Leibniz Institute of Plant Genetics and Crop Plant Research (Gatersleben, Germany).…”

Microspore embryogenesis in vitro: the role of stresses

“…Увеличилось количество белков, типичных для защиты от окислительного стресса, шаперонов и др. Функциональная классификация обнаруженных белков показала, что большую их часть составляют белки мета болизма (47 %), стрессовые (28 %) и запасные (9 %) белки (Krzewska et al, 2017).…”