Changes in distribution of zeatin and indole-3-acetic acid in cells during callus induction and organogenesis in vitro in immature embryo culture of wheat

Abstract

Plant organogenesis remains one of the most essential questions of plant developmental biology. Callus tissue in vitro is a valuable tool for the studies on hormonal aspects of plant organogenesis, especially its early events, and immunohistochemical analysis is one of the few approaches offering information on the localization and role of hormones during organ development. The localization of endogenous zeatin and indole-3-acetic acid was investigated during simultaneous bud and root formation in calluses derived from immature embryos of wheat (Triticum aestivum L.). Calluses were induced on Murashige and Skoog (MS) medium supplemented with 2.0 mg L −1 2,4-dichlorophenoxyacetic acid. To stimulate simultaneous bud and root formation, calluses were transferred onto MS medium supplemented with 0.2 mg L −1 kinetin and 0.2 mg L −1 indoleacetic acid. Strong immunostaining for both hormones was detected in proliferating callus tissue, in developing meristematic centers and meristematic zones (whose cells were shown to be involved in organ formation), and at the sites of shoot and root apex initiation. During further development, shoot apexes with leaf primordia were heavily immunostained for zeatin, while immunostaining for indole-3-acetic acid was more intense at the sites of leaf primordia initiation and incipient primordia themselves. In the developing roots, immunostaining for both hormones reached a maximum in the root apex and gradually declined with increasing distance from the apex. Cells of developing procambial strands were also strongly stained for both zeatin and indole-3-acetic acid. These data suggest considerable similarity between patterns of hormone distribution in organs in vitro and in vivo. Thus, callus culture is a convenient and useful model for the study of fundamental biological questions such as how hormones regulate development.

Gemmorhizogenesis as the Type of Organogenesis in Calli in Vitro During Biotechnoloical Investigations in Cereals

“…Иммуногистохимическими методами выявлено, что эндогенные гормоныцитокинины и ауксины -локализуются преимущественно в клетках апексов формирующихся в зародышевых каллусах пшеницы почек и корней (Seldimirova et al, 2016а). По мнению Н.Н.…”

THE HISTOLOGICAL STATUS OF THE WHEAT EMBRYO IN PLANTA AT THE DEVELOPMENTAL STAGE OPTIMAL FOR OBTAINING CALLUS IN ORDER TO STUDY IN MODEL STRESS CONDITIONS IN VITRO

“…Отдельная проблема в этой области -выявление особенностей экспланта, в условиях in vitro дающего начало морфогенному каллусу, тотипотентные клетки которого в ходе дальнейшего культивирования в модельных стрессовых условиях способны развиваться по различным путям морфогенеза, включая регенерацию стрессустойчивого растения. В качестве экспланта для получения in vitro морфогенного каллуса у злаков перспективно использование незрелого зародыша [Круглова, Катасонова, 2009; Митич и др., 2009; Plant Embryo Culture ..., 2011; Сельдимирова и др., 2011, 2017; Сельдимирова, Круглова, 2013; Thi-Bich, Duc-Thanh, 2014; Seldimirova et al, 2016]. При описании инокулируемого незрелого зародыша исследователи обычно указывают либо длину зародыша, либо сутки после опыления, на которые незрелый зародыш извлекался из зерновки (обзор [Круглова и др., 2018]); при этом, как правило, не указывается стадия развития зародыша, отсутствует его гистологическое описание.…”

Dynamics of the contents and distribution of ABA, auxins and aquaporins in developing caryopses of an ABA-deficient barley mutant and its parental cultivar

“…This goal can be achieved by taking immunohistochemical approaches using specific antibodies to hormones (Forestan et al ., 2010). Previously, we have detected auxins and cytokinins with the help of immunolocalization during the induction of morphogenesis in wheat callus (Seldimirova et al ., 2016). Immunohistochemical localization methods allow comparisons of the distribution of hormones and presumably hormone-regulated proteins between cells.…”