Morphogenetic Responses of Wheat Immature Embryo Culture Depending on Growing Conditions of Donor Plants

Abstract

The article considers the influence of growth conditions of donor plants on in vitro morphogenetic responses of immature embryos from spring bread wheat. The materials were 12 varieties of different ecological and geographical origin from the collection of Altai Research Institute of Agriculture (Barnaul, Russia). Donor plants were grown in the greenhouse and experimental field near Barnaul. Immature embryos with 1.3-1.5 mm in size isolated from seeds collected 14-16 days post anthesis were used as explants. The embryos were placed with the sculletum upwards on a sterile solid agar (0,8%) medium containing the inorganic components of Linsmaier & Skoog (LS), 3% sucrose, 2.0 mg l -1 2,4-dichlorophenoxyacetic acid (2,4-D) and incubated in the darkness at 26±1°C. Induced calli were subcultured after 25-30 days interval in fresh medium supplemented with 0.5 mg l -1 2,4-D and 0.5 mg l -1 kinetin and grown at 25±1 °C under a 16/8-h (day/night) photoperiod with a white fluorescent light. Embryogenic calli were transferred to LS medium containing 0.2 mg l -1 indole-3-acetic acid (IAA) for regeneration. A complete randomized block design with four replications per a genotype (60 embryos) was used. We stated by analysis of the components of phenotypic variance that genetic factor has been the leading one in all stages of tissue culture, but the highest effect (80%) was on plant regeneration. Interaction of the factors considered was competitive with genotypic influence on the stages of morphogenesis (28%) and callusogenesis (27%). Influence of "conditions" was statistically significant, but inferior to other factors. The greatest result was obtained for induction of morphogenic calli. Field conditions were the most favorable for genotypes with high callus and regeneration abilities. Culture responses of those with a low regeneration potential increased after growing donor plants in greenhouse vegetation in 3.4 -13.9 times. For most genotypes moderate temperature and precipitation as well as natural lighting before collecting material increased morphogenic competence of explants in wheat tissue culture. Users are permitted to copy, use, distribute, transmit, and display the work publicly and to make and distribute derivative works, in any digital medium for any responsible purpose, subject to proper attribution of authorship. содержащей 0,8% агара, 3% сахарозы, 2,0 мг/л дихлорфеноксиуксусной кислоты (2,4-Д), в темноте при температуре 26±1 °C. Через 25-30 дней каллусы переносили на дифференцирующую среду LS, дополненную 0,5 мг/л 2,4-Д и 0,5 мг/л кинетина и выращивали на свету с 16-часовом фотопериодом (день) и температуре 25±1°C. Морфогенные каллусы переносили на среду для регенерации, содержащую 0,2 мг/л индолил-3-уксусной кислоты (ИУК). Эксперимент выполнен в четырех повторениях по 60 эксплантов на генотип. Установлено, что генетический фактор является ведущим на всех стадиях культуры ткани с максимальным эффектом на этапе регенерации (80%). Доля фактора «взаимодействие генотип-условия» была примерно одинакова с генотипич...