doi: 10.15389/agrobiology.2019.3.557eng

UDC: 633.18:581.143.6



M.V. Ilyushko, M.V. Romashova

Primorskii Research Institute of Agriculture, 30, ul. Volozhenina, pos. Timityazevskii, Primorskii Krai, 692539 Russia, e-mai (✉ corresponding author),

Ilyushko M.V.
Romashova M.V.

Received September 25, 2018


The conditions for growing donor plants in androgegesis in vitro are considered from the point of view of the influence of physical factors on the plants (illumination, length of daylight, temperature, nutrition of plants). A priori it is believed that any period of the year (season) is suitable for plant tissue culture in vitro, and this is the main advantage of in vitro technologies compared to traditional ones. Seasonality is taken into account only when comparing donor plants grown in the field and under controlled conditions. However, we have not found reports on the study of using procedure of anther in vitro culture throughout the year. This paper is the first to show that, under uniform conditions of donor plants growing, the frequency of callus formation and regeneration of rice (Oryza sativa L.) plants from anthers in vitro culture differs depending on the month (season) when explants were collected. The aim of the studies was to study the seasonal dependence of in vitro androgenetic responses of O. sativa when growing donor plants in a climatic chamber conditions. The 5-9 plants of O. sativa subsp. japonica Kato, the Cascade variety, were planted monthly during a year and grown in a climatic chamber (at 24 °С and 21 °С, 15000 lux, 60 % humidity and photoperiod of 14 light/10 dark hours) to be the test anther donors. The anthers from naturally grown plants served as controls. As a result, the seasonal dependence of callus formation in the rice anther culture in vitro was revealed. The peak intensity of callus formation occurs in May and June (15.5-28.3 %). When growing donor plants under artificial conditions at both temperatures, in the best period for anther culture in vitro (May-June) callus formation was higher than in the control. The use of the climatic chamber makes it possible to obtain consistently high values of the intensity of callus formation in some months, which is not always possible in the natural conditions. The temperature of donor plant growing affects the frequency of rice callus formation and the regenerative capacity. The temperature which is lower than optimal for rice plants (21 °С) allows for different frequency of callus formation throughout the year, whereas the rice-comfortable temperature (24 °С) leads to a large seasonal dependence and callus formation from May to September and in December and January. At 21 °С vs. 24 °С, four times more calluses with green regenerants are formed (31.6 % vs. 8.8 %), with an increase in the fraction of doubled haploids up to 28.1 % and their number per callus up to 16.6. A moderate correlation was found between the share of calli with green rege-nerants and the average number of doubled haploids per callus (r = 0.59 at p = 0.05). This means that with an increase in the number of calli with green regenerants, which is observed at 21 °С, the total yield of doubled haploids also increases. As to the number of haploids, no such dependence was found. Thus, greenhouses and climatic chambers may serve not only for growing donor plants by researchers, but also for practical use of the most favorable periods for anthers culture technique.

Keywords: Oryza sativa L., androgenesis in vitro, callus formation, regeneration, doubled haploids, seasonality.



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