The Effect of Zinc, Copper, and Silver Ions on Oat (Avena sativa L.) Androgenesis

Oat (Avena sativa L.) cultivars ‘Bingo’ and ‘Chwat’ were used to compare the embryogenesis competence of another culture. Despite the embryo-like structures obtained from both tested cultivars, only ‘Chwat’ produced green plantlets, which confirmed the cultivar dependency. ‘Chwat’ produced the highest number of embryo-like structures and green plantlets (0.7/100 anthers and 0.1/100 anthers, respectively). The embryo-like structure formation also depended on cold pretreatment combined with Cu2+, Zn2+, or Ag+ ion supplementation, which was applied during the tiller pretreatment or added to the induction media. The highest number of embryo-like structures (2.1/100 anthers) were observed on anthers derived from the tillers kept in a 50% Hoagland medium with the addition of 10 µM of CuSO4. In turn, the induction media supplemented with the ions Cu2+, Zn2+, or Ag+ increased neither the number of embryo-like structures nor the green plantlet production compared to the control conditions. However, such ion applications turned out to be most effective when the induction medium was enriched with 25 µM of AgNO3 and left to obtain the highest number of embryo-like structures and green plantlets (0.8/100 anthers and 0.2/100 anthers, respectively). Therefore, more attention should be paid to the possibilities of adjusting the media nutrient composition, as this may be the only way to significantly increase the efficiency of this method.

Shoots formation from gynosinesis Cucumis sativus l.

Haploid plants achieve through androgenesis or gynogenesis. In gynogenesis method, the ovary or ovule are used as explants induct haploid plants. Female flower one day before flowering of Cucumis sativus L. are collected. Cold pretreatment of ovaries at 4°C up to 24 hours and culture under dark conditions. Significantly enhanced callus induction response is compared with cultures under 4-week cultured on CBM medium supplemented with various concentration of TDZ 0.01-0.04 mg/L. After 4 weeks, ovaries are transferred to medium with kinetin 0.05 – 0.20 mg/L. Then, ovaries were transferred to medium supplemented with BA: IAA 3:1. Finally, green ovaries were transferred to BA 1.5 mg/L and GA3 1.5 mg/L. The results showed that ovary induction has best affected on CBM with TDZ 0.03 mg/L with 11 callus/sample. Ovaries developed on kinetin 0.1 mg/L with 7.4 callus/sample. Ovaries become green and had leaves and roots formation on BA: IAA (3 mg/L: 1 mg/L). 11 plantlets were harvested from ovary culture after 12-week culture on CBM supplemented with BA 1.5 mg/L and GA3 1.5 mg/L.

Pengaruh Praperlakuan Dingin Antera terhadap Pembentukan Kalus Cabai Merah Keriting (Capsicum annuum L.) Pretreatment on the Callus Formation of Curly Red Chilli Pepper (Capsicum annuum L.)

The production of the double haploid plant in vitro through anther culture technique is a plant breeding technique used to obtain pure strain rapidly. A variety of pretreatment has been reported to induce callus and regenerate planlets efficiently. This study aims at describing the influence of cold anther pretreatment towards the callus formation of curly red chili pepper (Capsicum annuum L.). This research was conducted in the laboratory of Genetics and Breeding, Faculty of Agriculture and Animal Science, Universtas Islam Negeri Sultan Syarif Kasim Riau. The explants used are anther of local genotype of curly red chili pepper. The anthers are stored at low temperatures (4 °c) with different time intervals of 0, 24, 48 and 72 hours. The results showed that the percentage of highest callus formation was obtained at 24 and 72 hours length storage, ie 50%. Cold pretreatment of 72 hours anther storage results in a faster callus with a percentage of the highest yellowish white callus color of 17.65% and a compact structure. The cold pretreatment with 72 hours anther storage is the most optimal acceleration in the development stage of anther culture and induces te formation of curly red chili pepper (Capsicum annuum L.) local genotypes.

Influence of Initial Media and Duration of Cold Pretreatment on the Efficiency of Androgenesis in the Culture of Isolated Anthers of Winter Wheat (Triticum aestivum L.) Variety Irkutskaya

Haploid technologies are based on the regeneration of gamete cells to form whole plants. The main advantage of haploid technologies is the ability to quickly in one generation obtain homozygous plants with the required parameters. Nevertheless, the use of these technologies today is limited, what is associated with insufficient knowledge of the physiological and biochemical features of the application of this method. In this regard, this work is devoted to the study of the features of androgenesis in vitro in the culture of isolated anthers of winter wheat cultivar Irkutskaya and the determination of the methodological features of obtaining intact plants. In the autumn-winter period, donor plants were grown at the artificial climate station SIPPB SB RAS. In the spring-summer period, donor plants were grown in the field at the experimental plots of the Zalarinsky agroecological station (Tungui village). The plant material was selected at the booting stage. Anthers were isolated from 6-10 spikelets of the middle third of the spike. The features of androgenesis were assessed by the frequency of formation of calli and embryo-like structures to the total number of anthers and the frequency of formation of the total number of seedlings and the number of green seedlings to the number of embryo-like structures and calli. It was found that the optimal medium for the induction of androgenesis in the anther culture of this common wheat cultivar is the modification of medium N6 containing 9% sucrose, with the addition of glycine (2 mg/L), 2.4-D (1 mg/L) and NAA (1 mg/L). The maximum yield of embryoid-like structures was observed in the case of low-temperature pretreatment of donor plants at + 4 °C for 6-12 days. At the same time, the most intensive calli formation in the culture of isolated anthers occurred after 3-6 days of donor plant cold pretreatment. Regeneration of green plants took place on 190-2Cu medium with the addition of growth regulators (2.4-D, 0.5 mg/L; kinetin, 0.5 mg/L) with obligatory cold pretreatment of androclinic structures at +4 °C for 3-7 days. Obtained results indicate that low-temperature exposure at different stages, from pretreatment of donor ears to exposure to androclinic structures, is a key factor for winter wheat that promotes androgenesis in the culture of isolated anthers and the successful formation of intact plants.

Efficiency of spring barley haploid production in anther culture in vitro: comparison of basic and improved technologies

Aim. Evaluation of innovative methodological approaches elaborated on model genotypes for usability to increase frequencies of morphogenic structure induction and plant regeneration in anther culture in vitro in spring barley diverse material. Methods. Spikes isolated from F1 and F2 hybrids of four crosses were pretreated using an improved method (4ºC, 28 days), and anthers were inoculated onto nutrient media containing chemically modified starches D5-M and D5a-1 instead of agar. In control cut tillers were emerged in water and pretreated at 4ºC for 5 days. Anthers were cultivated on agar solidified medium. Results. Positive effects of the improved method of cold pretreatment and cultivation of anthers on media solidified with starches were confirmed. The advantage of new gelling agent D5a-1 was proved. Particularly, its usage resulted in a three-fold increase in the frequency of green plant regeneration. Conclusions. In order to increase spring barley androgenic haploid yield, combination of prolonged cold pretreatment with anther cultivation on media solidified with chemically modified starches instead of agar in an integrated technological process is reasonable. Keywords: Hordeum vulgare L., anther culture in vitro, cold pretreatment, starch, agar, embryo formation, plant regeneration.