RU2136144C1 - Barley seed mutagenic treatment method - Google Patents

Abstract

FIELD: genetics and selection, in particular, induction of mutations for creating barley selection basic material. SUBSTANCE: method involves exposing barley seeds to blue light with wave length of 455 plus and minus 10 nanometers and power density of 0.1 mW/sq cm for 60-120 min. Blue light is obtained from electric filament lamp and passed through interference light filter. Method allows yield of selection morphological and physiological mutations of second generation barley plants to be increased upon treatment by electromagnetic radiation. EFFECT: increased efficiency in creating basic material for barley selection. 1 tbl

Description

 The present invention belongs to the field of genetics and selection and can be used to induce mutations in the creation of source material for barley selection.

 Obtaining hereditary changes in agricultural plants is possible using x-ray [1] and gamma rays [2, 3], laser radiation [3,4], chemical mutagens (for example, nitrosoethylurea) [3], phytohormones [3,4,5] .

 Closest to the proposed invention is a method for mutagenic processing of barley using gamma radiation with single doses of 100 and 150 Gy [6].

 However, gamma radiation under the existing treatment regime suppresses field germination of seeds, resulting in significantly reduced survival of plants of the first generation (see table. 1).

 The inhibitory effect of gamma radiation affects the duration of the growing season, it increases by 4-7 days. Under the influence of gamma rays, productive bushiness, stem length, and seed set in a barley ear are reduced.

Among the morphological and physiological mutations in M _{2, the} predominant forms are those with a loose spike and its chaotic sterility, as well as families with a slow process of development and maturation.

The proposed method involves the processing of barley seeds with blue light obtained from an electric incandescent lamp through an interference filter. The wavelength of the light flux is 455 ± 10 nm, the power density is 0.1 mW / cm ² , the processing time is 60-120 minutes.

Blue light (SS) is a regulator of such vital processes in plants as phototropism, stomata movements, photomorphogenesis, etc. Plant reactions to SS are carried out both through the phytochrome system [7] and due to a receptor unrelated to phytochrome [8], which can be localized in the plasmalemma and in the membranes of intracellular organelles as part of the redox chain, which includes flavoprotein and cytochrome b ₅ .

 SS causes hyperpolarization of the membrane potential of plant cells, which leads to the redistribution of phytohormones. Under the influence of SS, the transport properties of cell membranes change. SS enhances the functional activity of nitrate reductase, whereby the synthesis of amino acids increases, while under the influence of SS, the absorption of nitrates by cells increases [9].

 Physiological reactions caused by blue light lead to deviations in the course of a number of processes important for the life of plants. Among these deviations may include disturbances in the functioning of enzymes of the DNA replication and repair system, whose work is closely related to the process of the occurrence of mutations [10].

 The following examples are provided to illustrate the method.

 Before the sowing, air-dried seeds of barley of the Dina variety were treated with blue light, the exposure duration was 5, 15, 30, 60 and 120 minutes. Untreated seeds served as control.

In each variant, M _{1 was} irradiated and sown 500 grains (125 pieces per plot of 1.0 m ² ).

 The treatment of barley seeds with blue light for 60 min significantly increased their field germination (see table. 2).

 Blue light did not significantly affect the dynamics of plant development in the year of exposure.

Mutagenic treatment had an ambiguous effect on barley productivity in M ₁ : with irradiation exposures of 30 and 120 min, a significant increase in grain weight from an ear was noted, and 5- and 60-minute exposures significantly reduced this trait (Table 3).

 In variants with irradiation, there was a tendency to increase productive bushiness, stem length and spike.

 In the second generation, starting from the seedling phase, chlorophyll mutations of barley were detected. They were found in all experimental variants (tab. 4).

 The largest spectrum of chlorophyll mutations was observed in the “blue light 120 minutes” variant. Mutations of the claroviridis type, differing from the original type by the light green color of plants, were found here; viridostriata with longitudinal green and light green stripes alternating on leaves; xanthotigrina, in which there was an alternation of transverse green and yellow stripes on the leaf blades; xanthoviridis having a normally colored leaf blade with a yellow top; as well as mutations characterized by a stepwise change in the color of the plant at the stage of formation of the 2nd - 3rd leaf - from green to yellow (viridoxanthoterminalis) and from green to white (viridoalboterminalis). In the “SS 30 minutes” variant, a chloina mutation was distinguished, characterized by a greenish-yellow color of plants, and in variants with irradiation durations of 15 and 60 minutes, mutations of the veridamaculata type, in which there were separate yellow and white spots with fuzzy edges on leaf blades.

 In addition to chlorophyll mutations, blue light induced other types of changes. These include deviations from the original variety in the shape of the bush (creeping and intermediate), the intensity of shoot formation, the length of the stem, the width of the leaf blades, the degree of wax formation, the time of tillering and ear formation, the length of the spike, the number of spikelets in the ear, and the weight of grain with spike, resistance to lodging and drooping of the spike and others (table. 5).

 The greatest number of types of morphological and physiological changes was found in the variant with the maximum dose of mutagen.

 Many of the distinguished families are characterized by a whole complex of mutant characters, where chlorophyll anomalies are combined, deviations in the morphology of various parts of the plant, and changes in the course of a number of life processes that affect the dynamics of the mutant, its flowering, and the formation of anthocyanins. This feature can be explained by the pleiotropic effect of the mutant gene or the simultaneous mutation of several genes.

 A number of selected barley mutants are of interest as starting material for selection (Table 6).

 Mutant 2-6 was obtained from seed treatment with blue light with an exposure of 5 minutes. Unlike the original Dina variety, this mutant more vigorously shrinks (2.2 times) and has a longer spike.

 The 3-3 mutant was created by exposing the barley seeds to blue light for 15 minutes. It has a chlorophyll anomaly viridoxanthoterminalis. Regarding the variety, Dina gives a significant decrease in the total and productive bushiness (2.7 and 3.8 times, respectively), stem length (by 27.3 cm), the number of spikelets and the weight of grain per ear.

 Mutant 4-12 is isolated in the “SS 30 minutes" variant. It has anthocyanin pigmentation of the ears and nerves of the grains, which is unusual for the original variety. It forms a long spike (8.2 cm), it has an increased number of spikelets and more grain weight per spike. Ripens three days later than control.

 Mutant 5-8 was obtained by irradiating barley seeds with blue light for 60 minutes. It carries a chlorophyll mutation viridoalboterminalis. Significantly exceeds control in total bushiness (1.6 times) and in spike length.

 Mutant 5-11 was selected in the SS 60 minutes variant. It is characterized by a significant reduction in the length of the stem relative to the original variety (by 11.5 cm), a long spike (7.6 cm), a large number of spikelets and the weight of grain per spike (1.16 g). The growing season lasts 4 days longer than the variety Dina.

 Mutant 6-29 was created by treating barley seeds with blue light for 120 minutes. Productive bushiness of the mutant is greater than that of the standard. The length of the spike and the mass of grain from it is much higher, ripening occurs 2 days later.

 All described mutants belong to the nutans variety.

 The relationship between the mutagenic efficacy of blue light and its dose can be established by analyzing the frequencies of morphological and physiological mutations in the second-generation experimental options (Table 7).

The treatment of barley seeds with blue light for 5 min induced 3.52% of mutant families, which significantly exceeded the background level of spontaneous mutagenesis recorded in the control. An increase in the exposure of mutagen to 15 and 30 min did not lead to a significant increase in the frequency of mutations, however, a significant increase in the frequency of hereditary changes was found when switching from a 30- to 60-minute exposure time. After exposure to barley seeds with blue light for 120 minutes, the mutation frequency was 7.67%, which is at the level of the “CC 60 minutes” variant (t _d = 0.86).

 Thus, the physical factor blue light proposed for seed treatment, in comparison with the prototype, is a new mutagenic factor for seeds and barley plants, which in the second generation provides a greater absolute yield of mutations and a wider range of selection-valuable hereditary changes.

List of references
1. Chaundhuri KL High yielding X-ray mutation of jute (Carchorus olitorius Linn) // Annu. rep. simitted to the Indian Cent. Jute Cummittee. 1948.P. 18 ... 21.

 2. Gustafsson A. Productive mutations indused in barley by ionizing radiations and chemical mutagens // Hereditas. 1963. Vol.50. P.211 ... 263.

 3. Kozachenko M. R., Manzyuk V. T. Obtaining mutants of spring barley by combining red laser radiation with chemical mutagens or penetrating radiation // Application of low-energy physical factors in biology and agriculture / Abstract. All-Union. scientific conf. Kirov, 1989. P.77 ... 78.

 4. Dudin G. P. Variability of barley under the influence of laser radiation and benzyladenine // Agricultural Radiobiology / Chisinau. S.-kh. institute Chisinau, 1990.P.23 ... 28.

 5. Vilensky E.R. Phytohormones and genetic homeostasis // 2 Congress of the All-Union. Society of Plant Physiologists, Minsk, September 24 ... 29. 1990 / Thes. doc. Part 2. M., 1992. S. 40.

 6. Purtova I.V. Creation of source material of spring barley using physical mutagenic factors, para-aminobenzoic and abscisic acids / Abstract. diss. for a job. student step. Cand. S.-kh. sciences. St. Petersburg, 1993.20 s. -prototype.

 7. Casal J.J., Smith H. Effects of blue light pretreatments on internode extension growth in mustard seedlings after the transition to darkness: analysis of the interaction with phytochrome // J. Exp. Bot. 1989. Vol. 40. N217. P.893 ... 899.

 8. Volkenburgh E. V., Cleland R.E., Watanabe M. Light-stimulated cell expansion in bean (Phaseolus vulgaris L.) leayes. II. Quantity and quality of light required // Planta. 1990. Vol.l82. Nl. P.77 ... 80.

 9. Field V.V., Salamatova T.S. Physiology of plant growth and development. L .: Publishing house Leningra. University, 1991.240s.

 10. Pitirimova M.A. Post-mutagenic recovery and mutational variability of barley // Application of low-energy physical factors in biology and agriculture / Abstract. All-Union. scientific conf. Kirov, 1989.S. 32 ... 33.

Claims (1)

A method for mutagenic treatment of barley seeds, including exposure to seeds with electromagnetic radiation, characterized in that the seeds are exposed to blue light with a wavelength of 455 ± 10 nm and a power density of 0.1 mW / cm ² for 60 to 120 minutes

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