RU1822683C - Method of induction of neoplasms and regenerant in anther culture in vitro in winter hexaploide triticale and wheat - Google Patents

Abstract

Usage: biotechnology, cultivation of tissue and plant cells, androgenesis. SUMMARY OF THE INVENTION: Neoplasms and regenerants of hexaploid triticale and wheat are obtained by growing donor plants, explanting anthers on a nutrient medium and further culturing, while seeds of donor plants are irradiated with radiation at a dose of 5-10 kP. 6 tab.

Description

Ј

The invention relates to biotechnology, in particular to genetics and selection, and can be used to accelerate the production of dihaploid lines, as well as to create starting material for plant breeding.

The purpose of the invention is to increase the induction of neoplasms and green regenerants.

This goal is achieved by the fact that in the method of inducing neoplasms and regenerates in the anther culture of winter hexaploid triticale and wheat, including growing donor plants and planting anthers on a nutrient prede, the seeds of donor plants are irradiated with / rays at a dose of 5-10 kP.

Compared with the prototype, the technical solution includes irradiating seeds of donor plants with γ rays at a dose of 5-10 kP,

which meets the criteria of the invention of novelty.

In the proposed method, the treatment of seeds of grain crops with γ rays is carried out when growing donor plants for anther culture in vitro in order to increase the yield of neoplasms and green plants.

Example. The seeds of the L160 homozygous line and the hybrid combination (Fj) GK340 of winter hexaploid triticale and winter wheat L1, 100 seeds of each sample were irradiated before sowing with γ-rays in doses of 5 kR-10 kR on the MD-y-1 installation M (cesium 137) with a capacity of 3000 R / min. As a control, unirradiated seeds were sown.

An ear with anthers at a one-nuclear stage was cut off and kept for 5 days at a temperature of + 4 ° С. Then mother00

Yu

hO O 00 00

Al was sterilized with 3% chlorocin and the anthers were planted not in Potato II medium; for each variant, at least 300 anthers were planted (Table 1).

After planting, the anthers were cultured in the dark with a temperature of 3 ° C until the appearance of neoplasms. The neoplasms were transferred onto a regeneration medium, which was used as P8 medium (modification of the Hamburg B-5 medium) (6) without hormonal additives:

Trace element / g

KMOZ2500

CaCl2 -2H2O150

MgS04 -7H20250

(NH4) 2S04134

NaH2P04 HzO150

Kl 0,0,75

NWO3.3.10

MnS04 H2010.00

ZnS04 -7H202.00

Na2Ma04 -2H2O0.25

CuS04 -5H200.025

CoCI2 -6H200.025

Na2 EDTA -2H2037.23

FeS04 -7H2027.80 Sucrose ZOOOO mg / l

Inoeitol100.0

NAA1 mg / L

Kinetin1 mg / L

Vitamin E0.1 mg / L

Agar 0.6

pH5.7 Stab Vitamins

Folic Acid 5 mg

Riboflavin 5 mg

Biotin 10 mg

10mg 10mg 10mg 20mg 0.015mg

0

5

0

5

0

5

Dissolve in 100 ml; to prepare 1 l of medium, take 5 ml of solution.

The results of analysis of variance presented confirm the positive effect of γ-radiation on the regeneration of green plants (Table 6).

Since there was a significant variation in the yield of plants depending on the genotype and radiation dose, it is recommended that the full range of given y-radiation doses be used to determine the most optimal for the genotype being tested.

Thus, the proposed method allows to increase the yield of neoplasms at a dose of 5 kP and the proportion of green rastenii among regenerants at doses of 5-10 kP depending on the genotype, which increases the efficiency of the method of producing haploid plants through anther culture.

Claims (1)

SUMMARY OF THE INVENTION A method for inducing neoplasms and regenerants in an anther culture in vitro in winter hexaploid triticale and wheat, including growing donor plants, explanting anthers on a nutrient medium and cultivating, wherein, in order to increase the frequency of induction of neoplasms and yield of green plants, donor seeds plants are irradiated with y-rays in a dose of 5-10 kP. Table 1 518226836 Table 2 The output of neoplasms at various doses of radiation to the seeds of donor plants,% The results of the analysis of variance on the yield of neoplasms during the cultivation of anthers Table 3 The share of green plants among the total number of regenerates depending on the radiation dose,% The yield of green regenerants depending on the dose of seed irradiation in the homozygous line of winter hexaploid triticale D160 Table in The results of the analysis of variance on the yield of green plants Table L Table 5