SU1701196A1 - Method for obtaining regenerated barley plants resistant to helminthosporiosis - Google Patents

Abstract

The invention relates to agricultural biotechnology and can be used in breeding practice in creating varieties of cereals resistant to gelmintosporioi. The aim of the invention is to increase the resistance of regenerated plants to the pathogen. The method consists in the fact that regenerants are obtained from callus grown on a nutrient medium Murashige-Skoog in the presence of the selective H.Sativum fungal toxin agent. 3 tab.

Description

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The invention relates to agricultural biotechnology and can be used in breeding practice for creating pathogen-resistant fungi that cause pathogens of varieties of cereals, in particular, barley.

The aim of the invention is to increase the resistance of regenerated plants to the pathogen.

The method involves the cultivation of callus tissue on Murashige and Skoog medium containing a selective agent, followed by the preparation of regenerants, with the Helmintosporlum satlvum toxin fungus being used as a selective agent.

The method is carried out as follows.

As explants that are planted on Murashige and Skoog callusogenic medium in test tubes with different concentrations of sterilized with the help of a bacterial filter of the toxins of the fungus N. satlvum, unripe embryos are used in barley varieties Good and Dean. Before planting, the explants were sterilized in a 5% solution of chloramine for 15 minutes. The explants are introduced in medium No. 1 with the H.sativum fungus toxin of the following composition: Mother solution No. 1, g: NH4N038.25

CMO9.5

MgS04-7H201.85

MnS04 -5H200,12

Zn4S04-4H200.043

CuSO4 -5H200,000125

Bring to 500 ml with distilled water, from where take 100 ml. The mother liquor Nfe 2, g: KN2OR040,85

NZVOZ0.031

4 o

YU

ABOUT

Kj. 0,00415

Na2Mo04 2H20 0.00125 (dissolved separately in 10 ml of warmed NaO and take 1 ml / l)

  bNaO 0.000125 (dissolved separately in 100 ml, from which 1 ml / l is taken).

Bring to 500 ml with distilled water and take 100 ml. Vitamins, g:

Glycine0.07

Vitamin Bi0.005

Vitamin Be0.005

Vitamin PP0.005

Dissolve in 100 ml of distilled H2U, from where 10 ml is taken.

2.4 Dichlorophenoxyacetic acid0.002 g

Iron chelate10 ml

Sucrose 25g

Ag.Ar7 g

, 332 g

All components are mixed and made up to a volume of 1 liter with distilled water.

The resulting callus is transplanted to medium No. 2 of the same composition, but without toxin. Then the cultivation of the surviving callus is carried out on an organic medium No. 3 of the following composition:

Solution number 1100 ml

Solution number 2100 ml

Chelate iron10ml

Vitamins 10ml

Yeast hydrolyzate1.0 g

CaCi20,332 g

IAA (indole acetic acid) 0.0005 g Kinetin 0.01 g Hyperellic acid 0.05 g Sucrose 15 g Agar 7 g and made up to 1 liter of distilled H20.

The explants are cultivated on Wed units 1 and 2 at illumination of 5-6 thousand lx, at T 26 ° C and a 16-hour photoperiod, and on medium No. 3 at T 15 + 17 ° C, illumination 10-15 thousand; lx, with a photoperiod of 16 h for 25-30 days on each medium.

The resulting regenerants in the phase of 2-3 leaves are planted in pots with sterile sand, then in the soil mixture: sod soil - sand - peat 1: 1: 1 m set in

growing chamber (T 2-4 ° C) for 56-60 days.

After rovization and deposition of regenerated plants, seeds from them are tested for resistance to helminthosporiosis.

EXAMPLE 1 A dose selection for administering a toxin to 1 callusogenic selective medium is determined empirically. For this

initially, the percentage of inhibition of the roots of one-day seedlings of seeds of the fungus toxin is established (Table 1). Based on a concentration that inhibits at least 30% of the growth of the central

root, select the dose for the introduction into the Cse --- selective callusogenic medium. Doses are selected emperically about 20-25 times less than that found in seedlings. The regeneration capacity of barley calluses is given in Table. 2

PRI me R 2 Regenerants obtained from surviving on the selective medium with the toxin of the fungus calluses after growing on an infectious background give a yield 1.6 times higher compared to the control.

The use of the proposed method for the production of barley-resistant plant regenerators of H.sativum fungus provides the following advantages in comparison with the known method:

the use of the toxin fungus as a selective factor is more economical, it does not contain associated impurities, the effect of which on the callus is manifested when using culture filtrate;

the introduction of certain doses of the fungus toxin into the Murashiga and Skoog medium allows the selection of regenerants from the minimum number of surviving calli.

Claims (1)

The invention of the method for producing regenerated barley-resistant plants for helminthosporioses, including the cultivation of callus tissue on the Murashige-Skoog medium in the presence of a selective agent with the subsequent production of regenerants, characterized in that, in order to increase the resistance of the regenerating plants to the pathogen, Helmintosporium sativum toxin is used as a selective agent. Table 1 The effect of phytotoxins of the fungus H.sativum on the length of the roots of one-day seedlings barley Table 2 Regenerative ability of barley samples on selective medium with toxin Table 3 Productivity indicators when grown on an infectious background of seeds obtained from the phytotoxin-resistant fungus H, sattvum regenerated plants