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Культура зародышевых мешков и зерновок кукурузы (англ. версия)

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Культура зародышевых мешков и зерновок кукурузы как биотехнологическая система получения зиготических зародышей для ускорения селекционного процесса

Culture of isolated embryo sacs and caryopses as a biotechnological system for the upgrowing of maize zygotic embryos in vitro for genetic transformation procedures

Technique of in vitro development of isolated plant female generative structures from early stages to maturity is little processing, but perspective experimental system for modeling the development and maturation of embryo sac, embryo, endosperm, seeds, for recovering key stages and limiting factors in formation of reproductive organs, which for many agricultural crops generate yield and seed productivity. The development of different isolated generative structures grown in vitro up to maturity is an important step in genetic transformation procedures with egg cells, sperms, zygotes, young embryos, in manipulations with single cells, in transferring organelles to gametes and zygotes.

The investigation established the possibility of updevelopment of maize embryo (53 %) in culture of isolated embryo sacs in in vitro conditions at the 7th day after pollinationwith genotype DК2/477-322хА22from the transition stage up to the beginning of organogenesis. Embryo sacs were cultured on artificial nutrient medium NBM (Mól, R et al., Planta 189: 213-217, 1993) with 90 g/l sucrose and 2 mg/l 6-benzylaminopurine.It was determined that the cultivation of embryo sacs with the aim to upgrow an embryo and to accumulate starch in endosperm should be continue more than 20 days and nights. The colour of cultured embryo sac can be considered a marker of embryo upgrowing and starch accumulation in endosperm.

Experimental data confirm the possibility of embryo developments (12 %) in culture of isolated maize caryopses at the 5th – 7th day after pollination and represent the production of viable embryos (65 %) germinating in regularly developed green plantlets. Caryopseswere cultured on artificial nutrient medium NBM (Mól, R et al., Planta 189: 213-217, 1993) with 90 g/l sucrose and 1 mg/l 6-benzylaminopurine. Isolated caryopses were taken from field donor plants of maize inbred DK366 and hybrid A22xDK307, DK2/477-322хА22. Genotype significantly affected both growth and development of caryopses in culture, especially starch accumulation in endosperm and covers and a frequency of developed embryos. The internal state of cultured caryopsescan be considered a marker of embryo upgrowing and starch accumulation in endosperm independently of genotype. Embryo development and caryopses growth were finished independently of genotype by the 20th day in culture. Soon after this term embryos should be isolated out of caryopses for germination and obtaining plantlets.

Источник: Liapustina О.V. Culture of isolated embryo sacs and caryopses as a biotechnological system for the upgrowing of maize zygotic embryos in vitro for genetic transformation procedures // 52nd Annual Maize Genetics Conference: Program and Abstracts (Congress Centre Riva del Garda (Trento), Italy, 18 – 21 march 2010) – Italy, 2010. – Р. 72.

Автор фото в статье: Ляпустина Е.В.

Перевод статьи на русский язык читайте Культура зародышевых мешков и зерновок кукурузы

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