GENETIC ENGINEERING AS A TOOL FOR MODIFICATION OF SEED STORAGE PROTEINS AND IMPROVEMENT OF NUTRITIONAL VALUE OF CEREAL GRAIN (review)

Elkonin, L. A.; Domanina, I. V.; Ital’yanskaya, Yu.V.

doi:10.15389/agrobiology.2016.1.17eng

Cited by 5 publications

(8 citation statements)

References 58 publications

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“…С развитием методов генетической инженерии возможности селекционеров и генетиков по созданию новых сортов и гибридов значительно расширились. Одним из перспективных направлений использования генетической инженерии в селекции является улучшение питательной ценности возделываемых куль-тур, а именно изменение содержания белка, крахмала, незаменимых аминокислот, по-давление синтеза аллергенов и различных веществ, обладающих антипитательными свойствами [1,2].…”

Section: Abstract: Kafirins Transgenic Plants In Vitro Protein Digeunclassified

“…1, треки 5-8). Следует отметить, что мучнистый тип эндосперма характерен для трансгенных растений ку-курузы и сорго с сайленсингом гамма-про-ламинов [2,7,8].…”

Section: результаты исследования и их обсуждениеunclassified

“…2), что свидетельствует о проявлении данного признака не только в искусствен-ных тепличных условиях, но и в естествен-ных условиях внешней среды. (1,3,5,7,9,11) и после обработки пепсином (2,4,6,8,10,12 1 s x -ошибка средней; 2 исходная нетрансгенная линия Желтозерное 10; **значимо отличается от исходной линии при p < 0,01, в соответствии с t-тестом Стьюдента.…”

Section: результаты исследования и их обсуждениеunclassified

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In Vitro Digestibility of Storage Endosperm Proteins of Transgenic Sorghum Plants Carrying Genetic Construct for Silencing of the Gamma-Kafirin Gene

Elkonin¹,

Ital’yanskaya²

2017

УСЕ (ACNR)

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Одним из перспективных направлений использования генетической инженерии в селекции является улучшение питательной ценности возделываемых культур. Это направление имеет особую актуальность для сорго -высокоурожайной засухоустойчивой злаковой культуры, отличающейся более низкой, по сравне-нию с другими злаками, питательной ценностью зерна, в основе которой лежит устойчивость его запасных белков (кафиринов) к протеолитическому расщеплению. В данной работе исследовали перевариваемость кафиринов у потомства трансгенных растений сорго (поколения Т 3 -Т 4 ), полученных посредством агробак-териальной трансформации со штаммом Agrobacterium tumefaciens GV3101, несущим генетическую кон-струкцию для РНК-сайленсинга гена гамма-кафирина. Муку, полученную из цельносмолотых зерновок, обрабатывали пепсином в условиях in vitro, образцы подвергали SDS-электрофорезу в ПААГ, для оценки количества непереваренного белка электрофореграммы сканировали и подвергали компьютерному анализу. Выявлено, что у растений из трех изученных семей перевариваемость белков эндосперма достигает 85-90 % (против 52-53 % у исходной нетрансгенной линии Желтозерное 10). Установлено, что данный признак проявляется у растений, выращиваемых как в искусственных тепличных условиях, так и в естественных условиях внешней среды. Выявлены растения, у которых высокая перевариваемость кафиринов сочетает-ся с формированием модифицированного эндосперма, в котором стекловидный слой представлен в виде тонкого «серпика», а также нормально развитого стекловидного эндосперма. Данный факт свидетельствует о возможности получения агрономически ценных линий сорго с высокой перевариваемостью кафиринов и твердым эндоспермом.Ключевые слова: кафирины, трансгенные растения, перевариваемость белка in vitro, сорго IN VITRO DIGESTIBILITY OF STORAGE ENDOSPERM PROTEINS OF TRANSGENIC SORGHUM PLANTS CARRYING GENETIC CONSTRUCT FOR SILENCING OF THE GAMMA-KAFIRIN GENE Elkonin L.A., Italyanskaya Yu.V. Agricultural Research Institute of the South-East Region, Saratov, e-mail: lelkonin@gmail.comImprovement of the nutritional value of cultivated crops is one of the promising areas for application of plant genetic engineering in plant breeding. This direction is of particular relevance for sorghum -a high-yielding drought-resistant cereal crop, which is characterized by a lower nutritional value of the grain, in comparison with other cereals. The basis of such a feature of sorghum grain is stability sorghum storage protein (kafirins) to proteolytic digestion. In this paper, we studied the digestibility of kafirins in offspring of transgenic sorghum plants (T 3 -T 4 generations) obtained by transformation with Agrobacterium strain carrying a genetic construct for the RNAsilencing of the gamma-kafirin gene. The flour obtained from the whole-ground kernels was treated with pepsin under in vitro conditions, after that the samples were subjected to SDS-PAGE electrophoresis, to evaluate the amount of undigested protein; the electrophoregrams were scanned and subjected to computer analysis. It was reveale...

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Section: Abstract: Kafirins Transgenic Plants In Vitro Protein Digeunclassified

Section: результаты исследования и их обсуждениеunclassified

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In Vitro Digestibility of Storage Endosperm Proteins of Transgenic Sorghum Plants Carrying Genetic Construct for Silencing of the Gamma-Kafirin Gene

Elkonin¹,

Ital’yanskaya²

2017

УСЕ (ACNR)

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“…To solve this problem, various genetic engineering technologies had been developed. These technologies allow the introduction of new genes and thereby modulate the synthesis of new proteins with higher nutritional value or in a highly specific way downregulate genes that control the synthesis of proteins with a low nutritional value or reducing the digestibility or assimilation of other proteins [3][4][5]. Genetic engineering techniques are quite promising for enrichment of cereal grain with essential amino acids, i.e., lysine, tryptophan and methionine [6].…”

Section: Introductionmentioning

confidence: 99%

“…Improving of sorghum genetic transformation technology [20,21] makes it possible to solve this problem by using RNA interference (RNAi) that allows targeted downregulation of individual genes. In recent years, RNAi technology has become widely used for changing the composition of the storage proteins and starch in different cereal species [3][4][5].…”

Section: Introductionmentioning

confidence: 99%

Development of Transgenic Sorghum Plants with Improved In Vitro Kafirin Digestibility

Elkonin¹,

Italyanskaya²,

Panin³

et al. 2017

Plant Engineering

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Improvement of nutritional value of crops is one of the main goals of plant biotechnology. These studies are extremely important for sorghum-a unique drought-tolerant cereal crop that is of special importance for sustainable grain production in the arid regions. The major cause of relatively low nutritive value of sorghum grain is the resistance of one of its seed storage proteins, γ-kafirin, to protease digestion. Using Agrobacteriummediated genetic transformation, we have obtained transgenic sorghum plants harboring a genetic construct for RNA interference (RNAi) silencing of the γ-kafirin gene. In T 1 generation, transgenic plants with modified endosperm texture were found. These plants had lowered level of the 28-kDa γ-kafirin protein and kafirin oligomers, which are formed by natural kafirin polymerization. In vitro protein digestibility analysis showed that the amount of undigested protein in transgenic plants was reduced by 2.9-3.2 times, in comparison with the original line, the digestibility index reached 85-88% (60% in the original line). HPLC analysis showed that total amino acid content in transgenic plants was reduced, while the lysine proportion was increased by 1.6-1.7 times. PCR analysis confirmed inheritance of the genetic construct up to T 4 generation.

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Manifestation of agronomically valuable traits in the progeny of a sorghum mutant carrying the genetic construct for RNA silencing of the γ-kafirin gene

Elkonin,

Borisenko,

Pylaev

et al. 2024

Vestn. VOGiS

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Improving the nutritional value of grain sorghum, a drought- and heat-tolerant grain crop, is an important task in the context of global warming. One of the reasons for the low nutritional value of sorghum grain is the resistance of its storage proteins (kafirins) to proteolytic digestion, which is due, among other things, to the structural organization of protein bodies, in which γ-kafirin, the most resistant to proteases, is located on the periphery, encapsulating more easily digested α-kafirins. The introduction of genetic constructs capable of inducing RNA silencing of the γ-kafirin (gKAF1) gene opens up prospects for solving this problem. Using Agrobacterium-mediated genetic transformation of immature embryos of the grain sorghum cv. Avans we have obtained a mutant with improved digestibility of endosperm proteins (up to 92 %) carrying a genetic construct for RNA silencing of the gKAF1 gene. The goal of this work was to study the stability of inheritance of the introduced genetic construct in T2–T4 generations, to identify the number of its copies, as well as to trace the manifestation of agronomically valuable traits in the offspring of the mutant. The mutant lines were grown in experimental plots in three randomized blocks. The studied lines were characterized by improved digestibility of kafirins, a modified type of endosperm, completely or partially devoid of the vitreous layer, an increased percentage of lysine (by 75 %), reduced plant height, peduncle length, 1000-grains weight, and grain yield from the panicle. In T2, a line with monogenic control of GA resistance was selected. qPCR analysis showed that in different T3 and T4 plants, the genetic construct was present in 2–4 copies. In T3, a line with a high digestibility of endosperm proteins (81 %) and a minimal decrease in agronomically valuable traits (by 5–7 %) was selected.

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GENETIC ENGINEERING AS A TOOL FOR MODIFICATION OF SEED STORAGE PROTEINS AND IMPROVEMENT OF NUTRITIONAL VALUE OF CEREAL GRAIN (review)

Abstract: So modern bread wheat is the result of crossing three species barriers, a kind of natural genetic engineering»

Cited by 5 publications

References 58 publications

In Vitro Digestibility of Storage Endosperm Proteins of Transgenic Sorghum Plants Carrying Genetic Construct for Silencing of the Gamma-Kafirin Gene

In Vitro Digestibility of Storage Endosperm Proteins of Transgenic Sorghum Plants Carrying Genetic Construct for Silencing of the Gamma-Kafirin Gene

Development of Transgenic Sorghum Plants with Improved In Vitro Kafirin Digestibility

Manifestation of agronomically valuable traits in the progeny of a sorghum mutant carrying the genetic construct for RNA silencing of the γ-kafirin gene

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