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21 results on '"Elkonin, Lev"'

4. Effect of sterile cytoplasms on content of absolutely dry matter in the grain sorghum hybrids

Author

Bychkova Vera and Elkonin Lev

Subjects
Microbiology, QR1-502, Physiology, QP1-981, Zoology, QL1-991
Abstract

Analysis of absolutely dry matter (ADM) accumulation is important indicator of productivity of crop plants. For three seasons, we investigated the effect of sterile cytoplasms on content of ADM in the F1 hybrids of grain sorghum obtained on the basis of two series of alloplasmic iso-nuclear CMS-lines: (1) with A3, A4, and 9E cytoplasms and (2) with 9E and M35-1A cytoplasms. For the first time, the effect of the type of sterile cytoplasm on content of ADM in the F1 sorghum hybrids was shown. In each season, A3 cytoplasm reduced ADM in the F1 hybrids at the “tillering – heading” stage, whereas 9E cytoplasm increased ADM at the “heading – complete maturity” stage. The most significant differences were observed under drought conditions. These data indicate the genetic influence of cytoplasm on assimilation capacity of sorghum hybrids and tolerance to drought stress.

Published
2022
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5. Inheritance of the genetic construct for RNA-silencing of the γ-kafirin gene (gKAF1) in the progeny of transgenic sorghum plants

Author

Borisenko Nataly, Elkonin Lev, and Kenzhegulov Odyssey

Subjects
Microbiology, QR1-502, Physiology, QP1-981, Zoology, QL1-991
Abstract

Sorghum is one of the most important cereal crops in drought regions of the Globe. However, a number of constrains, such as poor nutritional value compared to other cereals, which is caused by resistance of its grain storage proteins (kafirins) to protease digestion, makes it less popular. The use of modern genetic technologies, such as RNA interference, opens up prospects for solving this problem. In this paper, we describe the inheritance of the genetic construct for RNAi silencing of the γ-kafirin gene (gKAF1) in the progeny of the mutant Avans-1/18, that we obtained previously by Agrobacterium-mediated genetic transformation of immature embryos of commercial cv. Avans. Inheritance of the genetic construct was traced by PCR-analysis to the nos-promoter governing expression of the marker gene bar and to the ubi1-intron, which is part of the genetic construct for silencing. It was found that the genetic construct inherited in T1 and T2 generations of the mutant grown both in the growth chamber and in the field plot, and in the F1 hybrids of the mutant with CMS-lines A2 KVV-181, A2 KVV-114, A2 O-1237, and fertile lines Volzhskoe-4v, Volzhskoe-615, O-1237. The studied plants from T1 and T2 generations had significantly higher in vitro protein digestibility than original non-transgenic cv. Avans. The vitreous endosperm in these plants was reduced either completely or manifested a faint layer. These data demonstrated that genetic construct for gKAF1 silencing was not only stably integrated in genome of the mutant plants but also expressed and improved the digestibility of endosperm proteins. In addition, in T1 progeny we found the plant, PCR positive for nos-promoter but PCR-negative for ubi1-intron, that had vitreous endosperm and was characterized by poor in vitro protein digestibility. These data are an example of instability of the genetic construct for RNAi-silencing in genome of some transgenic plants; they show close correlation of the floury endosperm type and high in vitro protein digestibility.

Published
2022
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13. RNAi-Mutants of <em>Sorghum bicolor</em> (L.) Moench with Improved Digestibility of Seed Storage Proteins

Author

Elkonin, Lev A., Sarsenova, Saule Kh., Kenzhegulov, Odissey A., and Panin, Valery M.

Subjects
Technology & Engineering / Agriculture
Abstract

Modification of the composition of grain storage proteins is an intensively developing area of plant biotechnology, which is of particular importance for sorghum – high-yielding drought tolerant crop. Compared to other cereals, the majority of sorghum cultivars and hybrids are characterized by reduced nutritional value that is caused by a low content of essential amino acids in the seed storage proteins (kafirins), and resistance of kafirins to protease digestion. RNA interference (RNAi) by suppressing synthesis of individual kafirin subclasses may be an effective approach to solve this problem. In this chapter, we review published reports on RNAi silencing of the kafirin-encoding genes. In addition, we present new experimental data on phenotypic effects of RNAi-silencing of γ-KAFIRIN-1 gene in sorghum cv. Avans. To obtain RNAi mutants with γ-KAFIRIN-1 gene silencing we used Agrobacterium-mediated genetic transformation. Transgenic kernels had modified endosperm type with reduced vitreous layer and significantly improved in vitro protein digestibility (93% vs. 57%, according to the densitometry of SDS-PAGE patterns). SDS-PAGE of transgenic kernels showed lowered level of kafirins and appearance of globulin proteins, which were not observed in the original cultivar. For the first time, the cases of instability of inserted genetic construct were identified: elimination of ubi1-intron that is a constituent part of the genetic construct for RNAi silencing, or nos-promotor governing expression of the marker gene (bar) (in the RNAi mutants of cv. Zheltozernoe 10). The research findings presented in this chapter provide strong evidence that RNA interference can be used for improvement of the nutritional properties of sorghum grain.

Published
2018

15. Development of Transgenic Sorghum Plants with Improved In Vitro Kafirin Digestibility

Author

Elkonin, Lev A.

Subjects
Science / Life Sciences / Botany
Abstract

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 Agrobacterium‐mediated genetic transformation, we have obtained transgenic sorghum plants harboring a genetic construct for RNA interference (RNAi) silencing of the γ‐kafirin gene. In T1 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 T4 generation.

Published
2017

20. Effect of drought stress on male fertility restoration in A3CMS-inducing cytoplasm of sorghum

Author

Kozhemyakin, Valentin V., Elkonin, Lev A., and Dahlberg, Jeffery A.

Abstract

Use of cytoplasmic male sterility (CMS) in hybrid breeding requires effective male fertility-restoring lines. In sorghum, very few restoring lines that can restore fertility in A3CMS have been reported. To identify the reasons for this deficiency, F1and F2hybrids of an A3CMS line crossed with the line IS1112C, a donor of fertility-restoring (Rf) genes for A3cytoplasm, and testcrosses of fertile plants to A3CMS lines were grown under contrasting water availability regimes in dryland and irrigated field plots. In the irrigated plots the frequency of fertile plants in testcrosses was twice that in dryland plots (P<0.05). Fertile plants from the F2family grown in the irrigated plots showed significantly higher restoration ability than fertile plants from the same family grown in dryland plots. F3plants from the F2family grown in irrigated plots yielded on average a sixfold higher frequency of fertile plants in testcrosses than F3plants derived from dryland plots (P<0.01). Fertility of testcross hybrids correlated negatively with air vapor pressure deficit (VPD) at flowering (r=−0.96; P<0.01) suggesting that VPD is a trigger for downregulation of Rfgenes for A3cytoplasm.

Published
2017
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21. Development of sorghum mutants with improved in vitroprotein digestibility by CRISPR/Cas9 editing of kafirin genes

Author

Elkonin, Lev A., Gerashchenkov, Grigoriy A., Borisenko, Natalie V., Kenzhegulov, Odyssey A., Kh. Sarsenova, Saule, Rozhnova, Natalya A., and Panin, Valery M.

Abstract

Sorghum (Sorghum bicolor(L.) Moench) is a major world crop that is a reliable source of fodder and food grain in arid regions. However, unlike other cereals, sorghum grain has low nutritional value, owing mainly to the resistance of its storage proteins (kafirins) to protease digestion. Changing the composition of kafirins or their primary structure may address this problem. To induce mutations in kafirin-encoding genes that were expected to disturb their accumulation in endosperm cells, we used a genome-editing approach. By Agrobacterium-mediated genetic transformation of immature embryos of cv. Avans, we obtained 14 transgenic plants with genetic constructs for site-directed mutagenesis of the k1C5and gKAF1genes encoding 22 kDa α- and 28 kDa γ-kafirins, respectively. Sequencing of 5 regenerants obtained by using k1C5-addressing vector revealed two plants with mutations. T1progeny of these mutants had higher in vitrodigestibility of endosperm proteins (86%–92%), in comparison with the donor Avans (63%–67%). The kernels of these plants had a thick vitreous endosperm. A mutant with increased in vitroprotein digestibility and vitreous endosperm, carrying a mutation in the target sequence, was also obtained by use of the gKAF1-addressing vector. Thus, using genome editing technology, we have obtained mutants with improved kafirin digestibility that can be used in sorghum breeding.

Published
2023
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