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携带用于γ-高粱醇溶蛋白基因RNA沉默的遗传构建体的高粱突变体后代中农艺学上有价值性状的表现。

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

作者信息

Elkonin L A, Borisenko N V, Pylaev T E, Kenzhegulov O A, Sarsenova S Kh, Selivanov N Yu, Panin V M

机构信息

Federal Center of Agriculture Research of the South-East Region, Saratov, Russia.

Institute of Biochemistry and Physiology of Plants and Microorganisms, Saratov Federal Scientific Center of the Russian Academy of Sciences, Saratov, Russia Saratov State Medical University named after V.I. Razumovsky, Saratov, Russia.

出版信息

Vavilovskii Zhurnal Genet Selektsii. 2024 Feb;28(1):63-73. doi: 10.18699/vjgb-24-08.

Abstract

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.

摘要

提高高粱(一种耐旱耐热的谷类作物)的营养价值是全球变暖背景下的一项重要任务。高粱籽粒营养价值低的原因之一是其贮藏蛋白(高粱醇溶蛋白)对蛋白水解消化具有抗性,这尤其归因于蛋白体的结构组织,其中对蛋白酶最具抗性的γ-高粱醇溶蛋白位于外周,包裹着更易消化的α-高粱醇溶蛋白。引入能够诱导γ-高粱醇溶蛋白(gKAF1)基因RNA沉默的遗传构建体为解决这一问题开辟了前景。利用农杆菌介导的高粱品种阿万斯未成熟胚的遗传转化,我们获得了一个胚乳蛋白消化率提高(高达92%)的突变体,其携带用于gKAF1基因RNA沉默的遗传构建体。这项工作的目标是研究导入的遗传构建体在T2 - T4代中的遗传稳定性,确定其拷贝数,并追踪该突变体后代中农艺性状的表现。突变体系在三个随机区组的试验田中种植。所研究的品系具有高粱醇溶蛋白消化率提高、胚乳类型改变(完全或部分没有玻璃质层)、赖氨酸含量增加(75%)、株高降低、茎节长度、千粒重和穗粒产量降低等特征。在T2代,选择了一个对GA抗性由单基因控制的品系。qPCR分析表明,在不同的T3和T4植株中,遗传构建体以2 - 4个拷贝存在。在T3代,选择了一个胚乳蛋白消化率高(81%)且农艺性状下降最小(5 - 7%)的品系。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b188/10917670/e121c8e4feb2/VJGB-28-2408-Fig1.jpg

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