Silencing of TaCKX1 Mediates Expression of Other TaCKX Genes to Increase Yield Parameters in Wheat

Jabłoński, Bartosz; Ogonowska, Hanna; Szala, Karolina; Bajguz, Andrzej; Orczyk, Wacław; Nadolska‐Orczyk, Anna

doi:10.3390/ijms21134809

Cited by 30 publications

(72 citation statements)

References 74 publications

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“…Our results showing coordinated regulation of the GSKs in barley provide novel data on the mechanisms of BR homeostasis in plants. A similar data, concerning RNAi-based silencing of TaCKX1 and associated with this co-regulation of CKX paralogs, were reported by Jablonski et al [52]. The authors found that diverse levels of target gene silencing were associated with different models of co-expression with other TaCKX genes and parameters of yield-related traits in wheat.…”

Section: Discussionsupporting

confidence: 78%

Silencing of HvGSK1.1—A GSK3/SHAGGY-Like Kinase–Enhances Barley (Hordeum vulgare L.) Growth in Normal and in Salt Stress Conditions

Kloc

Dmochowska-Boguta

Zieleziński

et al. 2020

IJMS

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Glycogen synthase kinase 3 (GSK3) is a highly conserved kinase present in all eukaryotes and functions as a key regulator of a wide range of physiological and developmental processes. The kinase, known in land plants as GSK3/SHAGGY-like kinase (GSK), is a key player in the brassinosteroid (BR) signaling pathway. The GSK genes, through the BRs, affect diverse developmental processes and modulate responses to environmental factors. In this work, we describe functional analysis of HvGSK1.1, which is one of the GSK3/SHAGGY-like orthologs in barley. The RNAi-mediated silencing of the target HvGSK1.1 gene was associated with modified expression of its paralogs HvGSK1.2, HvGSK2.1, HvGSK3.1, and HvGSK4.1 in plants grown in normal and in salt stress conditions. Low nucleotide similarity between the silencing fragment and barley GSK genes and the presence of BR-dependent transcription factors’ binding sites in promoter regions of barley and rice GSK genes imply an innate mechanism responsible for co-regulation of the genes. The results of the leaf inclination assay indicated that silencing of HvGSK1.1 and the changes of GSK paralogs enhanced the BR-dependent signaling in the plants. The strongest phenotype of transgenic lines with downregulated HvGSK1.1 and GSK paralogs had greater biomass of the seedlings grown in normal conditions and salt stress as well as elevated kernel weight of plants grown in normal conditions. Both traits showed a strong negative correlation with the transcript level of the target gene and the paralogs. The characteristics of barley lines with silenced expression of HvGSK1.1 are compatible with the expected phenotypes of plants with enhanced BR signaling. The results show that manipulation of the GSK-encoding genes provides data to explore their biological functions and confirm it as a feasible strategy to generate plants with improved agricultural traits.

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Section: Discussionsupporting

confidence: 78%

Silencing of HvGSK1.1—A GSK3/SHAGGY-Like Kinase–Enhances Barley (Hordeum vulgare L.) Growth in Normal and in Salt Stress Conditions

Kloc

Dmochowska-Boguta

Zieleziński

et al. 2020

IJMS

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“…Some functions of these genes might also be predicted by known function of their orthologues in other cereal species. Silencing of HvCKX1 orthologous to TaCKX1 resulted in elevated grain number in these two, closely related species [9,11]. However, the same effect of increased grain number in rice was obtained by silencing or knocking-out OsCKX2 [30].…”

Section: Introductionmentioning

confidence: 92%

“…Only some TaCKX GFMs are functionally characterized. Silencing of TaCKX1 by RNAi influenced levels of expression of the other TaCKX GFMs, and increased content of most CK forms and GA 3 , which caused an increase of yield parameters such as a higher spike number, grain number, and grain yield, but lower thousand grain weight [11]. The effect of TaCKX2.4 gene silencing by RNAi increased grain number per spike [34].…”

Section: Introductionmentioning

confidence: 99%

TaCKX2.2 Genes Coordinate Expression of Other TaCKX Family Members, Regulate Phytohormone Content and Yield-Related Traits of Wheat

Jabłoński

Szala

Przyborowski

et al. 2021

IJMS

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TaCKX gene family members (GFMs) play essential roles in the regulation of cytokinin during wheat development and significantly influence yield-related traits. However, detailed function of most of them is not known. To characterize the role of TaCKX2.2 genes we silenced all homoeologous copies of both TaCKX2.2.1 and TaCKX2.2.2 by RNAi technology and observed the effect of silencing in 7 DAP spikes of T1 and T2 generations. The levels of gene silencing of these developmentally regulated genes were different in both generations, which variously determined particular phenotypes. High silencing of TaCKX2.2.2 in T2 was accompanied by slight down-regulation of TaCKX2.2.1 and strong up-regulation of TaCKX5 and TaCKX11, and expression of TaCKX1, TaCKX2.1, and TaCKX9 was comparable to the non-silenced control. Co-ordinated expression of TaCKX2.2.2 with other TaCKX GFMs influenced phytohormonal homeostasis. Contents of isoprenoid, active cytokinins, their conjugates, and auxin in seven DAP spikes of silenced T2 plants increased from 1.27 to 2.51 times. However, benzyladenine (BA) and abscisic acid (ABA) contents were significantly reduced and GA3 was not detected. We documented a significant role of TaCKX2.2.2 in the regulation of thousand grain weight (TGW), grain number, and chlorophyll content, and demonstrated the formation of a homeostatic feedback loop between the transcription of tested genes and phytohormones. We also discuss the mechanism of regulation of yield-related traits.

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“…Targeting cytokinin homeostasis through the application of exogenous compounds leads to a range of effects that are frequently inconsistent [ 68 ], which supports suggestions that cytokinin homeostasis is complex and tightly regulated. It is well known that targeting CKX with CKX-inhibiting PGRs has an effect on cytokinin homeostasis and/or cytokinin-associated processes [ 44 , 45 , 46 , 47 , 48 , 49 ], including INCYDE [ 34 , 51 , 52 , 54 , 56 ] and through targeting CKX in mutants and/or transgenic plants [ 36 , 37 , 38 , 39 , 41 , 42 , 69 ].…”

Section: Discussionmentioning

confidence: 99%

“…Inhibition of CKX has been suggested as a strategy for enhancing endogenous cytokinin to enhance growth [ 34 ] and yield [ 31 , 35 ]. This approach has been successful in enhancing yield components with ckx double mutants in Arabidopsis thaliana (Arabidopsis) [ 36 ] and oil seed rape [ 37 ], OsCKX2 loss-of-function mutants with rice [ 38 ], and RNAi gene silencing of HvCKX1 in barley [ 39 , 40 ], and TaCKX1 in wheat [ 41 ]. Notably, experiments with RNA-guided Cas9 ckx mutant lines in barley have shown that single CKX gene knockouts might be insufficient for enhancing yield [ 42 ].…”

Section: Introductionmentioning

confidence: 99%

Targeting Cytokinin Homeostasis in Rapid Cycling Brassica rapa with Plant Growth Regulators INCYDE and TD-K

Voorthuizen

Nisler

Song

et al. 2020

Plants

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Modifying the cytokinin content in plants is a means of improving plant productivity. Here, we report the development and biological activity of compound TD-K (1-(furan-2-ylmethyl)-3-(1,2,3-thiadiazol-5-yl)urea)which is related to thidiazuron. TD-K—which exhibited extremely high antisenescence activity in the wheat leaf bioassay—and INCYDE (2-chloro-6-(3-methoxyphenyl)aminopurine)—a plant growth regulator reported to inhibit cytokinin oxidase/dehydrogenase (CKX), an enzyme involved in the degradation of the plant hormone cytokinin—were selected for investigation of their effects on the model plant Rapid Cycling Brassica rapa (RCBr). We monitored the expression of BrCKX and isopentenyl transferase (BrIPT), which codes for the key cytokinin biosynthesis enzyme, in developing leaves following INCYDE and TD-K application. Growth room experiments revealed that INCYDE increased RCBr seed yield per plant, but only when applied multiple times and when grown in 5 mM KNO3. Expression in control leaves showed transient, high levels of expression of BrCKX and BrIPT at true leaf appearance. Following INCYDE application, there was a rapid and strong upregulation of BrCKX3, and a transient downregulation of BrIPT1 and BrIPT3. Interestingly, the upregulation of BrCKX3 persisted in a milder form throughout the course of the experiment (16 days). TD-K also upregulated BrCKX3. However, in contrast to INCYDE, this effect disappeared after two days. These results suggest that both compounds (CKX inhibitor and cytokinin TD-K) influenced cytokinin homeostasis in RCBr leaves, but with different mechanisms.

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Silencing of TaCKX1 Mediates Expression of Other TaCKX Genes to Increase Yield Parameters in Wheat

Cited by 30 publications

References 74 publications

Silencing of HvGSK1.1—A GSK3/SHAGGY-Like Kinase–Enhances Barley (Hordeum vulgare L.) Growth in Normal and in Salt Stress Conditions

Silencing of HvGSK1.1—A GSK3/SHAGGY-Like Kinase–Enhances Barley (Hordeum vulgare L.) Growth in Normal and in Salt Stress Conditions

TaCKX2.2 Genes Coordinate Expression of Other TaCKX Family Members, Regulate Phytohormone Content and Yield-Related Traits of Wheat

Targeting Cytokinin Homeostasis in Rapid Cycling Brassica rapa with Plant Growth Regulators INCYDE and TD-K

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