Genetic transformation of barley: limiting factors

Vyroubalová, Šárka; Šmehilová, Mária; Galuszka, Petr; Ohnoutková, Ludmila

doi:10.1007/s10535-011-0032-8

Cited by 12 publications

(5 citation statements)

References 102 publications

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“…S2e). Such abnormalities have already been described in several publications for transformant plants (reviewed by Vyroubalová et al 2011), but we found that the PITP and PI4K transformants showed this abnormality in a relatively higher rate. In the PI4K L6 line, 6.5% of the plants showed a pale leaf phenotype, while in the PI4K L4 line 6%, and in the PITP L11 line 5% of the plants showed albinism.…”

Section: Developmental Disorders Of the Transgenic Linessupporting

confidence: 80%

Overexpression of Two Upstream Phospholipid Signaling Genes Improves Cold Stress Response and Hypoxia Tolerance, but Leads to Developmental Abnormalities in Barley

et al. 2019

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Phosphatidylinositol transfer protein (PITP) and phosphatidylinositol 4-kinase (PI4K) are very upstream regulatory elements of the phospholipid signaling pathway in the signal transduction network. Unlike in animal systems, their role in stress signaling is poorly understood in plants. To study this area, PITP-and PI4K-overexpressing transgenic barley lines were developed. Morphological and developmental abnormalities were surveyed and characterized. It was revealed that the overexpression of the upstream signaling genes led to more phenotypic abnormalities than in other transgenic studies working with effector genes or even transcription factors. We hypothesize that this high level of abnormalities is the consequence of the modulation of the very upstream signal transduction pathway elements. On the other hand, we also revealed that overexpression of the PITP and PI4K genes increased stress tolerance during hypoxic cold stress, but not during salinity stress. Differences were also found in the level of frost tolerance between the transgenic overexpression plants and the recipient Golden Promise line. Molecular analysis showed that this improvement was not related to the most important cold responsive transcription factors, the CBF genes. We conclude that the transgenic method may be useful to prove the role of an upstream signaling element; however, due to the many developmental consequences that occur as side effects, it is a less advisable approach to achieve improved stress tolerance.

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Section: Developmental Disorders Of the Transgenic Linessupporting

confidence: 80%

Overexpression of Two Upstream Phospholipid Signaling Genes Improves Cold Stress Response and Hypoxia Tolerance, but Leads to Developmental Abnormalities in Barley

et al. 2019

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“…Genetic engineering is a useful approach for creating crop plants with desired qualities. Since cereal species have immense agricultural importance, there have been many recent advances in transformation techniques for monocot plants [1] allowing rapid development of novel transgenic varieties. Barley ( Hordeum vulgare L.) is a widely grown cereal with valuable traits, including high malting quality and nutritional value, which are exploited in brewing, distilling and the production of diverse human and animal foods.…”

Section: Introductionmentioning

confidence: 99%

Overexpression of Cytokinin Dehydrogenase Genes in Barley (Hordeum vulgare cv. Golden Promise) Fundamentally Affects Morphology and Fertility

et al. 2013

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Barley is one of the most important cereal crops grown worldwide. It has numerous applications, but its utility could potentially be extended by genetically manipulating its hormonal balances. To explore some of this potential we identified gene families of cytokinin dehydrogenases (CKX) and isopentenyl transferases, enzymes that respectively irreversibly degrade and synthesize cytokinin (CK) plant hormones, in the raw sequenced barley genome. We then examined their spatial and temporal expression patterns by immunostaining and qPCR. Two CKX-specific antibodies, anti-HvCKX1 and anti-HvCKX9, predominantly detect proteins in the aleurone layer of maturing grains and leaf vasculature, respectively. In addition, two selected CKX genes were used for stable, Agrobacterium tumefaciens-mediated transformation of the barley cultivar Golden Promise. The results show that constitutive overexpression of CKX causes morphological changes in barley plants and prevents their transition to flowering. In all independent transgenic lines roots proliferated more rapidly and root-to-shoot ratios were higher than in wild-type plants. Only one transgenic line, overexpressing CKX under the control of a promoter from a phosphate transporter gene, which is expressed more strongly in root tissue than in aerial parts, yielded progeny. Analysis of several T1-generation plants indicates that plants tend to compensate for effects of the transgene and restore CK homeostasis later during development. Depleted CK levels during early phases of development are restored by down-regulation of endogenous CKX genes and reinforced de novo biosynthesis of CKs.

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“…Plant transformation is essential for crop improvement and a wide range of basic research such as understanding gene functions and precision gene editing (Lawrenson et al 2015 ; Hinchliffe and Harwood 2019 ). Since the first transgenic barley was generated with microprojectile bombardment (Ritala et al 1994 ; Wan and Lemaux 1994 ), several barley transformation systems have been developed, but the Agrobacterium-mediated transformation with immature embryo explants by somatic embryogenesis is the most widely used procedures for genetic transformation in barley as it can yield higher transformation efficiency (Tingay et al 1997 ; Vyroubalová et al 2011 ). However, this transformation system only works for limited barley genotypes and failed to generate transgenic plants for many elite barley cultivars.…”

Section: Introductionmentioning

confidence: 99%

Agrobacterium-mediated transfer of the Fusarium graminearum Tri6 gene into barley using mature seed-derived shoot tips as explants

Gao,

Abdullah,

Baldwin

et al. 2024

Plant Cell Rep

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Key message We transferred the Tri6 gene into the elite barley GemCraft via new transformation method through shoot organogenesis and identified the rearrangements of transgenes and phenotypic variations in the transgenic plants. Abstract Despite its agronomic and economic importance, barley transformation is still very challenging for many elite varieties. In this study, we used direct shoot organogenesis to transform the elite barley cultivar GemCraft with the RNAi constructs containing Tri6 gene of Fusarium graminearum, which causes fusarium head blight (FHB). We isolated 4432 shoot tips and co-cultured these explants with Agrobacterium tumefaciens. A total of 25 independent T0 transgenic plants were generated including 15 events for which transgene-specific PCR amplicons were observed. To further determine the presence of transgenes, the T1 progenies of all 15 T0 plants were analyzed, and the expected PCR products were obtained in 10 T1 lines. Droplet digital (dd) PCR analysis revealed various copy numbers of transgenes in the transgenic plants. We determined the insertion site of transgenes using long-read sequencing data and observed the rearrangements of transgenes. We found phenotypic variations in both T1 and T2 generation plants. FHB disease was evaluated under growth chamber conditions, but no significant differences in disease severity or deoxynivalenol accumulation were observed between two Tri6 transgenic lines and the wildtype. Our results demonstrate the feasibility of the shoot tip transformation and may open the door for applying this system for genetic improvement and gene function research in other barley genotypes.

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Genetic transformation of barley: limiting factors

Cited by 12 publications

References 102 publications

Overexpression of Two Upstream Phospholipid Signaling Genes Improves Cold Stress Response and Hypoxia Tolerance, but Leads to Developmental Abnormalities in Barley

Overexpression of Two Upstream Phospholipid Signaling Genes Improves Cold Stress Response and Hypoxia Tolerance, but Leads to Developmental Abnormalities in Barley

Overexpression of Cytokinin Dehydrogenase Genes in Barley (Hordeum vulgare cv. Golden Promise) Fundamentally Affects Morphology and Fertility

Agrobacterium-mediated transfer of the Fusarium graminearum Tri6 gene into barley using mature seed-derived shoot tips as explants

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