Journal articles: 'Transgenic barley'

1

Wang, Ming-Bo, David C. Abbott, Narayana M. Upadhyaya, John V. Jacobsen, and Peter M. Waterhouse. "Agrobacterium tumefaciens-mediated transformation of an elite Australian barley cultivar with virus resistance and reporter genes." Functional Plant Biology 28, no. 2 (2001): 149. http://dx.doi.org/10.1071/pp00103.

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Efficient transformation of barley cv. Schooner was achieved using Agrobacterium delivery, hygromycin or bialaphos selection and embryogenic callus. Using this system, transgenic plants were generated that contained either the green fluorescent protein gene, or transgenes derived from barley yellow dwarf (BYDV) and cereal yellow dwarf (CYDV) viruses. Many of these plants contained 1–3 transgene copies that were inherited in a simple Mendelian manner. Some plants containing BYDV and/or CYDV derived transgenes showed reduced virus symptoms and rates of viral replication when challenged with the appropriate virus. The ability to transform Schooner is a significant advance for the Australian barley industry, as this elite malting variety is, and has for the last 15 years been, the most widely grown barley variety in eastern Australia.

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Ohnoutková, Ludmila, and Tomáš Vlčko. "Homozygous Transgenic Barley (Hordeum vulgare L.) Plants by Anther Culture." Plants 9, no. 7 (July 20, 2020): 918. http://dx.doi.org/10.3390/plants9070918.

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Production of homozygous lines derived from transgenic plants is one of the important steps for phenotyping and genotyping transgenic progeny. The selection of homozygous plants is a tedious process that can be significantly shortened by androgenesis, cultivation of anthers, or isolated microspores. Doubled haploid (DH) production achieves complete homozygosity in one generation. We obtained transgenic homozygous DH lines from six different transgenic events by using anther culture. Anthers were isolated from T0 transgenic primary regenerants and cultivated in vitro. The ploidy level was determined in green regenerants. At least half of the 2n green plants were transgenic, and their progeny were shown to carry the transgene. The process of dihaploidization did not affect the expression of the transgene. Embryo cultures were used to reduce the time to seed of the next generation. The application of these methods enables rapid evaluation of transgenic lines for gene function studies and trait evaluation.

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Koev, Gennadiy, B. R. Mohan, S. P. Dinesh-Kumar, Kimberly A. Torbert, David A. Somers, and W. Allen Miller. "Extreme Reduction of Disease in Oats Transformed with the 5′ Half of the Barley Yellow Dwarf Virus-PAV Genome." Phytopathology® 88, no. 10 (October 1998): 1013–19. http://dx.doi.org/10.1094/phyto.1998.88.10.1013.

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Barley yellow dwarf viruses (BYDVs) are the most serious and widespread viruses of oats, barley, and wheat worldwide. Natural resistance is inadequate. Toward overcoming this limitation, we engineered virus-derived transgenic resistance in oat. Oat plants were transformed with the 5′ half of the BYDV strain PAV genome, which includes the RNA-dependent RNA polymerase gene. In experiments on T2- and T3-generation plants descended from the same transformation event, all BYDV-inoculated plants containing the transgene showed disease symptoms initially, but recovered, flowered, and produced seed. In contrast, all but one of the BYDV-PAV-inoculated nontransgenic segregants died before reaching 25 cm in height. Although all of the recovered transgenic plants looked similar, the amount of virus and viral RNA ranged from substantial to undetectable levels. Thus, the transgene may act either by restricting virus accumulation or by a novel transgenic tolerance phenomenon. This work demonstrates a strategy for genetically stable transgenic resistance to BYDVs that should apply to all hosts of the virus.

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Kwapata, Kingdom, Thang Nguyen, and Mariam Sticklen. "Genetic Transformation of Common Bean (Phaseolus vulgarisL.) with theGusColor Marker, theBarHerbicide Resistance, and the Barley (Hordeum vulgare)HVA1Drought Tolerance Genes." International Journal of Agronomy 2012 (2012): 1–8. http://dx.doi.org/10.1155/2012/198960.

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Five common bean (Phaseolus vulgarisL.) varieties including “Condor,” “Matterhorn,” “Sedona,” “Olathe,” and “Montcalm” were genetically transformed via the Biolistic bombardment of the apical shoot meristem primordium. Transgenes includedguscolor marker which visually confirmed transgenic events, thebarherbicide resistance selectable marker used forin vitroselection of transgenic cultures and which confirmed Liberty herbicide resistant plants, and the barley (Hordeum vulgare) late embryogenesis abundant protein (HVA1) which conferred drought tolerance with a corresponding increase in root length of transgenic plants. Research presented here might assist in production of betterP. vulgarisgermplasm.

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Oldach, Klaus H., Dirk Becker, and Horst Lörz. "Heterologous Expression of Genes Mediating Enhanced Fungal Resistance in Transgenic Wheat." Molecular Plant-Microbe Interactions® 14, no. 7 (July 2001): 832–38. http://dx.doi.org/10.1094/mpmi.2001.14.7.832.

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Three cDNAs encoding the antifungal protein Ag-AFP from the fungus Aspergillus giganteus, a barley class II chitinase and a barley type I RIP, all regulated by the constitutive Ubiquitin1 promoter from maize, were expressed in transgenic wheat. In 17 wheat lines, stable integration and inheritance of one of the three transgenes has been demonstrated over four generations. The formation of powdery mildew (Erysiphe graminis f. sp. tritici) or leaf rust (Puccinia recondita f. sp. tritici) colonies was significantly reduced on leaves from afp or chitinase II- but not from rip I-expressing wheat lines compared with non-transgenic controls. The increased resistance of afp and chitinase II lines was dependent on the dose of fungal spores used for inoculation. Heterologous expression of the fungal afp gene and the barley chitinase II gene in wheat demonstrated that colony formation and, thereby, spreading of two important biotrophic fungal diseases is inhibited approximately 40 to 50% at an inoculum density of 80 to 100 spores per cm2.

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Yao, Qing A., Ecaterina Simion, Manilal William, Joan Krochko, and Ken J. Kasha. "Biolistic transformation of haploid isolated microspores of barley (Hordeum vulgare L.)." Genome 40, no. 4 (August 1, 1997): 570–81. http://dx.doi.org/10.1139/g97-075.

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Transgenic barley plants were produced by the direct delivery of plasmid DNA into isolated microspores of barley cv. Igri using high velocity microprojectiles. The plasmid pAHC25 contained the uidA and bar genes, each under the control of a maize Ubi1 promoter. Bombarded microspores were cultured and selected on solid medium containing varying concentrations (2–5 mg/L) of the Basta herbicide active agent bialaphos. The effectiveness of selection with bialaphos depended on its interaction with the medium component glutamine. Six transgenic plants (R0) were obtained, and the presence of the uidA and bar genes and their integration into nuclear DNA in transformed R0 plants were confirmed by PCR and Southern blot analysis. Phosphinothricin acetyltransferase activity was observed in all six R0 transgenic plants, whereas none showed β-glucuronidase (GUS) activity in histochemical GUS assays. Two of the six R0 plants were haploid and sterile; one of them was trisomic and partially sterile; the remainder were diploid, but one of them was also sterile. Inheritance of the transgenes in progeny of three seed-producing transgenic plants was investigated. Southern blot analysis of genomic DNA from R1 plants showed that the introduced bar and uidA genes were hemizygous and stably cotransmitted to the R1 progeny derived from self-pollination. Analysis of Basta resistance and the integration of the bar gene by PCR analysis in R1 plants indicated that the bar gene was being inherited and expressed as a single dominant trait. Fluorescent in situ hybridization was performed on chromosomes of the trisomic plant to confirm the presence of transgenes in the genome.Key words: barley, microspore, biolistic transformation, bialaphos, haploid, FISH.

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7

Ritala, Anneli, Reino Aikasalo, Kristian Aspegren, Marjatta Salmenkallio-Marttila, Satu Akerman, Leena Mannonen, Ulrika Kurtén, Riitta Puupponen-Pimiä, Teemu H. Teeri, and Veli Kauppinen. "Transgenic barley by particle bombardment. Inheritance of the transferred gene and characteristics of transgenic barley plants." Euphytica 85, no. 1-3 (February 1995): 81–88. http://dx.doi.org/10.1007/bf00023933.

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8

Bekalu, Zelalem Eshetu, Claus Krogh Madsen, Giuseppe Dionisio, Inger Bæksted Holme, Lise Nistrup Jørgensen, Inge S. Fomsgaard, and Henrik Brinch-Pedersen. "Overexpression of Nepenthesin HvNEP-1 in Barley Endosperm Reduces Fusarium Head Blight and Mycotoxin Accumulation." Agronomy 10, no. 2 (February 1, 2020): 203. http://dx.doi.org/10.3390/agronomy10020203.

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Fusarium head blight (FHB) causes substantial losses of yield and quality in grains, both in the field and in post-harvest storage. To date, adequate natural genetic resistance is not available for the control of FHB. This study reports the cloning and overexpression of a barley (Hordeum vulgare L.) antifungal gene, nepenthesin 1 (HvNEP-1), in the endosperm of barley grains. Transgenic barley lines overexpressing HvNEP-1 substantially reduced FHB severity and disease progression after inoculation with Fusarium graminearum or Fusarium culmorum. The transgenic barley also showed reduced accumulation of the mycotoxin deoxynivalenol (DON) in grain, far below the minimum value allowable for food. Semi-field evaluation of four HvNEP-1 transgenic lines revealed substantial reduction of FHB severity and progression as compared with the control H. vulgare cultivar Golden promise (GP) plants. Our study demonstrated the utility of HvNEP-1 for the control of FHB in barley, and possibly other grains such as wheat and maize.

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9

Choi, H. W., P. G. Lemaux, and M. J. Cho. "Long-term stability of transgene expression driven by barley endosperm-specific hordein promoters in transgenic barley." Plant Cell Reports 21, no. 11 (July 1, 2003): 1108–20. http://dx.doi.org/10.1007/s00299-003-0630-9.

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10

Matthews, Peter R., Sarah Thornton, Frank Gubler, Rosemary White, and John V. Jacobsen. "Use of the green fluorescent protein to locate α-amylase gene expression in barley grains." Functional Plant Biology 29, no. 9 (2002): 1037. http://dx.doi.org/10.1071/fp02011.

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A green fluorescent protein (GFP) gene was cloned between the promoter and 3� regions from a barley high isoelectric point (pI) α-amylase gene, then inserted into barley. GFP fluorescence was used to locate and quantify expression of the transgene in barley grains following hydration. Light and confocal laser microscopy revealed fluorescence in the known regions of α-amylase synthesis in the scutellar epithelium, aleurone layer and embryonic axis. Fluorescence was quantified using a simple fluorescence assay, which showed induction of the transgene to mirror the induction of α-amylase in aleurone exposed to gibberellic acid. Expression from the transgene was also shown to be inhibited by abscisic acid, in the same way as expression of endogenous α-amylase genes. Overall, the transgenic grain revealed patterns of α-amylase expression before and after germination, and showed strong potential for further studies investigating both α-amylase production and transport of gibberellin in malting grain.

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Pospíšilová, Hana, Šárka Vyroubalová, Katarína Mrízová, Petr Galuszka, and Ivo Frébort. "Preparation of transgenic barley with improved quality." Journal of Biotechnology 150 (November 2010): 119. http://dx.doi.org/10.1016/j.jbiotec.2010.08.310.

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12

Mihalik, D., M. Gubisova, T. Klempova, M. Certik, K. Ondreickova, M. Hudcovicova, L. Klcova, et al. "Transgenic barley producing essential polyunsaturated fatty acids." Biologia plantarum 58, no. 2 (June 1, 2014): 348–54. http://dx.doi.org/10.1007/s10535-014-0406-9.

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13

Hensel, G., C. Marthe, A. Kusserow, A. Himmelbach, N. Borisjuk, S. Goedeke, E. Kapusi, et al. "Transgenic barley in applied research and biotechnology." Journal für Verbraucherschutz und Lebensmittelsicherheit 2, S1 (December 2007): 104. http://dx.doi.org/10.1007/s00003-007-0267-7.

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14

Jähne, A., D. Becker, R. Brettschneider, and H. Lörz. "Regeneration of transgenic, microspore-derived, fertile barley." Theoretical and Applied Genetics 89, no. 4 (October 1994): 525–33. http://dx.doi.org/10.1007/bf00225390.

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15

Luo, Bingbing, Man Xu, Limei Zhao, Peng Xie, Yi Chen, Wendy Harwood, Guohua Xu, Xiaorong Fan, and Anthony J. Miller. "Overexpression of the High-Affinity Nitrate Transporter OsNRT2.3b Driven by Different Promoters in Barley Improves Yield and Nutrient Uptake Balance." International Journal of Molecular Sciences 21, no. 4 (February 15, 2020): 1320. http://dx.doi.org/10.3390/ijms21041320.

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Improving nitrogen use efficiency (NUE) is very important for crops throughout the world. Rice mainly utilizes ammonium as an N source, but it also has four NRT2 genes involved in nitrate transport. The OsNRT2.3b transporter is important for maintaining cellular pH under mixed N supplies. Overexpression of this transporter driven by a ubiquitin promoter in rice greatly improved yield and NUE. This strategy for improving the NUE of crops may also be important for other cereals such as wheat and barley, which also face the challenges of nutrient uptake balance. To test this idea, we constructed transgenic barley lines overexpressing OsNRT2.3b. These transgenic barley lines overexpressing the rice transporter exhibited improved growth, yield, and NUE. We demonstrated that NRT2 family members and the partner protein HvNAR2.3 were also up-regulated by nitrate treatment (0.2 mM) in the transgenic lines. This suggests that the expression of OsNRT2.3b and other HvNRT2 family members were all up-regulated in the transgenic barley to increase the efficiency of N uptake and usage. We also compared the ubiquitin (Ubi) and a phloem-specific (RSs1) promoter-driven expression of OsNRT2.3b. The Ubi promoter failed to improve nutrient uptake balance, whereas the RSs1 promoter succeed in increasing the N, P, and Fe uptake balance. The nutrient uptake enhancement did not include Mn and Mg. Surprisingly, we found that the choice of promoter influenced the barley phenotype, not only increasing NUE and grain yield, but also improving nutrient uptake balance.

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Salvo-Garrido, H., S. Travella, T. Schwarzacher, W. A. Harwood, and J. W. Snape. "An efficient method for the physical mapping of transgenes in barley using in situ hybridization." Genome 44, no. 1 (February 1, 2001): 104–10. http://dx.doi.org/10.1139/g00-090.

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The genetic transformation of crops by particle bombardment and Agrobacterium tumefaciens systems have the potential to complement conventional plant breeding programmes. However, before deployment, transgenic plants need to be characterized in detail, and physical mapping is an integral part of this process. Therefore, it is important to have a highly efficient method for transgene detection by fluorescence in situ hybridization (FISH). This study describes a new approach, which provides efficient control of probe length and labelling, both of which play an important role in in situ hybridization of transgenes. The approach is based on reducing the size of the plasmid prior to labelling by nick translation, rather than using the whole or linearized plasmid, or varying the amounts of DNaseI in the nick translation mixture. This provided much more efficient labelling of the probe, which yielded optimal hybridization, minimal fluorescent background, and accurate physical location of the transgene.Key words: barley, transformation, FISH, transgene detection, probe design.

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Liu, Q., S. Salih, J. Ingersoll, R. Meng, L. Owens, and F. Hammerschlag. "084 Response of Transgenic `Royal Gala' Apple (Malus ×domestica Borkh.) Shoots, Containing the Modified Cecropin MB39 Gene, to Erwinia amylovora." HortScience 35, no. 3 (June 2000): 403B—403. http://dx.doi.org/10.21273/hortsci.35.3.403b.

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Transgenic `Royal Gala' apple (Malus × domestica Borkh.) shoots were obtained by Agrobacterium-mediated gene transfer using the plasmid binary vector pGV-osm-AC with a T-DNA encoding a chimeric gene consisting of a secretory sequence from barley-amylase joined to the modified cecropin MB39 coding sequence. Shoots were placed under the control of a wound-inducible, osmotin promoter from tobacco. The integration of the cecropin MB39 gene into apple was confirmed by Southern blot analysis. The transformation efficiency was 1.5% when internodes from etiolated shoots were used as explants and 2% when leaf explants were used. Both non- and transgenic tetraploid plants were produced by treatment of leaf explants with colchicine at 25 mg·L-1, and polyploidy was confirmed by flow cytometry. Of the diploid transgenics, three of seven were significantly more resistant to Erwinia amylovora than the non-transgenic `Royal Gala' control. Also, in one instance, a tetraploid transgenic was significantly more resistant than the diploid shoot from which it was derived.

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Shim, Youn-Seb, K. Peter Pauls, and Ken J. Kasha. "Transformation of isolated barley (Hordeum vulgare L.) microspores: II. Timing of pretreatment and temperatures relative to results of bombardment." Genome 52, no. 2 (February 2009): 175–90. http://dx.doi.org/10.1139/g08-113.

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Based on paper I in this series, our goals in this paper were to determine the relationship between prebombardment pretreatments and temperatures, microspore cell cycle when bombarded, and the frequencies of homozygous and hemizygous transgenic progeny in barley ( Hordeum vulgare L.). Of the 104 fluorescent plants selected when using the GFP fluorescence transgene, 28 were albino and 76 plants were green. Thirty-one green plants were confirmed to be transgenic; the others were either transient green fluorescent protein expression or selected due to autofluorescence. Of the 31 plants, 23 came from embryos expressing a high level of fluorescence during selection and eight from 51 plants exhibiting a low level of fluorescence. Of the two pretreatments used to induce embryogenesis, 24 of 31 plants were from the cold pretreatment for 21 days (C) versus seven from the 4 day cold plus mannitol pretreatment. Following pretreatment, the microspores were subjected to a high-osmotic period (0.5 mol/L mannitol plus sorbitol) of 4 h prebombardment and 18 h postbombardment at either 25 or 4 °C. Of the 31 transgenic plants, 19 were produced following the 25 °C 4 h prebombardment. Sixteen of the 19 were doubled haploid plants (seven being homozygous for the transgene) and the other three plants were haploid. Of the remaining 12 plants recovered following the 4 h 4 °C prebombardment treatment, nine were haploid and three were doubled haploid plants, two of the latter being homozygous for the transgene. All 12 haploid plants obtained were treated with colchicine and produced homozygous transgenic doubled haploids. Of the two promoters compared, 30 plants had the actin promoter and only one had the 35S promoter. The use of arabinogalactan protein in the culture medium was very beneficial, giving rise to 29 of the 31 plants. The best procedure for obtaining transgenic barley plants from this study was pretreatment C, leaving the cultures at either 4 or 25 °C during the 4 h prebombardment high-osmotic period, using the actin promoter and having arabinogalactan protein in the microspore culture medium. With this procedure, the transgenic frequency was improved 8- to10-fold over previous reports on bombardment of microspores. It yielded about one transgenic plant per Petri dish and is comparable with Agrobacterium frequencies on structures derived from microspores.

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Gruber, Benjamin D., Emmanuel Delhaize, Alan E. Richardson, Ute Roessner, Richard A. James, Susan M. Howitt, and Peter R. Ryan. "Characterisation of HvALMT1 function in transgenic barley plants." Functional Plant Biology 38, no. 2 (2011): 163. http://dx.doi.org/10.1071/fp10140.

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HvALMT1 from barley (Hordeum vulgare L.) encodes a protein capable of facilitating the transport of malate and other organic anions when expressed in Xenopus oocytes. The HvALMT1 gene is primarily expressed in guard cells of stomata, in regions behind the root apex and at lateral root junctions. We investigated the function of HvALMT1 in planta by overexpressing it in barley under the control of a constitutive promoter. Transgenic plants expressing HvALMT1 at levels four to 9-fold greater than controls showed reduced growth and plants showing the highest expression failed to set seed. Although measurements of conductance indicated that stomatal function was not totally impaired in the transgenic plants the time taken for the stomata to close in response to low light was significantly longer compared with controls. Elemental and metabolomic analyses of the transgenic barley shoots revealed that the concentration of calcium and levels of ascorbate, serine, threonine and pentanoate were consistently greater (2- to 14-fold) in plants that overexpressed HvALMT1, whereas whole-shoot tissue levels of fumarate were significantly lower (60–85% reduction). Transgenic plants also showed significantly greater efflux of malate and succinate from their roots than control plants. Efflux of these organic anions occurred independently of Al3+ and conferred greater Al3+ resistance in solution culture and in acidic soil. These results are consistent with HvALMT1 contributing to anion homeostasis in the cytosol and osmotic adjustment by transporting organic anions out of the cell or by sequestering them into cytosolic vesicles.

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Kosová, K., J. Chrpová, and V. Šíp. "Recent advances in breeding of cereals for resistance to barley yellow dwarf virus." Czech Journal of Genetics and Plant Breeding 44, No. 1 (March 28, 2008): 1–10. http://dx.doi.org/10.17221/6/2008-cjgpb.

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The review focuses on recent progress in the breeding of small grain cereals (barley, wheat, oats) for resistance to the barley yellow dwarf virus (BYDV). First, the symptomatology of barley yellow dwarf (BYD) disease is briefly described and the genome of BYDV, its serotypes and mechanisms of its replication and translation in host plants are characterized. Great attention is paid to the description of resistance genes and sources of BYDV resistance that are currently used in some breeding programmes of barley, wheat and oats. In barley, the introduction of the Ryd2 gene into high-yielding cultivars is still desirable. An example of recent success reached in a European programme aimed at a pyramiding of resistance genes is the registration of the Italian feeding barley cultivar Doria, carrying resistance genes Ryd2, rym4 and Rdg1. The release of this cultivar resulted from the cooperation between EICR, Fiorenzuola d’Arda and CRI in Prague-Ruzyně in the field of virus resistance. Finally, some experiments employing transgenic techniques in the construction of resistant plants are mentioned. In conclusion, the advantages and disadvantages of classical breeding methods using crossing and transgenic techniques are compared and newly arising approaches are discussed.

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Escudero-Martinez, Carmen, Patricia A. Rodriguez, Shan Liu, Pablo A. Santos, Jennifer Stephens, and Jorunn I. B. Bos. "An aphid effector promotes barley susceptibility through suppression of defence gene expression." Journal of Experimental Botany 71, no. 9 (January 28, 2020): 2796–807. http://dx.doi.org/10.1093/jxb/eraa043.

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Abstract Aphids secrete diverse repertoires of effectors into their hosts to promote the infestation process. While ‘omics’ approaches facilitated the identification and comparison of effector repertoires from a number of aphid species, the functional characterization of these proteins has been limited to dicot (model) plants. The bird cherry-oat aphid Rhopalosiphum padi is a pest of cereal crops, including barley. Here, we extend efforts to characterize aphid effectors with regard to their role in promoting susceptibility to the R. padi–barley interaction. We selected three R. padi effectors based on sequence similarity to previously characterized Myzus persicae effectors and assessed their subcellular localization, expression, and role in promoting plant susceptibility. Expression of R. padi effectors RpC002 and Rp1 in transgenic barley lines enhanced plant susceptibility to R. padi but not M. persicae, for which barley is a poor host. Characterization of Rp1 transgenic barley lines revealed reduced gene expression of plant hormone signalling genes relevant to plant–aphid interactions, indicating that this effector enhances susceptibility by suppressing plant defences in barley. Our data suggest that some aphid effectors specifically function when expressed in host species, and feature activities that benefit their corresponding aphid species.

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Bregitzer, P., S. E. Halbert, and P. G. Lemaux. "Somaclonal variation in the progeny of transgenic barley." Theoretical and Applied Genetics 96, no. 3-4 (March 1998): 421–25. http://dx.doi.org/10.1007/s001220050758.

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Kamenarova, K., K. Gecheff, M. Stoyanova, Y. Muhovski, H. Anzai, and A. Atanassov. "Production of Recombinant Human Lactoferin in Transgenic Barley." Biotechnology & Biotechnological Equipment 21, no. 1 (January 2007): 18–27. http://dx.doi.org/10.1080/13102818.2007.10817407.

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Furtado, Agnelo, Robert J. Henry, and Alessandro Pellegrineschi. "Analysis of promoters in transgenic barley and wheat." Plant Biotechnology Journal 7, no. 3 (April 2009): 240–53. http://dx.doi.org/10.1111/j.1467-7652.2008.00394.x.

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Zombori, Zoltán, Bettina Nagy, Róbert Mihály, János Pauk, András Cseri, László Sass, Gábor Horváth V., and Dénes Dudits. "RING-Type E3 Ubiqitin Ligase Barley Genes (HvYrg1–2) Control Characteristics of Both Vegetative Organs and Seeds as Yield Components." Plants 9, no. 12 (December 2, 2020): 1693. http://dx.doi.org/10.3390/plants9121693.

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Previously, studies on RING-type E3 ubiquitin ligases in cereals were preferentially focused on GW2 genes primarily controlling seed parameters in rice and wheat. Here we report cloning two HvYrg genes from barley that share significant homology with rice GW2 gene. In antisense genotypes efficiency of gene silencing varied between genes and transgenic lines: ASHvYrg1: 30–50% and ASHvYrg2: 20–27%. Reduced activity of both genes altered shoot system with increasing number of side shoots. Changes in leaf width, weight, or plant weight and height reached significant levels in some transgenic lines. Lowering expression of the two barley HvYrg genes caused opposite responses in spike development. Plants with ASHvYrg1 gene construct showed earlier heading time and prolonged grain-filling period, while plants from ASHvYrg2 genotype flowered in delay. Digital imaging of root development revealed that down-regulation of HvYrg1 gene variant stimulated root growth, while ASHvYrg2 plants developed reduced root system. Comparison of seed parameters indicated an increase in thousand grain weight accompanied with longer and wider seed morphology. In summary we conclude that in contrast to inhibition of GW2 genes in rice and wheat plants, down-regulation of the barely HvYrg genes caused substantial changes in vegetative organs in addition to alteration of seed parameters.

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Partridge-Telenko, D. E., J. Hu, D. M. Livingstone, B. B. Shew, P. M. Phipps, and E. A. Grabau. "Sclerotinia Blight Resistance in Virginia-Type Peanut Transformed with a Barley Oxalate Oxidase Gene." Phytopathology® 101, no. 7 (July 2011): 786–93. http://dx.doi.org/10.1094/phyto-10-10-0266.

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Transgenic peanut lines expressing oxalate oxidase, a novel enzyme to peanut, were evaluated for resistance to Sclerotinia blight in naturally infested fields over a 5-year period. Area under the disease progress curve (AUDPC) for transgenic lines in single rows planted with seed from single-plant selections averaged 78, 83, and 90% lower than nontransgenic parents in 2004, 2005, and 2006, respectively. In addition, AUDPC in 14 transgenic lines planted with bulked seed in two-row plots averaged 81% lower compared with nontransgenic parents in 2005 and 86% lower in 16 transgenic lines in 2006. Six transgenic lines yielded 488 to 1,260 kg/ha greater than nontransgenic parents in 2005, and 10 lines yielded 537 to 2,490 kg/ha greater in 2006. Fluazinam (0.58 kg a.i./ha) fungicide sprays in 2008 and 2009 reduced AUDPC in transgenic and nontransgenic lines but AUDPC was lowest in transgenic lines. Without fluazinam, yields of transgenic lines averaged 1,133 to 1,578 kg/ha greater than nontransgenic lines in 2008 and 1,670 to 2,755 kg/ha greater in 2009. These results demonstrated that the insertion of barley oxalate oxidase in peanut conveyed a high level of resistance to Sclerotinia blight, and negated the need for costly fungicide sprays.

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Burbridge, E., S. K. Rasmussen, F. Bernier, B. K. Kristensen, P. F. McCabe, and P. J. Dix. "Altered Activity of Peroxidase and Oxalate Oxidase Influences Lignification in Transgenic Tobacco." Open Plant Science Journal 8, no. 1 (June 27, 2014): 1–8. http://dx.doi.org/10.2174/1874294701408010001.

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Peroxidase and hydrogen peroxide both play important roles in the final stages of the lignification pathway. Peroxidase, in the presence of H2O2 catalyses the oxidation of monolignols to give lignin. In order to examine this process we looked at lignification in transgenic tobacco plants expressing a barley peroxidase gene, HvPrx8, either alone or in combination with a wheat germin gene, g.f 2.8, which encodes oxalate oxidase, thereby providing a source of H2O2. Elevated activity of the antioxidant ascorbate peroxidase was found in plants expressing oxalate peroxidase and was greatly increased by co-expression with the barley peroxidase, although the latter had no effect when expressed alone. An increase was observed in the oxidation of the lignin monomer, syringaldazine in cell lines over-expressing barley peroxidase, while a decrease was observed in double transformants. Plants over-expressing barley peroxidase have elevated levels of lignin deposition compared to that of wild type tobacco plants. Over-expression of the individual enzymes was also shown to enhance heat-induced programmed cell death (PCD) in cell suspension cultures, an effect which was greatly reduced in the double-expressing lines.

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Mirzaee, Malihe, Edita Holásková, Alžbeta Mičúchová, David J. Kopečný, Zhila Osmani, and Ivo Frébort. "Long-Lasting Stable Expression of Human LL-37 Antimicrobial Peptide in Transgenic Barley Plants." Antibiotics 10, no. 8 (July 23, 2021): 898. http://dx.doi.org/10.3390/antibiotics10080898.

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Antimicrobial peptides play a crucial role in the innate immune system of multicellular organisms. LL-37 is the only known member of the human cathelicidin family. As well as possessing antibacterial properties, it is actively involved in various physiological responses in eukaryotic cells. Accordingly, there is considerable interest in large-scale, low-cost, and microbial endotoxin-free production of LL-37 recombinant peptides for pharmaceutical applications. As a heterologous expression biofactory, we have previously obtained homologous barley (Hordeum vulgare L.) as an attractive vehicle for producing recombinant human LL-37 in the grain storage compartment, endosperm. The long-term stability of expression and inheritance of transgenes is necessary for the successful commercialization of recombinant proteins. Here, we report the stable inheritance and expression of the LL-37 gene in barley after six generations, including two consecutive seasons of experimental field cultivation. The transgenic plants showed normal growth and remained fertile. Based on the bacteria viability test, the produced peptide LL-37 retained high antibacterial activity.

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Christensen, Anders B., Hans Thordal-Christensen, Grit Zimmermann, Torben Gjetting, Michael F. Lyngkjær, Robert Dudler, and Patrick Schweizer. "The Germinlike Protein GLP4 Exhibits Superoxide Dismutase Activity and Is an Important Component of Quantitative Resistance in Wheat and Barley." Molecular Plant-Microbe Interactions® 17, no. 1 (January 2004): 109–17. http://dx.doi.org/10.1094/mpmi.2004.17.1.109.

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Germinlike proteins (GLP) are encoded in plants by a gene family with proposed functions in plant development and defense. Genes of GLP subfamily 4 of barley (HvGLP4, formerly referred to as HvOxOLP) and the wheat orthologue TaGLP4 (formerly referred to as TaGLP2a) were previously found to be expressed in pathogen-attacked epidermal tissue of barley and wheat leaves, and the corresponding proteins are proposed to accumulate in the apoplast. Here, the role of HvGLP4 and TaGLP4 in the defense of barley and wheat against Blumeria graminis (DC.) E. O. Speer, the cereal powdery mildew fungus, was examined in an epidermal transient expression system and in transgenic Arabidopsis thaliana plants overexpressing His-tagged HvGLP4. Leaf extracts of transgenic Arabidopsis overexpressing HvGLP4 contained a novel His-tagged protein with superoxide dismutase activity and HvGLP4 epitopes. Transient overexpression of TaGLP4 and HvGLP4 enhanced resistance against B. graminis in wheat and barley, whereas transient silencing by RNA interference reduced basal resistance in both cereals. The effect of GLP4 overexpression or silencing was strongly influenced by the genotype of the plant. The data suggest that members of GLP subfamily 4 are components of quantitative resistance in both barley and wheat, acting together with other, as yet unknown, plant components.

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Ritala, A., A. M. Nuutila, R. Aikasalo, V. Kauppinen, and J. Tammisola. "Measuring Gene Flow in the Cultivation of Transgenic Barley." Crop Science 42, no. 1 (2002): 278. http://dx.doi.org/10.2135/cropsci2002.0278.

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Ritala, A., A. M. Nuutila, R. Aikasalo, V. Kauppinen, and J. Tammisola. "Measuring Gene Flow in the Cultivation of Transgenic Barley." Crop Science 42, no. 1 (January 2002): 278–85. http://dx.doi.org/10.2135/cropsci2002.2780.

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32

Higuchi, Kyoko, Michiko Takahashi, Hiromi Nakanishi, Shinji Kawasaki, Naoko K. Nishizawa, and Satoshi Mori. "Analysis of transgenic rice containing barley nicotianamine synthase gene." Soil Science and Plant Nutrition 47, no. 2 (June 2001): 315–22. http://dx.doi.org/10.1080/00380768.2001.10408395.

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Horvath, H., J. Huang, O. Wong, E. Kohl, T. Okita, C. G. Kannangara, and D. von Wettstein. "The production of recombinant proteins in transgenic barley grains." Proceedings of the National Academy of Sciences 97, no. 4 (February 4, 2000): 1914–19. http://dx.doi.org/10.1073/pnas.030527497.

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34

Salmenkallio-Marttila, M., K. Aspegren, S. Åkerman, U. Kurtén, L. Mannonen, A. Ritala, T. H. Teeri, and V. Kauppinen. "Transgenic barley (Hordeum vulgare L.) by electroporation of protoplasts." Plant Cell Reports 15, no. 3-4 (December 1995): 301–4. http://dx.doi.org/10.1007/bf00193741.

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35

Ritala, Anneli, Kristian Aspegren, Ulrika Kurt�n, Marjatta Salmenkallio-Marttila, Leena Mannonen, Riitta Hannus, Veli Kauppinen, Teemu H. Teeri, and Tor-Magnus Enari. "Fertile transgenic barley by particle bombardment of immature embryos." Plant Molecular Biology 24, no. 2 (January 1994): 317–25. http://dx.doi.org/10.1007/bf00020170.

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36

Mrízová, Katarína, Edita Holasková, M. Tufan Öz, Eva Jiskrová, Ivo Frébort, and Petr Galuszka. "Transgenic barley: A prospective tool for biotechnology and agriculture." Biotechnology Advances 32, no. 1 (January 2014): 137–57. http://dx.doi.org/10.1016/j.biotechadv.2013.09.011.

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Luan, Haiye, Baojian Guo, Huiquan Shen, Yuhan Pan, Yi Hong, Chao Lv, and Rugen Xu. "Overexpression of Barley Transcription Factor HvERF2.11 in Arabidopsis Enhances Plant Waterlogging Tolerance." International Journal of Molecular Sciences 21, no. 6 (March 13, 2020): 1982. http://dx.doi.org/10.3390/ijms21061982.

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Waterlogging stress significantly affects the growth, development, and productivity of crop plants. However, manipulation of gene expression to enhance waterlogging tolerance is very limited. In this study, we identified an ethylene-responsive factor from barley, which was strongly induced by waterlogging stress. This transcription factor named HvERF2.11 was 1158 bp in length and encoded 385 amino acids, and mainly expressed in the adventitious root and seminal root. Overexpression of HvERF2.11 in Arabidopsis led to enhanced tolerance to waterlogging stress. Further analysis of the transgenic plants showed that the expression of AtSOD1, AtPOD1 and AtACO1 increased rapidly, while the same genes did not do so in non-transgenic plants, under waterlogging stress. Activities of antioxidant enzymes and alcohol dehydrogenase (ADH) were also significantly higher in the transgenic plants than in the non-transgenic plants under waterlogging stress. Therefore, these results indicate that HvERF2.11 plays a positive regulatory role in plant waterlogging tolerance through regulation of waterlogging-related genes, improving antioxidant and ADH enzymes activities.

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Li, Huanpeng, Jiaojiao Wu, Xiaofeng Shang, Miaomiao Geng, Jing Gao, Shuqing Zhao, Xiumei Yu, et al. "WRKY Transcription Factors Shared by BTH-Induced Resistance and NPR1-Mediated Acquired Resistance Improve Broad-Spectrum Disease Resistance in Wheat." Molecular Plant-Microbe Interactions® 33, no. 3 (March 2020): 433–43. http://dx.doi.org/10.1094/mpmi-09-19-0257-r.

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In Arabidopsis, both pathogen invasion and benzothiadiazole (BTH) treatment activate the nonexpresser of pathogenesis-related genes 1 (NPR1)-mediated systemic acquired resistance, which provides broad-spectrum disease resistance to secondary pathogen infection. However, the BTH-induced resistance in Triticeae crops of wheat and barley seems to be accomplished through an NPR1-independent pathway. In the current investigation, we applied transcriptome analysis on barley transgenic lines overexpressing wheat wNPR1 (wNPR1-OE) and knocking down barley HvNPR1 (HvNPR1-Kd) to reveal the role of NPR1 during the BTH-induced resistance. Most of the previously designated barley chemical-induced (BCI) genes were upregulated in an NPR1-independent manner, whereas the expression levels of several pathogenesis-related (PR) genes were elevated upon BTH treatment only in wNPR1-OE. Two barley WRKY transcription factors, HvWRKY6 and HvWRKY70, were predicted and further validated as key regulators shared by the BTH-induced resistance and the NPR1-mediated acquired resistance. Wheat transgenic lines overexpressing HvWRKY6 and HvWRKY70 showed different degrees of enhanced resistance to Puccinia striiformis f. sp. tritici pathotype CYR32 and Blumeria graminis f. sp. tritici pathotype E20. In conclusion, the transcriptional changes of BTH-induced resistance in barley were initially profiled, and the identified key regulators would be valuable resources for the genetic improvement of broad-spectrum disease resistance in wheat. [Formula: see text] Copyright © 2020 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

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Hensel, Goetz, Christine Kastner, Sylwia Oleszczuk, Jan Riechen, and Jochen Kumlehn. "Agrobacterium-Mediated Gene Transfer to Cereal Crop Plants: Current Protocols for Barley, Wheat, Triticale, and Maize." International Journal of Plant Genomics 2009 (June 21, 2009): 1–9. http://dx.doi.org/10.1155/2009/835608.

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The development of powerful “omics” technologies has enabled researchers to identify many genes of interest for which comprehensive functional analyses are highly desirable. However, the production of lines which ectopically express recombinant genes, or those in which endogenous genes are knocked down via stable transformation, remains a major bottleneck for the association between genetics and gene function in monocotyledonous crops. Methods of effective DNA transfer into regenerable cells of immature embryos from cereals by means of Agrobacterium tumefaciens have been modified in a stepwise manner. The effect of particular improvement measures has often not been significantly evident, whereas their combined implementation has resulted in meaningful advances. Here, we provide updated protocols for the Agrobacterium-mediated generation of stably transgenic barley, wheat, triticale and maize. Based upon these methods, several hundred independent transgenic lines have been delivered, with efficiencies of inoculated embryos leading to stably transgenic plants reaching 86% in barley, 10% in wheat, 4% in triticale, and 24% in maize.

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40

Cho, M. J., H. W. Choi, B. B. Buchanan, and P. G. Lemaux. "Inheritance of tissue-specific expression of barley hordein promoter-uidA fusions in transgenic barley plants." Theoretical and Applied Genetics 98, no. 8 (June 1999): 1253–62. http://dx.doi.org/10.1007/s001220051191.

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41

Aksoy, Emre, Amir Maqbool, and Buasimuhan Abudureyimu. "Arpa Nikotinamin Sentaz1 (HvNAS1) Genini Yüksek Seviyede İfade Eden Arabidopsis thaliana Bitkileri Demir Eksikliğine Dayanıklılık Gösterir." Turkish Journal of Agriculture - Food Science and Technology 8, sp1 (December 12, 2020): 70–79. http://dx.doi.org/10.24925/turjaf.v8isp1.70-79.3974.

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Iron (Fe) is an important trace mineral for plant development, and plants grown in Fe deficiency experience yield losses due to the leaf chlorosis. In addition to agronomic measures that can be taken to minimize these losses, new plant genotypes can be developed effectively through genetic engineering. While dicots such as Arabidopsis thaliana use a reduction-based strategy to uptake high amounts of iron from the rhizosphere, the chelation strategy has evolved in Gramineous plants including barley (Hordeum vulgare). In this study, barley NICOTIANAMINE SYNTHASE1 (HvNAS1) gene, which is responsible for the production of nicotianamine that can complex with iron, was cloned and expressed at a constitutive high level in Arabidopsis plants. The expression levels of Arabidopsis genes encoding for the proteins involved in iron uptake increased together with HvNAS1 in the T3 Arabidopsis plants. Moreover, the root lengths, root and stem fresh weights, ferric chelate reductase enzyme activities of the plants also increased in the transgenic Arabidopsis plants under Fe deficiency. In addition, significant increases in iron and zinc levels were determined in the roots and shoots of transgenic Arabidopsis plants. As a result, transgenic Arabidopsis plants overexpressing the barley HvNAS1 gene can take up more iron from the rhizosphere and carry this iron to the shoots. This study demonstrates the power of genetic engineering to develop Arabidopsis plants overexpressing the HvNAS1 gene and therefore tolerate iron deficiency.

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Gomez-Sanchez, Andrea, M. Estrella Santamaria, Pablo Gonzalez-Melendi, Aleksandra Muszynska, Christiane Matthess, Manuel Martinez, and Isabel Diaz. "Repression of barley cathepsins, HvPap-19 and HvPap-1, differentially alters grain composition and delays germination." Journal of Experimental Botany 72, no. 9 (January 17, 2021): 3474–85. http://dx.doi.org/10.1093/jxb/erab007.

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Abstract During barley germination, cysteine proteases are essential in the mobilization of storage compounds providing peptides and amino acids to sustain embryo growth until photosynthesis is completely established. Knockdown barley plants, generated by artificial miRNA, for the cathepsins B- and F-like HvPap-19 and HvPap-1 genes, respectively, showed less cysteine protease activities and consequently lower protein degradation. The functional redundancy between proteases triggered an enzymatic compensation associated with an increase in serine protease activities in both knockdown lines, which was not sufficient to maintain germination rates and behaviour. Concomitantly, these transgenic lines showed alterations in the accumulation of protein and carbohydrates in the grain. While the total amount of protein increased in both transgenic lines, the starch content decreased in HvPap-1 knockdown lines and the sucrose concentration was reduced in silenced HvPap-19 grains. Consequently, phenotypes of HvPap-1 and HvPap-19 artificial miRNA lines showed a delay in the grain germination process. These data demonstrate the potential of exploring the properties of barley proteases for selective modification and use in brewing or in the livestock feeding industry.

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Adem, Getnet D., Stuart J. Roy, Yuqing Huang, Zhong-Hua Chen, Feifei Wang, Meixue Zhou, John P. Bowman, Paul Holford, and Sergey Shabala. "Expressing Arabidopsis thaliana V-ATPase subunit C in barley (Hordeum vulgare) improves plant performance under saline condition by enabling better osmotic adjustment." Functional Plant Biology 44, no. 12 (2017): 1147. http://dx.doi.org/10.1071/fp17133.

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Salinity is a global problem affecting agriculture that results in an estimated US$27 billion loss in revenue per year. Overexpression of vacuolar ATPase subunits has been shown to be beneficial in improving plant performance under saline conditions. Most studies, however, have not shown whether overexpression of genes encoding ATPase subunits results in improvements in grain yield, and have not investigated the physiological mechanisms behind the improvement in plant growth. In this study, we constitutively expressed Arabidopsis Vacuolar ATPase subunit C (AtVHA-C) in barley. Transgenic plants were assessed for agronomical and physiological characteristics, such as fresh and dry biomass, leaf pigment content, stomatal conductance, grain yield, and leaf Na+ and K+ concentration, when grown in either 0 or 300 mM NaCl. When compared with non-transformed barley, AtVHA-C expressing barley lines had a smaller reduction in both biomass and grain yield under salinity stress. The transgenic lines accumulated Na+ and K+ in leaves for osmotic adjustment. This in turn saves energy consumed in the synthesis of organic osmolytes that otherwise would be needed for osmotic adjustment.

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Jamahari, Azreena, Wong Ling-Chie, Fan Xioalai, Liu Qiaoquan, Leong Sui Sien, Fauziah Abu Bakar, Amy Halimah Rajaee, Patricia King Jie Hung, Hairul Azman Roslan, and Wong Sie Chuong. "CHARACTERISATION OF HORDEUM VULGARE CELLULOSE SYNTHASE-LIKE F6 PROMOTER VIA TRANSGENE EXPRESSION IN RICE." Malaysian Journal of Science 40, no. 2 (June 30, 2021): 61–86. http://dx.doi.org/10.22452/mjs.vol40no2.6.

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Beta-glucan in cereal crops is known as a functional food, which can reduce cardiovascular diseases by lowering blood cholesterol levels. However, beta-glucan content is relatively low in rice grains, despite being relatively abundant in barley and oat grains. Taking advantage of rice as the staple food for Asians, increasing beta-glucan content in rice for their consumption may help to reduce cardiovascular-related diseases among them. Previous attempts in increasing beta-glucan content in rice via transgene expression of beta-glucan synthase genes from barley into rice were unsuccessful due to the use of non-tissue specific as well as constitutively expressing promoter. The current transgenic expression study was performed to characterise the promoter of beta-glucan synthase gene in barley using beta-glucuronidase (GUS) reporter gene. Two fragments of HvCslF6 promoter (2771 bp and 1257 bp) were successfully fused with GUS reporter gene and integrated into rice plants, demonstrated that the promoter was functional in the heterologous plant system. The presence of blue GUS staining was observed on the leaf, root, stem, and grain of the transgenic rice regardless of the promoter length used and stayed functional up to the next generation. GUS qualitative analysis confirmed that the shorter promoter length generated a stronger GUS activity in comparison to the longer one. This indicated that the presence of repressor elements in between the -2771 bp and -1257 bp regions. The preliminary results shed light on the strong promoter activity in the rice endosperm tissue. It can become an alternative to the collection of plant promoters that can be used for grain quality improvement and biofortification.

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Baah, J., T. A. Scott, L. M. Kawchuk, J. D. Armstrong, L. B. Selinger, K. J. Cheng, and T. A. McAllister. "Feeding value in broiler chicken diets of a potato expressing a β-glucanase gene from Fibrobacter succinogenes." Canadian Journal of Animal Science 82, no. 1 (March 1, 2002): 111–13. http://dx.doi.org/10.4141/a01-007.

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Including a transgenic potato cultivar as a source of β-glucanase in CDC Silky barley-based diets for broiler chickens at 0.6 kg t–1 improved (P < 0.05) feed conversion by 8.8%; at 1.2 kg t–1 it reduced (P < 0.05) ileal digesta viscosity by 42%. With improved level and/or activity of expression, transformed potato may have potential as an enzyme additive. Key words: Barley, broilers, β-glucanase, performance, transformed potato

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Nagahatenna, Dilrukshi S. K., Jingwen Tiong, Everard J. Edwards, Peter Langridge, and Ryan Whitford. "Altering Tetrapyrrole Biosynthesis by Overexpressing Ferrochelatases (Fc1 and Fc2) Improves Photosynthetic Efficiency in Transgenic Barley." Agronomy 10, no. 9 (September 11, 2020): 1370. http://dx.doi.org/10.3390/agronomy10091370.

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Ferrochelatase (FC) is the terminal enzyme of heme biosynthesis. In photosynthetic organisms studied so far, there is evidence for two FC isoforms, which are encoded by two genes (FC1 and FC2). Previous studies suggest that these two genes are required for the production of two physiologically distinct heme pools with only FC2-derived heme involved in photosynthesis. We characterised two FCs in barley (Hordeum vulgare L.). The two HvFC isoforms share a common catalytic domain, but HvFC2 additionally contains a C-terminal chlorophyll a/b binding (CAB) domain. Both HvFCs are highly expressed in photosynthetic tissues, with HvFC1 transcripts also being abundant in non-photosynthetic tissues. To determine whether these isoforms differentially affect photosynthesis, transgenic barley ectopically overexpressing HvFC1 and HvFC2 were generated and evaluated for photosynthetic performance. In each case, transgenics exhibited improved photosynthetic rate (Asat), stomatal conductance (gs) and carboxylation efficiency (CE), showing that both FC1 and FC2 play important roles in photosynthesis. Our finding that modified FC expression can improve photosynthesis up to ~13% under controlled growth conditions now requires further research to determine if this can be translated to improved yield performance under field conditions.

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Oraby, Hesham F., Callista B. Ransom, Alexandra N. Kravchenko, and Mariam B. Sticklen. "Barley HVA1 Gene Confers Salt Tolerance in R3 Transgenic Oat." Crop Science 45, no. 6 (November 2005): 2218–27. http://dx.doi.org/10.2135/cropsci2004-0605.

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Kovalchuk, Nataliya, Wei Jia, Omid Eini, Sarah Morran, Tatiana Pyvovarenko, Stephen Fletcher, Natalia Bazanova, et al. "Optimization ofTaDREB3gene expression in transgenic barley using cold-inducible promoters." Plant Biotechnology Journal 11, no. 6 (March 16, 2013): 659–70. http://dx.doi.org/10.1111/pbi.12056.

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Gatford, Keith T., Zainuddin Basri, Jane Edlington, Julia Lloyd, Javed A. Qureshi, Richard Brettell, and Geoffrey B. Fincher. "Gene flow from transgenic wheat and barley under field conditions." Euphytica 151, no. 3 (September 29, 2006): 383–91. http://dx.doi.org/10.1007/s10681-006-9160-1.

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50

Funatsuki, H., H. Kuroda, M. Kihara, P. A. Lazzeri, E. Müller, H. Lörz, and I. Kishinami. "Fertile transgenic barley generated by direct DNA transfer to protoplasts." Theoretical and Applied Genetics 91, no. 5 (October 1995): 707–12. http://dx.doi.org/10.1007/bf00220947.

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Journal articles on the topic 'Transgenic barley'

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