Immature pollen-derived doubled haploid formation in barley cv. Golden Promise as a tool for transgene recombination

Coronado, Marı́a-José; Hensel, Goetz; Broeders, Sylvia; Otto, Ingrid; Kumlehn, Jochen

doi:10.1007/s11738-005-0063-x

Cited by 44 publications

(53 citation statements)

References 45 publications

(20 reference statements)

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“…Afterward, four to six T1 events were preliminarily used for molecular characterization. Homozygous material was generated via embryogenic pollen cultures (Coronado et al, 2005), and obtained homozygous plants were analyzed in depth. Two HvPap-1 over-expressing lines (OE Pap1: 919 and 937), two HvPap-1 silencing lines (KD Pap1: 1130 and 1175), and two Icy-2 silencing lines (KD Icy2: 1318 and 1399) were selected based on transgene copy number, transcript, and protein content for further studies (Supplemental Figs.…”

Section: Resultsmentioning

confidence: 99%

HvPap-1 C1A Protease and HvCPI-2 Cystatin Contribute to Barley Grain Filling and Germination

et al. 2016

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Proteolysis is an essential process throughout the mobilization of storage proteins in barley (Hordeum vulgare) grains during germination. It involves numerous types of enzymes, with C1A Cys proteases the most abundant key players. Manipulation of the proteolytic machinery is a potential way to enhance grain yield and quality, and it could influence the mobilization of storage compounds along germination. Transgenic barley plants silencing or over-expressing the cathepsin F-like HvPap-1 Cys protease show differential accumulation of storage molecules such as starch, proteins, and free amino acids in the grain. It is particularly striking that the HvPap-1 artificial microRNA lines phenotype show a drastic delay in the grain germination process. Alterations to the proteolytic activities in the over-expressing and knock-down grains associated with changes in the level of expression of several C1A peptidases were also detected. Similarly, down-regulating cystatin Icy-2, one of the proteinaceous inhibitors of the cathepsin F-like protease, also has important effects on grain filling. However, the ultimate physiological influence of manipulating a peptidase or an inhibitor cannot be always predicted, since the plant tries to compensate the modified proteolytic effects by modulating the expression of some other peptidases or their inhibitors.

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Section: Resultsmentioning

confidence: 99%

HvPap-1 C1A Protease and HvCPI-2 Cystatin Contribute to Barley Grain Filling and Germination

et al. 2016

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“…Microspores were isolated and purified following the procedures of Coronado et al (2005). The initial densities of microspores in the induction medium were 1.5 × 10 5 per ml.…”

Section: Methodsmentioning

confidence: 99%

The effect of copper on plant regeneration in barley microspore culture

Makowska¹,

Oleszczuk²,

Zimny³

2017

CZECH J GENET PLANT

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Makowska K., Oleszczuk S., Zimny J. (2017): The effect of copper on plant regeneration in barley microspore culture. Czech J. Genet. Plant Breed., 53: 17-22.Isolated microspore culture is an excellent system for the production of doubled haploids in many crops, including barley. In a more traditional barley anther culture method copper sulphate is known to enhance plant regeneration. Here we report that one hundred times higher concentration of copper sulphate in the isolated microspore culture of two spring barley genotypes compared to the standard content in the induction medium resulted in a 34% increase of total plant regeneration. Detailed analysis of plant regeneration showed that additional supplementation of copper sulphate increased not only the regeneration of green plants but also proportionately that of albino plants. Hence, the results from two studied genotypes do not support an assumption that the addition of copper reduces albinism in barley microspore culture.

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“…Two independent homozygous lines (T 3 generation) harboring a single insertion of the transgene were used for the experiments. For the pLea:ABA89OH RNAi construct, doubled haploid plants were produced using embryogenic pollen cultures generated from primary transgenic plants following the protocol previously described by Coronado et al (2005).…”

Section: Transformation Of Barley and Production Of Homozygous Linesmentioning

confidence: 99%

Abscisic Acid Flux Alterations Result in Differential Abscisic Acid Signaling Responses and Impact Assimilation Efficiency in Barley under Terminal Drought Stress

et al. 2014

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Abscisic acid (ABA) is a central player in plant responses to drought stress. How variable levels of ABA under short-term versus long-term drought stress impact assimilation and growth in crops is unclear. We addressed this through comparative analysis, using two elite breeding lines of barley (Hordeum vulgare) that show senescence or stay-green phenotype under terminal drought stress and by making use of transgenic barley lines that express Arabidopsis (Arabidopsis thaliana) 9-cis-epoxycarotenoid dioxygenase (AtNCED6) coding sequence or an RNA interference (RNAi) sequence of ABA 89-hydroxylase under the control of a droughtinducible barley promoter. The high levels of ABA and its catabolites in the senescing breeding line under long-term stress were detrimental for assimilate productivity, whereas these levels were not perturbed in the stay-green type that performed better. In transgenic barley, drought-inducible AtNCED expression afforded temporal control in ABA levels such that the ABA levels rose sooner than in wild-type plants but also subsided, unlike as in the wild type , to near-basal levels upon prolonged stress treatment due to down-regulation of endogenous HvNCED genes. Suppressing of ABA catabolism with the RNA interference approach of ABA 89-hydroxylase caused ABA flux during the entire period of stress. These transgenic plants performed better than the wild type under stress to maintain a favorable instantaneous water use efficiency and better assimilation. Gene expression analysis, protein structural modeling, and protein-protein interaction analyses of the members of the PYRABACTIN RESISTANCE1/PYRABACTIN RESISTANCE1-LIKE/REGULATORY COMPONENT OF ABA RECEPTORS, TYPE 2C PROTEIN PHOSPHATASE Sucrose nonfermenting1-related protein kinase2, and ABA-INSENSITIVE5/ABA-responsive element binding factor family identified specific members that could potentially impact ABA metabolism and stress adaptation in barley.Drought compromises grain yield in cereals, especially when the stress occurs during postanthesis (Boyer and Westgate, 2004;Sreenivasulu et al., 2007).In some crops, selection for and use of stay-green types has been promoted as a means for combating drought stress susceptibility (Thomas and Howarth, 2000). However, the molecular basis of stay-green phenotype in barley (Hordeum vulgare) has not been explored in detail. As the flag leaf is the principal source organ, sustaining its photosynthetic activity under postanthesis drought stress is considered a strategy to mitigate the yield penalty. Water use efficiency (WUE) is the amount of biomass (carbon) accumulated per unit of water, hence an important trait under water-limited conditions (Condon and Richards, 1992;Rebetzke et al., 2002;Richards et al., 2002). Breeding for WUE is considered important for developing drought-tolerant crops (Blum, 1996;Richards, 1996;Richards et al., 2002). Genetic loci that control transpiration efficiency have been identified (Teulat et al., 2002;Hall et al., 2005;Juenger et al., 2005). In Arabidopsis (Arabidopsis tha...

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Immature pollen-derived doubled haploid formation in barley cv. Golden Promise as a tool for transgene recombination

Cited by 44 publications

References 45 publications

HvPap-1 C1A Protease and HvCPI-2 Cystatin Contribute to Barley Grain Filling and Germination

HvPap-1 C1A Protease and HvCPI-2 Cystatin Contribute to Barley Grain Filling and Germination

The effect of copper on plant regeneration in barley microspore culture

Abscisic Acid Flux Alterations Result in Differential Abscisic Acid Signaling Responses and Impact Assimilation Efficiency in Barley under Terminal Drought Stress

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