Introduction and constitutive expression of a rice chitinase gene in bread wheat using biolistic bombardment and the bar gene as a selectable marker

Chen, W. P.; Gu, Xin; Liang, G. H.; Muthukrishnan, Subbaratnam; Chen, P. D.; Liu, D. J.; Gill, Bikram S.

doi:10.1007/s001220051022

Cited by 62 publications

(34 citation statements)

References 44 publications

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“…Standard deviations of mean values are indicated as error bars. On the y-axis, 'wt' stands for wild type (Chen et al 1998). The transformation efficiency achieved in our experiments is comparable to previous reports (2.4% by Bourdon et al 2004;4.0-7.5% by Chugh and Khurana 2003;0.6-3.1% by Tosi et al 2004).…”

Section: Discussionsupporting

confidence: 89%

Expression of the lipid transfer protein Ace-AMP1 in transgenic wheat enhances antifungal activity and defense responses

2006

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To enhance fungal disease resistance, wheat plants (cv. Bobwhite) were engineered to constitutively express the potent antimicrobial protein Ace-AMP1 from Allium cepa, driven by a maize ubiquitin promoter along with its first intron. The bar gene was used for selection of putative transformants on medium containing phosphinothricin (PPT). Transgene inheritance, integration and stability of expression were confirmed over two generations by PCR, Southern, northern and western blot analyses, respectively. The levels of Ace-AMP1 in different transgenic lines correlated with the transcript levels of the transgene. Up to 50% increase in resistance to Blumeria graminis f. sp. tritici was detected in detached leaf assays. In ears of transgenic wheat inoculated with Neovossia indica, Ace-AMP1 intensified expression of defense-related genes. Elevated levels of salicylic acid and of transcripts of phenylalanine ammonia lyase (PAL), glucanase (PR2) and chitinase (PR3) in the transgenic plants indicated manifestation of systemic acquired resistance (SAR).

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Section: Discussionsupporting

confidence: 89%

Expression of the lipid transfer protein Ace-AMP1 in transgenic wheat enhances antifungal activity and defense responses

2006

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“…Transformation: Tissue culture, particle bombardment of immature embryos, and initial screening for the bar gene were done following previously described methods (Altpeter et al 1996;Chen et al 1998;Huang et al 2003). A 5.8-kb subclone containing the WAP2 gene and its native promoter from the LDN 5A BAC was ligated into the pSMART vector (Lucigen, Middleton, WI).…”

Section: Plant Materials and Southern Analysismentioning

confidence: 99%

Molecular Characterization of the Major Wheat Domestication Gene Q

et al. 2006

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The Q gene is largely responsible for the widespread cultivation of wheat because it confers the freethreshing character. It also pleiotropically influences many other domestication-related traits such as glume shape and tenacity, rachis fragility, spike length, plant height, and spike emergence time. We isolated the Q gene and verified its identity by analysis of knockout mutants and transformation. The Q gene has a high degree of similarity to members of the AP2 family of transcription factors. The Q allele is more abundantly transcribed than q, and the two alleles differ for a single amino acid. An isoleucine at position 329 in the Q protein leads to an abundance of homodimer formation in yeast cells, whereas a valine in the q protein appears to limit homodimer formation. Ectopic expression analysis allowed us to observe both silencing and overexpression effects of Q. Rachis fragility, glume shape, and glume tenacity mimicked the q phenotype in transgenic plants exhibiting post-transcriptional silencing of the transgene and the endogenous Q gene. Variation in spike compactness and plant height were associated with the level of transgene transcription due to the dosage effects of Q. The q allele is the more primitive, and the mutation that gave rise to Q occurred only once leading to the world's cultivated wheats. W HEAT, rice, and maize are the three major cereal crops that provide most of the calories consumed by humans. Bread (common) wheat (Triticum aestivum L., 2n ¼ 6x ¼ 42, AABBDD genomes) arose 8000-10,000 years ago (for review see Nesbitt and Samuel 1996;Feldman 2001) from the spontaneous hybridization of the tetraploid wheat T. turgidum L. (2n ¼ 4x ¼ 28, AABB genomes) with the diploid goatgrass Aegilops tauschii Coss. (2n ¼ 2x ¼ 14, DD genomes) (Kihara 1944;McFadden and Sears 1946). Domestication of wheat resulted from mutations that gave rise to traits such as soft glumes, a nonfragile rachis, and the free-threshing character.The Q gene governs the free-threshing character and square spike phenotype. In addition, Q pleiotropically affects a repertoire of other characters important for domestication such as rachis fragility (Leighty and

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“…Western analysis was conducted as described by Chen et al (1998) to demonstrate the expression of the chitinase gene.…”

Section: Western Analysismentioning

confidence: 99%

Agrobacterium-mediated transformation of American ginseng with a rice chitinase gene

Punja

2002

Plant Cell Rep

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Transformation of American ginseng (Panax quinquefolius L.) with Agrobacterium strain LBA4404 containing a rice chitinase gene under control of the maize ubiquitin1 promoter and the phosphinothricin acetyltransferase (bar) and hygromycin phosphotransferase (hpt) genes as selectable markers is described. Epicotyl explants from 2-to 3-week-old ginseng seedlings were pre-cultured for 5-7 days on MS medium supplemented with 10 µM α-naphthaleneacetic acid and 9 µM 2,4-dichlorophenoxyacetic acid (ND medium) prior to Agrobacterium infection. The explants were either immersed in a bacterial suspension for 20 min or received a 10-µl or 15-µl droplet of bacteria. A co-culture period of 3 or 4 days was provided on ND medium with or without acetosyringone and ascorbic acid. Selection for transformed calli was conducted on ND medium containing 20 mg/l phosphinothricin or 100 mg/l hygromycin over a 10-month period. A maximum callusing frequency of 27.7% was achieved on selection medium when explants were infected by the droplet method and co-cultured on ND medium without acetosyringone and ascorbic acid. Almost 90% of the 32 lines that survived selection were shown to be transformed. Immersion of explants reduced the callusing frequency to 9.3%. The presence of the transgenes was detected by Southern hybridization and polymerase chain reaction (PCR) analysis. The expression of the chitinase gene was demonstrated by reverse transcription PCR and Western analysis. One hundred and two ginseng plantlets were recovered from 11 confirmed transgenic lines. The transgene integration in plantlets of two lines was demonstrated by Southern analysis. This is the first report of Agrobacterium-mediated transformation of this important medicinal plant.

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Introduction and constitutive expression of a rice chitinase gene in bread wheat using biolistic bombardment and the bar gene as a selectable marker

Cited by 62 publications

References 44 publications

Expression of the lipid transfer protein Ace-AMP1 in transgenic wheat enhances antifungal activity and defense responses

Expression of the lipid transfer protein Ace-AMP1 in transgenic wheat enhances antifungal activity and defense responses

Molecular Characterization of the Major Wheat Domestication Gene Q

Agrobacterium-mediated transformation of American ginseng with a rice chitinase gene

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