Constitutive over-expression of two wheat pathogenesis-related genes enhances resistance of tobacco plants to Phytophthora nicotianae

Fiocchetti, Floriana; D'Amore, R.; Palma, Monica De; Bertini, Laura; Caruso, Carla; Caporale, Carlo; Testa, Antonino; Cristinzio, Gennaro; Saccardo, F.; Tucci, Marina

doi:10.1007/s11240-007-9306-4

Cited by 13 publications

(9 citation statements)

References 47 publications

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“…Difference between the level of transgene expression at RNA and its protein product accumulation was observed in the current study. Similar results were reported previously (Fiocchetti et al 2008).…”

Section: Discussionsupporting

confidence: 93%

Ectopic overexpression of peach GDP-d-mannose pyrophosphorylase and GDP-d-mannose-3′,5′-epimerase in transgenic tobacco

Imai

Ban

Yamamoto

et al. 2012

Plant Cell Tiss Organ Cult

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We created transgenic tobacco overexpressing peach GDP-D-mannose pyrophosphorylase (PpGMPH) and GDP-D-mannose-3 0 ,5 0 -epimerase (PpGME), which are involved in the main L-ascorbate (Asc) biosynthetic pathway in plants. Despite an apparent increase in enzymatic activity in the PpGMPH-overexpressing primary transgenic (T0) lines, no significant increase in Asc pool size was observed in leaves. For PpGME-overexpressing T0 lines, Asc pool size was also unchanged, although transcriptional overexpression was confirmed. To reinforce the biosynthetic pathway from D-mannose-1-phosphate to GDP-L-galactose, we further created PpGMPH-and PpGME-double overexpressing crossed transgenic (dOx T1) lines. Transcriptional overexpression of PpGMPH and PpGME transgenes was confirmed, and the Asc content in the dOx T1 lines was slightly elevated in young leaves, albeit not significantly. Western blot analyses revealed protein amounts of GMPH and GME were similar among dOx T1, parental T0 and non-transgenic (SR1) lines in three tissue types tested: leaves, flower buds and immature fruits. Moreover, a similar amount of GMPH and GME proteins were detected in young leaves (YL) and old leaves (OL), although Asc content in YL is roughly double as that in OL. In vitro assay using recombinant PpGME suggested the equilibrium of GME reaction is unfavorable to forward Asc biosynthesis at this step. Finally, to investigate substrate availability in relation to Asc pool size, we added exogenous GDP-D-mannose (5 mM) or L-galactose (5 mM) to tobacco leaf discs; the former resulted in no increase in Asc content, whereas a significant increase was observed in the latter, not only in the dOx T1 lines, but also in the original line SR1. Similar treatments were also conducted under high light, which resulted in a further increase in the increment of Asc content in the L-galactose-fed discs irrespective of transgene expression level. Taken together, both L-galactose availability and light rather than upper biosynthetic gene expression levels of GMPH/GME are critical determinant of Asc content in tobacco leaves, indicating complex modulation of this compound.Keywords L-ascorbate Á GDP-D-mannose-3 0 ,5 0 -epimerase Á GDP-D-mannose pyrophosphorylase Á Overexpression Á Transgenic tobacco Abbreviations Asc L-Ascorbate

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Section: Discussionsupporting

confidence: 93%

Ectopic overexpression of peach GDP-d-mannose pyrophosphorylase and GDP-d-mannose-3′,5′-epimerase in transgenic tobacco

Imai

Ban

Yamamoto

et al. 2012

Plant Cell Tiss Organ Cult

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“…Durable resistance can be achieved by the expression and synergistic action of plant defense genes (Ouyang et al 2003;Fiocchetti et al 2008). The co-expression of plant defense genes appears to be a preferred route to engineering protection against Fusarium wilt (Punja 2001;Sarowar et al 2006;Fiocchetti et al 2008). Resistance to the fungal diseases was improved in transgenic tobacco and tomato plants that expressed the basic tobacco b-1, 3-glucanase and bean chitinase genes (Lan et al 2000;Ouyang et al 2003;Chen et al 2009).…”

Section: Introductionmentioning

confidence: 99%

Transgenic tomato plants expressing a wheat endochitinase gene demonstrate enhanced resistance to Fusarium oxysporum f. sp. lycopersici

Girhepuje

Shinde

2010

Plant Cell Tiss Organ Cult

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Various chitinases have been shown to inhibit the growth of fungal pathogens in in vitro as well as in planta conditions. chi194, a wheat chitinases gene encoding a 33-kDa chitinase protein, was overexpressed in tomato plants (cv. Pusa Ruby) under the control of maize ubiquitin 1 promoter. The integration of transgene in tomato plants was confirmed with polymerase chain reaction (PCR) and Southern blot analysis. The inheritance of the transgene in T 1 and T 2 generations were shown by molecular analysis and the hygromycin sensitivity test. The broad range of chitinase activity was observed among the transgenic lines in T 0 and a similar range was retained in the T 1 and T 2 generations. Most importantly, the transgenic tomato lines with high chitinase activity were found to be highly resistant to the fungal pathogen Fusarium oxysporum f. sp. lycopersici. Thus, the results demonstrated that the expression of the wheat endochitinase chi194 in tomato plants confers resistance against Fusarium wilt disease caused by the fungal pathogen Fusarium oxysporum f. sp. lycopersici.

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“…The PR-4 role in defense has been confirmed by recent evidence of an in planta activity 93 . In addition to a protective role against fungal pathogens, PR-4 may play a role in the response of the developing grain to elevated temperatures.…”

Section: Barwin (Barley Wound Induced Protein) (Pr-4)mentioning

confidence: 78%

A Review: Biological and Technological Functions of Barley Seed Pathogenesis-Related Proteins (PRs)

Gorjanović

2009

Journal of the Institute of Brewing

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The mature barley seed proteome is dominated by proteins involved in stress responses, including the Pathogenesis-Related proteins (PRs). PRs are currently classified into 17 families. The biochemistry of the PRs families, whose members are constitutively present in barley seed, is surveyed in this paper. These proteins are assumed to protect dormant and germinating seeds against pathogenic microorganisms and pests. The current knowledge on PRs structural and functional properties is summarized in an attempt to illustrate their importance in barley seed innate resistance. At the same time, a platform to understand PRs technological function in cereal foods and in beverage processing and quality is provided.

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Constitutive over-expression of two wheat pathogenesis-related genes enhances resistance of tobacco plants to Phytophthora nicotianae

Cited by 13 publications

References 47 publications

Ectopic overexpression of peach GDP-d-mannose pyrophosphorylase and GDP-d-mannose-3′,5′-epimerase in transgenic tobacco

Ectopic overexpression of peach GDP-d-mannose pyrophosphorylase and GDP-d-mannose-3′,5′-epimerase in transgenic tobacco

Transgenic tomato plants expressing a wheat endochitinase gene demonstrate enhanced resistance to Fusarium oxysporum f. sp. lycopersici

A Review: Biological and Technological Functions of Barley Seed Pathogenesis-Related Proteins (PRs)

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