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Enhanced methanol production in plants provides broad spectrum insect resistance.

Dixit S, Upadhyay SK, Singh H, Sidhu OP, Verma PC, K C - PLoS ONE (2013)

Bottom Line: In-planta bioassay on transgenic lines showed up to 99 and 75% reduction in the population multiplication of plant sap sucking pests Myzus persicae (aphid) and Bemisia tabaci (whitefly), respectively.Confocal microscopy showed no deformities in cellular integrity, structure and density of stomata and trichomes of transgenic plants compared to WT.Cell wall enzyme transcript levels were comparable with WT.

View Article: PubMed Central - PubMed

Affiliation: CSIR-National Botanical Research Institute, Council of Scientific and Industrial Research, Lucknow, Uttar Pradesh, India ; Academy of Scientific and Innovative Research (AcSIR), Anusandhan Bhawan, 2-Rafi Marg, New Delhi, India.

ABSTRACT
Plants naturally emit methanol as volatile organic compound. Methanol is toxic to insect pests; but the quantity produced by most of the plants is not enough to protect them against invading insect pests. In the present study, we demonstrated that the over-expression of pectin methylesterase, derived from Arabidopsis thaliana and Aspergillus niger, in transgenic tobacco plants enhances methanol production and resistance to polyphagous insect pests. Methanol content in the leaves of transgenic plants was measured using proton nuclear spectroscopy (1H NMR) and spectra showed up to 16 fold higher methanol as compared to control wild type (WT) plants. A maximum of 100 and 85% mortality in chewing insects Helicoverpa armigera and Spodoptera litura larvae was observed, respectively when fed on transgenic plants leaves. The surviving larvae showed less feeding, severe growth retardation and could not develop into pupae. In-planta bioassay on transgenic lines showed up to 99 and 75% reduction in the population multiplication of plant sap sucking pests Myzus persicae (aphid) and Bemisia tabaci (whitefly), respectively. Most of the phenotypic characters of transgenic plants were similar to WT plants. Confocal microscopy showed no deformities in cellular integrity, structure and density of stomata and trichomes of transgenic plants compared to WT. Pollen germination and tube formation was also not affected in transgenic plants. Cell wall enzyme transcript levels were comparable with WT. This study demonstrated for the first time that methanol emission can be utilized for imparting broad range insect resistance in plants.

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Assay of transgenic plants for PME activity.(A) Percent enrichment of PME activity in different transgenic plants over wild type (WT). (B) & (C) In-gel PME activity analysis of different transgenic lines expressing AnPME (An 1-5) & AtPME (At 1-5). Arrow head shows desired PME bands in transgenic plants.
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pone-0079664-g003: Assay of transgenic plants for PME activity.(A) Percent enrichment of PME activity in different transgenic plants over wild type (WT). (B) & (C) In-gel PME activity analysis of different transgenic lines expressing AnPME (An 1-5) & AtPME (At 1-5). Arrow head shows desired PME bands in transgenic plants.

Mentions: PME activity of transgenic and WT plants was compared by measuring its activity in young leaves of 12 week old plants by titration method and in-gel assay. We observed enhance PME activity in both types of transgenic plants in comparison to WT. Transgenic plants of AnPME and AtPME showed up to 42% and 39% increased PME activity, respectively (Figure 3A). Activity was further confirmed by in-gel assay. Several PME bands were observed in WT plants which are native PME’s of plants. However, AtPME and AnPME expressing transgenic lines showed an additional PME activity band at ~36 kDa (expected size of both PME), which was absent in WT plants (Figure 3, B and C).


Enhanced methanol production in plants provides broad spectrum insect resistance.

Dixit S, Upadhyay SK, Singh H, Sidhu OP, Verma PC, K C - PLoS ONE (2013)

Assay of transgenic plants for PME activity.(A) Percent enrichment of PME activity in different transgenic plants over wild type (WT). (B) & (C) In-gel PME activity analysis of different transgenic lines expressing AnPME (An 1-5) & AtPME (At 1-5). Arrow head shows desired PME bands in transgenic plants.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3818224&req=5

pone-0079664-g003: Assay of transgenic plants for PME activity.(A) Percent enrichment of PME activity in different transgenic plants over wild type (WT). (B) & (C) In-gel PME activity analysis of different transgenic lines expressing AnPME (An 1-5) & AtPME (At 1-5). Arrow head shows desired PME bands in transgenic plants.
Mentions: PME activity of transgenic and WT plants was compared by measuring its activity in young leaves of 12 week old plants by titration method and in-gel assay. We observed enhance PME activity in both types of transgenic plants in comparison to WT. Transgenic plants of AnPME and AtPME showed up to 42% and 39% increased PME activity, respectively (Figure 3A). Activity was further confirmed by in-gel assay. Several PME bands were observed in WT plants which are native PME’s of plants. However, AtPME and AnPME expressing transgenic lines showed an additional PME activity band at ~36 kDa (expected size of both PME), which was absent in WT plants (Figure 3, B and C).

Bottom Line: In-planta bioassay on transgenic lines showed up to 99 and 75% reduction in the population multiplication of plant sap sucking pests Myzus persicae (aphid) and Bemisia tabaci (whitefly), respectively.Confocal microscopy showed no deformities in cellular integrity, structure and density of stomata and trichomes of transgenic plants compared to WT.Cell wall enzyme transcript levels were comparable with WT.

View Article: PubMed Central - PubMed

Affiliation: CSIR-National Botanical Research Institute, Council of Scientific and Industrial Research, Lucknow, Uttar Pradesh, India ; Academy of Scientific and Innovative Research (AcSIR), Anusandhan Bhawan, 2-Rafi Marg, New Delhi, India.

ABSTRACT
Plants naturally emit methanol as volatile organic compound. Methanol is toxic to insect pests; but the quantity produced by most of the plants is not enough to protect them against invading insect pests. In the present study, we demonstrated that the over-expression of pectin methylesterase, derived from Arabidopsis thaliana and Aspergillus niger, in transgenic tobacco plants enhances methanol production and resistance to polyphagous insect pests. Methanol content in the leaves of transgenic plants was measured using proton nuclear spectroscopy (1H NMR) and spectra showed up to 16 fold higher methanol as compared to control wild type (WT) plants. A maximum of 100 and 85% mortality in chewing insects Helicoverpa armigera and Spodoptera litura larvae was observed, respectively when fed on transgenic plants leaves. The surviving larvae showed less feeding, severe growth retardation and could not develop into pupae. In-planta bioassay on transgenic lines showed up to 99 and 75% reduction in the population multiplication of plant sap sucking pests Myzus persicae (aphid) and Bemisia tabaci (whitefly), respectively. Most of the phenotypic characters of transgenic plants were similar to WT plants. Confocal microscopy showed no deformities in cellular integrity, structure and density of stomata and trichomes of transgenic plants compared to WT. Pollen germination and tube formation was also not affected in transgenic plants. Cell wall enzyme transcript levels were comparable with WT. This study demonstrated for the first time that methanol emission can be utilized for imparting broad range insect resistance in plants.

Show MeSH
Related in: MedlinePlus