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Enhanced monoterpene emission in transgenic orange mint (Mentha × piperita f. citrata) overexpressing a tobacco lipid transfer protein (NtLTP1)

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Abstract

Main conclusion

Overexpression of the tobacco lipid transfer protein (NtLTP1) gene in transgenic orange mint resulted in enhanced accumulation of monoterpenes in the cavity of head cells of glandular trichomes, which resulted in enhanced emission of monoterpenes from transgenic orange mints.

Abstract

Plants in the genus Mentha (Lamiaceae) produce volatile oils that accumulate in peltate glandular trichomes in the aerial parts of plants. A lipid transfer protein (NtLTP1) in tobacco showed glandular trichome-specific expression and supported the secretion of diterpenoid lipids from head cells of glandular trichomes (Choi et al., Plant J 70:480–491,2012). Here, we constructed transgenic orange mint (Mentha × piperita f. citrata) overexpressing the tobacco NtLTP1 gene via Agrobacterium-mediated transformation. Transgenic lines of orange mint overexpressing NtLTP1 were confirmed by genomic PCR and RT-PCR. Immunoblotting analysis using an NtLTP1 polyclonal antibody showed clear dark spots at the position of the lipid exudates from tobacco glandular trichomes and the squeezed out lipids from the glandular trichomes of transgenic orange mint. Heads of glandular trichomes in transgenic plants overexpressing the NtLTP1 gene showed a larger diameter than those of the wild-type control. The enhanced size of trichome heads in transgenic orange mint was confirmed by scanning electron microscopy. Volatile components were extracted from wild-type and transgenic orange mint by solid-phase microextraction (SPME) and analyzed by headspace–gas chromatography–mass spectrometry (HS/GC/MS). Linalyl acetate was the most abundant component among the eleven identified monoterpenes in the volatile compounds extracted from both the wild-type and transgenic lines of orange mint. Overexpression of NtLTP1 in transgenic orange mint plants resulted in enhanced emission of volatile monoterpenoids compared with that of volatile monoterpenoids in the wild-type control plants.

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Abbreviations

HS/SPME:

Headspace/solid-phase microextraction

HS–GC–MS:

Headspace–gas chromatography–mass spectrometry

LTP:

Lipid transfer protein

PCR:

Polymerase chain reaction

UHR-SEM:

Ultrahigh-resolution scanning electron microscopy

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Acknowledgements

This work was supported by the Rural Development Administration, Republic of Korea [Next-Generation Bio-Green 21 Program (PJ01344401 and PJ01369103)], and Kangwon National University.

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  1. Hwan-Su Hwang and Prakash Babu Adhikari contributed equally to this work.

Authors and Affiliations

  1. Division of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chuncheon, 200-701, Republic of Korea

    Hwan-Su Hwang, Prakash Babu Adhikari, Hye-Jeong Jo, Jung Yeon Han & Yong Eui Choi

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  1. Hwan-Su Hwang
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  2. Prakash Babu Adhikari
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  3. Hye-Jeong Jo
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Correspondence to Yong Eui Choi.

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425_2020_3447_MOESM1_ESM.docx

Supplementary file1 Supplemental Figure S1. DNA sequences of T-DNA regions of the binary vectors used in this experiment. (DOCX 19 kb)

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Hwang, HS., Adhikari, P.B., Jo, HJ. et al. Enhanced monoterpene emission in transgenic orange mint (Mentha × piperita f. citrata) overexpressing a tobacco lipid transfer protein (NtLTP1). Planta 252, 44 (2020). https://doi.org/10.1007/s00425-020-03447-6

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