TY  - CONF
AU  - Aničić, Neda
AU  - Matekalo, Dragana
AU  - Skorić, Marijana
AU  - Pećinar, Ilinka
AU  - Brkušanin, Miloš
AU  - Nestorović Živković, Jasmina
AU  - Dmitrović, Slavica
AU  - Božunović, Jelena
AU  - Dajić Stevanović, Zora
AU  - Schulz, Hartwig
AU  - Mišić, Danijela
PY  - 2018
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4389
AB  - The majority of species belonging to genus Nepeta (fam. Lamiaceae) produce and accumulate nepetalactones as the main secondary metabolites. Nepetalactones are iridoid monoterpenoids recognized for their wide spectrum of high-value bioactive properties, namely antimicrobial, cytostatic, anti-inflammatory, phytotoxic, etc. However, biosynthetic pathway of these compounds in plants remains to a great extent unexplored. Most probably they derive through general iridoid biosynthetic pathway leading to nepetalactol, a common intermediate of all plant-produced iridoids, that is formed in a reaction catalyzed by iridoid synthase (IS, EC 1.3.1.99). Present investigation entails homology based degenerative primer approach which led to the identification of two promising iridoid synthase candidates (NrIS1 and NrIS2) in leaves of Nepeta rtanjensis Diklić & Milojević, rich in trans,cis-nepetalactone (trans,cis-NL) and dehydronepetalactone (DNL). Comparative UHPLC/qqqMS profiling of nepetalactones in detached trichomes and abraded leaves, complemented with Raman spectroscopy and dichloromethane dipping experiment highlighted trichomes as the main site of their accumulation. This was further supported by the significantly high trichome/abraded leaves expression ratio of NrIS1 (~31) and NrIS2 (~25), which strongly indicated the involvement of these genes in nepetalactone biosynthesis. High correlation between NrIS1 expression profile and trans,cis-NL and DNL contents in five developmental stages of N. rtanjensis leaves confirmed developmental regulation of nepetalactone biosynthesis. Concentrations of trans,cis-NL and DNL were 22-fold and 80-fold lower in the oldest leaves investigated than in the youngest leaves, respectively. In addition, the expression pattern of the NrIS1 followed the same trend exhibiting 7.4-fold higher expression in the oldest leaves compared to the youngest ones. The present study on spatial and developmental regulation of the nepetalactones production and accumulation in N. rtanjensis leaves, as well as the identification of two IS candidate genes is a proof-of-the-concept research that serves as a cornerstone for future studies on elucidation of the nepetalactones biosynthesis.
PB  - Copenhagen: Department of Plant and Environmental Sciences, University of Copenhagen
C3  - Plant Biology Europe 2018 Conference: Abstract Book; 2018 Jun 18-21; Copenhagen, Denmark
T1  - Nepetalactones profiling and putative iridoid synthase expression analysis indicate trichome specific localization and developmental regulation of iridoids biosynthesis in leaves of Nepeta rtanjensis
SP  - 326
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4389
ER  - 

@conference{
author = "Aničić, Neda and Matekalo, Dragana and Skorić, Marijana and Pećinar, Ilinka and Brkušanin, Miloš and Nestorović Živković, Jasmina and Dmitrović, Slavica and Božunović, Jelena and Dajić Stevanović, Zora and Schulz, Hartwig and Mišić, Danijela",
year = "2018",
abstract = "The majority of species belonging to genus Nepeta (fam. Lamiaceae) produce and accumulate nepetalactones as the main secondary metabolites. Nepetalactones are iridoid monoterpenoids recognized for their wide spectrum of high-value bioactive properties, namely antimicrobial, cytostatic, anti-inflammatory, phytotoxic, etc. However, biosynthetic pathway of these compounds in plants remains to a great extent unexplored. Most probably they derive through general iridoid biosynthetic pathway leading to nepetalactol, a common intermediate of all plant-produced iridoids, that is formed in a reaction catalyzed by iridoid synthase (IS, EC 1.3.1.99). Present investigation entails homology based degenerative primer approach which led to the identification of two promising iridoid synthase candidates (NrIS1 and NrIS2) in leaves of Nepeta rtanjensis Diklić & Milojević, rich in trans,cis-nepetalactone (trans,cis-NL) and dehydronepetalactone (DNL). Comparative UHPLC/qqqMS profiling of nepetalactones in detached trichomes and abraded leaves, complemented with Raman spectroscopy and dichloromethane dipping experiment highlighted trichomes as the main site of their accumulation. This was further supported by the significantly high trichome/abraded leaves expression ratio of NrIS1 (~31) and NrIS2 (~25), which strongly indicated the involvement of these genes in nepetalactone biosynthesis. High correlation between NrIS1 expression profile and trans,cis-NL and DNL contents in five developmental stages of N. rtanjensis leaves confirmed developmental regulation of nepetalactone biosynthesis. Concentrations of trans,cis-NL and DNL were 22-fold and 80-fold lower in the oldest leaves investigated than in the youngest leaves, respectively. In addition, the expression pattern of the NrIS1 followed the same trend exhibiting 7.4-fold higher expression in the oldest leaves compared to the youngest ones. The present study on spatial and developmental regulation of the nepetalactones production and accumulation in N. rtanjensis leaves, as well as the identification of two IS candidate genes is a proof-of-the-concept research that serves as a cornerstone for future studies on elucidation of the nepetalactones biosynthesis.",
publisher = "Copenhagen: Department of Plant and Environmental Sciences, University of Copenhagen",
journal = "Plant Biology Europe 2018 Conference: Abstract Book; 2018 Jun 18-21; Copenhagen, Denmark",
title = "Nepetalactones profiling and putative iridoid synthase expression analysis indicate trichome specific localization and developmental regulation of iridoids biosynthesis in leaves of Nepeta rtanjensis",
pages = "326",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4389"
}

Aničić, N., Matekalo, D., Skorić, M., Pećinar, I., Brkušanin, M., Nestorović Živković, J., Dmitrović, S., Božunović, J., Dajić Stevanović, Z., Schulz, H.,& Mišić, D.. (2018). Nepetalactones profiling and putative iridoid synthase expression analysis indicate trichome specific localization and developmental regulation of iridoids biosynthesis in leaves of Nepeta rtanjensis. in Plant Biology Europe 2018 Conference: Abstract Book; 2018 Jun 18-21; Copenhagen, Denmark
Copenhagen: Department of Plant and Environmental Sciences, University of Copenhagen., 326.
https://hdl.handle.net/21.15107/rcub_ibiss_4389

Aničić N, Matekalo D, Skorić M, Pećinar I, Brkušanin M, Nestorović Živković J, Dmitrović S, Božunović J, Dajić Stevanović Z, Schulz H, Mišić D. Nepetalactones profiling and putative iridoid synthase expression analysis indicate trichome specific localization and developmental regulation of iridoids biosynthesis in leaves of Nepeta rtanjensis. in Plant Biology Europe 2018 Conference: Abstract Book; 2018 Jun 18-21; Copenhagen, Denmark. 2018;:326.
https://hdl.handle.net/21.15107/rcub_ibiss_4389 .

Aničić, Neda, Matekalo, Dragana, Skorić, Marijana, Pećinar, Ilinka, Brkušanin, Miloš, Nestorović Živković, Jasmina, Dmitrović, Slavica, Božunović, Jelena, Dajić Stevanović, Zora, Schulz, Hartwig, Mišić, Danijela, "Nepetalactones profiling and putative iridoid synthase expression analysis indicate trichome specific localization and developmental regulation of iridoids biosynthesis in leaves of Nepeta rtanjensis" in Plant Biology Europe 2018 Conference: Abstract Book; 2018 Jun 18-21; Copenhagen, Denmark (2018):326,
https://hdl.handle.net/21.15107/rcub_ibiss_4389 .

TY  - JOUR
AU  - Aničić, Neda
AU  - Matekalo, Dragana
AU  - Skorić, Marijana
AU  - Pećinar, Ilinka
AU  - Brkušanin, Miloš
AU  - Nestorović Živković, Jasmina
AU  - Dmitrović, Slavica
AU  - Dajić Stevanović, Zora
AU  - Schulz, Hartwig
AU  - Mišić, Danijela
PY  - 2018
UR  - https://www.sciencedirect.com/science/article/pii/S0926669018302413?via%3Dihub
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/3022
AB  - Nepetalactones are accumulated as the main secondary metabolites in the majority of Nepeta species; however, their biosynthetic pathway remains to a great extent unknown, as well as their relationship to the overall physiology of these plants. Nepetalactones most certainly arise through general iridoid biosynthetic pathway leading to nepetalactol, a common precursor of all plant-derived iridoids that is formed in a reaction catalysed by iridoid synthase (IS, EC 1.3.1.99). In this paper, we aimed to localize the sites of nepetalactone biosynthesis and accumulation by comparatively analysing the nepetalactones content and the expression of putative iridoid synthase (NrIS) in leaves of Nepeta rtanjensis Diklić & Milojević, a trans,cis-nepetalactone and dehydronepetalactone rich species. Another aim was to give an insight into the developmental regulation of nepetalactone biosynthesis in leaves of cultivated N. rtanjensis plants. Knowledge on the spatial-, and developmental- regulation of nepetalactone metabolism could greatly benefit potential growers by enabling them to adequately plan and manage plant cultivation, harvesting, and handling and thus increase yield of bioactive compounds. The expression profile of putative NrIS1 is highly correlated with trans,cis-nepetalactone and dehydronepetalactone content/ratio in leaves of different developmental stages. The results strongly indicate that developmental regulation of nepetalactone biosynthesis in leaves resides largely at the level of gene expression, and suggest an important role of NrIS1. Young leaves exhibited the most intensive nepetalactone biosynthesis and accumulation, but also its conversion to dehydronepetalactone. Thus, the pattern of NL accumulation in leaves can be explained by both the rate of its biosynthesis and metabolic degradation. The gene expression patterns of NrIS1 and NrIS2 transcripts in trichomes and abraded leaves revealed trichomes as the main site of their activity. UHPLC/qqqMS profiling of nepetalactones in trichomes and abraded leaves, complemented with Raman spectroscopy and dichloromethane dipping experiment, supported glandular trichome-specific localization of nepetalactone biosynthesis and accumulation.
T2  - Industrial Crops and Products
T1  - Trichome-specific and developmentally regulated biosynthesis of nepetalactones in leaves of cultivated Nepeta rtanjensis plants
VL  - 117
DO  - 10.1016/J.INDCROP.2018.03.019
SP  - 347
EP  - 358
ER  - 

@article{
author = "Aničić, Neda and Matekalo, Dragana and Skorić, Marijana and Pećinar, Ilinka and Brkušanin, Miloš and Nestorović Živković, Jasmina and Dmitrović, Slavica and Dajić Stevanović, Zora and Schulz, Hartwig and Mišić, Danijela",
year = "2018",
abstract = "Nepetalactones are accumulated as the main secondary metabolites in the majority of Nepeta species; however, their biosynthetic pathway remains to a great extent unknown, as well as their relationship to the overall physiology of these plants. Nepetalactones most certainly arise through general iridoid biosynthetic pathway leading to nepetalactol, a common precursor of all plant-derived iridoids that is formed in a reaction catalysed by iridoid synthase (IS, EC 1.3.1.99). In this paper, we aimed to localize the sites of nepetalactone biosynthesis and accumulation by comparatively analysing the nepetalactones content and the expression of putative iridoid synthase (NrIS) in leaves of Nepeta rtanjensis Diklić & Milojević, a trans,cis-nepetalactone and dehydronepetalactone rich species. Another aim was to give an insight into the developmental regulation of nepetalactone biosynthesis in leaves of cultivated N. rtanjensis plants. Knowledge on the spatial-, and developmental- regulation of nepetalactone metabolism could greatly benefit potential growers by enabling them to adequately plan and manage plant cultivation, harvesting, and handling and thus increase yield of bioactive compounds. The expression profile of putative NrIS1 is highly correlated with trans,cis-nepetalactone and dehydronepetalactone content/ratio in leaves of different developmental stages. The results strongly indicate that developmental regulation of nepetalactone biosynthesis in leaves resides largely at the level of gene expression, and suggest an important role of NrIS1. Young leaves exhibited the most intensive nepetalactone biosynthesis and accumulation, but also its conversion to dehydronepetalactone. Thus, the pattern of NL accumulation in leaves can be explained by both the rate of its biosynthesis and metabolic degradation. The gene expression patterns of NrIS1 and NrIS2 transcripts in trichomes and abraded leaves revealed trichomes as the main site of their activity. UHPLC/qqqMS profiling of nepetalactones in trichomes and abraded leaves, complemented with Raman spectroscopy and dichloromethane dipping experiment, supported glandular trichome-specific localization of nepetalactone biosynthesis and accumulation.",
journal = "Industrial Crops and Products",
title = "Trichome-specific and developmentally regulated biosynthesis of nepetalactones in leaves of cultivated Nepeta rtanjensis plants",
volume = "117",
doi = "10.1016/J.INDCROP.2018.03.019",
pages = "347-358"
}

Aničić, N., Matekalo, D., Skorić, M., Pećinar, I., Brkušanin, M., Nestorović Živković, J., Dmitrović, S., Dajić Stevanović, Z., Schulz, H.,& Mišić, D.. (2018). Trichome-specific and developmentally regulated biosynthesis of nepetalactones in leaves of cultivated Nepeta rtanjensis plants. in Industrial Crops and Products, 117, 347-358.
https://doi.org/10.1016/J.INDCROP.2018.03.019

Aničić N, Matekalo D, Skorić M, Pećinar I, Brkušanin M, Nestorović Živković J, Dmitrović S, Dajić Stevanović Z, Schulz H, Mišić D. Trichome-specific and developmentally regulated biosynthesis of nepetalactones in leaves of cultivated Nepeta rtanjensis plants. in Industrial Crops and Products. 2018;117:347-358.
doi:10.1016/J.INDCROP.2018.03.019 .

Aničić, Neda, Matekalo, Dragana, Skorić, Marijana, Pećinar, Ilinka, Brkušanin, Miloš, Nestorović Živković, Jasmina, Dmitrović, Slavica, Dajić Stevanović, Zora, Schulz, Hartwig, Mišić, Danijela, "Trichome-specific and developmentally regulated biosynthesis of nepetalactones in leaves of cultivated Nepeta rtanjensis plants" in Industrial Crops and Products, 117 (2018):347-358,
https://doi.org/10.1016/J.INDCROP.2018.03.019 . .