Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Cloning and Functional Analysis of NtHDR in ‘Jinzhanyintai’ of Narcissus tazetta var. chinensis M.Roem

Xiaomeng Hu
,
Na Zhang
ORCID logo ,
Keke Fan
,
Yanting Chang
,
Wenbo Zhang
,
Yayun Deng
,
Shuhua Wang
,
Tao Hu
* ORCID logo ,
Yanjun Ma
* ORCID logo
Version 1 : Received: 17 July 2023 / Approved: 17 July 2023 / Online: 18 July 2023 (08:42:05 CEST)

A peer-reviewed article of this Preprint also exists.

Hu, X.; Zhang, N.; Fan, K.; Chang, Y.; Zhang, W.; Deng, Y.; Wang, S.; Hu, T.; Ma, Y. Cloning and Functional Analysis of NtHDR in ‘Jinzhanyintai’ of Narcissus tazetta var. chinensis M.Roem. Horticulturae 2023, 9, 904. Hu, X.; Zhang, N.; Fan, K.; Chang, Y.; Zhang, W.; Deng, Y.; Wang, S.; Hu, T.; Ma, Y. Cloning and Functional Analysis of NtHDR in ‘Jinzhanyintai’ of Narcissus tazetta var. chinensis M.Roem. Horticulturae 2023, 9, 904.

Abstract

The key enzyme, 1-hydroxy-2-methyl-2-(E)-butenyl-4-pyrophosphate reductase HDR, responsible for the final step in the MEP pathway, was successfully cloned from the petals of Narcissus tazetta var. chinensis, herein referred to as NtHDR (accession number OQ739816). NtHDR exhibits a full-length coding sequence spanning 1380 base pairs, encoding a total of 460 amino acids. Through phylogenetic tree analysis, it has been determined that NtHDR shared the closest evolutionary relationship with monocotyledons, specifically Asparagus officinalis and Zingiber officinale. Utilizing quantitative real-time PCR analysis, it was observed that NtHDR exhibited significant expression in both petals and corona, with expression levels varying throughout the flowering process of Narcissus. Specifically, in petals, NtHDR expression demonstrated a pattern of initial increase followed by subsequent decrease, while in corona, it consistently increased. Subsequent subcellular localization experiments indicated that NtHDR was localized within the chloroplasts. To investigate the functional impact of NtHDR, stable transformation was performed, where NtHDR was introduced into Nicotiana benthamiana. The resulting transgenic N. benthamiana flowers were analyzed using solid-phase microextraction coupled with gas chromatography–mass spectrometry (SPME-GC-MS) to characterize their volatile components. The analysis revealed the presence of the monoterpene compound linalool, as well as the phenylpropanoids benzyl alcohol and phenylethanol, within the floral fragrance components of the transgenic N.benthamiana plants. However, these compounds were absent in the floral fragrance components of wild-type tobacco plants, thus highlighting the impact of NtHDR transformation on the floral scent profile.

Keywords

Chinese Narcissus ‘Jinzhanyintai’; floral scent; HDR; GC-MS; transgenic Nicotiana benthamiana

Subject

Biology and Life Sciences, Horticulture

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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