Overexpression of the Ib-rolB/C gene perturbs biosynthesis of caffeoylquinic acid derivatives in transgenic calli of sweet potato

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Ipomoea batatas is a root crop widely cultivated in South America, Africa, and Asia. This is also a source of caffeoylquinic acid derivatives (CQAs) with potential health-promoting benefits. Sweet potato genome carries two separate cellular T-DNA (cT-DNA) regions (IbT-DNA1 and IbT-DNA2). Especially, IbT-DNA2 contains five ORFs homologous to the Agrobacterium rhizogenes T-DNA, namely ORF13, ORF14, ORF17n, ORF18/ORF17n, and RolB/RolC proteins [1]. Unfortunately, presently there is insufficient information on IbT-DNA2 genes’ function in the physiological processes of sweet potatoes.

In this study, expressional levels of the IbT-DNA2 genes and the effect of Ib-rolB/C overexpression were examined using I. batatas cell culture. We discovered that I. batatas cT-DNA genes were not expressed in callus, and abiotic stresses and chemical elicitors affected their transcriptional levels weakly. Additionally, two Ib-rolBC-transgenic cell lines have been established though Agrobacterium-mediated transformation of I. batatas callus cells. Overexpression of Ib-rolB/C gene reduced biomass accumulation of transgenic cell lines by 1.2–1.6 times and increased the CQAs content by 1.5–1.9-fold. To justify the metabolic fluctuations, the study also looked into the expression patterns of the major biosynthetic genes, namely bPAL, IbC4H, Ib4CL, IbHCT, and IbHQT. The obtained data demonstrated that the overexpression of the Ib-rolB/C reduced the IbPAL transcript but considerable increase in the transcript levels of the IbHQT. We propose that this result was obtained through as-yet-uncharacterized signaling pathways activated by RolB/RolC.

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Ipomoea batatas is a root crop widely cultivated in South America, Africa, and Asia. This is also a source of caffeoylquinic acid derivatives (CQAs) with potential health-promoting benefits. Sweet potato genome carries two separate cellular T-DNA (cT-DNA) regions (IbT-DNA1 and IbT-DNA2). Especially, IbT-DNA2 contains five ORFs homologous to the Agrobacterium rhizogenes T-DNA, namely ORF13, ORF14, ORF17n, ORF18/ORF17n, and RolB/RolC proteins [1]. Unfortunately, presently there is insufficient information on IbT-DNA2 genes’ function in the physiological processes of sweet potatoes.

In this study, expressional levels of the IbT-DNA2 genes and the effect of Ib-rolB/C overexpression were examined using I. batatas cell culture. We discovered that I. batatas cT-DNA genes were not expressed in callus, and abiotic stresses and chemical elicitors affected their transcriptional levels weakly. Additionally, two Ib-rolBC-transgenic cell lines have been established though Agrobacterium-mediated transformation of I. batatas callus cells. Overexpression of Ib-rolB/C gene reduced biomass accumulation of transgenic cell lines by 1.2–1.6 times and increased the CQAs content by 1.5–1.9-fold. To justify the metabolic fluctuations, the study also looked into the expression patterns of the major biosynthetic genes, namely bPAL, IbC4H, Ib4CL, IbHCT, and IbHQT. The obtained data demonstrated that the overexpression of the Ib-rolB/C reduced the IbPAL transcript but considerable increase in the transcript levels of the IbHQT. We propose that this result was obtained through as-yet-uncharacterized signaling pathways activated by RolB/RolC.

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Об авторах

Elena Vasyutkina

Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far East Branch of Russian Academy of Sciences

Email: levina@biosoil.ru
SPIN-код: 9009-7848

Researcher, Laboratory of Bionanotechnology and Biomedicine

Россия, Vladivostok

Yulia Yugay

Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far East Branch of Russian Academy of Sciences

Email: yuya1992@mail.ru
SPIN-код: 1530-4110

Junior Researcher, Laboratory of Bionanotechnology and Biomedicine

Россия, Vladivostok

Valeria Grigorchuk

Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far East Branch of Russian Academy of Sciences

Email: kera1313@mail.ru
SPIN-код: 3373-9623

Researcher, Laboratory of Bionanotechnology and Biomedicine

Россия, Vladivostok

Olga Grishchenko

Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far East Branch of Russian Academy of Sciences

Email: grishchenkoov@gmail.com
SPIN-код: 3030-7430

Researcher, Laboratory of Bionanotechnology and Biomedicine

Россия, Vladivostok

Varvara Stepochkina

Far Eastern Federal University

Email: vdkislitsyna@gmail.com

PhD Student, Advanced Engineering School “Institute of Biotechnology, Bioengineering and Food Systems”

Россия, Vladivostok

Yury Shkryl

Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far East Branch of Russian Academy of Sciences

Автор, ответственный за переписку.
Email: yn80@mail.ru
SPIN-код: 3814-8026

PhD, Main Researcher, Laboratory of Bionanotechnology and Biomedicine

Россия, Vladivostok

Список литературы

  1. Kyndt T, Quispe D, Zhai H, et al. The genome of cultivated sweet potato contains Agrobacterium T-DNAs with expressed genes: an example of a naturally transgenic food crop. PNAS USA. 2015;112(18):5844–5849. doi: 10.1073/pnas.1419685112

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