Transmission of Engineered Plastids in Sugarcane, a C<sub>4 </sub>Monocotyledonous Plant, Reveals that Sorting of Preprogrammed Progenitor Cells Produce Heteroplasmy

We report here plastid transformation in sugarcane using biolistic transformation and embryogenesis-based regeneration approaches. Somatic embryos were developed from unfurled leaf sections, containing preprogrammed progenitor cells, to recover transformation events on antibiotic-containing regenera...

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Main Authors: Ghulam Mustafa, Muhammad Sarwar Khan
Format: Article
Language:English
Published: MDPI AG 2021-12-01
Series:Plants
Subjects:
Online Access:https://www.mdpi.com/2223-7747/10/1/26
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spelling doaj-a333ff86b71f4e9e863e9c0cfaafb57e2020-12-25T00:04:51ZengMDPI AGPlants2223-77472021-12-0110262610.3390/plants10010026Transmission of Engineered Plastids in Sugarcane, a C<sub>4 </sub>Monocotyledonous Plant, Reveals that Sorting of Preprogrammed Progenitor Cells Produce HeteroplasmyGhulam Mustafa0Muhammad Sarwar Khan1Center of Agricultural Biochemistry and Biotechnology (CABB), University of Agriculture, University Road, Faisalabad P.O. Box 38040, PakistanCenter of Agricultural Biochemistry and Biotechnology (CABB), University of Agriculture, University Road, Faisalabad P.O. Box 38040, PakistanWe report here plastid transformation in sugarcane using biolistic transformation and embryogenesis-based regeneration approaches. Somatic embryos were developed from unfurled leaf sections, containing preprogrammed progenitor cells, to recover transformation events on antibiotic-containing regeneration medium. After developing a proficient regeneration system, the FLARE-S (fluorescent antibiotic resistance enzyme, spectinomycin and streptomycin) expression cassette that carries species-specific homologous sequence tails was used to transform plastids and track gene transmission and expression in sugarcane. Plants regenerated from streptomycin-resistant and genetically confirmed shoots were subjected to visual detection of the fluorescent enzyme using a fluorescent stereomicroscope, after genetic confirmation. The resultant heteroplasmic shoots remained to segregate on streptomycin-containing MS medium, referring to the unique pattern of division and sorting of cells in C<sub>4 </sub>monocotyledonous compared to C<sub>3 </sub>monocotyledonous and dicotyledonous plants since in sugarcane bundle sheath and mesophyll, cells are distinct and sort independently after division. Hence, the transformation of either mesophyll or bundle sheath cells will develop heteroplasmic transgenic plants, suggesting the transformation of both types of cells. Whilst developed transgenic sugarcane plants are heteroplasmic, and selection-based regeneration protocol envisaging the role of division and sorting of cells in the purification of transplastomic demands further improvement, the study has established many parameters that may open up exciting possibilities to express genes of agricultural or pharmaceutical importance in sugarcane.https://www.mdpi.com/2223-7747/10/1/26plastid transformationsugarcaneunfurled leavesstreptomycinheteroplasmymesophyll and bundle sheath cells
collection DOAJ
language English
format Article
sources DOAJ
author Ghulam Mustafa
Muhammad Sarwar Khan
spellingShingle Ghulam Mustafa
Muhammad Sarwar Khan
Transmission of Engineered Plastids in Sugarcane, a C<sub>4 </sub>Monocotyledonous Plant, Reveals that Sorting of Preprogrammed Progenitor Cells Produce Heteroplasmy
Plants
plastid transformation
sugarcane
unfurled leaves
streptomycin
heteroplasmy
mesophyll and bundle sheath cells
author_facet Ghulam Mustafa
Muhammad Sarwar Khan
author_sort Ghulam Mustafa
title Transmission of Engineered Plastids in Sugarcane, a C<sub>4 </sub>Monocotyledonous Plant, Reveals that Sorting of Preprogrammed Progenitor Cells Produce Heteroplasmy
title_short Transmission of Engineered Plastids in Sugarcane, a C<sub>4 </sub>Monocotyledonous Plant, Reveals that Sorting of Preprogrammed Progenitor Cells Produce Heteroplasmy
title_full Transmission of Engineered Plastids in Sugarcane, a C<sub>4 </sub>Monocotyledonous Plant, Reveals that Sorting of Preprogrammed Progenitor Cells Produce Heteroplasmy
title_fullStr Transmission of Engineered Plastids in Sugarcane, a C<sub>4 </sub>Monocotyledonous Plant, Reveals that Sorting of Preprogrammed Progenitor Cells Produce Heteroplasmy
title_full_unstemmed Transmission of Engineered Plastids in Sugarcane, a C<sub>4 </sub>Monocotyledonous Plant, Reveals that Sorting of Preprogrammed Progenitor Cells Produce Heteroplasmy
title_sort transmission of engineered plastids in sugarcane, a c<sub>4 </sub>monocotyledonous plant, reveals that sorting of preprogrammed progenitor cells produce heteroplasmy
publisher MDPI AG
series Plants
issn 2223-7747
publishDate 2021-12-01
description We report here plastid transformation in sugarcane using biolistic transformation and embryogenesis-based regeneration approaches. Somatic embryos were developed from unfurled leaf sections, containing preprogrammed progenitor cells, to recover transformation events on antibiotic-containing regeneration medium. After developing a proficient regeneration system, the FLARE-S (fluorescent antibiotic resistance enzyme, spectinomycin and streptomycin) expression cassette that carries species-specific homologous sequence tails was used to transform plastids and track gene transmission and expression in sugarcane. Plants regenerated from streptomycin-resistant and genetically confirmed shoots were subjected to visual detection of the fluorescent enzyme using a fluorescent stereomicroscope, after genetic confirmation. The resultant heteroplasmic shoots remained to segregate on streptomycin-containing MS medium, referring to the unique pattern of division and sorting of cells in C<sub>4 </sub>monocotyledonous compared to C<sub>3 </sub>monocotyledonous and dicotyledonous plants since in sugarcane bundle sheath and mesophyll, cells are distinct and sort independently after division. Hence, the transformation of either mesophyll or bundle sheath cells will develop heteroplasmic transgenic plants, suggesting the transformation of both types of cells. Whilst developed transgenic sugarcane plants are heteroplasmic, and selection-based regeneration protocol envisaging the role of division and sorting of cells in the purification of transplastomic demands further improvement, the study has established many parameters that may open up exciting possibilities to express genes of agricultural or pharmaceutical importance in sugarcane.
topic plastid transformation
sugarcane
unfurled leaves
streptomycin
heteroplasmy
mesophyll and bundle sheath cells
url https://www.mdpi.com/2223-7747/10/1/26
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