A Transformation and Genome Editing System for Cassava Cultivar SC8

Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 ( Crantz cv. SC8) is one of the main locally planted cultivars. In this study, an efficient transformation system for cassava SC8 mediated with strain LBA4404 was presented for the first time. Ca...

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Published inGenes Vol. 13; no. 9; p. 1650
Main Authors Wang, Ya-Jie, Lu, Xiao-Hua, Zhen, Xing-Hou, Yang, Hui, Che, Yan-Nian, Hou, Jing-Yi, Geng, Meng-Ting, Liu, Jiao, Hu, Xin-Wen, Li, Rui-Mei, Guo, Jian-Chun, Yao, Yuan
Format Journal Article
LanguageEnglish
Published Switzerland MDPI AG 01.09.2022
MDPI
Subjects
Acetosyringone
Agrobacterium
Antibiotics
Cassava
Cell size
CRISPR
CRISPR/Cas9
Crop diseases
Cultivars
Efficiency
efficient transformation
Embryos
Equipment and supplies
friable embryogenic calli
Gene Editing
Genes
Genetic aspects
Genetic engineering
Genetic transformation
Genomes
homozygous
Manihot - genetics
Manihot esculenta
Mutation
Protocol
SC8
Starch - metabolism
Transformation, Genetic
China
Africa
homozygous
SC8
efficient transformation
CRISPR/Cas9
cassava
friable embryogenic calli
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Abstract Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 ( Crantz cv. SC8) is one of the main locally planted cultivars. In this study, an efficient transformation system for cassava SC8 mediated with strain LBA4404 was presented for the first time. Cassava friable embryogenic calli (FECs) were transformed through the binary vector pCAMBIA1304 harboring and fused genes driven by the promoter. The transformation efficiency was increased in the conditions of strain cell infection density (OD = 0.65), 250 µM acetosyringone induction, and -cultivation with wet FECs for 3 days in dark. Based on the optimized transformation protocol, approximately 120-140 independent transgenic lines per mL settled cell volume (SCV) of FECs were created by gene transformation in approximately 5 months, and 45.83% homozygous mono-allelic mutations of the gene with a promoter-driven CRISPR/Cas9 system were generated. This study will open a more functional avenue for the genetic improvement of cassava SC8.
AbstractList Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 (Manihot esculenta Crantz cv. SC8) is one of the main locally planted cultivars. In this study, an efficient transformation system for cassava SC8 mediated with Agrobacterium strain LBA4404 was presented for the first time. Cassava friable embryogenic calli (FECs) were transformed through the binary vector pCAMBIA1304 harboring GUS- and GFP-fused genes driven by the CaMV35S promoter. The transformation efficiency was increased in the conditions of Agrobacterium strain cell infection density (OD[sub.600] = 0.65), 250 µM acetosyringone induction, and agro-cultivation with wet FECs for 3 days in dark. Based on the optimized transformation protocol, approximately 120–140 independent transgenic lines per mL settled cell volume (SCV) of FECs were created by gene transformation in approximately 5 months, and 45.83% homozygous mono-allelic mutations of the MePDS gene with a YAO promoter-driven CRISPR/Cas9 system were generated. This study will open a more functional avenue for the genetic improvement of cassava SC8.
Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 ( Manihot esculenta Crantz cv. SC8) is one of the main locally planted cultivars. In this study, an efficient transformation system for cassava SC8 mediated with Agrobacterium strain LBA4404 was presented for the first time. Cassava friable embryogenic calli (FECs) were transformed through the binary vector pCAMBIA1304 harboring GUS- and GFP- fused genes driven by the CaMV35S promoter. The transformation efficiency was increased in the conditions of Agrobacterium strain cell infection density (OD 600 = 0.65), 250 µM acetosyringone induction, and agro -cultivation with wet FECs for 3 days in dark. Based on the optimized transformation protocol, approximately 120–140 independent transgenic lines per mL settled cell volume (SCV) of FECs were created by gene transformation in approximately 5 months, and 45.83% homozygous mono-allelic mutations of the MePDS gene with a YAO promoter-driven CRISPR/Cas9 system were generated. This study will open a more functional avenue for the genetic improvement of cassava SC8.
Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 ( Crantz cv. SC8) is one of the main locally planted cultivars. In this study, an efficient transformation system for cassava SC8 mediated with strain LBA4404 was presented for the first time. Cassava friable embryogenic calli (FECs) were transformed through the binary vector pCAMBIA1304 harboring and fused genes driven by the promoter. The transformation efficiency was increased in the conditions of strain cell infection density (OD = 0.65), 250 µM acetosyringone induction, and -cultivation with wet FECs for 3 days in dark. Based on the optimized transformation protocol, approximately 120-140 independent transgenic lines per mL settled cell volume (SCV) of FECs were created by gene transformation in approximately 5 months, and 45.83% homozygous mono-allelic mutations of the gene with a promoter-driven CRISPR/Cas9 system were generated. This study will open a more functional avenue for the genetic improvement of cassava SC8.
Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 (Manihot esculenta Crantz cv. SC8) is one of the main locally planted cultivars. In this study, an efficient transformation system for cassava SC8 mediated with Agrobacterium strain LBA4404 was presented for the first time. Cassava friable embryogenic calli (FECs) were transformed through the binary vector pCAMBIA1304 harboring GUS- and GFP-fused genes driven by the CaMV35S promoter. The transformation efficiency was increased in the conditions of Agrobacterium strain cell infection density (OD600 = 0.65), 250 µM acetosyringone induction, and agro-cultivation with wet FECs for 3 days in dark. Based on the optimized transformation protocol, approximately 120–140 independent transgenic lines per mL settled cell volume (SCV) of FECs were created by gene transformation in approximately 5 months, and 45.83% homozygous mono-allelic mutations of the MePDS gene with a YAO promoter-driven CRISPR/Cas9 system were generated. This study will open a more functional avenue for the genetic improvement of cassava SC8.
Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 (Manihot esculenta Crantz cv. SC8) is one of the main locally planted cultivars. In this study, an efficient transformation system for cassava SC8 mediated with Agrobacterium strain LBA4404 was presented for the first time. Cassava friable embryogenic calli (FECs) were transformed through the binary vector pCAMBIA1304 harboring GUS- and GFP-fused genes driven by the CaMV35S promoter. The transformation efficiency was increased in the conditions of Agrobacterium strain cell infection density (OD600 = 0.65), 250 µM acetosyringone induction, and agro-cultivation with wet FECs for 3 days in dark. Based on the optimized transformation protocol, approximately 120-140 independent transgenic lines per mL settled cell volume (SCV) of FECs were created by gene transformation in approximately 5 months, and 45.83% homozygous mono-allelic mutations of the MePDS gene with a YAO promoter-driven CRISPR/Cas9 system were generated. This study will open a more functional avenue for the genetic improvement of cassava SC8.Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 (Manihot esculenta Crantz cv. SC8) is one of the main locally planted cultivars. In this study, an efficient transformation system for cassava SC8 mediated with Agrobacterium strain LBA4404 was presented for the first time. Cassava friable embryogenic calli (FECs) were transformed through the binary vector pCAMBIA1304 harboring GUS- and GFP-fused genes driven by the CaMV35S promoter. The transformation efficiency was increased in the conditions of Agrobacterium strain cell infection density (OD600 = 0.65), 250 µM acetosyringone induction, and agro-cultivation with wet FECs for 3 days in dark. Based on the optimized transformation protocol, approximately 120-140 independent transgenic lines per mL settled cell volume (SCV) of FECs were created by gene transformation in approximately 5 months, and 45.83% homozygous mono-allelic mutations of the MePDS gene with a YAO promoter-driven CRISPR/Cas9 system were generated. This study will open a more functional avenue for the genetic improvement of cassava SC8.
Audience Academic
Author Zhen, Xing-Hou
Hou, Jing-Yi
Geng, Meng-Ting
Che, Yan-Nian
Yao, Yuan
Liu, Jiao
Li, Rui-Mei
Wang, Ya-Jie
Guo, Jian-Chun
Lu, Xiao-Hua
Yang, Hui
Hu, Xin-Wen
AuthorAffiliation 2 Key Laboratory for Biology and Genetic Resources of Tropical Crops of Hainan Province, Hainan Institute for Tropical Agricultural Resources, Haikou 571101, China
1 Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China
3 School of Life Sciences, Hainan University, Haikou 570228, China
4 San Yan Research Institute, Chinese Academy of Tropical Agricultural Sciences & Hainan Yazhou Bay Seed Lab, Sanya 572025, China
AuthorAffiliation_xml – name: 1 Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou 571101, China
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Keywords homozygous
SC8
efficient transformation
CRISPR/Cas9
cassava
friable embryogenic calli
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SSID ssj0000402005
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Snippet Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 ( Crantz cv. SC8) is one of the main locally planted...
Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 (Manihot esculenta Crantz cv. SC8) is one of the main...
Cassava starch is a widely used raw material for industrial production. South Chinese cassava cultivar 8 ( Manihot esculenta Crantz cv. SC8) is one of the main...
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StartPage 1650
SubjectTerms Acetosyringone
Agrobacterium
Antibiotics
Cassava
Cell size
CRISPR
CRISPR/Cas9
Crop diseases
Cultivars
Efficiency
efficient transformation
Embryos
Equipment and supplies
friable embryogenic calli
Gene Editing
Genes
Genetic aspects
Genetic engineering
Genetic transformation
Genomes
homozygous
Manihot - genetics
Manihot esculenta
Mutation
Protocol
SC8
Starch - metabolism
Transformation, Genetic
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Title A Transformation and Genome Editing System for Cassava Cultivar SC8
URI https://www.ncbi.nlm.nih.gov/pubmed/36140817
https://www.proquest.com/docview/2716540132/abstract/
https://www.proquest.com/docview/2717690250/abstract/
https://pubmed.ncbi.nlm.nih.gov/PMC9498335
https://doaj.org/article/68c6f7b848134b42a8a6c8a7f2611540
Volume 13
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