Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells

Vitis vinifera cv. Chardonnay suspension cultures were established from proembryogenic mass and employed for optimizing Agrobacterium-mediated transformation system. One-factor-at-a-time experiment revealed that OD600 of Agrobacterium, time of inoculation, co-cultivation, and cell-drying before inoc...

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Main Authors: Jiao WU, Ya-li ZHANG, Rong-rong HE, Lei ZHU, Chao-xia WANG, Xia XU, Jiang LU
Format: Article
Language:English
Published: KeAi Communications Co., Ltd. 2012-03-01
Series:Journal of Integrative Agriculture
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Online Access:http://www.sciencedirect.com/science/article/pii/S2095311912600239
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author Jiao WU
Ya-li ZHANG
Rong-rong HE
Lei ZHU
Chao-xia WANG
Xia XU
Jiang LU
author_facet Jiao WU
Ya-li ZHANG
Rong-rong HE
Lei ZHU
Chao-xia WANG
Xia XU
Jiang LU
author_sort Jiao WU
collection DOAJ
description Vitis vinifera cv. Chardonnay suspension cultures were established from proembryogenic mass and employed for optimizing Agrobacterium-mediated transformation system. One-factor-at-a-time experiment revealed that OD600 of Agrobacterium, time of inoculation, co-cultivation, and cell-drying before inoculation significantly affected the transformation efficiency which reached maximum 21.5% at the following conditions: 0.8 of OD600, 25 min of inoculation, 2 d of co-cultivation, and 10 min of cell drying. Response surface methodology experiments based on a five-level, four-factor central-composite rotatable design were then used to optimize these selected factors. The optimized conditions for Chardonnay grape transformation were: 0.8711 of OD600, 28.9 min of inoculation, 2.25 d of co-cultivation and 11.76 min of cell drying. After optimization, transformation efficiency was 26.2% and there were no interactions among different factors.
format Article
id doaj-art-418600b8edb44db08df8159e4a486476
institution Kabale University
issn 2095-3119
language English
publishDate 2012-03-01
publisher KeAi Communications Co., Ltd.
record_format Article
series Journal of Integrative Agriculture
spelling doaj-art-418600b8edb44db08df8159e4a4864762025-08-20T03:58:21ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192012-03-0111338739610.1016/S2095-3119(12)60023-9Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic CellsJiao WU0Ya-li ZHANG1Rong-rong HE2Lei ZHU3Chao-xia WANG4Xia XU5Jiang LU6College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R. China; WU Jiao, Tel: +86-10-62737465College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R. China; ZHANG Ya-li, Tel: +86-10-62737465College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R. ChinaCollege of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R. ChinaCollege of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R. ChinaCenter for Viticulture and Small Fruit Research, Florida A&M University, Tallahassee 32317, USACollege of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, P.R. China; Center for Viticulture and Small Fruit Research, Florida A&M University, Tallahassee 32317, USA; Correspondence LU Jiang, Tel: +1-850-412-7393, Fax: +1-850-412-7464Vitis vinifera cv. Chardonnay suspension cultures were established from proembryogenic mass and employed for optimizing Agrobacterium-mediated transformation system. One-factor-at-a-time experiment revealed that OD600 of Agrobacterium, time of inoculation, co-cultivation, and cell-drying before inoculation significantly affected the transformation efficiency which reached maximum 21.5% at the following conditions: 0.8 of OD600, 25 min of inoculation, 2 d of co-cultivation, and 10 min of cell drying. Response surface methodology experiments based on a five-level, four-factor central-composite rotatable design were then used to optimize these selected factors. The optimized conditions for Chardonnay grape transformation were: 0.8711 of OD600, 28.9 min of inoculation, 2.25 d of co-cultivation and 11.76 min of cell drying. After optimization, transformation efficiency was 26.2% and there were no interactions among different factors.http://www.sciencedirect.com/science/article/pii/S2095311912600239transformationChardonnaysuspension culturesresponse surface methodology
spellingShingle Jiao WU
Ya-li ZHANG
Rong-rong HE
Lei ZHU
Chao-xia WANG
Xia XU
Jiang LU
Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells
Journal of Integrative Agriculture
transformation
Chardonnay
suspension cultures
response surface methodology
title Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells
title_full Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells
title_fullStr Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells
title_full_unstemmed Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells
title_short Optimization of Transformation Efficiency of Suspension Cultured Vitis vinifera cv. Chardonnay Embryogenic Cells
title_sort optimization of transformation efficiency of suspension cultured vitis vinifera cv chardonnay embryogenic cells
topic transformation
Chardonnay
suspension cultures
response surface methodology
url http://www.sciencedirect.com/science/article/pii/S2095311912600239
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