A Rapid Method for Obtaining the Transgenic Roots of Crassulaceae Plants

Crassulaceae plants are valued for their horticultural, ecological, and economic significance, but their genetic improvement is hindered by the absence of efficient and stable genetic transformation methods. Therefore, the development of a tailored genetic transformation method is crucial for enhanc...

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Main Authors: Lan Zhou, Yulu Yang, Anket Sharma, Vijay Pratap Singh, Durgesh Kumar Tripathi, Wona Ding, Muhammad Junaid Rao, Bingsong Zheng, Xiaofei Wang
Format: Article
Language:English
Published: MDPI AG 2024-10-01
Series:Plants
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Online Access:https://www.mdpi.com/2223-7747/13/21/3024
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author Lan Zhou
Yulu Yang
Anket Sharma
Vijay Pratap Singh
Durgesh Kumar Tripathi
Wona Ding
Muhammad Junaid Rao
Bingsong Zheng
Xiaofei Wang
author_facet Lan Zhou
Yulu Yang
Anket Sharma
Vijay Pratap Singh
Durgesh Kumar Tripathi
Wona Ding
Muhammad Junaid Rao
Bingsong Zheng
Xiaofei Wang
author_sort Lan Zhou
collection DOAJ
description Crassulaceae plants are valued for their horticultural, ecological, and economic significance, but their genetic improvement is hindered by the absence of efficient and stable genetic transformation methods. Therefore, the development of a tailored genetic transformation method is crucial for enhancing the progress of the genetic improvement of Crassulaceae plants. The results indicate that, in the transformation experiments conducted on <i>Kalanchoe tetraphylla</i>, the K599 strain exhibited the highest transformation efficiency (76.67%), while C58C1 was least efficient (21.43%). An acetosyringone concentration of 100 μM was optimal for the hairy root transformation, and the immersion method yielded the highest efficiency. Additionally, the Silwet L-77 concentration significantly influenced the transformation efficiency, with 0.05% leading to a decrease. Upon four Crassulaceae species, notable differences were observed, with <i>K. tetraphylla</i> exhibiting the highest efficiency of 100%, and <i>Sedum alfredii</i> displaying the lowest efficiency of 5%. The <i>RUBY</i> reporter gene offers a more distinct advantage over <i>GFP</i> in observing the transformation effects. This study developed a simple, feasible, and cost-effective method for obtaining transgenic roots from leaves of Crassulaceae. The methodology provides technical support for the genetic improvement and gene function research of Crassulaceae plants.
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spelling doaj-art-1c7d6f5be7164f4ba40f829ae242c33d2025-08-20T02:13:14ZengMDPI AGPlants2223-77472024-10-011321302410.3390/plants13213024A Rapid Method for Obtaining the Transgenic Roots of Crassulaceae PlantsLan Zhou0Yulu Yang1Anket Sharma2Vijay Pratap Singh3Durgesh Kumar Tripathi4Wona Ding5Muhammad Junaid Rao6Bingsong Zheng7Xiaofei Wang8State Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, ChinaState Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, ChinaState Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, ChinaState Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, ChinaCrop Nanobiology and Molecular Biology Laboratory, Amity Institute of Organic Agriculture (AIOA), Amity University, Noida 201303, Uttar Pradesh, IndiaCollege of Science and Technology, Ningbo University, Ningbo 315300, ChinaState Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, ChinaState Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, ChinaState Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, ChinaCrassulaceae plants are valued for their horticultural, ecological, and economic significance, but their genetic improvement is hindered by the absence of efficient and stable genetic transformation methods. Therefore, the development of a tailored genetic transformation method is crucial for enhancing the progress of the genetic improvement of Crassulaceae plants. The results indicate that, in the transformation experiments conducted on <i>Kalanchoe tetraphylla</i>, the K599 strain exhibited the highest transformation efficiency (76.67%), while C58C1 was least efficient (21.43%). An acetosyringone concentration of 100 μM was optimal for the hairy root transformation, and the immersion method yielded the highest efficiency. Additionally, the Silwet L-77 concentration significantly influenced the transformation efficiency, with 0.05% leading to a decrease. Upon four Crassulaceae species, notable differences were observed, with <i>K. tetraphylla</i> exhibiting the highest efficiency of 100%, and <i>Sedum alfredii</i> displaying the lowest efficiency of 5%. The <i>RUBY</i> reporter gene offers a more distinct advantage over <i>GFP</i> in observing the transformation effects. This study developed a simple, feasible, and cost-effective method for obtaining transgenic roots from leaves of Crassulaceae. The methodology provides technical support for the genetic improvement and gene function research of Crassulaceae plants.https://www.mdpi.com/2223-7747/13/21/3024Crassulaceae<i>Agrobacterium</i>-mediated transformation<i>GFP</i> reporter<i>RUBY</i> reportertransformation system
spellingShingle Lan Zhou
Yulu Yang
Anket Sharma
Vijay Pratap Singh
Durgesh Kumar Tripathi
Wona Ding
Muhammad Junaid Rao
Bingsong Zheng
Xiaofei Wang
A Rapid Method for Obtaining the Transgenic Roots of Crassulaceae Plants
Plants
Crassulaceae
<i>Agrobacterium</i>-mediated transformation
<i>GFP</i> reporter
<i>RUBY</i> reporter
transformation system
title A Rapid Method for Obtaining the Transgenic Roots of Crassulaceae Plants
title_full A Rapid Method for Obtaining the Transgenic Roots of Crassulaceae Plants
title_fullStr A Rapid Method for Obtaining the Transgenic Roots of Crassulaceae Plants
title_full_unstemmed A Rapid Method for Obtaining the Transgenic Roots of Crassulaceae Plants
title_short A Rapid Method for Obtaining the Transgenic Roots of Crassulaceae Plants
title_sort rapid method for obtaining the transgenic roots of crassulaceae plants
topic Crassulaceae
<i>Agrobacterium</i>-mediated transformation
<i>GFP</i> reporter
<i>RUBY</i> reporter
transformation system
url https://www.mdpi.com/2223-7747/13/21/3024
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