Optimization of High-Efficiency Tissue Culture Regeneration Systems in Gray Poplar

A series of tissue culture regeneration protocols were conducted on gray poplar (<i>P. tremula × P. alba</i>) to select the most efficient callus induction medium, adventitious shoot differentiation medium, shoot elongation medium and rooting medium, which laid the foundation for the opt...

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Main Authors: Huanhuan Li, Hang Wang, Lianke Guan, Zihui Li, Hua Wang, Jie Luo
Format: Article
Language:English
Published: MDPI AG 2023-09-01
Series:Life
Subjects:
Online Access:https://www.mdpi.com/2075-1729/13/9/1896
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author Huanhuan Li
Hang Wang
Lianke Guan
Zihui Li
Hua Wang
Jie Luo
author_facet Huanhuan Li
Hang Wang
Lianke Guan
Zihui Li
Hua Wang
Jie Luo
author_sort Huanhuan Li
collection DOAJ
description A series of tissue culture regeneration protocols were conducted on gray poplar (<i>P. tremula × P. alba</i>) to select the most efficient callus induction medium, adventitious shoot differentiation medium, shoot elongation medium and rooting medium, which laid the foundation for the optimization of genetic transformation technology for gray poplar. The results showed that the Woody Plant Medium (WPM) supplemented with 0.10 mg L<sup>−1</sup> kinetin (KT) and 1.00 mg L<sup>−1</sup> 2,4-dichlorophenoxyacetic acid (2,4-D) was the most suitable medium for callus induction. The callus induction rates of different tissues were greater than 85.7%. The optimal adventitious shoot differentiation medium was the WPM supplemented with 0.02 mg L<sup>−1</sup> thidiazuron (TDZ), and the adventitious shoot differentiation rates of young tissues were 22.2–41.9%. The optimal direct differentiation medium was the Murashige and Skoog (MS) medium supplemented with 0.20 mg L<sup>−1</sup> 6-benzylaminopurine (6-BA), 0.10 mg L<sup>−1</sup> indole butyric acid (IBA) and 0.001 mg L<sup>−1</sup> TDZ, and the differentiation rate of adventitious shoots was greater than 94%. The best shoot elongation medium for adventitious shoots was the MS medium with 0.10 mg L<sup>−1</sup> naphthylacetic acid (NAA). After 45 days of cultivation in the MS medium with 0.10 mg L<sup>−1</sup> NAA, the average plant height was 1.8 cm, and the average number of elongated adventitious shoots was 11 per explant. The 1/2 MS medium with 0.10 mg L<sup>−1</sup> NAA showed the best performance for rooting, and later, shoot growth. The direct shoot induction pathway can induce adventitious shoots much faster than the indirect adventitious shoot induction pathway can, and the time cost via the direct adventitious shoot induction pathway can be shortened by 2–6 weeks compared to that of the indirect shoot induction pathway.
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spelling doaj.art-8ce698a3e5824b44ad211347666c0d832023-11-19T11:37:48ZengMDPI AGLife2075-17292023-09-01139189610.3390/life13091896Optimization of High-Efficiency Tissue Culture Regeneration Systems in Gray PoplarHuanhuan Li0Hang Wang1Lianke Guan2Zihui Li3Hua Wang4Jie Luo5College of Horticulture and Forestry Science, Hubei Engineering Technology Research Center for Forestry Information, Huazhong Agricultural University, Wuhan 430070, ChinaCollege of Horticulture and Forestry Science, Hubei Engineering Technology Research Center for Forestry Information, Huazhong Agricultural University, Wuhan 430070, ChinaCollege of Horticulture and Forestry Science, Hubei Engineering Technology Research Center for Forestry Information, Huazhong Agricultural University, Wuhan 430070, ChinaCollege of Horticulture and Forestry Science, Hubei Engineering Technology Research Center for Forestry Information, Huazhong Agricultural University, Wuhan 430070, ChinaCollege of Horticulture and Forestry Science, Hubei Engineering Technology Research Center for Forestry Information, Huazhong Agricultural University, Wuhan 430070, ChinaCollege of Horticulture and Forestry Science, Hubei Engineering Technology Research Center for Forestry Information, Huazhong Agricultural University, Wuhan 430070, ChinaA series of tissue culture regeneration protocols were conducted on gray poplar (<i>P. tremula × P. alba</i>) to select the most efficient callus induction medium, adventitious shoot differentiation medium, shoot elongation medium and rooting medium, which laid the foundation for the optimization of genetic transformation technology for gray poplar. The results showed that the Woody Plant Medium (WPM) supplemented with 0.10 mg L<sup>−1</sup> kinetin (KT) and 1.00 mg L<sup>−1</sup> 2,4-dichlorophenoxyacetic acid (2,4-D) was the most suitable medium for callus induction. The callus induction rates of different tissues were greater than 85.7%. The optimal adventitious shoot differentiation medium was the WPM supplemented with 0.02 mg L<sup>−1</sup> thidiazuron (TDZ), and the adventitious shoot differentiation rates of young tissues were 22.2–41.9%. The optimal direct differentiation medium was the Murashige and Skoog (MS) medium supplemented with 0.20 mg L<sup>−1</sup> 6-benzylaminopurine (6-BA), 0.10 mg L<sup>−1</sup> indole butyric acid (IBA) and 0.001 mg L<sup>−1</sup> TDZ, and the differentiation rate of adventitious shoots was greater than 94%. The best shoot elongation medium for adventitious shoots was the MS medium with 0.10 mg L<sup>−1</sup> naphthylacetic acid (NAA). After 45 days of cultivation in the MS medium with 0.10 mg L<sup>−1</sup> NAA, the average plant height was 1.8 cm, and the average number of elongated adventitious shoots was 11 per explant. The 1/2 MS medium with 0.10 mg L<sup>−1</sup> NAA showed the best performance for rooting, and later, shoot growth. The direct shoot induction pathway can induce adventitious shoots much faster than the indirect adventitious shoot induction pathway can, and the time cost via the direct adventitious shoot induction pathway can be shortened by 2–6 weeks compared to that of the indirect shoot induction pathway.https://www.mdpi.com/2075-1729/13/9/1896poplarregeneration in vitrotissue culturephytohormoneproliferation
spellingShingle Huanhuan Li
Hang Wang
Lianke Guan
Zihui Li
Hua Wang
Jie Luo
Optimization of High-Efficiency Tissue Culture Regeneration Systems in Gray Poplar
Life
poplar
regeneration in vitro
tissue culture
phytohormone
proliferation
title Optimization of High-Efficiency Tissue Culture Regeneration Systems in Gray Poplar
title_full Optimization of High-Efficiency Tissue Culture Regeneration Systems in Gray Poplar
title_fullStr Optimization of High-Efficiency Tissue Culture Regeneration Systems in Gray Poplar
title_full_unstemmed Optimization of High-Efficiency Tissue Culture Regeneration Systems in Gray Poplar
title_short Optimization of High-Efficiency Tissue Culture Regeneration Systems in Gray Poplar
title_sort optimization of high efficiency tissue culture regeneration systems in gray poplar
topic poplar
regeneration in vitro
tissue culture
phytohormone
proliferation
url https://www.mdpi.com/2075-1729/13/9/1896
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AT zihuili optimizationofhighefficiencytissuecultureregenerationsystemsingraypoplar
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