An efficient leaf regeneration and genetic transformation system for Pyrus betulifolia

Abstract Pears rank as the third most produced fruit in China, with widespread cultivation throughout the country. Asexual propagation, primarily through grafting onto Pyrus betulifolia rootstocks, is the predominant method of cultivation. However, the absence of an efficient genetic transformation...

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Main Authors: Zhihui Yu, Yuqi Wang, Manyu Zhang, Qiushi Liu, Qi Pan, Tianzhong Li, Shengnan Wang
Format: Article
Language:English
Published: Springer 2025-02-01
Series:Horticulture Advances
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Online Access:https://doi.org/10.1007/s44281-024-00060-5
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author Zhihui Yu
Yuqi Wang
Manyu Zhang
Qiushi Liu
Qi Pan
Tianzhong Li
Shengnan Wang
author_facet Zhihui Yu
Yuqi Wang
Manyu Zhang
Qiushi Liu
Qi Pan
Tianzhong Li
Shengnan Wang
author_sort Zhihui Yu
collection DOAJ
description Abstract Pears rank as the third most produced fruit in China, with widespread cultivation throughout the country. Asexual propagation, primarily through grafting onto Pyrus betulifolia rootstocks, is the predominant method of cultivation. However, the absence of an efficient genetic transformation system for P. betulifolia significantly hinders genetic enhancement efforts for pear rootstocks. In this study, TDL39, a P. betulifolia genotype, was identified for its remarkable regenerative capacity. Leaf strips derived from 60-day-old TDL39 seedlings were cultured abaxial side up on a regeneration medium consisting of NN69, 3.0 mg/L thidiazuron (TDZ), 0.1 mg/L indole-3-butyric acid (IBA), 30 g/L sucrose, and 6.6 g/L agar. The strips were kept in the dark for 28 days and subsequently exposed to light for 30 days, resulting in the formation of adventitious shoots with an average regeneration rate of 81.5%. Employing this regeneration protocol, transgenic P. betulifolia plants overexpressing the Green Fluorescent Protein (GFP) gene were successfully generated, with 15 mg/L kanamycin (Kan) used for selection. The highest transformation efficiency achieved was 4.2%. This study successfully establishes a regeneration system for P. betulifolia and facilitates the production of transgenic plants, thereby advancing molecular breeding and the functional characterization of P. betulifolia and other pear genotypes.
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spelling doaj-art-495a34d46f884515a3d43bd3f212828c2025-08-20T02:16:40ZengSpringerHorticulture Advances2948-11042025-02-013111510.1007/s44281-024-00060-5An efficient leaf regeneration and genetic transformation system for Pyrus betulifoliaZhihui Yu0Yuqi Wang1Manyu Zhang2Qiushi Liu3Qi Pan4Tianzhong Li5Shengnan Wang6College of Horticulture, China Agricultural UniversityCollege of Horticulture, China Agricultural UniversityCollege of Horticulture, China Agricultural UniversityCollege of Horticulture, China Agricultural UniversityCollege of Horticulture, China Agricultural UniversityCollege of Horticulture, China Agricultural UniversityCollege of Horticulture, China Agricultural UniversityAbstract Pears rank as the third most produced fruit in China, with widespread cultivation throughout the country. Asexual propagation, primarily through grafting onto Pyrus betulifolia rootstocks, is the predominant method of cultivation. However, the absence of an efficient genetic transformation system for P. betulifolia significantly hinders genetic enhancement efforts for pear rootstocks. In this study, TDL39, a P. betulifolia genotype, was identified for its remarkable regenerative capacity. Leaf strips derived from 60-day-old TDL39 seedlings were cultured abaxial side up on a regeneration medium consisting of NN69, 3.0 mg/L thidiazuron (TDZ), 0.1 mg/L indole-3-butyric acid (IBA), 30 g/L sucrose, and 6.6 g/L agar. The strips were kept in the dark for 28 days and subsequently exposed to light for 30 days, resulting in the formation of adventitious shoots with an average regeneration rate of 81.5%. Employing this regeneration protocol, transgenic P. betulifolia plants overexpressing the Green Fluorescent Protein (GFP) gene were successfully generated, with 15 mg/L kanamycin (Kan) used for selection. The highest transformation efficiency achieved was 4.2%. This study successfully establishes a regeneration system for P. betulifolia and facilitates the production of transgenic plants, thereby advancing molecular breeding and the functional characterization of P. betulifolia and other pear genotypes.https://doi.org/10.1007/s44281-024-00060-5Pyrus betulifolia BungeLeaf regenerationGenetic transformation
spellingShingle Zhihui Yu
Yuqi Wang
Manyu Zhang
Qiushi Liu
Qi Pan
Tianzhong Li
Shengnan Wang
An efficient leaf regeneration and genetic transformation system for Pyrus betulifolia
Horticulture Advances
Pyrus betulifolia Bunge
Leaf regeneration
Genetic transformation
title An efficient leaf regeneration and genetic transformation system for Pyrus betulifolia
title_full An efficient leaf regeneration and genetic transformation system for Pyrus betulifolia
title_fullStr An efficient leaf regeneration and genetic transformation system for Pyrus betulifolia
title_full_unstemmed An efficient leaf regeneration and genetic transformation system for Pyrus betulifolia
title_short An efficient leaf regeneration and genetic transformation system for Pyrus betulifolia
title_sort efficient leaf regeneration and genetic transformation system for pyrus betulifolia
topic Pyrus betulifolia Bunge
Leaf regeneration
Genetic transformation
url https://doi.org/10.1007/s44281-024-00060-5
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