Efficient induction of tetraploids via adventitious bud regeneration and subsequent phenotypic variation in Acacia melanoxylon

Abstract Background acacia melanoxylon is an important species for establishing pulpwood plantations due to its high application value in engineered wood products. However, the lack of a well-established in vitro regeneration system has severely constrained its industrial-scale propagation and the i...

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Main Authors: Shenxiu Jiang, Yufei Xia, Aoyu Ling, Jianghai Shu, Kairan You, Shun Wang, Dingju Zhan, Bingshan Zeng, Jun Yang, Xiangyang Kang
Format: Article
Language:English
Published: BMC 2025-08-01
Series:Plant Methods
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Online Access:https://doi.org/10.1186/s13007-025-01426-0
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author Shenxiu Jiang
Yufei Xia
Aoyu Ling
Jianghai Shu
Kairan You
Shun Wang
Dingju Zhan
Bingshan Zeng
Jun Yang
Xiangyang Kang
author_facet Shenxiu Jiang
Yufei Xia
Aoyu Ling
Jianghai Shu
Kairan You
Shun Wang
Dingju Zhan
Bingshan Zeng
Jun Yang
Xiangyang Kang
author_sort Shenxiu Jiang
collection DOAJ
description Abstract Background acacia melanoxylon is an important species for establishing pulpwood plantations due to its high application value in engineered wood products. However, the lack of a well-established in vitro regeneration system has severely constrained its industrial-scale propagation and the induction of tetraploids. Results In this study, using the superior A. melanoxylon clone SR3, an in vitro regeneration system using a bud-bearing stem segment was established. A DKW medium supplemented with 0.5 mg/L 6-BA, 0.1 mg/L IAA, and 0.2 mg/L NAA was determined as the optimal differentiation medium. Adding 0.5 mg/L IBA and 0.25 mg/L NAA to the 1/2 MS medium produced a higher rooting percentage and root number. To determine the optimal timing for tetraploid induction in A. melanoxylon, morphological, cytological, and flow cytometric analyses were conducted on the swollen tissue at the base of the bud-bearing stem segment. On the 5th day of preculture, white callus tissue was observed, characterized by vigorous cell division and the highest G2/M-phase cell content in the adventitious bud primordia. After colchicine treatment, the tetraploid induction efficiency on the 5th day of preculture was significantly higher compared to the 4th or 6th day. The highest induction rate of 12.26 ± 0.80% was achieved with 100 mg/L colchicine for 72 h on the 5th day of preculture. Furthermore, tetraploid A. melanoxylon exhibited morphological traits such as reduced plant height, leaf number, and stomatal density. Conclusions This study establishes a stable and effective method for in vitro tetraploid induction in A. melanoxylon, providing theoretical and technical support for polyploid breeding and laying the groundwork for subsequent triploid development.
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spelling doaj-art-e5fd34a2af804ef0a6d7cc429ab1efde2025-08-24T11:31:53ZengBMCPlant Methods1746-48112025-08-0121111510.1186/s13007-025-01426-0Efficient induction of tetraploids via adventitious bud regeneration and subsequent phenotypic variation in Acacia melanoxylonShenxiu Jiang0Yufei Xia1Aoyu Ling2Jianghai Shu3Kairan You4Shun Wang5Dingju Zhan6Bingshan Zeng7Jun Yang8Xiangyang Kang9State Key Laboratory of Tree Genetics and Breeding, National Engineering Research Center of Tree Breeding and Ecological Restoration, College of Biological Sciences and Biotechnology, Beijing Forestry UniversityState Key Laboratory of Tree Genetics and Breeding, National Engineering Research Center of Tree Breeding and Ecological Restoration, College of Biological Sciences and Biotechnology, Beijing Forestry UniversityState Key Laboratory of Tree Genetics and Breeding, National Engineering Research Center of Tree Breeding and Ecological Restoration, College of Biological Sciences and Biotechnology, Beijing Forestry UniversityState Key Laboratory of Tree Genetics and Breeding, National Engineering Research Center of Tree Breeding and Ecological Restoration, College of Biological Sciences and Biotechnology, Beijing Forestry UniversityState Key Laboratory of Tree Genetics and Breeding, National Engineering Research Center of Tree Breeding and Ecological Restoration, College of Biological Sciences and Biotechnology, Beijing Forestry UniversityState Key Laboratory of Tree Genetics and Breeding, National Engineering Research Center of Tree Breeding and Ecological Restoration, College of Biological Sciences and Biotechnology, Beijing Forestry UniversityGuangxi Bagui R&D Institute for Forest Tree and Flower BreedingState Key Laboratory of Tree Genetics and Breeding, Key Laboratory of State Forestry and Grassland Administration on Tropical Forestry, Research Institute of Tropical Forestry, Chinese Academy of ForestryState Key Laboratory of Tree Genetics and Breeding, National Engineering Research Center of Tree Breeding and Ecological Restoration, College of Biological Sciences and Biotechnology, Beijing Forestry UniversityState Key Laboratory of Tree Genetics and Breeding, National Engineering Research Center of Tree Breeding and Ecological Restoration, College of Biological Sciences and Biotechnology, Beijing Forestry UniversityAbstract Background acacia melanoxylon is an important species for establishing pulpwood plantations due to its high application value in engineered wood products. However, the lack of a well-established in vitro regeneration system has severely constrained its industrial-scale propagation and the induction of tetraploids. Results In this study, using the superior A. melanoxylon clone SR3, an in vitro regeneration system using a bud-bearing stem segment was established. A DKW medium supplemented with 0.5 mg/L 6-BA, 0.1 mg/L IAA, and 0.2 mg/L NAA was determined as the optimal differentiation medium. Adding 0.5 mg/L IBA and 0.25 mg/L NAA to the 1/2 MS medium produced a higher rooting percentage and root number. To determine the optimal timing for tetraploid induction in A. melanoxylon, morphological, cytological, and flow cytometric analyses were conducted on the swollen tissue at the base of the bud-bearing stem segment. On the 5th day of preculture, white callus tissue was observed, characterized by vigorous cell division and the highest G2/M-phase cell content in the adventitious bud primordia. After colchicine treatment, the tetraploid induction efficiency on the 5th day of preculture was significantly higher compared to the 4th or 6th day. The highest induction rate of 12.26 ± 0.80% was achieved with 100 mg/L colchicine for 72 h on the 5th day of preculture. Furthermore, tetraploid A. melanoxylon exhibited morphological traits such as reduced plant height, leaf number, and stomatal density. Conclusions This study establishes a stable and effective method for in vitro tetraploid induction in A. melanoxylon, providing theoretical and technical support for polyploid breeding and laying the groundwork for subsequent triploid development.https://doi.org/10.1186/s13007-025-01426-0Acacia treeBud-bearing stem segmentAdventitious budDevelopmental stage observationTetraploid inductionMorphological variation
spellingShingle Shenxiu Jiang
Yufei Xia
Aoyu Ling
Jianghai Shu
Kairan You
Shun Wang
Dingju Zhan
Bingshan Zeng
Jun Yang
Xiangyang Kang
Efficient induction of tetraploids via adventitious bud regeneration and subsequent phenotypic variation in Acacia melanoxylon
Plant Methods
Acacia tree
Bud-bearing stem segment
Adventitious bud
Developmental stage observation
Tetraploid induction
Morphological variation
title Efficient induction of tetraploids via adventitious bud regeneration and subsequent phenotypic variation in Acacia melanoxylon
title_full Efficient induction of tetraploids via adventitious bud regeneration and subsequent phenotypic variation in Acacia melanoxylon
title_fullStr Efficient induction of tetraploids via adventitious bud regeneration and subsequent phenotypic variation in Acacia melanoxylon
title_full_unstemmed Efficient induction of tetraploids via adventitious bud regeneration and subsequent phenotypic variation in Acacia melanoxylon
title_short Efficient induction of tetraploids via adventitious bud regeneration and subsequent phenotypic variation in Acacia melanoxylon
title_sort efficient induction of tetraploids via adventitious bud regeneration and subsequent phenotypic variation in acacia melanoxylon
topic Acacia tree
Bud-bearing stem segment
Adventitious bud
Developmental stage observation
Tetraploid induction
Morphological variation
url https://doi.org/10.1186/s13007-025-01426-0
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