Genotype-dependent induction of embryogenic callus and programmed cell death in Korean pine

Abstract Background The strong genotype dependence of embryogenic callus (EC) induction limits the high-frequency regeneration of Korean pine (Pinus koraiensis) via somatic embryogenesis (SE). Methods We examined the morphological and histocytological features of various callus types using external...

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Main Authors: Yan Liang, Jianghe Li, Xin Xu, Qingzhu Yang, Meiling Gao, Yang Chen, Chunyu Guan
Format: Article
Language:English
Published: BMC 2025-06-01
Series:BMC Plant Biology
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Online Access:https://doi.org/10.1186/s12870-025-06787-1
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author Yan Liang
Jianghe Li
Xin Xu
Qingzhu Yang
Meiling Gao
Yang Chen
Chunyu Guan
author_facet Yan Liang
Jianghe Li
Xin Xu
Qingzhu Yang
Meiling Gao
Yang Chen
Chunyu Guan
author_sort Yan Liang
collection DOAJ
description Abstract Background The strong genotype dependence of embryogenic callus (EC) induction limits the high-frequency regeneration of Korean pine (Pinus koraiensis) via somatic embryogenesis (SE). Methods We examined the morphological and histocytological features of various callus types using external morphology, microstructure, and ultrastructure analyses. To assess their embryogenic potential, we conducted somatic embryogenesis (SE) experiments. Additionally, we investigated programmed cell death (PCD) events associated with genotype-dependent embryogenic callus (EC) formation in Korean pine by performing TUNEL assays, detecting caspase-3 and vacuolar processing enzyme (VPE) activity, and measuring H2O2 and O2 •− levels. Results Obvious cell death was observed in the cell ultrastructure of LEC and NEC induced by recalcitrant genotypes. In LEC cells, we found typical characteristics of autolytic PCD, such as vacuole fusion, vacuolation of cells, invagination and rupture of the plasma membrane, few cytoplasm and organelles. In the NEC, degrading nucleus, withered cytoplasm and invagination and rupture of the plasma membrane were detected in early stage, but vacuole fusion was not observed. In addition, we observed DNA fragmentation into small fragments in LEC and NEC. Significant changes in the activities of caspase-3 and VPE, reactive oxygen species (ROS) levels, especially for H2O2, have been identified in LEC. However, in NEC, only the significant changes in the O2 •− level and the caspase-3 activity were observed. Conclusion PCD was present in NEC and LEC cells in recalcitrant genotypes of Korean pine, which may be responsible for the loss or reduction of embryonic capacity of induced Korean pine callus. Our novel findings on PCD help elucidate the underlying causes of the difficulty in inducing EC from recalcitrant genotypes of Korean pine from a novel perspective.
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spelling doaj-art-7b3e7b700ff34ebf8317b115dff51ef22025-08-20T03:46:08ZengBMCBMC Plant Biology1471-22292025-06-0125111910.1186/s12870-025-06787-1Genotype-dependent induction of embryogenic callus and programmed cell death in Korean pineYan Liang0Jianghe Li1Xin Xu2Qingzhu Yang3Meiling Gao4Yang Chen5Chunyu Guan6School of Life Sciences and A & F, Qiqihar UniversitySchool of Life Sciences and A & F, Qiqihar UniversitySchool of Life Sciences and A & F, Qiqihar UniversitySchool of Life Sciences and A & F, Qiqihar UniversitySchool of Life Sciences and A & F, Qiqihar UniversitySchool of Life Sciences and A & F, Qiqihar UniversitySchool of Life Sciences and A & F, Qiqihar UniversityAbstract Background The strong genotype dependence of embryogenic callus (EC) induction limits the high-frequency regeneration of Korean pine (Pinus koraiensis) via somatic embryogenesis (SE). Methods We examined the morphological and histocytological features of various callus types using external morphology, microstructure, and ultrastructure analyses. To assess their embryogenic potential, we conducted somatic embryogenesis (SE) experiments. Additionally, we investigated programmed cell death (PCD) events associated with genotype-dependent embryogenic callus (EC) formation in Korean pine by performing TUNEL assays, detecting caspase-3 and vacuolar processing enzyme (VPE) activity, and measuring H2O2 and O2 •− levels. Results Obvious cell death was observed in the cell ultrastructure of LEC and NEC induced by recalcitrant genotypes. In LEC cells, we found typical characteristics of autolytic PCD, such as vacuole fusion, vacuolation of cells, invagination and rupture of the plasma membrane, few cytoplasm and organelles. In the NEC, degrading nucleus, withered cytoplasm and invagination and rupture of the plasma membrane were detected in early stage, but vacuole fusion was not observed. In addition, we observed DNA fragmentation into small fragments in LEC and NEC. Significant changes in the activities of caspase-3 and VPE, reactive oxygen species (ROS) levels, especially for H2O2, have been identified in LEC. However, in NEC, only the significant changes in the O2 •− level and the caspase-3 activity were observed. Conclusion PCD was present in NEC and LEC cells in recalcitrant genotypes of Korean pine, which may be responsible for the loss or reduction of embryonic capacity of induced Korean pine callus. Our novel findings on PCD help elucidate the underlying causes of the difficulty in inducing EC from recalcitrant genotypes of Korean pine from a novel perspective.https://doi.org/10.1186/s12870-025-06787-1Pinus koraiensisGenotype-dependent responseEmbryogenic callusLow embryogenic callusNon-embryogenic callusProgrammed cell death
spellingShingle Yan Liang
Jianghe Li
Xin Xu
Qingzhu Yang
Meiling Gao
Yang Chen
Chunyu Guan
Genotype-dependent induction of embryogenic callus and programmed cell death in Korean pine
BMC Plant Biology
Pinus koraiensis
Genotype-dependent response
Embryogenic callus
Low embryogenic callus
Non-embryogenic callus
Programmed cell death
title Genotype-dependent induction of embryogenic callus and programmed cell death in Korean pine
title_full Genotype-dependent induction of embryogenic callus and programmed cell death in Korean pine
title_fullStr Genotype-dependent induction of embryogenic callus and programmed cell death in Korean pine
title_full_unstemmed Genotype-dependent induction of embryogenic callus and programmed cell death in Korean pine
title_short Genotype-dependent induction of embryogenic callus and programmed cell death in Korean pine
title_sort genotype dependent induction of embryogenic callus and programmed cell death in korean pine
topic Pinus koraiensis
Genotype-dependent response
Embryogenic callus
Low embryogenic callus
Non-embryogenic callus
Programmed cell death
url https://doi.org/10.1186/s12870-025-06787-1
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