Somatic Embryogenesis and Genetic Transformation of <i>Caragana intermedia</i>

<i>Caragana intermedia</i> is a perennial shrub species in the genus <i>Caragana</i> (Fabaceae), demonstrating remarkable stress resistance and adaptability. However, research on its somatic embryogenesis (SE) and genetic transformation techniques remains limited. In this stu...

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Main Authors: Ju Tian, Jialei Zhu, Xiaohan Deng, Xu Zhu, Ruigang Wang, Guojing Li
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Plants
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Online Access:https://www.mdpi.com/2223-7747/14/10/1545
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author Ju Tian
Jialei Zhu
Xiaohan Deng
Xu Zhu
Ruigang Wang
Guojing Li
author_facet Ju Tian
Jialei Zhu
Xiaohan Deng
Xu Zhu
Ruigang Wang
Guojing Li
author_sort Ju Tian
collection DOAJ
description <i>Caragana intermedia</i> is a perennial shrub species in the genus <i>Caragana</i> (Fabaceae), demonstrating remarkable stress resistance and adaptability. However, research on its somatic embryogenesis (SE) and genetic transformation techniques remains limited. In this study, we established an SE system by utilizing immature cotyledons isolated from young <i>C. intermedia</i> seeds. Our findings demonstrated that the immature cotyledons at 6–7 weeks after flowering (WAF) were the best explants for SE. The optimal embryo induction medium consisted of an MS basal medium supplemented with 5 mg/L α-naphthaleneacetic acid (NAA), 3 mg/L 6-benzylaminopurine (6-BA), 30 g/L sucrose, 7 g/L agar, and 500 mg/L hydrolyzed casein. Cotyledon-stage embryos germinated on a half-strength MS medium, exhibiting a 34.36% germination rate. Based on the SE system, we developed a preliminary genetic transformation system using the <i>RUBY</i> reporter gene, which successfully generated transgenic calli and cotyledon-stage embryos. The establishment of the SE system is expected to shorten breeding cycles, facilitate propagation of superior cultivars, and support large-scale industrial applications in <i>C. intermedia</i>. Furthermore, the stable transformation system provides a platform for molecular breeding and gene function verification.
format Article
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institution Kabale University
issn 2223-7747
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publishDate 2025-05-01
publisher MDPI AG
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spelling doaj-art-52012d427bd7428bbb3d4c37d7820d272025-08-20T03:47:54ZengMDPI AGPlants2223-77472025-05-011410154510.3390/plants14101545Somatic Embryogenesis and Genetic Transformation of <i>Caragana intermedia</i>Ju Tian0Jialei Zhu1Xiaohan Deng2Xu Zhu3Ruigang Wang4Guojing Li5Inner Mongolia Key Laboratory of Plants Adversity Adaptation and Genetic Improvement in Cold and Arid Regions, Inner Mongolia Agricultural University, Hohhot 010018, ChinaSchool of Biology and Agriculture, Shaoguan University, Shaoguan 512005, ChinaInner Mongolia Key Laboratory of Plants Adversity Adaptation and Genetic Improvement in Cold and Arid Regions, Inner Mongolia Agricultural University, Hohhot 010018, ChinaInner Mongolia Key Laboratory of Plants Adversity Adaptation and Genetic Improvement in Cold and Arid Regions, Inner Mongolia Agricultural University, Hohhot 010018, ChinaInner Mongolia Key Laboratory of Plants Adversity Adaptation and Genetic Improvement in Cold and Arid Regions, Inner Mongolia Agricultural University, Hohhot 010018, ChinaInner Mongolia Key Laboratory of Plants Adversity Adaptation and Genetic Improvement in Cold and Arid Regions, Inner Mongolia Agricultural University, Hohhot 010018, China<i>Caragana intermedia</i> is a perennial shrub species in the genus <i>Caragana</i> (Fabaceae), demonstrating remarkable stress resistance and adaptability. However, research on its somatic embryogenesis (SE) and genetic transformation techniques remains limited. In this study, we established an SE system by utilizing immature cotyledons isolated from young <i>C. intermedia</i> seeds. Our findings demonstrated that the immature cotyledons at 6–7 weeks after flowering (WAF) were the best explants for SE. The optimal embryo induction medium consisted of an MS basal medium supplemented with 5 mg/L α-naphthaleneacetic acid (NAA), 3 mg/L 6-benzylaminopurine (6-BA), 30 g/L sucrose, 7 g/L agar, and 500 mg/L hydrolyzed casein. Cotyledon-stage embryos germinated on a half-strength MS medium, exhibiting a 34.36% germination rate. Based on the SE system, we developed a preliminary genetic transformation system using the <i>RUBY</i> reporter gene, which successfully generated transgenic calli and cotyledon-stage embryos. The establishment of the SE system is expected to shorten breeding cycles, facilitate propagation of superior cultivars, and support large-scale industrial applications in <i>C. intermedia</i>. Furthermore, the stable transformation system provides a platform for molecular breeding and gene function verification.https://www.mdpi.com/2223-7747/14/10/1545cotyledonplant tissue cultureembryogenic callusmorphogenesisGMO
spellingShingle Ju Tian
Jialei Zhu
Xiaohan Deng
Xu Zhu
Ruigang Wang
Guojing Li
Somatic Embryogenesis and Genetic Transformation of <i>Caragana intermedia</i>
Plants
cotyledon
plant tissue culture
embryogenic callus
morphogenesis
GMO
title Somatic Embryogenesis and Genetic Transformation of <i>Caragana intermedia</i>
title_full Somatic Embryogenesis and Genetic Transformation of <i>Caragana intermedia</i>
title_fullStr Somatic Embryogenesis and Genetic Transformation of <i>Caragana intermedia</i>
title_full_unstemmed Somatic Embryogenesis and Genetic Transformation of <i>Caragana intermedia</i>
title_short Somatic Embryogenesis and Genetic Transformation of <i>Caragana intermedia</i>
title_sort somatic embryogenesis and genetic transformation of i caragana intermedia i
topic cotyledon
plant tissue culture
embryogenic callus
morphogenesis
GMO
url https://www.mdpi.com/2223-7747/14/10/1545
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AT jialeizhu somaticembryogenesisandgenetictransformationoficaraganaintermediai
AT xiaohandeng somaticembryogenesisandgenetictransformationoficaraganaintermediai
AT xuzhu somaticembryogenesisandgenetictransformationoficaraganaintermediai
AT ruigangwang somaticembryogenesisandgenetictransformationoficaraganaintermediai
AT guojingli somaticembryogenesisandgenetictransformationoficaraganaintermediai