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Direct somatic embryogenesis and related gene expression networks in leaf explants of Hippeastrum ‘Bangkok Rose’

Horticultural Plant Journal 2024年第2期10卷 556-572页

作者： Jingjue Zeng Yi Deng Shahid Iqbal Jiarui Zhang Kunlin Wu Guohua Ma Lin Li Guangyi Dai Rufang Deng Lin Fang Songjun Zeng Guangdong Provincial Key Laboratory of Applied Botany South China Botanical GardenChinese Academy of SciencesGuangzhouGuangdong 510650China University of the Chinese Academy of Sciences Beijing 100049China South China National Botanical Garden GuangzhouGuangdong 510650China Zhaoqing Branch Center of Guangdong Laboratory for Lingnan Modern Agricultural Science and Technology ZhaoqingGuangdong 526000China Horticultural Science Department North Florida Research and Education CenterUniversity of Florida/IFASQuincyFL 32351USA Public Laboratory of South China Botanical Garden Chinese Academy of SciencesGuangzhouGuangdong 510650China Key Laboratory of South China Agricultural Plant Molecular Analysis and Gene Improvement South China Botanical GardenChinese Academy of SciencesGuangzhouGuangdong 510650China

Hippeastrum, a highly diverse genus in the Amaryllidaceae family, is a valuable ornamental bulbous flowering plant. somatic embryogenesis(SE) is an efficient method for mass production of Hippeastrum plantlets. Previous studies have been devoted to the in vitro propagation of Hippeastrum, but the SE and its regulatory networks are rarely reported. In this study, we established a direct SE method of Hippeastrum Bangkok Rose' using leaf bases as explants. MS supplemented with 1.00 mg·L^(-1)NAA +1.00 mg·L^(-1)KT + 0.25 mg·L^(-1)TDZ was the optimal medium for SE. Histological observations showed that the bipolar somatic embryo originated from the epidermal cell layer and underwent initiation,globular, scutellar and coleoptile stages. During SE, endogenous hormones of IAA, CTK, ABA, and SA were highly accumulated. Transcriptomic analysis revealed the genes encoding auxin biosynthesis/metabolic enzymes and efflux carriers were induced, while the auxin receptor of TIR1 and ARF transcriptional repressor of Aux/IAA were down-regulated and up-regulated, respectively, leading to suppression of auxin signaling. In contrast, cytokine signaling was promoted at the early stage of SE, as biosynthesis, transport, and signaling components were *** stress-related genes were up-regulated at the early or late stages of SE. Chromatin remodeling could also be dynamically regulated via distinct expression enzymes that control histone methylation and acetylation during SE. Moreover, key SE regulators, including WOXs and SERKs were highly expressed along with SE. Overall, the present study provides insights into the SE regulatory mechanisms of the Hippeastrum.

关键词： Hippeastrum Tissue culture somatic embryogenesis Gene regulation

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Genotypic effects on accelerated propagation of oil palm breeding materials selected(Elaeis guineensis jacq.)using somatic embryogenesis

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Oil Crop Science 2024年第2期9卷 111-120页

作者： Retno Puji Astari Mohammad Basyuni Luthfi A.M. Siregar Revandy I.M. Damanike Deni Arifiyanto Dadang Affandi Indra Syahputra Doctoral Program of Agricultural Sciences Faculty of AgricultureUniversitas Sumatera UtaraMedan20155North SumatraIndonesia PT.Socfin Indonesia Jl.K.L.Yos Sudarso No.106Medan20244North SumatraIndonesia Department of Forestry Faculty of ForestryUniversitas Sumatera UtaraJl.Tri Dharma Ujung No.1Medan20155North SumatraIndonesia Center of Excellence for Mangrove Universitas Sumatera UtaraMedan20155Indonesia Department of Agrotechnology Faculty of AgricultureUniversitas Sumatera UtaraMedan20155North SumatraIndonesia

Vegetable oil production from oil palm(Elaeis guineensis Jacq.)is an important industry due to the rising demand every *** somatic embryogenesis culture can propagate oil palm duplicate as parent plant,which can be selected as breeding material to produce new planting germplasm with high production or disease *** study aims to evaluate the genotypic effect of somatic embryogenesis,while immature leaflets were employed as *** culture used embryo induction medium based on Murashige and Skoog(MS)modifications that contained 5 mg/L Naphthalene Acetic acid(NAA)and 0.5 mg/L Benzyl Amino Purine(BAP).The genotypic effect was statistically significant in the percentage of callus induction,producing somatic embryos,and germination *** this study,we successfully cloned thirteen oil palm genotypes(GE-02,GE-03,GE-06,GE-07,GE-09,GE-23,GE-24,GE-27,GE-28,GE-32,GE-33,GE-34,and GE-35),with the highest number of somatic embryos formed on GE-27 with a percentage of 70.1%.The cloning was successful in accelerating the propagation of oil palm for materials breeding programs to create new varieties with high production and disease *** is necessary to observation the performance of these clones in the field in terms of mantle flower appearance.

关键词： somatic embryogenesis Oil palm Genotypic effect Support breeding

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somatic embryogenesis and Plant Regeneration from Two Recalcitrant Genotypes of Gossypium hirsutum L.

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Agricultural Sciences in China 2006年第5期5卷 323-329页

作者： WANG Yan-xia WANG Xing-fen MA Zhi-ying ZHANG Gui-yin HAN Gai-ying Hebei Agricultural University/Key Laboratory of Crop Germplasm Resources of Hebei Baoding 071001 P.R. China

An improved protocol has been developed for somatic embryogenesis and plant regeneration of recalcitrant cotton cultivars. High callus frequencies and embryogenic tissue were developed in MSB medium supplemented with gradient concentrations of KT and 2,4-D, their concentration decreasing from 0.1 to 0.01 mg·L^-1. somatic embryos were successfully incubated in 1/2 macronutrient MSB suspension supplemented with 0.5 g· L^-1 glutamine and 0.5 g·L^-1 asparagine. Decrease in macronutrient concentration of MSB significantly alleviated browning and was beneficial to suspension cells. Transformation of somatic embryos into plants was induced in MSB medium supplemented with 3% sucrose, 0.5 g·L^-1 glutamine, 0.5 g·L^-1 asparagine, and 6.0 g·L^-1 agar. The effect of sucrose as carbohydrate was better than that of glucose for plant germination. Using this protocol, regenerated plantlets from the CCRI521 and Zhongzhi86-6 reached to as much as 19.6 and 18.5% somatic embryos, respectively.

关键词： Gossypium hirsutum L. recalcitrant genotypes plant regeneration somatic embryogenesis effect of macronutrients suspension culture

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Reactive oxygen species, nitric oxide and plant cell death associated with caspase-like protease activity during somatic embryogenesis in Fraxinus mandshurica

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Journal of Forestry Research 2022年第3期33卷 1005-1017页

作者： Hao Wang Dongyan Zhang Juan Luis Fernández-Lorenzo Hailong Shen Ling Yang State Key Laboratory of Tree Genetics and Breeding School of ForestryNortheast Forestry UniversityHarbin150040People’s Republic of China Crop Production Department High Polytechnique School of EngineeringSantiago de Compostela UniversityLugoSpain

Programmed cell death occurs in browning explants of Fraxinus mandshurica during somatic embryogenesis, but the underlying mechanism is unclear. In this study, single cotyledons of zygotic embryos of F. mandshurica were used as explants. Mitochondrial structure and function, caspase-3-like protease activity, hydrogen peroxide metabolism, and nitric oxide accumulation induced by high concentrations of sucrose and plant growth regulators were studied. The results show that plant growth regulators induced somatic embryogenesis and also promoted explant browning. High sucrose concentrations had similar effects. High concentrations of sucrose and plant growth regulators led to the accumulation of hydrogen peroxide and nitric oxide which induced changes in mitochondrial structure and function such as modifications in mitochondrial morphology, increased membrane permeability, decreased membrane potential, and the release of cytochrome c into the cytoplasm. An increase in caspase-3-like protease activity triggered programmed cell death in some browning explant cells. During somatic embryogenesis there were increased activities of superoxide dismutase, peroxidase, and catalase, which are associated with hydrogen peroxide metabolism and jointly maintain reactive oxygen species levels. Intracellular nitric oxide synthase and nitrate reductase activities were not significantly correlated with nitric oxide content. Instead, intracellular nitric oxide may be derived from non-enzymatic reactions. Our results indicate that hydrogen peroxide and nitric oxide may function as signals, playing key roles in somatic embryogenesis and programmed cell death of explant cells of F. mandshurica. The interaction between nitric oxide and reactive oxygen species determines the occurrence of programmed cell death in explant cells;somatic embryogenesis and programmed cell death are positively regulated by hydrogen peroxide. However, the regulation of nitric oxide is complex.

关键词： Manchurian ash somatic embryogenesis Programmed cell death Mitochondria Reactive oxide species Nitric oxide

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somatic embryogenesis and Plantlet Regeneration in Callus Cultures Derived from Mature Zygotic Embryos of Slash Pine

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Journal of Forestry Research 1997年第2期8卷 83-86页

作者：唐巍欧阳藩郭忠琛 Chinese Academy of Sciences State Key Laboratory of Biochemical Engineering Institute of Chemical Metallurgy Beijing P.R. China (GRID:grid.9227.e) (ISNI:***) Chinese Academy of Sciences Institute of Botany Beijing (GRID:grid.9227.e) (ISNI:***)

White, translucent, and mucilaginous embryogenic callus was initiated in cultured mature zygotic embryoexplants of two different seed sources of slash pine(Pinus elliottii) on several culture media containing auxin and *** proembryos were induced on media containing 2,4-D and BA. Maturatiol1 was successfully achieved on mediumsupplemented with ABA. somatic embryos germinated into regeneration plantlets On DCR medium containing activated charcoal. HiStological observatiol1 and scanning electron inicroscopic observatiol1 showed that proembryos derived from em-bryonal suspensor mass(ESM) were formed on the surface or the inside of embryogenic callus, and the prolitbration of pro-embryos was mainly from clcavage polyembryos.

关键词： Pinus elliottii somatic embryogenesis Plantet regeneration

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Relationship between H_(2)O_(2) accumulation and NO synthesis during osmotic stress:promoted somatic embryogenesis of Fraxinus mandshurica

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Journal of Forestry Research 2021年第3期32卷 917-925页

作者： Ling Yang Hanyang Guo Yingying Liu Dongyan Zhang Hongnan Liu Hailong Shen State Key Laboratory of Tree Genetics and Breeding Northeast Forestry UniversityHarbin 150040People’s Republic of China

Osmotic stress promotes somatic embryogenesis of Fraxinus mandshurica,which leads to accumulation of reactive oxygen species(ROS).The single pieces of cotyledons of *** were used as explants to induce somatic embryogenesis in osmotic-stress ***,the hydrogen peroxide H_(2)O_(2) content of explanted cells was varied by adding exogenous H_(2)O_(2) or catalase solution to assess the effects of the exogenous H_(2)O_(2)on somatic embryogenesis,intracellular H_(2)O_(2)accumulation,and the relationship between signaling mediated by ROS or reactive nitrogen *** results revealed that exogenous H_(2)O_(2)(100?300μmol L^(–1))increased the number of somatic *** 60th day of exogenous H_(2)O_(2)(200μmol L^(–1))treatment,the number of somatic embryos of explants treated,which was 136.54%,was higher than the ***,exogenous H_(2)O_(2)(100μmol L^(–1))significantly increased the intracellular H_(2)O_(2)content and enhanced the activities of superoxidase dismutase and ***,exogenous H_(2)O_(2)(100μmol L^(–1))activated the intracellular non-enzymatic pathway for nitric oxide(NO)*** somatic embryogenesis in broadleaf trees increases with the change of endogenic ROS content,and depends on the upregulation of antioxidant *** H_(2)O_(2)and NO,as signaling molecules,were found to be involved in the process of somatic embryogenesis in broadleaf *** the process of exogenous H_(2)O_(2)promoting somatic embryogenesis,NO synthesis depended on non-enzymatic *** results provide a scientific basis for resolving the mechanism by which ROS levels are regulated during somatic embryogenesis of broadleaf trees and establish a reasonable and efficient technology system for regulating somatic embryogenesis of trees.

关键词： Fraxinus mandshurica somatic embryogenesis Hydrogen peroxide Nitric oxide Osmotic stress

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Plantlet regeneration through somatic embryogenesis in Nordmann's fir (Abies nordmanniana)

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Journal of Forestry Research 2016年第6期27卷 1219-1228页

作者： Katarzyna Nawrot-Chorabik Department of Forest Pathology Mycology and Tree Physiology University of Agriculture in Krakow

I studied the influence of various combinations of auxin and cytokinin concentrations, and the increased content of zinc and enzymatic casein hydrolizate in SH medium on initiation and proliferation of embryogenic callus of Abies nordmanniana （Steven） Spach. Addition- ally, the effect of ABA, PEG-4000 and different wave- lengths on the maturation of somatic embryos was tested. The use of optimum composition of modified SH medium with BA, KIN and 2.4-D while simultaneously ensuring appropriate external conditions resulted in 15.5 % embryogenesis. Finally, satisfactory results of microprop- agation of A. nordmanniana by somatic embryogenesis were obtained providing seven lines of embryogenic callus with high proliferation capacity. Those lines gave properly developed seedlings in white LED light with a wavelength of 400-700 nm, preceded by eight-week vernalization treatment of the callus. This paper may provide a protocol by which all stages of somatic embryogenesis of A. nord- manniana can be carded out, including the preceding 24-h seed disinfection with NaOCl and PVP, which resulted in 100 % frequency of uninfected zygotic embryos that were capable of starting embryogenesis.

关键词： Abies In vitro - LED light Polyvinylpyrrolidone somatic embryogenesis

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Indirect somatic embryogenesis and regeneration of Fraxinus mandshurica plants via callus tissue

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Journal of Forestry Research 2021年第4期32卷 1613-1625页

作者： Yang Liu Cheng Wei Hao Wang Xiao Ma Hailong Shen Ling Yang State Key Laboratory of Tree Genetics and Breeding Northeast Forestry UniversityHarbin 150040People’s Republic of China State Forestry and Grassland Administration Engineering Technology Research Center of Native Tree Species Harbin 150040People’s Republic of China

somatic embryogenesis of Fraxinus mandshurica has the problems of low somatic embryo(SE) yield,unsynchronized SE development,and a high percentage of deformed *** aimed to improve *** SE production by synchronizing SE development,improving SE quality,and inducing root formation to obtain complete regenerated *** of immature zygotic embryos of *** were induced to form callus and then *** SE induction percentage from explants differed among 32 mother trees,and the one with the highest SE induction percentage(29.8%) was used for further *** highest callus induction percentage was94.2% on 1/2-strength Murashige and Skoog medium(MS1/2)supplemented with 0.15 mg·L^(-1) naphthalene acetic *** highest callus proliferation coefficient(240.5) was obtained on McCown's Woody Plant Medium containing 0.1 mg·L^(-1)6-benzyl adenine and 0.15 mg·L^(-1) 2,4-dichlorophenoxyacetic *** highest number of SEs(1020.5 g~(-1) fresh weight) was obtained on MS1/2 medium supplemented with1 mg·L^(-1) *** highest number of cotyledon embryos(397/g fresh weight) was obtained by incubating materials on medium containing 1 mg·L^(-1) abscisic acid and then applying a drying *** cotyledon embryos were milky white,uniformly sized(average length 4.7 mm),and 80% of them were *** SE rooting percentage on 1/2 MS medium containing 0.01 mg·L^(-1) NAA was 37.5%.Overall,the germination percentage of SEs was 26.4%,and complete regenerated plants were obtained after transplanting and *** results provide more possibilities for the preservation and breeding of ***.

关键词： Fraxinus mandshurica somatic embryogenesis Callus induction Cell differentiation Plant regeneration

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Selection of culture conditions for callus induction and proliferation by somatic embryogenesis of Pinus koraiensis

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Journal of Forestry Research 2021年第2期32卷 483-491页

作者： Fang Gao Chunxue Peng Hao Wang Hailong Shen Ling Yang State Key Laboratory of Tree Genetics and Breeding School of ForestryNortheast Forestry UniversityHarbin 150040People’s Republic of China

The induction and proliferation of embryogenic callus are key steps for large-scale propagation of somatic embryogenesis pathway and long-term preservation of coniferous *** can be induced from immature embryos of Korean pine(Pinus koraiensis *** Zucc.;Pinaceae)as explants,but there are problems,such as low proliferation efficiency,loss of embryogenicity,poor vigor;thus,best conditions for proliferation and culture of immature embryos of Korean pine are not yet *** solve the problems with somatic embryogenesis of Korean pine and determine the best culture conditions for callus induction and proliferation,we varied hormone concentration,subculture cycle of proliferation and other plant growth regulators combinations in media to induce callus formation by megagametophytes of three Korean pine families at different developmental stages,then analyzed the effects on embryogenic callus retention and cell proliferation using a quadratic regression orthogonal rotation *** results showed that the family origin and collection date of explants significantly affected callus induction(induction rate reached 1.67%).Embryogenic maintenance and callus proliferation were best on DCR medium supplemented with 0.25 mg L^(−1)6-benzyl adenine,1 mg L^(−1)naphthaleneacetic acid,30 g L^(−1)sucrose,500 mg L^(−1),L-glutamine,500 mg L^(−1)casein hydrolysis and 6.5 g L^(−1)*** addition,the combination of 2,4-dichlorophenoxyacetic acid+6-benzyl adenine also had a better proliferative effect on *** effects of different combinations of growth regulators on callus proliferation efficiency were significantly *** to new medium every 13–15 days not only maintained robust callus vigor,but also yielded a larger proliferation *** techniques and conditions for embryogenic callus induction and proliferation of Korean determined here will serve as a foundation for establishing a large-scale system for somatic embryogenesis and propagation of

关键词： Korean pine somatic embryogenesis Embryogenic callus Megagametophytes Embryonic maintenance

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Characterization of Gh SERK2 and its expression associated with somatic embryogenesis and hormones level in Upland cotton

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Journal of Integrative Agriculture 2018年第3期17卷 517-529页

作者： LIU Zheng-jie ZHAO Yan-peng ZENG Ling-he ZHANG Yuan WANG Yu-mei HUA Jin-ping College of Agronomy and Biotechnology China Agricultural University/Key Laboratory of Crop Heterosis and Utilization Ministry of Education/Beijing Key Laboratory of Crop Genetic Improvement United States Department of Agriculture - Agricultural Research Service (USDA-ARS) Crop Genetics Research Unit Research Institute of Cash Crop Hubei Academy of Agricultural Sciences

somatic embryogenesis （SE） is one of the most important steps during regeneration of cotton, but the molecular mechanism of SE remains unclear. somatic EMBRYOGENSIS RECEPTOR KINASE （SERK） gene is known to function in SE. A homolog GhSERK2 （accession number： JF430801） was cloned from Upland cotton and characterized for its functions in SE. GhSERK2 expressed in different tissues and showed higher expression level in floral organs than vegetative ones with the highest levels in ovule and anther. GhSERK2 expressed during SE with a high level at globular embryos stage. Upon treatment with indole-3-butytic acid （IBA）, the transcription level of GhSERK2 was induced and promoted SE subsequently. A 2-day treatment of 2,4-dichlorophenoxyacetic acid （2,4-D） induced the expression of GhSERK2, but treatments of 2,4-D for longer periods sharply inhibited the GhSERK2 transcription level of embryogenic callus （EC）. The levels of hormones, including 3-indoleacetic acid （IAA）, abscisic acid （ABA）, and brassinosteroid （BR）, were increased in the initial calli induced from the over-expression of GhSERK2 cotton. Our results indicated that GhSERK2 expression was associated with induction of SE and closely related to hormone levels during tissue culture in Upland cotton, and the gene might play an important role in regeneration of cotton.

关键词： GhSERK2 somatic embryogenesis hormone level expression pattern Upland cotton (Gossypium hirsutum L.)

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