Transcriptome and metabolome analyses reveal the efficiency of in vitro regeneration by TDZ pretreatment in mulberry

Luo, Yiwei; Han, Yuanxiang; Wei, Wuqi; Han, Yue; Yuan, Jianglian; He, Ningjia

doi:10.1016/j.scienta.2022.111678

Cited by 10 publications

(5 citation statements)

References 68 publications

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“…In this study, we also found that WPM medium was more suitable for somatic embryogenesis in A. trifoliata (a perennial woody vine) than MS medium. In addition, TDZ is believed to have stronger effects on plant growth and development in vitro and has been reported to be effective in the induction of various morphogenic responses, including somatic embryogenesis [30,33,35]. In this study, the SEs were induced with the best results on the medium containing 1.0 mg L −1 of TDZ and 1.0 mg L −1 of 6-BA.…”

Section: Factors Affecting Se Inductionmentioning

confidence: 71%

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Somatic Embryogenesis Induction and Genetic Stability Assessment of Plants Regenerated from Immature Seeds of Akebia trifoliate (Thunb.) Koidz

Zhang

Cao

Wang

et al. 2023

Forests

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Akebia trifoliata is a perennial woody plant with considerable potential in nutrition, food, and health, and the production of seedlings with high quality is critical for its economic utilization. Plant regeneration through somatic embryogenesis is a powerful alternative for propagating many plant species. In this study, a simple and practicable protocol was developed for plant regeneration from immature seeds of A. trifoliata via somatic embryogenesis, and the genetic stability of regenerated plants was also assessed. In the somatic embryo (SE) induction stage, the highest frequency of somatic embryogenesis (35.2%) was observed on the WPM medium containing 1.0 mg L−1 of thidiazuron (TDZ) and 1.0 mg L−1 of 6-benzyladenine (6-BA). The concentration of 6-BA was optimized at 1.0 mg L−1 for the proliferation and maturation of the induced SEs, and the combination of 2.0 mg L−1 of indole-3-butyric acid (IBA) and 0.5 mg L−1 of TDZ was the most responsive for root development and plant growth. The leaf morphological characteristics greatly varied among the established plants, and they could be grouped into three plant types, namely the normal type, Type Ι, and Type ΙΙ. Remarkable differences in the number, size, shape, and color of the leaflets were observed among the three types, while their ploidy level was the same via flow cytometry analysis. The Type ΙΙ and the Type Ι plants had the highest and the lowest net photosynthesis rate, transpiration rate, and stomatal conductance among the three groups, respectively, and both had a smaller size of stomatal guard cells than the normal type. Simple sequence repeat (SSR) analysis detected that 41 bands (43.62%) were different from those observed in the wild, indicating a high degree of polymorphism between the regenerants and their donor parent. The obtained plants might hold potential for future genetic improvement and breeding in A. trifoliata, and the established regeneration protocol might serve as a foundation for in vitro propagation and germplasm preservation of this crop.

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Section: Factors Affecting Se Inductionmentioning

confidence: 71%

“…It has been documented that nutrient medium and PGRs have a great influence on SEs' induction [30,33]. To support the growth and development of the explants in tissue culture, WPM medium is frequently used for woody plants, whereas MS medium is often applied in herbaceous plants [34].…”

Section: Factors Affecting Se Inductionmentioning

confidence: 99%

Somatic Embryogenesis Induction and Genetic Stability Assessment of Plants Regenerated from Immature Seeds of Akebia trifoliate (Thunb.) Koidz

Zhang

Cao

Wang

et al. 2023

Forests

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“…Deepening our knowledge of the molecular mechanisms underlying hormone and stress responses in mulberry, along with identifying novel genes pivotal in these processes, continues to be imperative and holds significant potential for advancing mulberry breeding. With the advancement of sequencing technologies, high-quality genomic sequences of several mulberry species have been made publicly available (He et al 2013;Jiao et al 2020;Xuan et al 2022), as well as numerous global gene expression profiles (Liu et al 2021;Luo et al 2023). This progress has bestowed a wealth of information for predicting genes associated with hormones and stress regulations in mulberry.…”

Section: Introductionmentioning

confidence: 99%

Efficient mesophyll-derived protoplast manipulation system as a versatile tool for characterization of genes responding to multiple stimuli in mulberry

Zhou,

Lei,

Hou

et al. 2024

Plant Cell Tiss Organ Cult

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Versatile protoplast manipulation systems have been established in a variety of plant species for gene functional studies, especially on physiological perceptions and responses to hormones, metabolites, environmental stimuli. However, efficient protoplast-based systems are still lacking for the economically important woody plant mulberry, of which functional genomics research is hindered by its long life cycle and recalcitrance to genetic transformation. In this study, we set up sterile cultures for a hybrid mulberry resource and established an efficient protocol for isolating mesophyll-derived protoplast via optimizing the starting materials and maceration conditions. With sufficient viable protoplasts, polyethylene glycol (PEG)-mediated transient gene expression has been carried out and a transformation efficiency of nearly 50 % was achieved through optimizing the incubation strategy. The transient gene expression system was validated by subcellular localization assays of the mulberry nuclear marker gene MaHTR6 and chloroplast marker gene MaTRX. Subsequently, the system was used for functional characterization of putative mulberry cytokinin response factors (MaCRFs) in response to phytohormone and environmental stimuli. Combined the established system with RT-qPCR to test the CRFs transcriptional regulation, we found that among the three MaCRFs, conserved with Arabidopsis CRF2, MaCRF1 turned out to function primarily as a response factor to cytokinin and auxin. While MaCRF2 and MaCRF3 respond not only to phytohormones but also to environmental stresses. In summary, we have established an efficient mesophyll-derived protoplast manipulation system in mulberry, thereby facilitating transient function analysis of genes in response to multiple external stimuli.

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“…With the rapid development of high-throughput sequencing and mass spectrometry technology, an abundance of data from transcriptomics and metabolomics has been produced, offering valuable insights into the mechanisms of callus regeneration. Transcriptome studies make it possible to e ciently and quickly analyze differentially expressed genes and functionally annotate regulatory pathways [22]. Integrated transcriptomic and endogenous hormone content analysis insighted uncovering regeneration mechanism in Osmanthus fragrans calli [23].…”

Section: Introductionmentioning

confidence: 99%

Integrated transcriptomic and endogenous hormones analyses revealed the molecular mechanism of light and auxin for the regeneration of callus tissue in seashore paspalum

Jiang,

Hu,

Sun

et al. 2024

Preprint

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Background: Seashore paspalum (Paspalum vaginatum O. Swartz) is a halophyte known for its exceptional salt tolerance and ecological adaptability. It is an excellent candidate for studying salt tolerance mechanisms and screening salt tolerance genes. However, the difficulties with callus tissue regeneration and the influence of genotype during cultivation provide a significant obstacle to the process of molecular breeding employing genetic transformation and gene editing techniques in seashore paspalum. Results: To elucidate the molecular mechanism of callus regeneration in seashore paspalum, this study analyzed the content of endogenous hormones and investigated the effects of light, KT, and genotype on callus regeneration; Through transcriptome analysis between different treatments, the molecular mechanisms were explored. Under light conditions, almost all callus tissues of genotype I could produce regenerated green buds, but genotype II could not regenerate. A total of 106.2 Gb clean readings were obtained from 12 cDNA sample libraries in four regeneration states (CK, KT-D, KT-L, and KT-L-NR). The Pearson correlation coefficients, principal component analysis, and DEG hierarchical clustering heatmap analysis results indicated good intra-group repeatability and reliable data. The specific expression genes induced by different genotypes (KT-L-NR vs KT-L) (3083) were significantly higher than those in other groups by Venn plot analysis. A total of 73 endogenous hormone substances were quantitatively detected in all samples. KEGG enrichment analysis showed that all comparison groups significantly enriched differentially changed hormones (DCHs) in diterpenoid biosynthesis and plant hormone signal transduction pathways. In KT-L, GA5 and GA51 were significantly higher than those in other groups, while GA20 and GA29 were significantly lower. KT-L-NR showed noticeably higher levels of GA3, GA20, and GA29, which could be a contributing cause to the incapacity of callus regeneration. The expression level of GA2ox (Pavag03G280900. v3.1) was very high, significantly negatively regulating GA51. In KT-L, the content of ABA and JA were the lowest and significantly lower than that in KT-L-NR. The content of indole-3-acetic acid (IAA) in KT-L and KT-L-NR were significantly higher than that in CK and KT-D, indicating that light played an important role in synthesizing of IAA, which was beneficial for the regeneration of callus tissue. Conclusions: This is the first report on callus regeneration mechanisms of seashore paspalum by combined transcriptome and endogenous hormone profiling. The results will improve the understanding of molecular mechanisms and the effects of endogenous hormones, and provide new insights to address the issue of genotype dependence in callus regeneration.

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Transcriptome and metabolome analyses reveal the efficiency of in vitro regeneration by TDZ pretreatment in mulberry

Cited by 10 publications

References 68 publications

Somatic Embryogenesis Induction and Genetic Stability Assessment of Plants Regenerated from Immature Seeds of Akebia trifoliate (Thunb.) Koidz

Somatic Embryogenesis Induction and Genetic Stability Assessment of Plants Regenerated from Immature Seeds of Akebia trifoliate (Thunb.) Koidz

Efficient mesophyll-derived protoplast manipulation system as a versatile tool for characterization of genes responding to multiple stimuli in mulberry

Integrated transcriptomic and endogenous hormones analyses revealed the molecular mechanism of light and auxin for the regeneration of callus tissue in seashore paspalum

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