Analysis and review of trichomes in plants

Background Rosette leaf trichomes of Arabidopsis thaliana have been broadly used to study cell development, cell differentiation and, more recently, cell wall biogenesis. However, trichome-specific biochemical or -omics analyses require a proper separation of trichomes from residual plant tissue. Thus, different strategies were proposed in the past for trichome isolation, which mostly rely on harsh conditions and suffer from low yield, thereby limiting the spectrum of downstream analyses. Results To take trichome-leaf separation to the next level, we revised a previously proposed method for isolating A. thaliana trichomes by optimizing the mechanical and biochemical specifications for trichome release. We additionally introduced a density gradient centrifugation step to remove residual plant debris. We found that prolonged, yet mild seedling agitation increases the overall trichome yield by more than 60% compared to the original protocol. We noticed that subsequent density gradient centrifugation further visually enhances trichome purity, which may be advantageous for downstream analyses. Gene expression analysis by quantitative reverse transcriptase-polymerase chain reaction validated a substantial enrichment upon purification of trichomes by density gradient centrifugation. Histochemical and biochemical investigation of trichome cell wall composition indicated that unlike the original protocol gentle agitation during trichome release largely preserves trichome integrity. We used enriched and density gradient-purified trichomes for proteomic analysis in comparison to trichome-depleted leaf samples and present a comprehensive reference data set of trichome-resident and -enriched proteins. Collectively we identified 223 proteins that are highly enriched in trichomes as compared to trichome-depleted leaves. We further demonstrate that the procedure can be applied to retrieve diverse glandular and non-glandular trichome types from other plant species. Conclusions We provide an advanced method for the isolation of A. thaliana leaf trichomes that outcompetes previous procedures regarding yield and purity. Due to the large amount of high-quality trichomes our method enabled profound insights into the so far largely unexplored A. thaliana trichome proteome. We anticipate that our protocol will be of use for a variety of downstream analyses, which are expected to shed further light on the biology of leaf trichomes in A. thaliana and possibly other plant species.

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“…6 C). This finding is in good agreement with the prevailing opinion that trichomes are not photosynthetically active in A. thaliana [ 60 , 61 ].…”

Section: Discussionsupporting

confidence: 92%

An advanced method for the release, enrichment and purification of high-quality Arabidopsis thaliana rosette leaf trichomes enables profound insights into the trichome proteome

et al. 2022

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“…The molecular mechanisms involved in this process include a regulatory network composed of numerous transcription factors and other proteins, such as MYBs, bHLHs, WD40 repeat proteins, HD-ZIP, and C2H2 zinc finger proteins (Hülskamp, 2004;Pan et al, 2015;Doroshkov et al, 2019). Similar types of proteins also regulate the development of trichomes in plants such as cotton, tobacco, tomato, and A. annua (Chalvin et al, 2020;Wang et al, 2021). Among these, C2H2 zinc finger proteins play important roles in various stages of trichome development in different plants.…”

Section: Regulation Of Trichome Development By C2h2 Zinc Finger Proteinsmentioning

confidence: 99%

Advances in the Regulation of Epidermal Cell Development by C2H2 Zinc Finger Proteins in Plants

Han

Qiao

et al. 2021

Front. Plant Sci.

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Plant epidermal cells, such as trichomes, root hairs, salt glands, and stomata, play pivotal roles in the growth, development, and environmental adaptation of terrestrial plants. Cell fate determination, differentiation, and the formation of epidermal structures represent basic developmental processes in multicellular organisms. Increasing evidence indicates that C2H2 zinc finger proteins play important roles in regulating the development of epidermal structures in plants and plant adaptation to unfavorable environments. Here, we systematically summarize the molecular mechanism underlying the roles of C2H2 zinc finger proteins in controlling epidermal cell formation in plants, with an emphasis on trichomes, root hairs, and salt glands and their roles in plant adaptation to environmental stress. In addition, we discuss the possible roles of homologous C2H2 zinc finger proteins in trichome development in non-halophytes and salt gland development in halophytes based on bioinformatic analysis. This review provides a foundation for further study of epidermal cell development and abiotic stress responses in plants.

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“…6C, G). The apparent increase seems likely to be a secondary effect of VIGS, as trichomes are known to be involved in pathogen response (Wang et al, 2021). Overall, these results demonstrate that silencing of CvFUL in C. violacea produces modified phenotypes in both valve and replum tissue.…”

Section: Resultsmentioning

confidence: 99%

Virus‐induced gene silencing as a tool for functional studies in Cleome violacea

et al. 2021

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Premise Cleomaceae is emerging as a promising family to investigate a wide range of phenomena, such as C 4 photosynthesis and floral diversity. However, functional techniques are lacking for elucidating this diversity. Herein, we establish virus‐induced gene silencing (VIGS) as a method of generating functional data for Cleome violacea , bolstering Cleomaceae as a model system. Methods We leveraged the sister relationship of Cleomaceae and Brassicaceae by using constructs readily available for Arabidopsis thaliana to provide initial information about the feasibility of VIGS in C . violacea . We then developed endogenous constructs to optimize VIGS efficiency and viability for fruit development. Results PHYTOENE DESATURASE was successfully downregulated in C . violacea using both heterologous and endogenous constructs. The endogenous construct had the highest degree of downregulation, with many plants displaying strong photobleaching. FRUITFULL ‐treated plants were also successfully downregulated, with a high rate of survival but less effective silencing; only a small percentage of survivors showed a strong phenotype. Discussion Our optimized VIGS protocol in C . violacea enables functional gene analyses at different developmental stages. Additionally, C . violacea is amenable to heterologous knockdown, which suggests that a first pass using non‐endogenous constructs is a possible route to test additional species of Cleomaceae.

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Analysis and review of trichomes in plants

Cited by 117 publications

References 137 publications

An advanced method for the release, enrichment and purification of high-quality Arabidopsis thaliana rosette leaf trichomes enables profound insights into the trichome proteome

An advanced method for the release, enrichment and purification of high-quality Arabidopsis thaliana rosette leaf trichomes enables profound insights into the trichome proteome

Advances in the Regulation of Epidermal Cell Development by C2H2 Zinc Finger Proteins in Plants

Virus‐induced gene silencing as a tool for functional studies in Cleome violacea

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