Establishing an efficient protoplast transient expression system for investigation of floral thermogenesis in aroids

Maekawa, Haruhiko; Otsubo, Miyabi; Sato, Mitsuhiko; Takahashi, Tomoko; Mizoguchi, Koichiro; Koyamatsu, Daiki; Inaba, Tomoki; Ito‐Inaba, Yasuko

doi:10.1007/s00299-021-02806-1

Cited by 5 publications

(7 citation statements)

References 63 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Using a recently developed protoplast‐based transient expression (PTE) system (Maekawa et al, 2022), we carried out subcellular localization analyses of SrSTP1a, SrSTP1b, and SrSTP14, using GFP fusion proteins in leaf protoplasts prepared from S. renifolius (Figure 4). The fluorescence of FM4‐64 was used to label the plasma membrane.…”

Section: Resultsmentioning

confidence: 99%

“…ScHXT5 transcripts were amplified from the cDNAs using primer pairs: PMA1‐ScHXT5‐F1 and ScHXT5‐Lin‐R1. Those amplified fragments and the GFP genes amplified from the pUC 35S::GFP construct (Maekawa et al, 2022), were mixed and inserted into the pDRf1 vector (Addgene), which was digested with Bam HI and Xho I using In‐Fusion HD cloning system (Takara). The resulting DNA inserts were sequenced.…”

Section: Methodsmentioning

confidence: 99%

“…SrSTP1 and SrSTP14 transcripts were amplified from the cDNAs using primer pairs: 35S‐SrSTP1‐F1 and SrSTP1‐Lin‐R1 for SrSTP1 , and 35S‐SrSTP14‐F1 and SrSTP14‐Lin‐R1 for SrSTP14 . The GFP gene was amplified from the pUC 35S::GFP construct, as previously described (Maekawa et al, 2022). To generate the 35S::SrSTP1‐GFP and 35S::SrSTP14‐GFP constructs, the two PCR products were mixed and cloned into the Xba I site of pBI221‐luc+ (Maekawa et al, 2022) using an In‐Fusion HD Cloning Kit w/Cloning Enhancer (Clontech Laboratories, Inc., Mountain View, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

“…So far, at least 90 plant species have been documented as thermogenic. Among them, several aroids have extensive heat‐producing ability (Maekawa et al, 2022) as they can increase their internal temperature more than 20°C above the surrounding air temperature. Thermogenesis in the reproductive organs of these plants is positively correlated to their oxygen consumption rate (Seymour, 2010).…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Molecular characterization of SrSTP14, a sugar transporter from thermogenic skunk cabbage, and its possible role in developing pollen

Koyamatsu,

Otsubo,

Ohira

et al. 2023

Physiologia Plantarum

Self Cite

View full text Add to dashboard Cite

In floral thermogenesis, sugars play an important role not only as energy providers but also as growth and development facilitators. Yet, the mechanisms underlying sugar translocation and transport in thermogenic plants remain to be studied. Asian skunk cabbage (Symplocarpus renifolius) is a species that can produce durable and intense heat in its reproductive organ, the spadix. Significant morphological and developmental changes in the stamen are well‐characterized in this plant. In this study, we focused on the sugar transporters (STPs), SrSTP1 and SrSTP14, whose genes were identified by RNA‐seq as the upregulated STPs during thermogenesis. Real‐time PCR confirmed that mRNA expression of both STP genes was increased from the pre‐thermogenic to the thermogenic stage in the spadix, where it is predominantly expressed in the stamen. SrSTP1 and SrSTP14 complemented the growth defects of a hexose transporter‐deficient yeast strain, EBY4000, on media containing 0.02, 0.2, and 2% (w/v) glucose and galactose. Using a recently developed transient expression system in skunk cabbage leaf protoplasts, we revealed that SrSTP1 and SrSTP14‐GFP fusion proteins were mainly localized to the plasma membrane. To dig further into the functional analysis of SrSTPs, tissue‐specific localization of SrSTPs was investigated by in situ hybridization. Using probes for SrSTP14, mRNA expression was observed in the microspores within the developing anther at the thermogenic female stage. These results indicate that SrSTP1 and SrSTP14 transport hexoses (e.g., glucose and galactose) at the plasma membrane and suggest that SrSTP14 may play a role in pollen development through the uptake of hexoses into pollen precursor cells.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Molecular characterization of SrSTP14, a sugar transporter from thermogenic skunk cabbage, and its possible role in developing pollen

Koyamatsu,

Otsubo,

Ohira

et al. 2023

Physiologia Plantarum

Self Cite

View full text Add to dashboard Cite

show abstract

“…Thermogenic plants are defined as plants that can increase temperature in their reproductive organs (e.g., inflorescences, flowers, and cones) to at least 0.5°C above ambient temperature (Maekawa et al, 2022). Based on this definition, at least 90 plant species have been reported as thermogenic, half of which are gymnosperm cycads (Ito‐Inaba et al, 2019; Tang, 1987), and the remaining half are angiosperms comprising mostly aroids (Araceae) and a few species of Nymphaeaceae, Magnoliacea, and Nelumbonaceae (Seymour, 2010).…”

Section: Introductionmentioning

confidence: 99%

Potential contribution of floral thermogenesis to cold adaptation, distribution pattern, and population structure of thermogenic and non/slightly thermogenic Symplocarpus species

Sato

Matsuo

Otsuka

et al. 2023

Ecology and Evolution

Self Cite

View full text Add to dashboard Cite

The genus Symplocarpus in basal Araceae includes both thermogenic and non/ slightly thermogenic species that prefer cold environments. If floral thermogenesis of Symplocarpus contributes to cold adaptation, it would be expected that thermogenic species have a larger habitat than non/slightly thermogenic species during an ice age, leading to increased genetic diversity in the current population. To address this question, potential distribution in past environment predicted by ecological niche modeling (ENM), genetic diversity, and population structure of chloroplast and genome-wide single nucleotide polymorphisms were compared between thermogenic Symplocarpus renifolius and non/slightly thermogenic Symplocarpus nipponicus. ENM revealed that the distribution of S. nipponicus decreased, whereas that of S. renifolius expanded in the Last Glacial Maximum. Phylogeographic analyses have shown that the population structures of the two species were genetically segmented and that the genetic diversity of S. renifolius was higher than that of S. nipponicus. The phylogenetic relationship between chloroplast and nuclear DNA is topologically different in the two species, which may be due to the asymmetric gene flow ubiquitously observed in plants. The results of this study imply that floral thermogenesis of Symplocarpus contributes to expanding the distribution during an ice age, resulting in increased genetic diversity due to cold adaptation.

show abstract

Plant Tissue Culture and Crop Improvement

Aslam,

Bibi,

Bibi

et al. 2023

Climate-Resilient Agriculture, Vol 2

View full text Add to dashboard Cite

Establishing an efficient protoplast transient expression system for investigation of floral thermogenesis in aroids

Cited by 5 publications

References 63 publications

Molecular characterization of SrSTP14, a sugar transporter from thermogenic skunk cabbage, and its possible role in developing pollen

Molecular characterization of SrSTP14, a sugar transporter from thermogenic skunk cabbage, and its possible role in developing pollen

Potential contribution of floral thermogenesis to cold adaptation, distribution pattern, and population structure of thermogenic and non/slightly thermogenic Symplocarpus species

Plant Tissue Culture and Crop Improvement

Contact Info

Product

Resources

About