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When P<jats:sub>i<\/jats:sub> in chloroplasts is insufficient, the increase in the CO<jats:sub>2<\/jats:sub> assimilation rate (<jats:italic>A<\/jats:italic>) with an increase in CO<jats:sub>2<\/jats:sub> level is restricted, whereas <jats:italic>A<\/jats:italic> per unit total protein level moderately decreases under low-to-normal CO<jats:sub>2<\/jats:sub> levels. Some phosphate transporters (PHT) are localized in the chloroplast envelope; however, their contribution to the maintenance of P<jats:sub>i<\/jats:sub> homeostasis for CO<jats:sub>2<\/jats:sub> assimilation has rarely been reported. In this study, we generated transgenic rice plants with RNAi-suppressed <jats:italic>PHT4;4<\/jats:italic>, one of the two genes of chloroplast envelope-localized PHT, and examined the changes in the characteristics of CO<jats:sub>2<\/jats:sub> assimilation. In three transgenic lines, the mRNA levels of <jats:italic>PHT4;4<\/jats:italic> decreased by approximately 80% without a notable decrease in total leaf-P levels or total leaf-N levels, which is thought to approximately correspond to total protein levels. <jats:italic>A<\/jats:italic> in the transgenic plants tended to be slightly lower than that in the wild-type plants, irrespective of the CO<jats:sub>2<\/jats:sub> level, and typically increased to saturation with increasing CO<jats:sub>2<\/jats:sub> levels. <jats:italic>A<\/jats:italic> per unit total leaf-N level in transgenic plants tended to be slightly lower than that in wild-type plants. These results indicate that substantial <jats:italic>PHT4;4<\/jats:italic> suppression caused slight symptoms of P<jats:sub>i<\/jats:sub>-limited CO<jats:sub>2<\/jats:sub> assimilation. Therefore, PHT4;4 is suggested to be involved in the maintenance of chloroplast P<jats:sub>i<\/jats:sub> homeostasis for healthy CO<jats:sub>2<\/jats:sub> assimilation, although its contribution is minor.<\/jats:p>","DOI":"10.1007\/s10265-025-01638-4","type":"journal-article","created":{"date-parts":[[2025,4,18]],"date-time":"2025-04-18T11:54:26Z","timestamp":1744977266000},"page":"667-677","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Effects of suppression of phosphate transporter 4;4 on CO2 assimilation in rice"],"prefix":"10.1007","volume":"138","author":[{"given":"Ryosei","family":"Harada","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takaya","family":"Sugimoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuki","family":"Takegahara-Tamakawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8789-5289","authenticated-orcid":false,"given":"Amane","family":"Makino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8029-9385","authenticated-orcid":false,"given":"Yuji","family":"Suzuki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,4,18]]},"reference":[{"key":"1638_CR1","doi-asserted-by":"publisher","first-page":"4493","DOI":"10.1016\/j.cub.2022.08.032","volume":"32","author":"N Bouain","year":"2022","unstructured":"Bouain N, Cho H, Sandhu J, Tuiwong P, Prom-u-thai C, Zheng L et al (2022) Plant growth stimulation by high CO2 depends on phosphorus homeostasis in chloroplasts. 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