{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,13]],"date-time":"2026-09-13T05:04:45Z","timestamp":1789275885130,"version":"build-2803163510"},"reference-count":24,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T00:00:00Z","timestamp":1789084800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Central Public-interest Scientific Institution Basal Research Fund for Chinese Academy of Tropical Agricultural","award":["1630062026004"],"award-info":[{"award-number":["1630062026004"]}]},{"name":"Central Public-interest Scientific Institution Basal Research Fund for Chinese Academy of Tropical Agricultural","award":["1630062022001"],"award-info":[{"award-number":["1630062022001"]}]},{"name":"Natural Science Foundation of Hainan Provincial","award":["325QN425"],"award-info":[{"award-number":["325QN425"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["32360608"],"award-info":[{"award-number":["32360608"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"award":["32360608"],"award-info":[{"award-number":["32360608"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Hainan Provincial Key R&D Program Projects","award":["ZDYF2026XDNY138"],"award-info":[{"award-number":["ZDYF2026XDNY138"]}]},{"name":"Safe Conservation of Tropical Fruit Tree Germplasm Resources","award":["22260030"],"award-info":[{"award-number":["22260030"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Quantitative real-time PCR (qRT-PCR) is a widely used technique for quantifying gene expression. However, accurate data normalization requires stable reference genes that exhibit constant expression levels across different experimental conditions. To date, no suitable reference genes have been validated for atemoya fruits (Annona cherimola Mill \u00d7 A. squamosa L.). This study aimed to evaluate the expression stability of candidate reference genes in atemoya across distinct tissues, postharvest storage stages, and various treatment conditions. To this end, we designed specific primers for six reference genes: ubiquitin carrier-like protein (UBC), actin 7 (ACT7), actin 11 (ACT11), elongation factor 1\u03b1 (EF1\u03b1), 18S ribosomal RNA (18S), and \u03b2-tubulin (TUB). Total RNA was extracted from atemoya tissues (e.g., petals, young fruits, and roots), for pulp across three post-harvest time points (0, 2, and 4 days) and under different conditions: 28 \u00b0C, 15 \u00b0C, ethylene, or 1-MCP treatment. The geNorm tool was subsequently used to identify the most stable reference transcripts under each experimental condition. ACT7 was the most stable reference gene across atemoya tissues, while UBC ranked first under all postharvest conditions tested (28 \u00b0C storage, 15 \u00b0C cold storage, ethylene, and 1-MCP). Furthermore, the suitability of these reference genes under ethylene and 1-MCP treatments was validated by examining the expression patterns of AaPG and AaERF. Collectively, the combined use of ACT7, UBC, and 18S proved to be the most reliable normalization strategy across all experimental conditions. This validated multi-gene approach provides a robust foundation for future investigations into gene expression and the postharvest ripening mechanisms of atemoya.<\/jats:p>","DOI":"10.3390\/ijms27188114","type":"journal-article","created":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T17:14:59Z","timestamp":1789146899000},"page":"8114","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Identification and Validation of Reliable Reference Genes for Gene Expression Studies in Postharvest Atemoya Pulp (Annona cherimola Mill \u00d7 A. squamosa L.) Subjected to Different Preservation Treatments"],"prefix":"10.3390","volume":"27","author":[{"given":"Xueyu","family":"Zhang","sequence":"first","affiliation":[{"name":"Key Laboratory of Tropical Fruit Biology, Ministry of Agriculture and Rural Affairs, South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Zhanjiang 524091, China"},{"name":"State Key Laboratory of Tropical Crop Breeding, South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572024, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"}]},{"given":"Qinyi","family":"Gao","sequence":"additional","affiliation":[{"name":"Key Laboratory of Tropical Fruit Biology, Ministry of Agriculture and Rural Affairs, South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Zhanjiang 524091, China"},{"name":"State Key Laboratory of Tropical Crop Breeding, South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572024, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"}]},{"given":"Ying","family":"Zhou","sequence":"additional","affiliation":[{"name":"Key Laboratory of Tropical Fruit Biology, Ministry of Agriculture and Rural Affairs, South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Zhanjiang 524091, China"},{"name":"State Key Laboratory of Tropical Crop Breeding, South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572024, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"methodology","vocabulary":"credit"}]},{"given":"Hanzhou","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Tropical Fruit Biology, Ministry of Agriculture and Rural Affairs, South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Zhanjiang 524091, China"},{"name":"State Key Laboratory of Tropical Crop Breeding, South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572024, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"validation","vocabulary":"credit"}]},{"given":"Zhihui","family":"Chen","sequence":"additional","affiliation":[{"name":"Key Laboratory of Tropical Fruit Biology, Ministry of Agriculture and Rural Affairs, South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Zhanjiang 524091, China"},{"name":"State Key Laboratory of Tropical Crop Breeding, South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572024, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"validation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5814-8695","authenticated-orcid":false,"given":"Jingjing","family":"Chen","sequence":"additional","affiliation":[{"name":"Key Laboratory of Tropical Fruit Biology, Ministry of Agriculture and Rural Affairs, South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Zhanjiang 524091, China"},{"name":"State Key Laboratory of Tropical Crop Breeding, South Subtropical Crops Research Institute, Chinese Academy of Tropical Agricultural Sciences, Sanya 572024, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"writing-review-editing","vocabulary":"credit"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1016\/j.bjp.2015.07.006","article-title":"Alkaloids Isolated from the Leaves of Atemoya (Annona cherimola \u00d7 Annona squamosa)","volume":"25","author":"Costa","year":"2015","journal-title":"Rev. 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