Functional Analysis of Transcription Factor CsbHLH3 in Regulating Citric Acid Metabolism of Citrus Fruit

doi:10.16420/j.issn.0513-353x.2022-0205

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (5): 947-958.doi: 10.16420/j.issn.0513-353x.2022-0205

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Functional Analysis of Transcription Factor CsbHLH3 in Regulating Citric Acid Metabolism of Citrus Fruit

GUO Jing, LIAO Manyu, JIN Yan, MA Xiaochuan, ZHANG Fen, LU Xiaopeng, DENG Ziniu, SHENG Ling()

College of Horticulture，National Center for Citrus Improvement Changsha，Hunan Agricultural University，Changsha 410128，China

Received:2022-06-09 Revised:2023-01-19 Online:2023-05-25 Published:2023-05-31
Contact: SHENG Ling E-mail:shengling0629@163.com

Abstract

Abstract:

Transcription factor CsbHLH3 plays an important role in regulating citrate metabolism in citrus fruits，but the specific mechanism remains unclear. In this study，the CsbHLH3 gene was cloned，and transient transformation in tobacco leaves demonstrated the subcellular localization of CsbHLH3 protein in the nucleus. CsbHLH3 overexpression tomato fruit showed a significant increase in the expression of CsbHLH3 while a significant decrease in the content of citrate relative to the wide type. Quantitative analysis revealed that the overexpression of CsbHLH3 induced the up-regulation of SlACO3b and SlPH8. On the contrary，transient suppression of the CsbHLH3 gene in Bingtangcheng leaves resulted in a significantly higher content of citrate than that of the control. In addition，the suppression of CsbHLH3 expression down-regulated the expression of the CsACO2，CsACLα1，CsACLα2 and CsGABP genes in the citrate degradation pathway and proton pump genes CsPH8，CsVHP2 and CsVHP3. Moreover，CsbHLH3 was further confirmed to target the promoter of PH8 and GABP genes to regulate their expression through Y1H and EMSA. These results suggested that the transcription factor CsbHLH3 negatively regulates the citrate level by regulating the expression of several genes related to citrate metabolism，including PH8 and the degradation gene GABP.

Key words: citrus, citrate, CsbHLH3, gene function, regulation

CLC Number:

S666.1

GUO Jing, LIAO Manyu, JIN Yan, MA Xiaochuan, ZHANG Fen, LU Xiaopeng, DENG Ziniu, SHENG Ling. Functional Analysis of Transcription Factor CsbHLH3 in Regulating Citric Acid Metabolism of Citrus Fruit[J]. Acta Horticulturae Sinica, 2023, 50(5): 947-958.

Figures/Tables 7

References 35

[1]	Butelli E, Licciardello C, Ramadugu C, Durand-Hulak M, Celant A, Recupero G R, Froelicher Y, Martin C. 2019. Noemi controls production of flavonoid pigments and fruit acidity and illustrates the domestication routes of modern citrus varieties. Current Biology, 29 (1):158-164. doi: S0960-9822(18)31539-2 pmid: 30581020
[2]	Cercós M, Soler G, Iglesias D J, Gadea J, Forment J, Talón M. 2006. Global analysis of gene expression during development and ripening of citrus fruit flesh. A proposed mechanism for citric acid utilization. Plant Molecular Biology, 62 (4):513-527. doi: 10.1007/s11103-006-9037-7 URL
[3]	Chen Dan-dan. 2016. cis-Regulatory elements analysis and interacting protein screening of CsGABP promoter in Citrus[M. D. Dissertation]. Wuhan: Huazhong Agricultural University. (in Chinese)
	谌丹丹. 2016. 柑橘CsGABP启动子作用元件分析及互作蛋白筛选[硕士论文]. 武汉: 华中农业大学.
[4]	Chen M, Jiang Q, Yin X R, Lin Q, Chen J Y, Allan A C, Xu C J, Chen K S. 2012. Effect of hot air treatment on organic acid- and sugar-metabolism in Ponkan(Citrus reticulata)fruit. Scientia Horticulturae, 147:118-125. doi: 10.1016/j.scienta.2012.09.011 URL
[5]	Degu A, Hatew B, Nunes-Nesi A, Shlizerman L, Zur N, Katz E, Fernie A R, Blumwald E, Sadka A. 2011. Inhibition of aconitase in citrus fruit callus results in a metabolic shift towards amino acid biosynthesis. Planta, 234 (3):501-513. doi: 10.1007/s00425-011-1411-2 pmid: 21528417
[6]	Etienne A, Génard M, Lobit P, Mbeguié-A-Mbéguié D, Bugaud C. 2013. What controls fleshy fruit acidity? A review of malate and citrate accumulation in fruit cells. Journal of Experimental Botany, 64 (6):1451-1469. doi: 10.1093/jxb/ert035 pmid: 23408829
[7]	Etienne C, Moing A, Dirlewanger E, Raymond P, Monet R, Rothan C. 2002. Isolation and characterization of six peach cDNAs encoding key proteins in organic acid metabolism and solute accumulation:involvement in regulating peach fruit acidity. Physiologia Plantarum, 114 (2):259-270. doi: 10.1034/j.1399-3054.2002.1140212.x URL
[8]	Guillet C, Just D, Bénard N, Destrac-Irvine A, Baldet P, Hernould M, Causse M, Raymond P, Rothan C. 2002. A fruit-specific phosphoenolpyruvate carboxylase is related to rapid growth of tomato fruit. Planta, 214 (5):717-726. doi: 10.1007/s00425-001-0687-z pmid: 11882940
[9]	Guo L X, Shi C Y, Liu X, Ning D Y, Jing L F, Yang H, Liu Y Z. 2016. Citrate accumulation-related gene expression and/or enzyme activity analysis combined with metabolomics provide a novel insight for an orange mutant. Scientific Reports, 6 (1):1-12. doi: 10.1038/s41598-016-0001-8
[10]	Gupta K J, Shah J K, Brotman Y, Jahnke K, Willmitzer L, Kaiser W M, Bauwe H, Igamberdiev A U. 2012. Inhibition of aconitase by nitric oxide leads to induction of the alternative oxidase and to a shift of metabolism towards biosynthesis of amino acids. Journal of Experimental Botany, 63 (4):1773-1784. doi: 10.1093/jxb/ers053 pmid: 22371326
[11]	Hu X M, Shi C Y, Liu X, Jin L F, Liu Y Z, Peng S A. 2015. Genome-wide identification of citrus ATP-citrate lyase genes and their transcript analysis in fruits reveals their possible role in citrate utilization. Molecular Genetics, 290 (1):29-38.
[12]	Huang D, Zhao Y, Cao M., Qiao L, Zheng Z L. 2016. Integrated systems biology analysis of transcriptomes reveals candidate genes for acidity control in developing fruits of sweet orange(Citrus sinensis L. Osbeck). Frontiers in Plant Science, 7:486.
[13]	Katz E, Boo K H, Kim H Y, Eigenheer R A, Phinney B S, Shulaev V, Negre-Zakharov F, Sadka A, Blumwald E. 2011. Label-free shotgun proteomics and metabolite analysis reveal a significant metabolic shift during citrus fruit development. Journal of Experimental Botany, 62 (15):5367-5384. doi: 10.1093/jxb/err197 pmid: 21841177
[14]	Krebs M, Beyhl D, Görlich E, Al-Rasheid K A, Marten I, Stierhof Y D, Hedrich R, Schumacher K. 2010. Arabidopsis V-ATPase activity at the tonoplast is required for efficient nutrient storage but not for sodium accumulation. Proceedings of the National Academy of Sciences, 107 (7):3251-3256.
[15]	Li J, Yang H, Peer W A, Richter G, Blakeslee J, Bandyopadhyay A, Titapiwantakun B, Undurraga S, Khodakovskaya M, Richards E L, Krizek B, Murphy A S, Gilroy S, Gaxiola R. 2005. Arabidopsis H⁺-PPase AVP 1 regulates auxin-mediated organ development. Science, 310 (5745):121-125. doi: 10.1126/science.1115711 URL
[16]	Li S J, Yin X R, Wang W L, Liu X F, Zhang B, Chen K S. 2017. Citrus Cit NAC62 cooperates with Cit WRKY1 to participate in citric acid degradation via up-regulation of Cit Aco3. J Exp Bot, 68:3419-3426. doi: 10.1093/jxb/erx187 URL
[17]	Liang Fang-fei, Wang Xiao-rong, Deng Li-li, Zeng Kai-fang, Yao Shi-xiang. 2018. Research advances in sugar and acid metabolism of postharvest citrus fruit. Food and Fermentation Industries, 44 (10):268-274. (in Chinese)
	梁芳菲, 王小容, 邓丽莉, 曾凯芳, 姚世响. 2018. 采后柑橘果实糖酸代谢研究进展. 食品与发酵工业, 44 (10):268-274.
[18]	Liu X, Hu X M, Jin L F, Shi C Y, Liu Y Z, Peng S A. 2014. Identification and transcript analysis of two glutamate decarboxylase genes, CsGAD1 and CsGAD2,reveal the strong relationship between CsGAD1 and citrate utilization in citrus fruit. Molecular Biology Reports, 41 (9):6253-6262. doi: 10.1007/s11033-014-3506-x URL
[19]	Lopez-Bucio J, Nieto-Jacobo M F, Ramırez-Rodrıguez V, Herrera-Estrella L. 2000. Organic acid metabolism in plants:from adaptive physiology to transgenic varieties for cultivation in extreme soils. Plant Science, 160 (1):1-13. doi: 10.1016/S0168-9452(00)00347-2 URL
[20]	Lu X P, Cao X J, Li F F, Li J, Xiong J, Long G Y, Cao S Y, Xie S X. 2016. Comparative transcriptome analysis reveals a global insight into molecular processes regulating citrate accumulation in sweet orange(Citrus sinensis). Physiologia Plantarum, 158 (4):463-482. doi: 10.1111/ppl.12484 URL
[21]	Lu Xiao-peng, Li Fei-fei, Xie Shen-xi. 2018. Citrate accumulation in citrus fruit:a molecular perspective. Journal of Fruit Science, 35 (1):118-127. (in Chinese)
	卢晓鹏, 李菲菲, 谢深喜. 2018. 柑橘果实柠檬酸积累调控基因研究进展. 果树学报, 35 (1):118-127.
[22]	Ma Xiao-qian. 2020. Effects of citrate content on the storage performance of‘Bingtang’Sweet Orange and expression analysis of genes in citrate metabolism[M. D. Dissertation]. Changsha: Hunan Agricultural University. (in Chinese)
	马晓倩. 2020. 柠檬酸含量对冰糖橙果实贮藏性能的影响及其代谢相关基因分析[硕士论文]. 长沙: 湖南农业大学.
[23]	Sadka A, Dahan E, Cohen L, Marsh K B. 2000. Aconitase activity and expression during the development of lemon fruit. Physiologia Plantarum, 108 (3):255-262. doi: 10.1034/j.1399-3054.2000.108003255.x URL
[24]	Sheng L, Shen D D, Yang W, Zhang M F, Zeng Y L, Xu J, Deng X X, Cheng Y J. 2017. GABA pathway rate-limit citrate degradation in postharvest citrus fruit evidence from HB pumelo(Citrus grandis)× fairchild(Citrus reticulata)hybrid population. Journal of Agricultural and Food Chemistry, 65 (8):1669-1676. doi: 10.1021/acs.jafc.6b05237 pmid: 28150945
[25]	Sheng Ling. 2017. Mechanism of gamma-aminobutyric acid shunt regulating citratemetabolism in citrus fruit[Ph. D. Dissertation]. Wuhan: Huazhong Agricultural University. (in Chinese)
	盛玲. 2017. GABA支路调控柑橘果实柠檬酸代谢的机理研究[博士论文]. 武汉: 华中农业大学.
[26]	Shi C Y, Hussain S B, Yang H, Bai Y X, Khan M A, Liu Y Z. 2019. CsPH8,a P-type proton pump gene,plays a key role in the diversity of citric acid accumulation in Citrus fruits. Plant Science, 289:110-288.
[27]	Shi C Y, Song R Q, Hu X M, Liu X, Jin L F, Liu Y Z. 2015. Citrus PH5-like H⁺-ATPase genes:and transcript analysis to investigate their possible relationship with citrate accumulation in fruits. Frontiers in Plant Science, 6,135.
[28]	Shimada T, Nakano R, Shulaev V, Sadka A, Blumwald E. 2006. Vacuolar citrate/H⁺ symporter of citrus juice cells. Planta, 224 (2):472-480. doi: 10.1007/s00425-006-0223-2 URL
[29]	Sinclair W B. 1984. The biochemistry and physiology of the lemon and other citrus fruits. University of California System Division of Agriculture and Natural.
[30]	Strazzer P, Spelt C E, Li S, Bliek M, Federici C T, Roose M L, Koes R, Quattrocchio F M. 2019. Hyperacidification of citrus fruits by a vacuolar proton-pumping P-ATPase complex. Nature Communications, 10 (1):1-11. doi: 10.1038/s41467-018-07882-8
[31]	Verweij W, Spelt C, Di Sansebastiano G P, Vermeer J, Reale L, Ferranti F, Koes R, Quattrocchio F. 2008. An H⁺ P-ATPase on the tonoplast determines vacuolar pH and flower colour. Nature Cell Biology, 10 (12):1456-1462. doi: 10.1038/ncb1805 pmid: 18997787
[32]	Zeng Yike, Shi Ying, Chen Siyi, Li Guojing, Huang Xianbiao, Xie Zongzhou, Li Chunlong, Guo Dayong, Liu Jihong. 2022. Effects of film mulching on improving fruit quality of ponkan and possible mechanisms. Acta Horticulturae Sinica, 49 (11):2419-2430. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-0720
	曾译可, 石莹, 陈思怡, 李国敬, 黄先彪, 谢宗周, 李春龙, 郭大勇, 刘继红. 2022. 地面覆膜提升椪柑果实品质的效果和可能机制探究. 园艺学报, 49 (2):2419-2430.
[33]	Zhang G, Xie S. 2014. Influence of water stress on the citric acid metabolism related gene expression in the ponkan fruits. Agricultural Sciences, 5 (14):1513. doi: 10.4236/as.2014.514162 URL
[34]	Zhao Yong, Zhu Hongju, Yang Dongdong, Gong Chengsheng, Liu Wenge. 2022. Research progress of citric acid metabolism in the fruit. Acta Horticulturae Sinica, 49 (12):2579-2596. (in Chinese) doi: 10.16420/j.issn.0513-353x.2021-0709
	赵永, 朱红菊, 杨东东, 龚成胜, 刘文革. 2022. 果实柠檬酸代谢研究进展. 园艺学报, 49 (12):2579-2596. doi: 10.16420/j.issn.0513-353x.2021-0709
[35]	Zheng Li-jing, Nie Ji-yun, Li Ming-qiang, Kang Yan-ling, Kuang Li-xue, Ye Meng-liang. 2015. Study on screening of taste evaluation indexes for apple. Scientia Agricultura Sinica, 48 (14):2796-2805. (in Chinese) doi: 10.3864/j.issn.0578-1752.2015.14.011
	郑丽静, 聂继云, 李明强, 康艳玲, 匡立学, 叶孟亮. 2015. 苹果风味评价指标的筛选研究. 中国农业科学, 48 (14):2796-2805. doi: 10.3864/j.issn.0578-1752.2015.14.011

引物名称	正向引物（5′-3′）	反向引物（5′-3′）
Primer name	Forward	Reverse
p1300-CsbHLH3	GAGAACACGGGGGACGAGCTCATGGCTGAGAAATTTTGGACAAAGG	GCCCTTGCTCACCATGGTACCTTTAGAAAGGGCAGCAAGTAGT
RNAi-CsbHLH3	AAAAAGCAGGCTAGCTGTCCTCTGGACGAC	GAAAGCTGGGTTGATTCGTATAGATTCATCATA
Adapter-attB	GGGACAAGTTTGTACAAAAAAGCAGGCT;	GGGACCACTTTGTACAAGAAAGCTGGGT
P35s-F	CCCACCCACGAGGAGCAT
CsbHLH3（qRT）	TTACTGGCATGCTTCAAACTTGA	TGGCCATCTCCCCAAATTAG
Actin（qRT）	CCAAGCAGCATGAAGATCAA	ATCTGCTGGAAGGTGCTGAG
CsCS1（qRT）	GGTGCCCCCAATATTAACAA	AGAGCTCGGTCCCATATCAA
CsCS2（qRT）	ACTGGTGTATGGATGCGACA	TCTTCGTCTTGTGGCATTTG
CsPEPC1（qRT）	GTGCGATCCCGTCTATCTGT	AAGGCTCAAGGCCACTTTTT
CsPEPC2（qRT）	GGCATGCAAAACACTGGTTA	CATGTTCATTACGGCTTGGA
CsPEPC3（qRT）	GAACAATGACGGACACAACG	TGGACTCGCTTCCAACTTCT
CsACO1（qRT）	GGCAAGTCATTCACATGCGTT	TGAAGAAGTAGACCCCGGTTGA
CsACO2（qRT）	GGCAATGATGAAGTGATGGCT	GTTGGAACATGGACCGTCTTT
CsACO3（qRT）	TGCAGCAATGAGGTACAAGGC	TCACACCCAGAAGCATTGGAC
CsACLα1（qRT）	GATACTGTTGGAGACTTGGG	GCTCTCTTACGACCATCAGG
CsACLα2（qRT）	TACAGTGGAGCACCCAACGA	CCTTCAGGGCTTGGATTATG
CsACLβ（qRT）	GAGGAGATAACAGAGACAAA	AACAAAGAGCCCATTCAGAT
CsGABP（qRT）	GTTGAGTGTTCCATCTCATA	GAACATGGCCTGAATCAACA
CsPH8（qRT）	CCGTGAAGGAATTGATTTGG	CCATGACAATGGATTCCACA
CsVHP2（qRT）	TGAGCCACAGAATCAGAGAGAGAA	GCACCAACAATCAAACCAATAAAC
CsVHP3（qRT）	CCCTGCACATACAACACAG	TGCTGACTCCTTTCCTTGCT
Sl Actin（qRT）	CACCATTGGGTCTGAGCGAT	GGGCGACAACCTTGATCTTC
Sl CS（qRT）	TTGGGGAACATCACAGTTG	TGATGGCACCTTTCCTGTT
Sl PEPC1（qRT）	TGTGAACCTGAACCCGACT	GTCCCCTATTCGGGACTTC
Sl PEPC2（qRT）	TATCACTACTTTAAATGTCTGC	TCAAGGATACATGATTCTTAAT
Sl ACO3a（qRT）	GCCGCTTGCTTCAACTTCTAC	GACTCCACCTCGGCACAGA
Sl ACO3b（qRT）	TGGTGCTTATTGCTCTAGTGGGTA	CAACACCGTATCTCCACCTCA
Sl GAD2（qRT）	CTTTGATCTTCTCCGTCGTTG	ATATCGAGACGCGAAAGTCG
Sl GAD3（qRT）	CAGGACGTTTCAATATAATC	CCTACGGAGGGTCTCAGAG
Sl GDH（qRT）	AGCACGACAATGCACGAGGG	ATATTGGCGACCGCTGTCTTCC
Sl PEPCK（qRT）	AGACGAAACCACTGAGGACGA	CATTCACAAACACCTTCTCCAA
Sl GABA-T（qRT）	CAGCACAAGCTTGACGATGG	TGGTGATTACTGGTTAAGGG
Sl SSADH（qRT）	TCTCCGCTGAGGAGGGTAAACG	ACAAGCAAGAGCAGGGCCAACC
Sl PH8（qRT）	TCGGAAAGGCTGCACATCTT	CGCTTTTGAATCGGCCACAT

引物名称	正向引物（5′-3′）	反向引物（5′-3′）
Primer name	Forward	Reverse
p1300-CsbHLH3	GAGAACACGGGGGACGAGCTCATGGCTGAGAAATTTTGGACAAAGG	GCCCTTGCTCACCATGGTACCTTTAGAAAGGGCAGCAAGTAGT
RNAi-CsbHLH3	AAAAAGCAGGCTAGCTGTCCTCTGGACGAC	GAAAGCTGGGTTGATTCGTATAGATTCATCATA
Adapter-attB	GGGACAAGTTTGTACAAAAAAGCAGGCT;	GGGACCACTTTGTACAAGAAAGCTGGGT
P35s-F	CCCACCCACGAGGAGCAT
CsbHLH3（qRT）	TTACTGGCATGCTTCAAACTTGA	TGGCCATCTCCCCAAATTAG
Actin（qRT）	CCAAGCAGCATGAAGATCAA	ATCTGCTGGAAGGTGCTGAG
CsCS1（qRT）	GGTGCCCCCAATATTAACAA	AGAGCTCGGTCCCATATCAA
CsCS2（qRT）	ACTGGTGTATGGATGCGACA	TCTTCGTCTTGTGGCATTTG
CsPEPC1（qRT）	GTGCGATCCCGTCTATCTGT	AAGGCTCAAGGCCACTTTTT
CsPEPC2（qRT）	GGCATGCAAAACACTGGTTA	CATGTTCATTACGGCTTGGA
CsPEPC3（qRT）	GAACAATGACGGACACAACG	TGGACTCGCTTCCAACTTCT
CsACO1（qRT）	GGCAAGTCATTCACATGCGTT	TGAAGAAGTAGACCCCGGTTGA
CsACO2（qRT）	GGCAATGATGAAGTGATGGCT	GTTGGAACATGGACCGTCTTT
CsACO3（qRT）	TGCAGCAATGAGGTACAAGGC	TCACACCCAGAAGCATTGGAC
CsACLα1（qRT）	GATACTGTTGGAGACTTGGG	GCTCTCTTACGACCATCAGG
CsACLα2（qRT）	TACAGTGGAGCACCCAACGA	CCTTCAGGGCTTGGATTATG
CsACLβ（qRT）	GAGGAGATAACAGAGACAAA	AACAAAGAGCCCATTCAGAT
CsGABP（qRT）	GTTGAGTGTTCCATCTCATA	GAACATGGCCTGAATCAACA
CsPH8（qRT）	CCGTGAAGGAATTGATTTGG	CCATGACAATGGATTCCACA
CsVHP2（qRT）	TGAGCCACAGAATCAGAGAGAGAA	GCACCAACAATCAAACCAATAAAC
CsVHP3（qRT）	CCCTGCACATACAACACAG	TGCTGACTCCTTTCCTTGCT
Sl Actin（qRT）	CACCATTGGGTCTGAGCGAT	GGGCGACAACCTTGATCTTC
Sl CS（qRT）	TTGGGGAACATCACAGTTG	TGATGGCACCTTTCCTGTT
Sl PEPC1（qRT）	TGTGAACCTGAACCCGACT	GTCCCCTATTCGGGACTTC
Sl PEPC2（qRT）	TATCACTACTTTAAATGTCTGC	TCAAGGATACATGATTCTTAAT
Sl ACO3a（qRT）	GCCGCTTGCTTCAACTTCTAC	GACTCCACCTCGGCACAGA
Sl ACO3b（qRT）	TGGTGCTTATTGCTCTAGTGGGTA	CAACACCGTATCTCCACCTCA
Sl GAD2（qRT）	CTTTGATCTTCTCCGTCGTTG	ATATCGAGACGCGAAAGTCG
Sl GAD3（qRT）	CAGGACGTTTCAATATAATC	CCTACGGAGGGTCTCAGAG
Sl GDH（qRT）	AGCACGACAATGCACGAGGG	ATATTGGCGACCGCTGTCTTCC
Sl PEPCK（qRT）	AGACGAAACCACTGAGGACGA	CATTCACAAACACCTTCTCCAA
Sl GABA-T（qRT）	CAGCACAAGCTTGACGATGG	TGGTGATTACTGGTTAAGGG
Sl SSADH（qRT）	TCTCCGCTGAGGAGGGTAAACG	ACAAGCAAGAGCAGGGCCAACC
Sl PH8（qRT）	TCGGAAAGGCTGCACATCTT	CGCTTTTGAATCGGCCACAT

Functional Analysis of Transcription Factor CsbHLH3 in Regulating Citric Acid Metabolism of Citrus Fruit

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 7

References 35

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	ZHANG Lehuan, ZOU Changyu, WANG Zhaohao, YANG Wen, ZOU Xiuping, HE Yongrui, CHEN Shanchun, LONG Qin. Cloning and Expression Analysis of CsAOS1-2 in Responding to Citrus Canker Disease [J]. Acta Horticulturae Sinica, 2023, 50(6): 1355-1367.
[2]	CHEN Xinchen, ZHAO Huimin, WANG Sen, WU Simin, XIANG Lin, CHAN Zhulong, WANG Yanping. Analysis of Floral Bud Differentiation and Related Genes Expression Under Different Temperatures in Tulipa gesneriana [J]. Acta Horticulturae Sinica, 2023, 50(5): 1037-1047.
[3]	LÜ Ruoya, LI Yun, ZHENG Yongqin, DENG Xiaoling, ZHENG Zheng. The Distribution Pattern of Candidatus Liberibacter asiaticus in Fruit Pith [J]. Acta Horticulturae Sinica, 2023, 50(5): 1110-1117.
[4]	WANG Ping, SHENG Ling, YANG Jinpeng, ZHOU Linglei, JIN Yan, LUO Xuzhao, MA Xianfeng, DENG Ziniu. Evaluation of Resistance to Citrus Canker Disease in Hybrid Progeny of Red Pomelo and American Citron [J]. Acta Horticulturae Sinica, 2023, 50(4): 765-777.
[5]	ZOU Yunqian, LUO Qujuan, ZHANG Jin, XU Rangwei, CHENG Yunjiang. Coating Containing Shellac，Rosin Significantly Improves Commercial Value of Satsuma Mandarins and Lane Late Navel Orange During Shelf Life [J]. Acta Horticulturae Sinica, 2023, 50(4): 853-863.
[6]	TIAN Xiaocheng, ZHU Lingcheng, ZOU Hui, LI Baiyun, MA Fengwang, LI Mingjun. Research Progress on Accumulation Pattern and Regulation of Soluble Sugar in Fruit [J]. Acta Horticulturae Sinica, 2023, 50(4): 885-895.
[7]	LAI Hengxin, LI Wenguang, PENG Liangzhi, HE Yizhong, ZHU Panpan, YANG Wanyun, LING Lili, FU Xingzheng, CHUN Changpin, CAO Li. Quality Changes of On-tree Storage Fruit of Orah Mandarin（Citrus reticulata Blanco）During Spring and Summer Seasons [J]. Acta Horticulturae Sinica, 2023, 50(3): 485-494.
[8]	LIU Yunuo, CAO Ya, WANG Shuai, DU Meixia, ZHENG Lin, CHEN Shanchun, ZOU Xiuping. Expression Analysis of CsMYB41 and CsMYB63 Genes in Response to Citrus Canker [J]. Acta Horticulturae Sinica, 2023, 50(3): 495-507.
[9]	XUE Yuqian, LIU Zhiyong, SUN Kairong, ZHANG Xiuxin, LÜ Yingmin, XUE Jingqi. The Mechanism of Sugar Signal Involved in Regulating Re-flowering of Tree Peony Under Forcing Culture [J]. Acta Horticulturae Sinica, 2023, 50(3): 596-606.
[10]	YE Zimao, SHEN Wanxia, LIU Mengyu, WANG Tong, ZHANG Xiaonan, YU Xin, LIU Xiaofeng, ZHAO Xiaochun. Effect of R2R3-MYB Transcription Factor CitMYB21 on Flavonoids Biosynthesis in Citrus [J]. Acta Horticulturae Sinica, 2023, 50(2): 250-264.
[11]	ZHENG Qingbo, BAO Zeyang, LAN Qingqing, ZHOU Yuwen, ZHOU Yufei, ZHENG Caixia, LI Xu. Advances in Studies on Adventitious Root Formation by Juvenile- and Auxin-determined [J]. Acta Horticulturae Sinica, 2023, 50(2): 441-450.
[12]	JIANG Jingdong, WEI Zhuangmin, WANG Nan, ZHU Chenqiao, YE Junli, XIE Zongzhou, DENG Xiuxin, CHAI Lijun. Exploitation and Identification of Tetraploid Resources of Hongkong Kumquat（Fortunella hindsii） [J]. Acta Horticulturae Sinica, 2023, 50(1): 27-35.
[13]	DU Yuling, YANG Fan, ZHAO Juan, LIU Shuqi, LONG Chaoan. Antifungal Mechanisms of Sodium New Houttuyfonate Against Penicillium digitatum [J]. Acta Horticulturae Sinica, 2023, 50(1): 145-152.
[14]	LI Zhenxi, PAN Ruixuan, XU Meirong, ZHENG Zheng, DENG Xiaoling. Development of Duplex Real-time PCR Assay of‘Candidatus Liberibacter asiatics’ [J]. Acta Horticulturae Sinica, 2023, 50(1): 188-196.
[15]	SHAO Fengqing, LUO Xiurong, WANG Qi, ZHANG Xianzhi, WANG Wencai. Advances in Research of DNA Methylation Regulation During Fruit Ripening [J]. Acta Horticulturae Sinica, 2023, 50(1): 197-208.