Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (2): 355-366.doi: 10.16420/j.issn.0513-353x.2020-0377

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Cloning and Spatiotemporal Expression Analysis of PgWUS and PgBEL1 in Punica granatum

ZHAO Yujie, LIU Cuiyu, ZHAO Xueqing, WANG Yuying, YAN Ming, YUAN Zhaohe*()   

  1. College of Forestry,Nanjing Forestry University,Nanjing 210037,China
  • Received:2020-08-04 Revised:2020-11-01 Online:2021-02-25 Published:2021-03-09
  • Contact: YUAN Zhaohe


Based on the pomegranate genome database,PgWUS and PgBEL1,the homolog of WUS and BEL1 were identified and cloned by homologous cloning. The full-length CDS sequences of PgBEL1 and PgWUS were 1 851 bp and 936 bp,encoding 616 and 311 amino acids,respectively. The protein sequence multiple alignment and phylogenetic analysis suggested that PgWUS had the highest evolution relationship with WUS in Cucumis melo var. makuwa and C. sativus. There was high similarity of amino acid sequences in PgBEL1 and BEL1 of Eucatyptus grandis. Subcellular localization results showed that both PgBEL1 and PgWUS were located in the nucleus. Real-time quantitative PCR analysis suggested that the expression level of PgBEL1 in bisexual flowers was higher than that in functional male flowers at P1 P4 periods(bud vertical diameter < 12.0 mm). PgBEL1 was highly expressed in leaf,it was 3.2 times than that in calyx while 1.2 times than that in stem,respectively. PgBEL1 expression was low in shoot apex. The expression level of PgBEL1 in pistil was 1.16 times than that in stamen. At the P2 and P3 periods of pomegranate flower development that bud vertical diameter was 5.1-10.0 mm,the expression level of PgWUS in bisexual flowers was higher than that in functional male flowers. PgWUS expression was the lowest in calyx and was the highest in stem. The expression level of PgWUS in pistil was 1.5 times than that in stamen and 1.8 times than that in shoot apex. There was a significant difference in the expression levels of PgWUS in pistil and stamen.

Key words: pomegranate, WUS, BEL1, gene clone, expression analysis

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