TY - JOUR
T1 - Characterization and analysis of the promoter region of monodehydroascorbate reductase 4 (CpMDAR4) in papaya
AU - Zerpa-Catanho, Dessireé
AU - Clough, Steven J.
AU - Ming, Ray
N1 - Research funded by the National Science Foundation (NSF) Plant Genome Research Program Award DBI-1546890 to R.M. The Department of Plant Biology at the University of Illinois Urbana-Champaign gave D.Z.C a “Govindjee and Rajni Govindjee Award for Excellence in Biological Science” that provided $1000 that were used on this research.
Authors would like to thank Juan Liu and Ping Zhou from the Fujian Agriculture and Forestry University (FAFU), for providing raw transcriptome data for data analysis. Authors would like to thank Dr. Li-Qing Chen at the University of Illinois Urbana-Champaign, for providing the original pMDC162 vector used for transformation. Authors would like to thank Neha Pandey, Lulu Wang, Julie Nguyen and Yinghui Xin, for providing advice to improve the methods.
PY - 2022/12
Y1 - 2022/12
N2 - Key message: Differential spatial and temporal expression patterns due to regulatory cis-elements and two different isoforms are detected among CpMDAR4 alleles in papaya. Abstract: The aim of this research was to study the effects of cis-element differences between the X, Y and Yh alleles on the expression of CpMDAR4, a potential candidate gene for sex differentiation in papaya, using a transcriptional reporter system in a model species Arabidopsis thaliana. Possible effects of a retrotransposon insertion in the Y and Yh alleles on the transcription and expression of CpMDAR4 alleles in papaya flowers were also examined. When comparing promoters and cis-regulatory elements among genes in the non-recombining region of the sex chromosomes, paired genes exhibited differences. Our results showed that differences in the promoter sequences of the CpMDAR4 alleles drove the expression of a reporter gene to different flower tissues in Arabidopsis. β-glucuronidase staining analysis of T2 and T3 lines for constructs containing 5′ deletions of native Y and Yh allele promoters showed the loss of specific expression of the reporter gene in the anthers, confirming the existence and location of cis-regulatory element POLLEN1LELAT52. The expression analysis of CpMDAR4 alleles in papaya flowers also showed that all alleles are actively expressed in different flower tissues, with the existence of a shorter truncated isoform, with unknown function, for the Y and Yh alleles due to an LTR-RT insertion in the Y and Yh chromosomes. The observed expression patterns in Arabidopsis thaliana flowers and the expression patterns of CpMDAR4 alleles in papaya flowers suggest that MDAR4 might have a role on development of reproductive organs in papaya, and that it constitutes an important candidate for sex differentiation.
AB - Key message: Differential spatial and temporal expression patterns due to regulatory cis-elements and two different isoforms are detected among CpMDAR4 alleles in papaya. Abstract: The aim of this research was to study the effects of cis-element differences between the X, Y and Yh alleles on the expression of CpMDAR4, a potential candidate gene for sex differentiation in papaya, using a transcriptional reporter system in a model species Arabidopsis thaliana. Possible effects of a retrotransposon insertion in the Y and Yh alleles on the transcription and expression of CpMDAR4 alleles in papaya flowers were also examined. When comparing promoters and cis-regulatory elements among genes in the non-recombining region of the sex chromosomes, paired genes exhibited differences. Our results showed that differences in the promoter sequences of the CpMDAR4 alleles drove the expression of a reporter gene to different flower tissues in Arabidopsis. β-glucuronidase staining analysis of T2 and T3 lines for constructs containing 5′ deletions of native Y and Yh allele promoters showed the loss of specific expression of the reporter gene in the anthers, confirming the existence and location of cis-regulatory element POLLEN1LELAT52. The expression analysis of CpMDAR4 alleles in papaya flowers also showed that all alleles are actively expressed in different flower tissues, with the existence of a shorter truncated isoform, with unknown function, for the Y and Yh alleles due to an LTR-RT insertion in the Y and Yh chromosomes. The observed expression patterns in Arabidopsis thaliana flowers and the expression patterns of CpMDAR4 alleles in papaya flowers suggest that MDAR4 might have a role on development of reproductive organs in papaya, and that it constitutes an important candidate for sex differentiation.
KW - Carica papaya
KW - Cis-elements
KW - Isoforms
KW - Monodehydroascorbate reductase
KW - Promoter analysis
UR - http://www.scopus.com/inward/record.url?scp=85135328393&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85135328393&partnerID=8YFLogxK
U2 - 10.1007/s00497-022-00447-2
DO - 10.1007/s00497-022-00447-2
M3 - Article
C2 - 35920937
AN - SCOPUS:85135328393
SN - 2194-7953
VL - 35
SP - 233
EP - 264
JO - Plant Reproduction
JF - Plant Reproduction
IS - 4
ER -