Differential methylation and expression of HUA1 ortholog in three sex types of papaya

Juan Liu, Laura Chatham, Rishi Aryal, Qingyi Yu, Ray R Ming

Research output: Contribution to journalArticle

Abstract

Papaya is trioecious and an excellent system for studying sex determination and differentiation in plants. An ortholog of HUA1, CpHUA1, a gene controlling stamen and carpel development in Arabidopsis, was cloned and characterized in papaya. CpHUA1 consists of 12 exons with full genomic length of 19,313 bp in male AU9 and 19,312 bp in hermaphrodite SunUp, whereas the Arabidopsis HUA1 consists of 12 exons with full genomic length of 4300 bp. All the 324 SNPs between male and hermaphrodite varieties are in the 11th intron, which spans 8.5 kb. Quantitative RT-PCR revealed that CpHUA1 expression is highly elevated in carpels, suggesting that CpHUA1 may be involved in sex differentiation gene network. Southern blot analysis revealed a distinct restriction pattern in male AU9 compared to hermaphrodite Kapoho and SunUp, despite high DNA sequence identity and sharing of all but two EcoR I restriction sites in genomic CpHUA1 sequences of AU9 and SunUp. The methylation of cytosine at one restriction site in male but not in other two sex types may result in distinct restriction pattern of EcoR I in southern blot result. Bisulfite sequencing showed differential methylation of CpHUA1 among sex types, particularly the enrichment of sex-specific methylation in 9th and 11th intron. The methylation difference in cold stress induced male to hermaphrodite mutant mostly observed in the CHH context of CpHUA1, but no methylation difference detected in CHH context in other sex types, which may indicate the role of methylation in CHH context of CpHUA1 in temperature-related stress response and sex reversal.

Original languageEnglish (US)
Pages (from-to)99-106
Number of pages8
JournalPlant Science
Volume272
DOIs
StatePublished - Jul 2018

Fingerprint

Carica
papayas
methylation
Methylation
gender
Sex Differentiation
carpels
Southern Blotting
genomics
Arabidopsis
Introns
Southern blotting
exons
introns
Exons
bisulfites
sex reversal
Gene Regulatory Networks
sexual development
cytosine

Keywords

  • Carpel
  • CpHUA1 gene
  • Methylation
  • Papaya
  • Southern blot

ASJC Scopus subject areas

  • Genetics
  • Agronomy and Crop Science
  • Plant Science

Cite this

Differential methylation and expression of HUA1 ortholog in three sex types of papaya. / Liu, Juan; Chatham, Laura; Aryal, Rishi; Yu, Qingyi; Ming, Ray R.

In: Plant Science, Vol. 272, 07.2018, p. 99-106.

Research output: Contribution to journalArticle

Liu, Juan ; Chatham, Laura ; Aryal, Rishi ; Yu, Qingyi ; Ming, Ray R. / Differential methylation and expression of HUA1 ortholog in three sex types of papaya. In: Plant Science. 2018 ; Vol. 272. pp. 99-106.
@article{9ed9cae0f43e42f99dc08c5f58ce78ff,
title = "Differential methylation and expression of HUA1 ortholog in three sex types of papaya",
abstract = "Papaya is trioecious and an excellent system for studying sex determination and differentiation in plants. An ortholog of HUA1, CpHUA1, a gene controlling stamen and carpel development in Arabidopsis, was cloned and characterized in papaya. CpHUA1 consists of 12 exons with full genomic length of 19,313 bp in male AU9 and 19,312 bp in hermaphrodite SunUp, whereas the Arabidopsis HUA1 consists of 12 exons with full genomic length of 4300 bp. All the 324 SNPs between male and hermaphrodite varieties are in the 11th intron, which spans 8.5 kb. Quantitative RT-PCR revealed that CpHUA1 expression is highly elevated in carpels, suggesting that CpHUA1 may be involved in sex differentiation gene network. Southern blot analysis revealed a distinct restriction pattern in male AU9 compared to hermaphrodite Kapoho and SunUp, despite high DNA sequence identity and sharing of all but two EcoR I restriction sites in genomic CpHUA1 sequences of AU9 and SunUp. The methylation of cytosine at one restriction site in male but not in other two sex types may result in distinct restriction pattern of EcoR I in southern blot result. Bisulfite sequencing showed differential methylation of CpHUA1 among sex types, particularly the enrichment of sex-specific methylation in 9th and 11th intron. The methylation difference in cold stress induced male to hermaphrodite mutant mostly observed in the CHH context of CpHUA1, but no methylation difference detected in CHH context in other sex types, which may indicate the role of methylation in CHH context of CpHUA1 in temperature-related stress response and sex reversal.",
keywords = "Carpel, CpHUA1 gene, Methylation, Papaya, Southern blot",
author = "Juan Liu and Laura Chatham and Rishi Aryal and Qingyi Yu and Ming, {Ray R}",
year = "2018",
month = "7",
doi = "10.1016/j.plantsci.2018.04.001",
language = "English (US)",
volume = "272",
pages = "99--106",
journal = "Plant Science",
issn = "0168-9452",
publisher = "Elsevier Ireland Ltd",

}

TY - JOUR

T1 - Differential methylation and expression of HUA1 ortholog in three sex types of papaya

AU - Liu, Juan

AU - Chatham, Laura

AU - Aryal, Rishi

AU - Yu, Qingyi

AU - Ming, Ray R

PY - 2018/7

Y1 - 2018/7

N2 - Papaya is trioecious and an excellent system for studying sex determination and differentiation in plants. An ortholog of HUA1, CpHUA1, a gene controlling stamen and carpel development in Arabidopsis, was cloned and characterized in papaya. CpHUA1 consists of 12 exons with full genomic length of 19,313 bp in male AU9 and 19,312 bp in hermaphrodite SunUp, whereas the Arabidopsis HUA1 consists of 12 exons with full genomic length of 4300 bp. All the 324 SNPs between male and hermaphrodite varieties are in the 11th intron, which spans 8.5 kb. Quantitative RT-PCR revealed that CpHUA1 expression is highly elevated in carpels, suggesting that CpHUA1 may be involved in sex differentiation gene network. Southern blot analysis revealed a distinct restriction pattern in male AU9 compared to hermaphrodite Kapoho and SunUp, despite high DNA sequence identity and sharing of all but two EcoR I restriction sites in genomic CpHUA1 sequences of AU9 and SunUp. The methylation of cytosine at one restriction site in male but not in other two sex types may result in distinct restriction pattern of EcoR I in southern blot result. Bisulfite sequencing showed differential methylation of CpHUA1 among sex types, particularly the enrichment of sex-specific methylation in 9th and 11th intron. The methylation difference in cold stress induced male to hermaphrodite mutant mostly observed in the CHH context of CpHUA1, but no methylation difference detected in CHH context in other sex types, which may indicate the role of methylation in CHH context of CpHUA1 in temperature-related stress response and sex reversal.

AB - Papaya is trioecious and an excellent system for studying sex determination and differentiation in plants. An ortholog of HUA1, CpHUA1, a gene controlling stamen and carpel development in Arabidopsis, was cloned and characterized in papaya. CpHUA1 consists of 12 exons with full genomic length of 19,313 bp in male AU9 and 19,312 bp in hermaphrodite SunUp, whereas the Arabidopsis HUA1 consists of 12 exons with full genomic length of 4300 bp. All the 324 SNPs between male and hermaphrodite varieties are in the 11th intron, which spans 8.5 kb. Quantitative RT-PCR revealed that CpHUA1 expression is highly elevated in carpels, suggesting that CpHUA1 may be involved in sex differentiation gene network. Southern blot analysis revealed a distinct restriction pattern in male AU9 compared to hermaphrodite Kapoho and SunUp, despite high DNA sequence identity and sharing of all but two EcoR I restriction sites in genomic CpHUA1 sequences of AU9 and SunUp. The methylation of cytosine at one restriction site in male but not in other two sex types may result in distinct restriction pattern of EcoR I in southern blot result. Bisulfite sequencing showed differential methylation of CpHUA1 among sex types, particularly the enrichment of sex-specific methylation in 9th and 11th intron. The methylation difference in cold stress induced male to hermaphrodite mutant mostly observed in the CHH context of CpHUA1, but no methylation difference detected in CHH context in other sex types, which may indicate the role of methylation in CHH context of CpHUA1 in temperature-related stress response and sex reversal.

KW - Carpel

KW - CpHUA1 gene

KW - Methylation

KW - Papaya

KW - Southern blot

UR - http://www.scopus.com/inward/record.url?scp=85045574984&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85045574984&partnerID=8YFLogxK

U2 - 10.1016/j.plantsci.2018.04.001

DO - 10.1016/j.plantsci.2018.04.001

M3 - Article

C2 - 29807610

AN - SCOPUS:85045574984

VL - 272

SP - 99

EP - 106

JO - Plant Science

JF - Plant Science

SN - 0168-9452

ER -