Polyphenol oxidase genes in barley (Hordeum vulgare L.): functional activity with respect to black grain pigmentation

Polyphenol oxidase (PPO) is an oxidoreductase. In damaged plant tissues, it catalyzes enzymatic browning by oxidizing o-diphenols to highly reactive o-quinones, which polymerize producing heterogeneous dark polymer melanin. In intact tissues, functions of PPO are not well understood. The aim of the...

Full description

Bibliographic Details
Main Authors: Anastasiia Y. Glagoleva, Tat’jana V. Kukoeva, Elena K. Khlestkina, Olesya Y. Shoeva
Format: Article
Language:English
Published: Frontiers Media S.A. 2024-01-01
Series:Frontiers in Plant Science
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fpls.2023.1320770/full
_version_ 1827387688246837248
author Anastasiia Y. Glagoleva
Tat’jana V. Kukoeva
Elena K. Khlestkina
Elena K. Khlestkina
Olesya Y. Shoeva
author_facet Anastasiia Y. Glagoleva
Tat’jana V. Kukoeva
Elena K. Khlestkina
Elena K. Khlestkina
Olesya Y. Shoeva
author_sort Anastasiia Y. Glagoleva
collection DOAJ
description Polyphenol oxidase (PPO) is an oxidoreductase. In damaged plant tissues, it catalyzes enzymatic browning by oxidizing o-diphenols to highly reactive o-quinones, which polymerize producing heterogeneous dark polymer melanin. In intact tissues, functions of PPO are not well understood. The aim of the study was to investigate the barley PPO gene family and to reveal the possible involvement of Ppo genes in melanization of barley grain, which is controlled by the Blp1 gene. Based on known barley Ppo genes on chromosome 2H (Ppo1 and Ppo2), two additional genes—Ppo3 and Ppo4—were found on chromosomes 3H and 4H, respectively. These genes have one and two exons, respectively, contain a conserved tyrosinase domain and are thought to be functional. Comparative transcriptional analyzes of the genes in samples of developing grains (combined hulls and pericarp tissues) were conducted in two barley lines differing by melanin pigmentation. The genes were found to be transcribed with increasing intensity (while grains mature) independently from the grain color, except for Ppo2, which is transcribed only in black-grained line i:BwBlp1 accumulating melanin in grains. Analysis of this gene’s expression in detached hulls and pericarps showed its elevated transcription in both tissues in comparison with yellow ones, while it was significantly higher in hulls than in pericarp. Segregation analysis in two F2 populations obtained based on barley genotypes carrying dominant Blp1 and recessive ppo1 (I) and dominant Blp1 and recessive ppo1 and ppo2 (II) was carried out. In population I, only two phenotypic classes corresponding to parental black and white ones were observed; the segregation ratio was 3 black to 1 white, corresponding to monogenic. In population II, aside from descendants with black and white grains, hybrids with a gray phenotype — light hulls and dark pericarp — were observed; the segregation ratio was 9 black to 3 gray to 4 white, corresponding to the epistatic interaction of two genes. Most hybrids with the gray phenotype carry dominant Blp1 and a homozygous recessive allele of Ppo2. Based on transcription and segregation assays one may conclude involvement of Ppo2 but not Ppo1 in melanin formation in barley hulls.
first_indexed 2024-03-08T16:06:51Z
format Article
id doaj.art-de01f26c73b244d4a15e21b7eea81574
institution Directory Open Access Journal
issn 1664-462X
language English
last_indexed 2024-03-08T16:06:51Z
publishDate 2024-01-01
publisher Frontiers Media S.A.
record_format Article
series Frontiers in Plant Science
spelling doaj.art-de01f26c73b244d4a15e21b7eea815742024-01-08T05:44:49ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2024-01-011410.3389/fpls.2023.13207701320770Polyphenol oxidase genes in barley (Hordeum vulgare L.): functional activity with respect to black grain pigmentationAnastasiia Y. Glagoleva0Tat’jana V. Kukoeva1Elena K. Khlestkina2Elena K. Khlestkina3Olesya Y. Shoeva4Institute of Cytology and Genetics (ICG), Siberian Branch of Russian Academy of Sciences (SB RAS), Novosibirsk, RussiaInstitute of Cytology and Genetics (ICG), Siberian Branch of Russian Academy of Sciences (SB RAS), Novosibirsk, RussiaInstitute of Cytology and Genetics (ICG), Siberian Branch of Russian Academy of Sciences (SB RAS), Novosibirsk, RussiaN.I. Vavilov All-Russian Research Institute of Plant Genetic Resources (VIR), Saint Petersburg, RussiaInstitute of Cytology and Genetics (ICG), Siberian Branch of Russian Academy of Sciences (SB RAS), Novosibirsk, RussiaPolyphenol oxidase (PPO) is an oxidoreductase. In damaged plant tissues, it catalyzes enzymatic browning by oxidizing o-diphenols to highly reactive o-quinones, which polymerize producing heterogeneous dark polymer melanin. In intact tissues, functions of PPO are not well understood. The aim of the study was to investigate the barley PPO gene family and to reveal the possible involvement of Ppo genes in melanization of barley grain, which is controlled by the Blp1 gene. Based on known barley Ppo genes on chromosome 2H (Ppo1 and Ppo2), two additional genes—Ppo3 and Ppo4—were found on chromosomes 3H and 4H, respectively. These genes have one and two exons, respectively, contain a conserved tyrosinase domain and are thought to be functional. Comparative transcriptional analyzes of the genes in samples of developing grains (combined hulls and pericarp tissues) were conducted in two barley lines differing by melanin pigmentation. The genes were found to be transcribed with increasing intensity (while grains mature) independently from the grain color, except for Ppo2, which is transcribed only in black-grained line i:BwBlp1 accumulating melanin in grains. Analysis of this gene’s expression in detached hulls and pericarps showed its elevated transcription in both tissues in comparison with yellow ones, while it was significantly higher in hulls than in pericarp. Segregation analysis in two F2 populations obtained based on barley genotypes carrying dominant Blp1 and recessive ppo1 (I) and dominant Blp1 and recessive ppo1 and ppo2 (II) was carried out. In population I, only two phenotypic classes corresponding to parental black and white ones were observed; the segregation ratio was 3 black to 1 white, corresponding to monogenic. In population II, aside from descendants with black and white grains, hybrids with a gray phenotype — light hulls and dark pericarp — were observed; the segregation ratio was 9 black to 3 gray to 4 white, corresponding to the epistatic interaction of two genes. Most hybrids with the gray phenotype carry dominant Blp1 and a homozygous recessive allele of Ppo2. Based on transcription and segregation assays one may conclude involvement of Ppo2 but not Ppo1 in melanin formation in barley hulls.https://www.frontiersin.org/articles/10.3389/fpls.2023.1320770/fullBlack lemma and pericarp 1enzymatic browningepistasisgenetic segregationmelanin
spellingShingle Anastasiia Y. Glagoleva
Tat’jana V. Kukoeva
Elena K. Khlestkina
Elena K. Khlestkina
Olesya Y. Shoeva
Polyphenol oxidase genes in barley (Hordeum vulgare L.): functional activity with respect to black grain pigmentation
Frontiers in Plant Science
Black lemma and pericarp 1
enzymatic browning
epistasis
genetic segregation
melanin
title Polyphenol oxidase genes in barley (Hordeum vulgare L.): functional activity with respect to black grain pigmentation
title_full Polyphenol oxidase genes in barley (Hordeum vulgare L.): functional activity with respect to black grain pigmentation
title_fullStr Polyphenol oxidase genes in barley (Hordeum vulgare L.): functional activity with respect to black grain pigmentation
title_full_unstemmed Polyphenol oxidase genes in barley (Hordeum vulgare L.): functional activity with respect to black grain pigmentation
title_short Polyphenol oxidase genes in barley (Hordeum vulgare L.): functional activity with respect to black grain pigmentation
title_sort polyphenol oxidase genes in barley hordeum vulgare l functional activity with respect to black grain pigmentation
topic Black lemma and pericarp 1
enzymatic browning
epistasis
genetic segregation
melanin
url https://www.frontiersin.org/articles/10.3389/fpls.2023.1320770/full
work_keys_str_mv AT anastasiiayglagoleva polyphenoloxidasegenesinbarleyhordeumvulgarelfunctionalactivitywithrespecttoblackgrainpigmentation
AT tatjanavkukoeva polyphenoloxidasegenesinbarleyhordeumvulgarelfunctionalactivitywithrespecttoblackgrainpigmentation
AT elenakkhlestkina polyphenoloxidasegenesinbarleyhordeumvulgarelfunctionalactivitywithrespecttoblackgrainpigmentation
AT elenakkhlestkina polyphenoloxidasegenesinbarleyhordeumvulgarelfunctionalactivitywithrespecttoblackgrainpigmentation
AT olesyayshoeva polyphenoloxidasegenesinbarleyhordeumvulgarelfunctionalactivitywithrespecttoblackgrainpigmentation