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The effect of phytosulfokine alpha on haploid embryogenesis and gene expression of Brassica napus microspore cultures
ID
Mestinšek Mubi, Špela
(
Author
),
ID
Kunej, Urban
(
Author
),
ID
Vogrinčič, Valentin
(
Author
),
ID
Jakše, Jernej
(
Author
),
ID
Murovec, Jana
(
Author
)
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https://www.frontiersin.org/articles/10.3389/fpls.2024.1336519
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Abstract
Microspore embryogenesis (ME) is the most powerful tool for creating homozygous lines in plant breeding and molecular biology research. It is still based mainly on the reprogramming of microspores by temperature, osmotic and/or nutrient stress. New compounds are being sought that could increase the efficiency of microspore embryogenesis or even induce the formation of haploid embryos from recalcitrant genotypes. Among these, the mitogenic factor phytosulfokine alpha (PSK-α) is promising due to its broad spectrum of activity in vivo and in vitro. The aim of our study was to investigate the effect of PSK-α on haploid embryogenesis from microspores of oilseed rape (Brassica napus L., DH4079), one of the most important oil crops and a model plant for studying the molecular mechanisms controlling embryo formation. We tested different concentrations (0, 0.01, 0.1 and 1 µM) of the peptide and evaluated its effect on microspore viability and embryo regeneration after four weeks of culture. Our results showed a positive correlation between addition of PSK-α and cultured microspore viability and a positive effect also on the number of developed embryos. The analysis of transcriptomes across three time points (day 0, 2 and 4) with or without PSK-α supplementation (15 RNA libraries in total) unveiled differentially expressed genes pivotal in cell division, microspore embryogenesis, and subsequent regeneration. PCA grouped transcriptomes by RNA sampling time, with the first two principal components explaining 56.8% variability. On day 2 with PSK, 45 genes (15 up- and 30 down-regulated) were differentially expressed when PSK-α was added and their number increased to 304 by day 4 (30 up- and 274 down-regulated). PSK, PSKR, and PSI gene expression analysis revealed dynamic patterns, with PSK2 displaying the highest increase and overall expression during microspore culture at days 2 and 4. Despite some variations, only PSK1 showed significant differential expression upon PSK-α addition. Of 16 ME-related molecular markers, 3 and 15 exhibited significant differential expression in PSK-supplemented cultures at days 2 and 4, respectively. Embryo-specific markers predominantly expressed after 4 days of culture, with higher expression in medium without PSK, while on day 0, numerous sporophyte-specific markers were highly expressed.
Language:
English
Keywords:
PSK-α
,
PSK
,
haploids
,
doubled haploids
,
transcriptome
,
RNA-seq
,
mitogenic factor
,
differentially expressed genes
Work type:
Article
Typology:
1.01 - Original Scientific Article
Organization:
BF - Biotechnical Faculty
Publication status:
Published
Publication version:
Version of Record
Year:
2024
Number of pages:
13 str.
Numbering:
Vol. 15, art. 1336519
PID:
20.500.12556/RUL-154506
UDC:
577.2
ISSN on article:
1664-462X
DOI:
10.3389/fpls.2024.1336519
COBISS.SI-ID:
183606531
Publication date in RUL:
19.02.2024
Views:
315
Downloads:
42
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Record is a part of a journal
Title:
Frontiers in plant science
Shortened title:
Front. plant sci.
Publisher:
Frontiers Research Foundation
ISSN:
1664-462X
COBISS.SI-ID:
3011663
Licences
License:
CC BY 4.0, Creative Commons Attribution 4.0 International
Link:
http://creativecommons.org/licenses/by/4.0/
Description:
This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
Secondary language
Language:
Slovenian
Keywords:
žlahtnjenje rastlin
,
embriogeneza
,
haploidi
,
oljna ogrščica
,
tkivne kulture
,
kultura mikrospor
Projects
Funder:
ARIS - Slovenian Research and Innovation Agency
Project number:
P4-0077
Name:
Kmetijske rastline - genetika in sodobne tehnologije
Funder:
ARIS - Slovenian Research and Innovation Agency
Project number:
J4-9307
Name:
Preurejanje genomov izbranih vrst rodu Brassica s tehnologijo CRISPR/Cas9
Funder:
ARIS - Slovenian Research and Innovation Agency
Project number:
ARRS-MR-496
Name:
Mladi raziskovalci - ARRS
Funder:
ARIS - Slovenian Research and Innovation Agency
Project number:
IO-0022-0481-001
Name:
Infrastructural centre
Acronym:
IC RRC-AG
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