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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 BMC+Plant+Biol
2025 ; 25
(1
): 1449
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Molecular cloning and the expression profile of two calnexin genes - CNX1 and
CNX2 - during pollen development and pollen tube growth in Petunia
#MMPMID41131475
Suwi?ska A
; Was?g P
; Lenartowska M
; Tyburski J
; Lenartowski R
BMC Plant Biol
2025[Oct]; 25
(1
): 1449
PMID41131475
show ga
BACKGROUND: Calnexin (CNX) is a crucial chaperone of the endoplasmic reticulum
(ER) that participates in the folding and quality control of glycoproteins. In
plants, CNX contributes to multiple physiological processes, including growth,
development, and adaptation to abiotic stresses. Nevertheless, its specific
function in male gametophyte development and pollen tube growth remains poorly
understood. In this work, we report for the first time the molecular cloning of
two Petunia hybrida CNX homologs, PhCNX1 and PhCNX2, examine their expression
profiles during pollen development, germination, and tube elongation, and discuss
their potential functional involvement in these processes. RESULTS: We
successfully cloned and characterized full-length cDNAs of PhCNX1 and PhCNX2,
which encode proteins containing conserved motifs typical of the CNX/CRT family.
Through a combination of qRT-PCR, western blotting, fluorescence in situ
hybridization (FISH), and immunocytochemistry, we show that both genes are
expressed in Petunia anthers, germinating pollen grains, and elongating tubes.
During male gametophyte development, PhCNX1 shows peak expression at the
microspore stage, whereas PhCNX2 reaches its highest transcript levels in mature
pollen grains. Notably, in dry pollen, both genes exhibit a marked decrease in
transcript abundance. FISH analysis indicates that PhCNX mRNAs are detected in
both somatic and germline tissues but are predominantly localized to the
cytoplasm of tapetal cells from the microsporocyte to microspore stages.
Moreover, western blot analysis reveals a progressive increase in CNX protein
levels throughout anther development and its accumulation in dry pollen.
Immunocytochemical staining confirms CNX localization in all anther cell types,
with notable enrichment in tapetal cells and the cytoplasm of pollen grains prior
to anther dehiscence; this is further supported by immunogold labeling indicating
CNX localization within the ER. In germinating pollen and elongating tubes, PhCNX
transcripts accumulate in the cytoplasm near the apertures and along the tube
(except the clear zone), whereas CNX protein is concentrated in the ER-dense
subapical region. CONCLUSION: Our findings reveal that PhCNX1 and PhCNX2 are
dynamically regulated throughout the course of pollen development within the
anther, as well as during pollen germination and tube growth. This spatiotemporal
expression pattern supports the notion that CNX functions as a molecular
chaperone facilitating the high levels of protein synthesis essential for proper
male gametophyte maturation and the polarized growth of the pollen tube.
SUPPLEMENTARY INFORMATION: The online version contains supplementary material
available at 10.1186/s12870-025-07186-2.
free
free
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