Page 44 - 《水产学报》2026年第2期

2 期 水 产 学 报 50 卷

Tab. 6 Comprehensive ranking in females evaluated by RefFinder
recommended comprehensive rank
female
1 2 3 4 5
brain 18S ef-1γ ef-1α β-actin gapdh
optic lobe ef-1α gapdh 18S ef-1γ β-actin
heart ef-1α 18S gapdh ef-1γ β-actin
gill ef-1α ef-1γ gapdh 18S β-actin
branchial heart ef-1γ ef-1α 18S gapdh β-actin
intestine ef-1γ 18S ef-1α β-actin gapdh
stomach ef-1γ ef-1α 18S β-actin gapdh
caecus ef-1α β-actin gapdh 18S ef-1γ
kidney ef-1α ef-1γ 18S gapdh β-actin
liver ef-1γ ef-1α 18S gapdh β-actin
pancreas ef-1α ef-1γ 18S β-actin gapdh
white body ef-1γ 18S ef-1α gapdh β-actin
skin gapdh 18S ef-1α ef-1γ β-actin
ovary 18S ef-1γ ef-1α β-actin gapdh
nidamental gland β-actin ef-1α 18S ef-1γ gapdh
accessory nidamental gland ef-1α ef-1γ gapdh 18S β-actin
all tissues ef-1α ef-1γ 18S gapdh β-actin

male female
number of tissues number of tissues
0 5 10 15 0 4 8 12 16

18S 18S
ef-1α ef-1α
ef-1γ ef-1γ
β-actin β-actin

gapdh gapdh
rank-1 rank-2 rank-3 rank-1 rank-2 rank-3
rank-4 rank-5 rank-4 rank-5
(a) (b)
Fig. 9 The proportion of each gene across the five rankings

were optimal for accurate normalization. In the fell below 0.15 in the white body. In contrast, all four
remaining tissues (brain, intestine, stomach, caecus, V n/n+1 values quantified exceeded 0.15 in the optic
lobe, gill, branchial heart, caecus, kidney, liver, ovary,
kidney, skin, testis, and spermatophore), none of the
and nidamental gland. Likewise, the V values for the
2/3
V n/n+1 (V , V , V , and V ) values were greater
4/5
5/6
2/3
3/4
optic lobe and ovary were in close proximity to 0.15,
than 0.15. However, in these tissues, either the V or
2/3
as was the V value in the branchial heart. Consistent
4/5
V value was notably close to the 0.15 threshold.
3/4
with the male integrated analysis, none of the V n/n+1
Interestingly, the integrated analysis did not yield any
values in the female integrated analysis were found to
V value below 0.15, with the V value represent-
n/n+1 2/3 be below 0.15.
ing the minimum.
2.9 Impact of reference gene stability on qPCR
In females, the V value was less than 0.15 in
2/3
Results
the brain, heart, intestine, stomach, pancreas, skin, and
accessory nidamental gland (Fig. 10-b). The V value For initial validation and to justify the sub-
3/4
https://www.china-fishery.cn 中国水产学会主办 sponsored by China Society of Fisheries
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