Characteristics of main components in Phoebe bournei EOs after fertilization
The yield of EOs from leaves and twigs of P. bournei was 0.102% and 0.135% respectively. Only the yield of the leaf EO was decreased by the compost treatment (Table 1). The results of GC / MS showed that the components are mainly sesquiterpenes in EOs from P. bournei leaves and twigs (Fig. 2-A and B). As shown in Fig. 2-C and D, the dominant component in the leaves is (+)-calarene), and the relative content is more than 20%; the primary component in the twigs is (+) - δ-cadinene, with a relative content of more than 14%. The other main components (relative content > 3%) in EOs of P. bournei are as follows: δ - cadinene (leaf9.21%, twigs 14.54%), copaene (leaf 5.72%, twigs 12.70%), caryophyllene (leaf 3.09%, twigs 10.11%) and α- caryophyllene (leaf 3.46%, twigs 3.52%). (−) – Allo - aromadendrene (6.54%) was only detected in leaves, while β - eudesmol (6.20%), γ - maaliene (6.37%) and α - bergamotene (6.73%) were merely detected in twigs. After fertilization, the categories of components in P. bournei EOs were not changed significantly, but the content of some main components was changed. After the application of the compost, the content of (+) - calarene in the leaf EO was increased significantly, while the content of other main components was not changed significantly (Fig. 2-C); the contents of α - bergamotene and γ - maaliene in the twig EO were decreased significantly, and other main components were not significantly changed (Fig. 2-D). After the application of compound fertilizer the contents of (+) - calarene, (+) - δ - cadinene, (−) - allo aromadendrene and α- caryophyllene in the leaf EO were significantly decreased, only germacrene B was significantly increased (Fig. 2-C); the content of copaene in the twig EO was significantly increased, while the contents of β - eudesmol, γ - maaliene and α - bergamotene were significantly decreased (Fig. 2-D). In general, compost increased the content of the primary component in the leaf EO, but had no significant effects on other components, while compound fertilizer decreased the content of most main components in leaf and twig EOs.
Table 1 The yield of EOs from leaves and twigs of Phoebe bournei
Samples
|
Yield (%)
|
Compost
|
Compound fertilizer
|
Control
|
Leaves
|
0.089494±0.00313b
|
0.095891±0.01457a
|
0.112113±0.00548a
|
Twigs
|
0.128376±0.00629a
|
0.147014±0.03066a
|
0.134758±0.01255a
|
Note: 1) LCK --- leaf control group; LOF --- leaves treated with compost; LCF --- leaves treated with compound fertilizer; TCK --- twig control group; TOF --- twigs treated with compost; TCF --- twigs treated with compound fertilizer, the same below.
2) The letters a, b, c represents the significance of the difference.
Establishment and annotation of Phoebe bournei transcriptomes
The functional annotation of unigenes was conducted using BLASTx against public datebases (NR, Swiss-Prot, Pram, COG, GO and KEGG datebases). A total of 226819 unigenes were annotated from the public datebases. Different numbers of unigenes from all the libraries were annotated against NR, Swiss-Prot, Pram, COG, GO and KEGG database, respectively (Table 2). The results indicated an extensive coverage of Phoebe bournei transcriptomes. BLASTX results against NR database showed that the annotated unigenes (26615, 30.65%) in Phoebe bournei is largely similar to Quercus suber assembled unigenes (Fig. 3). The correlation coefficients of the unigenes expression in all the sample were between 0.78 ~ 1, indicating that the biological repeatability of this experiment was good, and sequencing data were accurate and reasonable (Fig. 4).
Table 2 The number of unigenes annotated against six databases
|
Total
|
NR
|
Swiss-Prot
|
Pfam
|
COG
|
GO
|
KEGG
|
Unigene number
|
226819
|
87595
|
64362
|
62401
|
10938
|
54370
|
42768
|
Percentage
|
100%
|
38.62%
|
28.38%
|
27.51%
|
4.82%
|
23.97%
|
18.86%
|
In comparison with the control many DEGs (different expression genes) were annotated in twigs and leaves after the treatment of compost and compound fertilizer. The total number of DEGs in twigs and leaves treated with the compost were significantly larger than that with the compound fertilizer. The number of DEGs in leaves was noteworthy more than that in twigs treated with the compost. However, the number of DEGs in twigs was significantly higher than that in leaves treated with the compound fertilizer. After the two fertilization treatments, there were many common DEGs in twigs and leaves, but the number of common DEGs in twigs was less than that in leaves (Fig. 5).
Among the DEGs the number of genes significantly up-regulated or down-regulated was as shown in Fig. 6 (p-ajust < 0.05). The number of up-regulated genes and down-regulated genes in twigs and leaves treated with the compost was more than those in twigs and leaves treated with compound fertilizer. The number of up-regulated and down-regulated genes in leaves was significantly higher than that in twigs treated with the compost. After adding the compound fertilizer the number of up-regulated genes was significantly higher than that of down-regulated genes in twigs; the number of up-regulated genes was lower than that of down-regulated genes in leaves; the number of up-regulated genes in twigs was more than that in leaves, and the number of the down-regulated genes in twigs was lower than that in leaves.
Gene ontology annotation and KEGG pathway analysis of Phoebe bournei
Phoebe bournei GO and KEGG classification
Knowledge of the DEGs function of P. bournei would be important for us to understand the changes in yields and components of P. bournei EOs. All unigenes were annotated and classified by web gene ontology annotation plot (WEGO). WEGO annotation results showed that all the assembled unigenes were classified into 54 functional categories, as shown in Fig. 7. Among all the GO functional categories the dominant categories were 'binding' and 'catalytic activity' (> 56%), followed by 'cellular process' and' metadata process'. In addition, "cell", "cellular part" and "membrane" also accounted for a high proportion. Out of the total identified unigenes 21713 unigenes (39.94%) were categorized as "metabolic process", among which 1343 unigenes were involved in “secondary metabolism process”.
The functional biological pathway of P. bournei was indenetified by 34368 unigenes mapped into the canonical pathways reference in KEGG using KOBAS (Fig. 8). Among them 674 unigenes were assigned as "terpenoid and polyketide metabolism", and 5459 genes were classified as "carbohydrate metabolism". The pathway hierarchical classification of the unigenes is shown in Fig. 8-B and Fig. 8-C. It can be seen from the above that the main components of P. bournei EOs are sesquiterpenes. Therefore, 185 unigene in terpenoid backbone biosynthesis (pathway ID: map00900) pathway, 36 unigene in sesquiterpenoid and triterpenoid biosynthesis (pathway ID: map00909) pathway and 11 unigenes in monoterpenoid biosynthesis (pathway ID: map00902) pathway would play an important role in the synthesis of P. bournei EOs.
Go functional annotation of DEGs
GO annotation was carried out on DEGs in twigs and leaves of P. bournei after fertilization. Significant differences were shown in the number of annotated DEGs in GO level 1 classification: cellular component (CC), molecular function (MF) and biological process (BP) (Table 3). DEGs in twigs were mainly involved in CC and MF after fertilization, while DEGs in leaves were assigned to CC, MF and BP, with the percentage of >50%.
According to GO level 2 classification of the DEGs (Fig. 9) the number of up-regulated and downregulated genes (p-ajust < 0.05) involved in the processes of 'binding', 'catalytic activity', 'cellular process', 'metallic process', 'cell', 'cellular part' and 'membrane' in twigs and leaves of P. bournei after the compost treatment were significantly higher than that after the compound fertilizer treatment. In other words, the effect of compost on the leaves was stronger than that on the twigs. For P. bournei leaves the number of up-regulated genes was more than that of down regulated genes after the compost treatment, while the number of down-regulated genes was more than that of up-regulated genes after the compound fertilizer treatment; for P. bournei twigs, the number of down-regulated genes involved in the processes of 'cellular process' and' molecular process' was more than that of up-regulated genes; the number of down regulated genes involved in 'cellular activity' and 'cellular part' and 'membrane' was more than that of up-regulated genes after the compost treatment; no differences were displayed in the number of up-regulated and down-regulated genes involved in ‘binding’ and ‘cell’. The number of down-regulated genes involved in the processes was smaller than that of up-regulated genes after the compound fertilizer. Number of DEGs participating in metabolic process was listed as follows: LCF_ Down (108), LCF_ Up (33), LOF_ Down (746), LOF_ Up (691), TCF_ Down (86), TCF_ Up (118), TOF-Down (271), TOF-Up (242).
Table 3 GO level 1 classification of DEGs in leaves and twigs of Phoebe bournei after fertilization
Groups
|
CC
|
BP
|
MF
|
DEGs number
|
Percentage (%)
|
DEGs number
|
Percentage (%)
|
DEGs number
|
Percentage (%)
|
NF_twig_vs_CF_twig_G
|
2093
|
66.78
|
1546
|
17.73
|
1589
|
50.7
|
NF_twig_vs_OF_twig_G
|
4022
|
26.85
|
3157
|
10.27
|
3393
|
41.82
|
NF_leaf_vs_CF_leaf_G
|
2064
|
74.27
|
1660
|
59.73
|
1451
|
52.21
|
NF_leaf_vs_OF_leaf_G
|
10296
|
62.57
|
8673
|
52.71
|
8107
|
49.27
|
Identifition of DEGs in KEGG pathways
The main components of EOs from P. bournei are sesquiterpenes and few monoterpenes (Fig. 2). The metabolic pathways related to their synthesis are terpenoid backbone biosynthesis (TBB), glycolysis / gluconeogenesis (GG), pentose phosphate pathway (PPP), sesquiterpenoid and triterpenoid biosynthesis (STB) and monoterpenoid biosynthesis (MB). Among them TBB can be regarded as the core metabolic pathway (Fig. 10). Fertilization has different effects on these metabolic pathways (Table 4). In the TBB pathway, 18 and 4 DEGs were identified in the leaves and twigs by the compost treatment, and 4 DEGs in the leaves and twigs after the compound fertilizer application. In the GG pathway, 96 and 26 DEGs were obtained in the leaves and twigs treated with the compost, and 17 and 23 DEGs in the leaves and twigs after the compound fertilizer application. In the PPP pathway, 55 and 7 DEGs were found in the leaves and twigs after the compost treatment, and 12 and 5 DEGs in the leaves and twigs by the compound fertilizer treatment. In these three metabolic pathways the number of DEGs in leaves and twigs by the compost treatment was significantly higher than that of the compound fertilizer treatment. In STB pathway only one DEG was obtained in the leaves and twigs by the compost treatment. In the MB pathway one and two DEGs were also identified in the leaves and twigs treated only with the compost. In general, the percentage of DEGs in twigs and leaves treated with the compost was relatively high in the five pathways (Table 4).
Table 4 The number of DEGs involved in sesquiterpenes biosynthesis pathway after fertilization.
Groups
|
Pathways
|
TBB
|
STB
|
MB
|
GG
|
PPP
|
DEGs
|
percentage/%
|
DEGs
|
percentage/%
|
DEGs
|
percentage/%
|
DEGs
|
percentage/%
|
DEGs
|
percentage/%
|
NF_leaf_vs_OF_leaf
|
18
|
9.73
|
1
|
2.78
|
1
|
9.09
|
96
|
6.34
|
55
|
9.89
|
NF_leaf_vs_CF_leaf
|
4
|
2.16
|
0
|
0.00
|
0
|
0.00
|
17
|
0.11
|
12
|
2.16
|
NF_twig_vs_OF_twig
|
4
|
2.16
|
1
|
2.78
|
2
|
18.18
|
26
|
0.09
|
7
|
1.26
|
NF_twig_vs_CF_twig
|
4
|
2.16
|
0
|
0.00
|
0
|
0.00
|
23
|
0.05
|
5
|
0.90
|
The number of significantly up- or down-regulated unigenes was different in the five pathways (Table 5). After the compost treatment up-regulated and down-regulated unigenes in GG and PP pathway were found, and the number of down-regulated unigenes was more than that of up-regulated unigenes. In TBB pathway the number of up-regulated unigenes was the same as that of down-regulated unigenes. A up-regulated unigene and a down-regulated unigene were identified in STB and MB pathways, respectively. The number of down-regulated genes was higher than that of up-regulated genes in leaves treated with the compound fertilizer. The number of up- or down- regulated unigenes in the leaves treated with the compound fertilizer was significantly smaller than that in the leaves with the compost.
For the twigs treated with the compost, only 3 and 2 up-regulated unigenes were enriched in TBB and MB pathways, and only one down-regulated unigene was identified in STB pathway. The number of down-regulated unigenes in GG and PPP pathways was larger than that of up-regulated unigenes. The number of up-regulated and down-regulated unigenes in most of the pathways in the twigs treated with the compost was significantly smaller than that in the leaves with the compost. For the twigs treated with the compound fertilizer, the up-regulated and down-regulated unigenes were involved in TBB, GG and PPP. In comparison with the compost, the effects of the compound fertilizer on P. bournei twigs were relatively weak.
In summary, the effect of the compost on P. bournei was much stronger than that of the compound fertilizer, especially on P. bournei leaves. The significantly up/down regulated unigenes might have contributed to the differences in sesquiterpene metabolism before and after the fertilization treatments.
Table 5 The number of up / down regulated unigenes in the five pathways (q-value < 0.05).
|
Groups
|
Pathways
|
TBB
|
STB
|
MB
|
GG
|
PPP
|
Up/Down
|
percentage/%
|
Up/Down
|
percentage/%
|
Up/Down
|
percentage/%
|
Up/Down
|
percentage/%
|
Up/Down
|
percentage/%
|
NF_leaf_vs_OF_leaf
|
5/5
|
27.78/27.78
|
1/0
|
5.56/0
|
0/1
|
0/5.56
|
41/48
|
42.71/50
|
20/26
|
36.36/47.27
|
NF_leaf_vs_CF_leaf
|
0/2
|
0/50
|
-
|
-
|
-
|
-
|
2/4
|
11.76/23.53
|
1/7
|
8.33/58.33
|
NF_twig_vs_OF_twig
|
3/0
|
75/0
|
0/1
|
0/25
|
2/0
|
50/0
|
10/15
|
38.46/57.69
|
2/5
|
28.57/71.43
|
NF_twig_vs_CF_twig
|
1/0
|
25/0
|
-
|
-
|
-
|
-
|
17/3
|
73.91/13.04
|
2/2
|
40/40
|
Identification of genes involved in biosynthesis of sesquiterpenoids.
TBB contains two metabolic pathways: mevalonate (MVA) pathway and 2C-methyl-D-erythritol 4-phosphate (MEP) pathway (Fig.10). MVA pathway is mainly responsible for the biosynthesis of sesquiterpenes and triterpenes, while MEP pathway is mainly responsible for the biosynthesis of monoterpenes [21]. The main components of P. bournei EOs are squiterpenes, and therefore, TBB is the core pathway of EO metabolite synthesis. 3-hydroxy-3-methyl-glutaryl-CoA (HMG CoA) and 1-deoxy-d-xylulose-5phosphate (DXP) are two key substrates for the synthesis of sesquiterpenes and monoterpenes. DXP is produced by 1-deoxy-D-xylulose-5-phosphate synthase (DXS) catalyzing glycraldehyde 3-phosphate (G3p) and pyruvate. DXS has been proved to be an important rate-limiting enzyme in MEP pathway [21,36]. DXS should be an important regulatory site for the sesquiterpenes biosynthesis. This indicated that DXS would play an important role in the anabolism pathway of P. bournei EO. The expression of genes regulating DXS in different branches varied with development, tissue type and environmental conditions. After the treatment of the compost significant changes are found in the expression of DXS genes in leaves and twigs. Three unigenes about DXS in the leaves were changed: one up-regulated unigene (TRINITY_DN56563_c0_g1) and two down-regulated unigenes (TRINITY_DN73974_c0_g2, TRINITY_DN60255_c0_g7); the unigene (TRINITY_DN70707_c0_g6) in the twigs may be a DXS gene. However, one down-regulated DXS unigene (TRINITY_DN73974_c0_g2) was found only in leaves after the compound fertilizer treatment. If an enzyme is regulated by two or more genes, these unigenes may belong to a multigene family or part of a larger gene [37]. The expression of DXS genes is usually divided into three categories (DXS1, DXS2, DXS3), while most belongs to DXS2 regulating plant secondary metabolite biosynthesis [36,38]. According to Swissprot Description (Table 6), only TRINITY_DN60255_c0_g7 is the unigene regulating DXS2, which is closely related to the metabolism of P. bournei EO. The expression of the unigene was not significantly changed in all twigs, but obviously decreased in leaves treated with the compost (Fig. 11). One unigene (TRINITY_DN73974_c0_g2), not being DXS2 gene, was enriched only in leaves after the compound fertilizer treatment.
A key step in MVA pathway is HMG-CoA synthesis by Hydroxymethylglutaryl-CoA synthase (HMGS) catalyzing Acetyl-CoA and Acetoacetyl-CoA [39]. HMGs is the first committed enzyme in the MVA pathway. The change of HMGS expression level has an important impact on the production of secondary metabolites [40]. The expression of two unigenes (TRINITY_DN79706_c2_g1, TRINITY_DN84575_c6_g2) enriched in P. bournei regulating HMGS was both up-regulated in twigs and leaves after the compost treatment and twigs after the compound fertilizer treatment (Fig.11 and Table 6). Here, the expression of TRINITY_DN79706_c2_g1 in leaves was similar to that in twigs after the compost treatment (Fig. 11).
Another important step in MVA pathway is mevalonate synthesis by 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR) catalyzing HMG-CoA. HMGR is also an important rate-limiting enzyme in MVA pathway, and the overexpression of HMGR can promote the synthesis of sesquiterpenes in plants [41]. HMGR is usually divided into HMGR1, HMGR2 and HMGR3 [42]. The overexpression of HMGR1 and HMGR3 can also have a positive effect on the synthesis of secondary metabolites [42-43]. The expression level of HMGR1 gene (TRINITY_DN83281_c2_g2) enriched in all the twigs was not significantly changed, but clearly decreased in the leaves by the compost treatment. This indicated that the compost treatment would reduce the EO yield of P. bournei leaves. Similarly, the expression of HMGR3 gene (TRINITY_DN76855_c1_g3) was clearly decreased in leaves treated with the organic and compound fertilizer, except in all the twigs (Fig.11). Both the two fertilization treatments, especially the compost treatment, to some extent, had a negative effect on the yield and the constituent of P. bournei EOs.
Farnesyl pyrophosphate synthase (FPPS) is an important branch-point enzyme, whose activity can change the direction of isoprene metabolism in the TBB pathway and plays an important role in isoprene metabolism [44-46]. The expression of FPPS gene (TRINITY_DN61719_c0_g1) was down-regulated in P. bournei leaves and twigs by the fertilization treatment, but the down-regulation of the expression was the most significant in the leaves by the compost treatment (Fig.11). The change in the expression of FPPS gene could be a main reason that the EOs of P. bournei twigs and leaves were varied in the main components. Geranylgeranyl diphosphate synthase (GGPPS) is also an important branch enzyme in terpenoid biosynthesis [40]. Contrary to FPPS, the expression of GGPPS gene (TRINITY_DN67962_c1_g3) was up-regulated in all the twigs and leaves, particularly in the leaves by the compost treatment. Therefore, changes in the expression of FPPS and GGPPS genes may be an important factor for the change of species or content of the components in P. bournei EOs.
The terminal products geranyl diphosphate (GPP) and farnesyl diphosphate (FPP), produced by Acetyl-CoA, Acetoacetyl-CoA, Pyruvate and D-Glyceraldehyde-3P going through MVA and MEP processes, are further catalysed by terpene syntheses (TPS) to form sesquiterpenes and monoterpenes (Fig. 10). Here, the expression of four unigenes (TRINITY_DN64565_c0_g4, TRINITY_DN83483_c1_g2, TRINITY_DN64925_c1_g1 and TRINITY_DN71731_c0_g2) about TPS was significantly changed in P. bournei. However, these sesquiterpenes (Germacrene D, (E, E)-farnesyl-P, terpineol and (+)-Neomenthol) produced by the regulation of the four genes were not the main components of P. bournei EOs.
In addition to TBB pathway, and the synthesis of volatile components and the yield in P. bournei EOs is also related to GG and PPP. The first four substrates (Acetyl-CoA, Acetoacetyl-CoA, Pyruvate and D-Glyceraldehyde-3P) in the TBB process are produced by the metabolism of GG and PPP (Fig. 10). After fertilization the expression of many genes regulating the two pathways was also affected to a great extent. From the above KEGG enrichment results, except for the twigs treated with the compound fertilizer, the number of down-regulated genes was higher than that of the up-regulated genes in other samples. These genes to some extent indirectly affected the yield and composition of P. bournei EOs.
Table 6 The information of DEGs in map00900, map00909 and map00902
|
Gene ID
|
Swissprot Description
|
Log2FC
|
Pvalue
|
Pajust
|
Up/ Down
|
Group
|
TRINITY_DN79706_c2_g1
|
Hydroxymethylglutaryl-CoA synthase
|
1.39516492
|
7.82E-03
|
4.90E-02
|
up
|
NF_leaf_vs_OF_leaf
|
1.707138136
|
3.68E-04
|
7.63E-03
|
up
|
NF_twig_vs_OF_twig
|
TRINITY_DN84575_c6_g2
|
Hydroxymethylglutaryl-CoA synthase
|
2.150896458
|
1.68E-03
|
2.41E-02
|
up
|
NF_twig_vs_OF_twig
|
2.493661561
|
2.26E-04
|
1.19E-02
|
up
|
NF_twig_vs_CF_twig
|
TRINITY_DN56563_c0_g1
|
1-deoxy-D-xylulose-5-phosphate synthase 1 (DXS1)
|
1.06898745
|
4.73E-04
|
5.48E-03
|
up
|
NF_leaf_vs_OF_leaf
|
TRINITY_DN73974_c0_g2
|
1-deoxy-D-xylulose-5-phosphate synthase 1 (DXS1)
|
-5.5884809
|
7.98E-05
|
1.26E-03
|
down
|
NF_leaf_vs_CF_leaf
|
-4.21307726
|
5.45E-06
|
1.31E-03
|
down
|
NF_leaf_vs_OF_leaf
|
TRINITY_DN60255_c0_g7
|
1-deoxy-D-xylulose 5-phosphate synthase 2 (DXS2)
|
-1.557267
|
2.84E-03
|
2.25E-02
|
down
|
NF_leaf_vs_OF_leaf
|
TRINITY_DN70707_c0_g6
|
Probable 1-deoxy-D-xylulose-5-phosphate synthase, chloroplastic
|
1.923531945
|
8.85E-07
|
5.23E-05
|
up
|
NF_twig_vs_OF_twig
|
TRINITY_DN83281_c2_g2
|
3-hydroxy-3-methylglutaryl-coenzyme A reductase 1 (HMGR1)
|
-1.92527724
|
9.50E-07
|
2.87E-05
|
down
|
NF_leaf_vs_OF_leaf
|
TRINITY_DN76855_c1_g3
|
3-hydroxy-3-methylglutaryl-coenzyme A reductase 3 (HMGR3)
|
-1.57011729
|
9.19E-06
|
2.04E-04
|
down
|
NF_leaf_vs_OF_leaf
|
-1.34758953
|
1.21E-03
|
4.35E-02
|
down
|
NF_leaf_vs_CF_leaf
|
TRINITY_DN61719_c0_g1
|
Farnesyl pyrophosphate synthase 2 (FPPS2)
|
-1.57943298
|
6.17E-05
|
1.02E-03
|
down
|
NF_leaf_vs_OF_leaf
|
TRINITY_DN64139_c1_g1
|
Probable phytol kinase 3
|
1.818575961
|
8.01E-08
|
3.31E-06
|
up
|
NF_leaf_vs_OF_leaf
|
TRINITY_DN75454_c1_g3
|
3 beta-hydroxysteroid dehydrogenase/Delta 5-->4-isomerase
|
1.019357385
|
2.79E-03
|
2.22E-02
|
up
|
NF_leaf_vs_OF_leaf
|
TRINITY_DN67962_c1_g3
|
Geranylgeranyl pyrophosphate synthase, chloroplastic (GGPPS)
|
1.132667099
|
1.63E-03
|
1.47E-02
|
up
|
NF_leaf_vs_OF_leaf
|
TRINITY_DN64565_c0_g4
|
TPSGD_VITVI/ Terpene synthase family (Pfam)
|
-2.39226398
|
3.13E-03
|
3.79E-02
|
down
|
NF_twig_vs_OF_twig
|
TRINITY_DN83483_c1_g2
|
Geraniol synthase, chloroplastic
|
1.966248415
|
4.55E-04
|
8.97E-03
|
up
|
NF_twig_vs_OF_twig
|
TRINITY_DN64925_c1_g1
|
Alpha-terpineol synthase, chloroplastic
|
-6.04327435
|
6.47E-06
|
1.52E-04
|
down
|
NF_leaf_vs_OF_leaf
|
TRINITY_DN71731_c0_g2
|
Salutaridine reductase
|
1.419760693
|
1.80E-04
|
4.35E-03
|
up
|
NF_twig_vs_OF_twig
|