Table 1.
Experimental treatment reagent formula.
| Solution | Control/mL | 0.1 mmol · L−1 Na2SeO3+ 15 mmol · L−1, NO3–, (0.1Se + 15N)/mL | 0.2 mmol · L−1 Na2SeO3 + 15 mmol · L−1, NO3– (0.2Se + 15N)/mL | 0.4 mmol · L−1 Na2SeO3 + 15 mmol · L−1, NO3–, (0.4Se + 15N)/mL | 0.1 mmol · L−1 Na2SO3 (0.1Se)/mL | 0.2 mmol · L−1 Na2SeO3 (0.2Se)/mL | 0.4 mmol · L−1 Na2SeO3 (0.4Se)/mL |
|---|---|---|---|---|---|---|---|
| 1.0 mol/L Ca(NO3)2 | 0 | 5 | 5 | 5 | 0 | 0 | 0 |
| 1.0 mol/L KNO3 | 0 | 5 | 5 | 5 | 0 | 0 | 0 |
| 0.5 mol/L K2SO4 | 5 | 0 | 0 | 0 | 5 | 5 | 5 |
| 1.0 mol/L CaCl2 | 5 | 0 | 0 | 0 | 5 | 5 | 5 |
| 1.0 mol/L MgSO4 | 2 | 2 | 2 | 2 | 2 | 2 | 2 |
| 1.0 mol/L KH2PO4 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
| 20.0 nimol/L FeSO4 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
| 20.0 nimol/L MnSO4 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
| 10.0 nimol/L ZnSO4 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
| 1.0 nimol/L Na2MoO4 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
| 0.1 mol/L H3BO3 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
| 0.01 mol/L CuSO4 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
| 0.01 mol/L Na2SeO3 | 0 | 10 | 20 | 30 | 10 | 20 | 30 |
| Deionised water | 981 | 971 | 961 | 951 | 971 | 961 | 951 |

Figure 1.
Effects of Na2SeO3 treatment on the growth and physiological characteristics of grape under various nitrogen conditions. (A) The growth status of grape plants; (B) the net growth of plant height, the net growth of stem thickness, root volume and internode length of grape; (C) leaf thickness, biomass, root shoot ratio and flavonoids content of grape; (D) starch content, soluble sugar content, nitrogen content and soluble protein content of grape. The results were shown as mean ± standard deviation (n = 4), and one-way ANOVA was used to compare the significant differences among the treatments. Different lowercase letters on the bar graph indicate significant differences at the p < 0.05 level. Control group is Control; 0.1 mmol · L−1 + 15 mmol · L−1 N is 0.1Se + 15 N; 0.2 mmol · L−1 + 15 mmol · L−1 N is 0.2Se + 15 N; 0.4 mmol · L−1 + 15 mmol · L−1 N is 0.4Se + 15 N; 0.1 mmol · L−1 Na2SeO3 is 0.1Se; 0.2 mmol · L−1 Na2SeO3 is 0.2Se; 0.4 mmol · L−1 Na2SeO3 is 0.4Se.
Table 2.
Grape transcriptome sequencing quality data treated with Na2SeO3 and nitrogen.
| Sample | Raw reads | Clean reads | Clean bases | Error rate (%) | Q20 (%) | Q30 (%) | GC content (%) |
|---|---|---|---|---|---|---|---|
| Control-1 | 44342452 | 43068728 | 6392908153 | 0.0252 | 97.92 | 93.97 | 46.14 |
| Control-2 | 51674990 | 50251708 | 7420078791 | 0.0255 | 97.81 | 93.73 | 46.66 |
| Control-3 | 58656668 | 56709132 | 8314120252 | 0.0251 | 97.96 | 94.14 | 46.80 |
| 0.2Se + 15N-1 | 61956434 | 60368950 | 8886922304 | 0.0252 | 97.94 | 94.10 | 46.57 |
| 0.2Se + 15N-2 | 53393402 | 51882436 | 7638583022 | 0.0256 | 97.77 | 93.68 | 46.64 |
| 0.2Se + 15N-3 | 51910586 | 50527188 | 7411722373 | 0.0249 | 98.04 | 94.32 | 46.82 |
| 0.2Se-1 | 43739408 | 42003504 | 6241771742 | 0.0258 | 97.69 | 93.44 | 46.06 |
| 0.2Se-2 | 45886300 | 44202816 | 6568402293 | 0.0256 | 97.80 | 93.66 | 45.69 |
| 0.2Se-3 | 46115382 | 44372272 | 6564671570 | 0.0257 | 97.73 | 93.54 | 45.94 |
1 Clean reads: The total number of entries in the sequencing data after quality control; Clean bases: Total amount of sequencing data after quality control; Error rate (%): The average error rate of sequencing bases corresponding to quality control data; Q20 and Q30 (%): respectively refer to the percentage of bases with sequencing quality above 99% and 99.9% in total bases; GC content (%): The percentage of the total number of G and C bases corresponding to the quality control data to the total number of bases.
Table 3.
Assembly quality for transcript and unigene of grape.
| Type | Unigene | Transcript |
|---|---|---|
| Total number | 89,421 | 147,924 |
| N50 length (bp) | 1954 | 2268 |
| BUSCO | C: 73.5%[S: 70.2%; D: 3.3%] | C: 89.7%[S: 64.2%; D: 25.5%] |
1 Total number, the number of sequence entries of the assembled unigene/transcript. N50 length, sort the assembled unigene/transcript in descending order of length, and accumulate the length of the transcript to half of the total length, corresponding to the length of the transcript. BUSCO, using BUSCO to evaluate assembly integrity score, the higher the score, the better the complete (C). Complete (C) represents the proportion of sequences that reach the desired length in the assembled sequence compared to the total BUSCO sequence, consisting of two parts. S represents a sequence that can align with one gene in the database, and D represents a sequence that can align with multiple genes in the database.
Table 4.
Grape comparison and statistics of sequencing data and assembly results treated with Na2SeO3 and nitrogen.
| Sample | Clean reads | Mapped reads | Mapped ratio (%) |
|---|---|---|---|
| Control-1 | 43068728 | 38170787 | 88.63 |
| Control-2 | 50251708 | 44199319 | 87.96 |
| Control-3 | 56709132 | 50304056 | 88.71 |
| 0.2Se + 15N-1 | 60368950 | 53347721 | 88.74 |
| 0.2Se + 15N-2 | 51882436 | 45851321 | 88.27 |
| 0.2Se + 15N-3 | 50527188 | 44810011 | 88.34 |
| 0.2Se-1 | 42003504 | 37275528 | 88.37 |
| 0.2Se-2 | 44202816 | 39017275 | 88.38 |
| 0.2Se-3 | 44372272 | 39199620 | 88.68 |

Figure 2.
Transcriptome analysis of grape leaves after Na2SeO3 treatment under various nitrogen conditions. (A) Correlation between samples; (B) unigene distribution and group overlap; (C) Venn diagram displaying the number of DEGs in each comparison group and group overlap; (D–F) The DEGs in 0.2Se + 15N versus Control, 0.2Se versus Control, and 0.2Se + 15N vs 0.2Se are plotted in a volcano plot. Control group is Control; 0.1 mmol · L−1 + 15 mmol · L−1 N is 0.1Se + 15 N; 0.2 mmol · L−1 + 15 mmol · L−1 N is 0.2Se + 15 N; 0.4 mmol · L−1 + 15 mmol · L−1 N is 0.4Se + 15 N; 0.1 mmol · L−1 Na2SeO3 is 0.1Se; 0.2 mmol · L−1 Na2SeO3 is 0.2Se; 0.4 mmol · L−1 Na2SeO3 is 0.4Se.

Figure 3.
GO pathway enrichment analysis of DEGs in grape after Na2SeO3 treatment under various nitrogen conditions. The numbers listed on the horizontal axis represent the top 40 GO-enriched entries in different comparison groups (Padjust < 0.05). Control group is Control; 0.2 mmol · L−1+ 15 mmol · L−1 N is 0.2 Se + 15 N; 0.2 mmol · L−1 Na2SeO3 is 0.2 Se. 1. Catalytic activity; 2. oxidoreductase activity; 3. DNA binding; 4. transferase activity, transferring acyl groups; 5. transferase activity, transferring acyl groups other than amino-acyl groups; 6. DNA-binding transcription factor activity; 7. tetrapyrrole binding; 8. heme binding; 9. lyase activity; 10. cytoskeletal protein binding; 11. extracellular region; 12. cell wall; 13. external encapsulating structure; 14. supramolecular complex; 15. plasmodesma; 16. anchoring junction; 17. cell-cell junction; 18. cell junction; 19. polymeric cytoskeletal fibre; 20. supramolecular polymer; 21. response to stimulus; 22. oxidation-reduction process; 23. response to stress; 24. transmembrane transport; 25. response to chemical; 26. cell cycle process; 27. response to abiotic stimulus; 28. lipid biosynthetic process; 29. small molecule biosynthetic process; 30. microtubule-based process; 31. response to oxygen-containing compound; 32. response to external stimulus; 33. ion transmembrane transport; 34. response to oxidative stress; 35. mitotic cell cycle process; 36. organic acid biosynthetic process; 37. carboxylic acid biosynthetic process; 38. regulation of cell cycle; 39. inorganic ion transmembrane transport; 40. cell wall organisation or biogenesis; 41. transferase activity, transferring glycosyl groups; 42. transferase activity, transferring hexosyl groups; 43. hydrolase activity, acting on glycosyl bonds; 44. UDP-glycosyltransferase activity; 45. oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen; 46. hydrolase activity, hydrolysing O-glycosyl compounds; 47. monooxygenase activity; 48. iron ion binding; 49. intrinsic component of membrane; 50. integral component of membrane; 51. membrane; 52. plasma membrane; 53. Golgi apparatus; 54. carbohydrate metabolic process; 55. catabolic process; 56. protein phosphorylation; 57. polysaccharide metabolic process; 58. cellular carbohydrate metabolic process; 59. cellular polysaccharide metabolic process; 60. external encapsulating structure organisation; 61. cell wall organisation; 62. apoplast; 63. organic substance catabolic process; 64. ion transport; 65. regulation of biological quality; 66. homeostatic process; 67. cation transport; 68. metal ion transport; 69. response to inorganic substance; 70. inorganic ion homeostasis.

Figure 4.
KEGG pathway enrichment analysis of DEGs in grape after Na2SeO3 treatment under various nitrogen conditions. The numbers listed on the horizontal axis represent the top 15 KEGG enrichment items in different treatment comparisons (Padjust < 0.05). Control group is Control; 0.2 mmol · L−1 + 15 mmol · L−1 N is 0.2 Se + 15 N; 0.2 mmol · L−1 Na2SeO3 is 0.2 Se. 1. Flavonoid biosynthesis; 2. phenylpropanoid biosynthesis; 3. starch and sucrose metabolism; 4. stilbenoid, diarylheptanoid, and gingerol biosynthesis; 5. glycine, serine, and threonine metabolism; 6. glycerolipid metabolism; 7. cysteine and methionine metabolism; 8. glutathione metabolism; 9. pentose and glucuronate interconversions; 10. monoterpenoid biosynthesis; 11. glycolysis/gluconeogenesis; 12. plant hormone signal transduction; 13. MAPK signalling pathway–plant; 14. circadian rhythm–plant; 15. plant-pathogen interaction; 16. glycerophospholipid metabolism; 17. amino sugar and nucleotide sugar metabolism; 18. tyrosine metabolism; 19. phagosome; 20. endocytosis; 21. protein processing in endoplasmic reticulum; 22. galactose metabolism; 23. oxidative phosphorylation.

Figure 5.
Key DEGs involved in flavonoid and phenylpropane biosynthesis in grape after Na2SeO3 treatment under various nitrogen conditions. CYP73A: nodulation receptor kinase; CYP75A: flavonoid 3′,5′-hydroxylase; CYP75B1: flavonoid 3′-monooxygenase; CHS/ST: stilbene synthase; PGT1: UDP-glycosyltransferase; E2.3.1.133: stemmadenine O-acetyltransferase; E2.1.1.104: probable caffeoyl-CoA O-methyltransferase; DFR: dihydroflavonol 4-reductase; F3H: naringenin,2-oxoglutarate 3-dioxygenase; LAR: leucoanthocyanidin reductase; ANS: leucoanthocyanidin dioxygenase; E3.2.1.21: beta-glucosidase; E1.11.1.7: lignin-forming anionic peroxidase; CYP73A: trans-cinnamate 4-monooxygenase; CAD: probable mannitol dehydrogenase; UGT72E: anthocyanidin 3-O-glucosyltransferase; E2.3.1.133: stemmadenine O-acetyltransferase; 4CL: 4-coumarate-CoA ligase; bglX: xylan 1,4-beta-xylosidase. *FDR < 0.05, where red represents significant upregulation and green represents significant downregulation. PAL, phenylalanine ammonia lyase.
Table 5.
Differential expression results of flavonoid biosynthesis.
| Gene name | Gene ID | KO name | 0.2Se + 15N versus Control | 0.2Se versus Control | 0.2Se versus 0.2se + 15N | |||
|---|---|---|---|---|---|---|---|---|
| Log2FC | FDR | Log2FC | FDRt | Log2FC | FDRt | |||
| Trans-cinnamate 4-monooxygenase | VIT_06s0004g08150 | CYP73A | –1.24 | 0.00489 | –0.13 | 0.51476 | 1.14 | 0.00104 |
| Chalcone synthase | VIT_14s0068g00930 | CHS | –1.65 | 0.03460 | –1.39 | 0.00004 | 1.28 | 0.01740 |
| Dihydroflavonol reductase | VIT_18s0001g12800 | DFR | –1.11 | 0.01526 | 0.36 | 0.02693 | –1.49 | 0.00002 |
| Stilbene synthase | VIT_16s0100g01000 | CHS | 2.44 | 0.00540 | –1.09 | 0.59531 | 3.67 | 0.00024 |
| Stilbene synthase | VIT_16s0100g00860 | ST | 2.15 | 0.00000 | –0.54 | 0.45448 | –0.27 | 0.10247 |
| Flavanone 3-hydroxylase | VIT_04s0023g03370 | F3H | –1.73 | 0.03047 | –0.45 | 0.01205 | 1.29 | 0.06844 |
| Glycosyltransferase | VIT_18s0041g00830 | PGT1 | 1.01 | 0.04337 | 0.90 | 0.09188 | 1.29 | 0.04528 |
| Flavonoid-3′-hydroxylase | VIT_17s0000g07200 | CYP75B1 | –1.20 | 0.03266 | 0.16 | 0.35881 | 1.38 | 0.00276 |
| Unnamed protein product | VIT_17s0000g04150 | LAR | –1.41 | 0.00198 | 0.11 | 0.55600 | –1.54 | 0.00039 |
| Stilbene synthase | VIT_16s0100g01040 | CHS | 4.44 | 0.03436 | 1.01 | 1.00000 | –3.39 | 0.06491 |
| Anthocyanidin synthase | VIT_02s0025g04720 | ANS | –1.57 | 0.01520 | –0.41 | 0.01691 | 1.18 | 0.03871 |
| Stilbene synthase | VIT_16s0100g00930 | CHS | 1.88 | 0.00301 | –0.61 | 0.60305 | 2.48 | 0.00000 |
| Flavonoid 3′,5′-hydroxylase | VIT_06s0009g02970 | CYP75A | –1.00 | 0.00000 | –1.43 | 0.00000 | 1.41 | 0.00252 |
| Flavonoid 3′,5′-hydroxylase | VIT_06s0009g02880 | CYP75A | –1.22 | 0.00346 | –1.98 | 0.00000 | –2.73 | 0.38401 |
| Anthocyanin acyltransferase | VIT_03s0017g00870 | E2.3.1.133 | –3.60 | 0.00000 | –4.76 | 0.00000 | –1.14 | 0.12038 |
| Stilbene synthase | VIT_16s0100g00940 | CHS | 1.82 | 0.00030 | –0.44 | 0.60887 | –2.25 | 0.00001 |
| Chalcone synthase isoform | VIT_14s0068g00920 | CHS | –1.23 | 0.03394 | –0.29 | 0.13223 | 0.96 | 0.038498 |
| Unnamed protein product | VIT_11s0016g02610 | E2.1.1.104 | –4.99 | 0.02208 | –3.75 | 0.07667 | 1.26 | 1.00000 |
| Stilbene synthase | VIT_16s0100g01010 | CHS | 1.62 | 0.00326 | –0.45 | 0.65109 | –2.05 | 0.00075 |
| Flavonoid 3′,5′-hydroxylase | VIT_06s0009g02860 | CYP75A | –1.03 | 0.03085 | –1.03 | 0.01032 | 0.02 | 0.94689 |
| Flavonoid 3′-monooxygenase | VIT_17s0000g07210 | CYP75B1 | –1.09 | 0.00124 | 0.33 | 0.28077 | 0.14 | 0.50203 |
| UDP-glycosyltransferase | VIT_18s0041g00800 | PGT1 | –5.41 | 0.00000 | –7.00 | 0.00000 | –1.62 | 1.00000 |
| Hypothetical protein | VIT_06s0009g02830 | CYP75A | –1.42 | 0.00052 | –1.07 | 0.00000 | –1.37 | 0.04739 |
| UDP-glycosyltransferase | VIT_18s0041g00930 | PGT1 | –3.30 | 0.03199 | –0.35 | 0.58657 | –1.09 | 0.00002 |
| Unnamed protein product | VIT_03s0038g01330 | E2.3.1.133 | 1.20 | 0.0000 | 0.10 | 0.66635 | 7.94 | 0.00000 |
| Stilbene synthase | VIT_16s0100g00920 | CHS | 1.57 | 0.03144 | –2.68 | 0.03760 | 0.92 | 0.07035 |
| Stilbene synthase | VIT_16s0100g00900 | ST | 1.68 | 0.02265 | –1.04 | 0.37134 | 2.71 | 0.00043 |
| Hypothetical protein | VIT_16s0100g00950 | ST | 2.31 | 0.04592 | –0.32 | 0.90820 | –2.62 | 0.03463 |
| Stilbene synthase | VIT_16s0100g00840 | CHS | 1.58 | 0.00000 | –1.22 | 0.07250 | –1.78 | 0.00000 |
| Chalcone synthase | VIT_05s0136g00260 | CHS | –1.48 | 0.04403 | 0.26 | 0.11752 | –1.92 | 0.05836 |
Table 6.
Differential expression results of phenylpropanoid biosynthesis.
| Gene name | Gene ID | KO name | 0.2Se + 15N versus Control | 0.2Se versus Control | 0.2Se versus 0.2se + 15N | |||
|---|---|---|---|---|---|---|---|---|
| Log2FC | FDR | Log2FC | FDR | Log2FC | FDR | |||
| Phenylalanine ammonia lyase | VIT_16s0039g01170 | PAL | 3.31 | 0.01280 | –0.38 | 1.00000 | –3.78 | 0.00447 |
| Uncharacterised protein | VIT_12s0055g00810 | E1.11.1.7 | –0.78 | 0.00031 | –2.28 | 0.00000 | –1.49 | 0.00000 |
| Phenylalanine ammonia lyase | VIT_16s0039g01100 | PAL | 2.40 | 0.03015 | 0.04 | 0.98827 | –2.37 | 0.03448 |
| Unnamed protein product | VIT_06s0004g01430 | E3.2.1.21 | 0.55 | 0.62314 | 3.13 | 0.00000 | 2.61 | 0.00000 |
| Phenylalanine ammonia lyase | VIT_16s0039g01120 | PAL | 2.05 | 0.13924 | –2.61 | 1.00000 | –4.62 | 0.00953 |
| Peroxidase | VIT_12s0055g00810 | E1.11.1.7 | –0.02 | 0.92531 | –2.00 | 0.00000 | –1.96 | 0.00000 |
| Trans-cinnamate 4-monooxygenase | VIT_06s0004g08150 | CYP73A | –1.24 | 0.00488 | –0.13 | 0.51475 | 1.14 | 0.00104 |
| Putative beta-glucosidase | VIT_19s0014g04750 | E3.2.1.21 | –1.46 | 0.00000 | –3.54 | 0.00000 | –2.06 | 0.00005 |
| Glycosyltransferase | VIT_16s0022g01970 | UGT72E | –1.15 | 0.02177 | –3.40 | 0.00000 | –2.23 | 0.00028 |
| Unnamed protein product | VIT_13s0064g01750 | E3.2.1.21 | –0.89 | 0.10202 | –1.99 | 0.00000 | –1.08 | 0.00000 |
| Probable cinnamyl alcohol dehydrogenase | VIT_18s0001g14910 | CAD | 0.06 | 0.84734 | –1.21 | 0.00000 | –1.25 | 0.00000 |
| Peroxidase | VIT_13s0067g02360 | E1.11.1.7 | –2.14 | 0.00000 | –4.01 | 0.00000 | –1.85 | 0.00000 |
| Peroxidase | VIT_10s0116g01780 | E1.11.1.7 | –0.63 | 0.14670 | –1.71 | 0.00000 | –1.06 | 0.00000 |
| Phenylalanine ammonia lyase | VIT_16s0039g01300 | PAL | 1.34 | 0.16736 | –1.43 | 0.36247 | –2.75 | 0.01649 |
| Phenylalanine ammonia lyase | VIT_11s0016g01520 | PAL | 0.61 | 0.55273 | –2.11 | 0.04852 | –2.70 | 0.00120 |
| Peroxidase | VIT_16s0100g00090 | E1.11.1.7 | 0.94 | 0.15751 | –1.77 | 0.00308 | –2.69 | 0.00000 |
| Phenylalanine ammonia lyase | VIT_16s0039g01110 | PAL | 1.78 | 0.11341 | –1.22 | 0.54672 | –2.99 | 0.01490 |
| Unnamed protein product | VIT_06s0004g01420 | E3.2.1.21 | –0.44 | 0.35440 | 0.82 | 0.12460 | 1.27 | 0.00000 |
| Peroxidase | VIT_08s0040g02200 | E1.11.1.7 | –1.02 | 0.08130 | 0.01 | 0.68626 | 1.06 | 0.04688 |
| Peroxidase | VIT_12s0055g00990 | E1.11.1.7 | 2.58 | 0.10764 | –2.51 | 1.00000 | –5.02 | 0.00337 |
| Cytochrome P450 CYP73A100 | VIT_11s0065g00350 | CYP73A | 0.11 | 0.88529 | –1.52 | 0.00240 | –1.62 | 0.00002 |
| Peroxidase | VIT_16s0022g02470 | E1.11.1.7 | 0.73 | 0.00275 | –1.76 | 0.00000 | –2.47 | 0.00000 |
| Berberine bridge enzyme | VIT_10s0003g05420 | K22395 | –0.63 | 0.26053 | –2.19 | 0.00000 | –1.54 | 0.00000 |
| Peroxidase | VIT_18s0072g00160 | E1.11.1.7 | –2.60 | 0.00004 | –0.34 | 0.06431 | 2.28 | 0.00000 |
| Peroxidase | VIT_07s0191g00050 | E1.11.1.7 | 0.01 | 0.98307 | 1.02 | 0.00000 | 1.03 | 0.00000 |
| Peroxidase | VIT_10s0003g00650 | E1.11.1.7 | –0.61 | 0.29475 | –2.91 | 0.00001 | –2.27 | 0.00104 |
| Peroxidase | VIT_12s0059g02420 | E1.11.1.7 | –0.69 | 0.05056 | –1.93 | 0.00000 | –1.22 | 0.00439 |
| Unnamed protein product | VIT_13s0064g01640 | E3.2.1.21 | –0.46 | 0.74547 | 1.16 | 0.15059 | 1.65 | 0.02022 |
| Unnamed protein product | VIT_11s0016g01640 | PAL | 2.70 | 0.12995 | –1.38 | 1.00000 | –4.04 | 0.03834 |
| Unnamed protein product | VIT_03s0038g01330 | E2.3.1.133 | 1.20 | 0.00000 | 0.10 | 0.66635 | –1.09 | 0.00000 |
| 4-coumarate – CoA ligase | VIT_02s0109g00250 | 4CL | 2.14 | 0.00939 | 0.06 | 0.97506 | –2.07 | 0.01062 |
| Uncharacterised protein | VIT_06s0004g06110 | bglX | -0.90 | 0.37906 | 0.31 | 0.22344 | 1.23 | 0.00000 |
| Peroxidase | VIT_12s0055g01010 | E1.11.1.7 | 1.89 | 0.32080 | –2.98 | 1.00000 | –4.86 | 0.02264 |
| Probable mannitol dehydrogenase | VIT_04s0044g00190 | CAD | 0.15 | 0.60019 | –1.12 | 0.00000 | –1.25 | 0.00000 |
| Probable cinnamyl alcohol dehydrogenase | VIT_03s0180g00250 | CAD | –3.13 | 0.00000 | –6.10 | 0.00000 | –2.98 | 0.01763 |

Figure 6.
Key DEGs involved in plant hormone signal transduction in grape after Na2SeO3 treatment under various nitrogen conditions. ARR-A: two-component response regulator; CYCD3: cyclin-D3-1; SAUR: auxin-responsive protein; AUX1: auxin transporter-like protein; CH3: probable indole-3-acetic acid-amido synthetase; PR1: basic form of pathogenesis-related protein; IAA: auxin-induced protein; PYL: abscisic acid receptor; GID1: gibberellin receptor; NPR1: BTB/POZ domain and ankyrin repeat-containing protein; TCH4: probable xyloglucan endotransglucosylase/hydrolase protein; IRAK4: receptor-like cytosolic serine/threonine-protein kinase; AHP: histidine-containing phosphotransfer protein; JAZ: jasmonic acid protein; ETR: ethylene receptor; DELLA: DELLA protein GAI; E2.4.1.207: probable xyloglucan endotransglucosylase/hydrolase protein; TGA: transcription factor; PP2C: protein phosphatase 2C. *FDR < 0.05, where red represents significant upregulation and green represents significant downregulation.
Table 7.
Differential expression results of plant hormone signal transduction.
| Gene name | Gene ID | KO name | 0.2Se + 15N versus Control | 0.2Se versus Control | 0.2Se versus 0.2se + 15N | |||
|---|---|---|---|---|---|---|---|---|
| Log2FC | FDR | Log2FC | Log2FC | FDR | Log2FC | |||
| Two-component response regulator | VIT_13s0067g03510 | ARR-A | –0.18 | 0.83317 | 2.04 | 0.00000 | 2.24 | 0.00000 |
| Cyclin-D3-1 | VIT_18s0001g09920 | CYCD3 | –0.05 | 0.89851 | –2.63 | 0.00000 | –2.56 | 0.00000 |
| Unnamed protein product | VIT_07s0129g01100 | CYCD3 | –0.79 | 0.00000 | –1.32 | 0.00000 | –0.51 | 0.25060 |
| Gibberellin receptor GID1B | VIT_07s0104g00930 | GID1 | –0.48 | 0.06023 | 1.42 | 0.00000 | 1.92 | 0.00000 |
| Regulatory protein NPR5 | VIT_08s0007g05740 | NPR1 | –0.86 | 0.34463 | –2.23 | 0.00791 | –1.35 | 0.23369 |
| Auxin-responsive protein SAUR71 | VIT_01s0146g00180 | SAUR | 1.37 | 0.00002 | 1.14 | 0.00003 | –0.22 | 0.54304 |
| Unnamed protein product | VIT_03s0038g02140 | AUX1 | –0.53 | 0.07953 | –2.14 | 0.00000 | –1.59 | 0.00000 |
| Basic form of pathogenesis-related protein | VIT_03s0088g00780 | PR1 | 1.42 | 0.00000 | 1.18 | 0.00099 | –0.22 | 0.29702 |
| Auxin transporter | VIT_13s0067g00330 | AUX1 | –1.64 | 0.00000 | –1.76 | 0.00000 | –0.11 | 0.69752 |
| Indole-3-acetic acid-amido synthetase | VIT_19s0014g04690 | GH3 | –0.65 | 0.17485 | –1.06 | 0.02835 | –0.40 | 0.52344 |
| Xyloglucan endotransglucosylase/hydrolase | VIT_11s0052g01200 | TCH4 | –1.45 | 0.00058 | –2.11 | 0.00000 | –0.64 | 0.22629 |
| Hypothetical protein DKX38 | VIT_03s0088g00910 | IRAK4 | 1.03 | 0.00000 | 1.32 | 0.00000 | 0.31 | 0.16875 |
| Auxin-induced protein 6B | VIT_03s0038g00940 | SAUR | –0.91 | 0.15417 | –1.17 | 0.04523 | –0.25 | 0.78689 |
| Histidine-containing phosphotransfer protein | VIT_04s0008g00210 | AHP | 6.05 | 0.00000 | 3.12 | 0.01971 | –2.92 | 0.00000 |
| Auxin transporter | VIT_18s0001g03540 | AUX1 | –1.34 | 0.00000 | –1.04 | 0.00000 | 0.31 | 0.07698 |
| Auxin-responsive protein SAUR36 | VIT_15s0048g00530 | SAUR | –0.34 | 0.12180 | 1.35 | 0.00000 | 1.71 | 0.00000 |
| Cyclin-D3-1 | VIT_03s0180g00040 | CYCD3 | –2.26 | 0.00000 | –2.19 | 0.00000 | 0.08 | 0.87093 |
| Basic form of pathogenesis-related protein | VIT_03s0097g00700 | PR1 | –0.27 | 0.93243 | –2.22 | 0.00000 | –1.94 | 0.28237 |
| Pathogenesis-related protein | VIT_03s0088g00810 | PR1 | 0.82 | 0.00000 | 1.21 | 0.00000 | 0.41 | 0.02667 |
| Two-component response regulator ORR9 isoform X1 | VIT_13s0067g03490 | ARR-A | 0.51 | 0.00025 | 1.05 | 0.00000 | 0.56 | 0.05015 |
| Basic form of pathogenesis-related protein | VIT_03s0088g00710 | PR1 | 0.47 | 0.84569 | 2.85 | 0.00775 | 2.40 | 0.00982 |
| Auxin-responsive protein IAA9 isoform X1 | VIT_11s0016g05640 | IAA | 0.52 | 0.41001 | 1.93 | 0.00000 | 1.43 | 0.000000 |
| Jasmonate-zim-domain protein | VIT_01s0146g00480 | JAZ | –0.38 | 0.37895 | –1.53 | 0.00028 | –1.13 | 0.00697 |
| Ethylene receptor | VIT_05s0049g00090 | ETR | –0.02 | 0.96813 | –1.35 | 0.00000 | –1.32 | 0.00000 |
| DELLA protein SLR1 | VIT_11s0016g04630 | DELLA | –0.49 | 0.00519 | –1.16 | 0.00000 | –0.65 | 0.00003 |
| Xyloglucan endotransglucosylase/hydrolase | VIT_11s0052g01270 | E2.4.1.207 | –0.87 | 0.26567 | –2.51 | 0.00023 | 1.62 | 0.08526 |
| Transcription factor TGA9 | VIT_06s0080g00360 | TGA | –0.05 | 0.94243 | 1.17 | 0.00036 | 1.24 | 0.00002 |
| Two-component response regulator ORR9 | VIT_13s0067g03430 | ARR-A | 0.53 | 0.76430 | 1.96 | 0.00000 | 1.45 | 0.00000 |
| Two-component response regulator ARR6 | VIT_01s0026g00940 | ARR-A | –0.07 | 0.86455 | 1.46 | 0.00000 | 1.55 | 0.00000 |
| Auxin-responsive protein SAUR32 | VIT_15s0048g02860 | SAUR | 0.59 | 0.32414 | 1.03 | 0.00000 | 0.46 | 0.11100 |
| Auxin-responsive protein SAUR50 | VIT_04s0023g03230 | SAUR | 0.14 | 0.87585 | 1.25 | 0.00574 | 1.13 | 0.00406 |
| Auxin-responsive protein | VIT_07s0141g00270 | IAA | 0.03 | 0.93143 | 1.42 | 0.00000 | 1.41 | 0.00000 |
| Auxin-responsive protein SAUR36 | VIT_02s0154g00010 | SAUR | 0.38 | 0.32725 | 1.02 | 0.00028 | 0.65 | 0.20123 |
| Unnamed protein product | VIT_02s0012g01270 | PYL | 0.58 | 0.24239 | –1.04 | 0.00037 | –1.61 | 0.00000 |
| Auxin-induced protein 6B | VIT_03s0038g00930 | SAUR | –2.16 | 0.00509 | –3.62 | 0.00014 | –1.45 | 0.34481 |
| Abscisic acid receptor PYL4 | VIT_13s0067g01940 | PYL | 0.89 | 0.01109 | –1.46 | 0.00418 | –2.34 | 0.00000 |
| Probable indole-3-acetic acid-amido synthetase | VIT_07s0129g00660 | GH3 | –0.13 | 1.00000 | 4.03 | 0.04928 | 4.18 | 0.03700 |
| Auxin-induced protein | VIT_08s0007g03120 | SAUR | –0.82 | 0.54502 | –2.97 | 0.04849 | –2.14 | 0.23987 |
| Unnamed protein product | VIT_06s0004g05460 | PP2C | –0.16 | 0.88910 | 1.24 | 0.00000 | 1.42 | 0.00771 |

Figure 7.
Key DEGs involved in starch and sucrose metabolism in grape after Na2SeO3 treatment under various nitrogen conditions. E3.2.1.21: beta-glucosidase; WAXY: granule-bound starch synthase; TPS: alpha-trehalose-phosphate synthase; bglX: xylan 1,4-beta-xylosidase; E3.2.1.4: endoglucanase; E2.7.1.4: fructokinase; E2.4.1.13: sucrose synthase; GN4: glucan endo-1,3-beta-glucosidase; malZ: alpha-glucosidase; AMY: alpha-amylase; INV: beta-fructofuranosidase; HK: hexokinase. * FDR < 0.05, where red represents significant upregulation and green represents significant downregulation.
Table 8.
Differential expression results of starch and sucrose metabolism.
| Gene name | Gene ID | KO name | 0.2Se + 15N versus Control | 0.2Se versus Control | 0.2Se versus 0.2se + 15N | |||
|---|---|---|---|---|---|---|---|---|
| Log2FC | FDR | Log2FC | FDR | Log2FC | FDR | |||
| Uncharacterised protein | VIT_06s0009g00810 | E3.2.1.21 | –0.78 | 0.30589 | –2.28 | 0.00000 | –1.49 | 0.00000 |
| Unnamed protein product | VIT_06s0004g01430 | E3.2.1.21 | 0.55 | 0.62314 | 3.13 | 0.00000 | 2.61 | 0.00000 |
| Hypothetical protein | VIT_06s0004g00720 | GN4 | 0.23 | 0.32455 | –0.98 | 0.09370 | –1.20 | 0.00000 |
| Starch synthase, chloroplastic/amyloplastic | VIT_02s0025g02790 | WAXY | 0.98 | 0.08222 | 0.27 | 0.15132 | 1.28 | 0.00238 |
| Alpha-trehalose-phosphate synthase | VIT_10s0003g02160 | TPS | –0.06 | 0.78358 | 0.92 | 0.27515 | 1.00 | 0.00000 |
| Endoglucanase | VIT_07s0005g00740 | E3.2.1.4 | –1.19 | 0.00000 | –2.27 | 0.00000 | –1.06 | 0.00000 |
| Alpha-glucosidase | VIT_10s0092g00240 | malZ | 0.79 | 0.32308 | –0.24 | 0.10826 | –1.01 | 0.00000 |
| Putative beta-glucosidase | VIT_19s0014g04750 | E3.2.1.21 | –1.46 | 0.00000 | –3.54 | 0.00000 | –2.06 | 0.00006 |
| Probable fructokinase-5 | VIT_18s0089g01230 | E2.7.1.4 | –0.06 | 0.88916 | –3.53 | 0.00000 | –2.01 | 0.00000 |
| Alpha-amylase | VIT_18s0001g00560 | AMY | –0.82 | 0.73105 | 0.63 | 0.58606 | 1.48 | 0.00000 |
| Beta-fructofuranosidase, soluble isoenzyme I isoform X1 | VIT_16s0022g00670 | INV | –0.35 | 0.19816 | –2.12 | 0.00000 | –1.75 | 0.00000 |
| Unnamed protein product | VIT_13s0064g01750 | E3.2.1.21 | –0.89 | 0.41109 | –1.99 | 0.00000 | –1.07 | 0.00000 |
| Putative alpha,alpha-trehalose-phosphate synthase | VIT_01s0026g00280 | TPS | 1.37 | 0.00000 | –0.43 | 0.00279 | –1.07 | 0.00000 |
| Phosphotransferase | VIT_09s0002g03390 | HK | 0.97 | 0.48252 | –2.70 | 0.23476 | –3.66 | 0.027023 |
| Sucrose synthase | VIT_11s0016g00470 | E2.4.1.13 | –0.39 | 0.23845 | –1.80 | 0.00000 | –1.40 | 0.00000 |
| Endoglucanase | VIT_04s0008g02010 | E3.2.1.4 | 1.03 | 0.08715 | –0.14 | 0.88231 | –1.15 | 0.04076 |
| Sucrose synthase | VIT_17s0053g00700 | E2.4.1.13 | 0.20 | 0.36444 | –1.60 | 0.00000 | –1.78 | 0.00000 |
| Probable fructokinase | VIT_05s0102g00710 | E2.7.1.4 | –0.50 | 0.18347 | –1.62 | 0.00000 | –1.11 | 0.00000 |
| Probable alpha,alpha-trehalose-phosphate synthase | VIT_12s0028g01670 | TPS | –0.03 | 0.96391 | –1.21 | 0.00261 | –1.16 | 0.00280 |
| Unnamed protein product | VIT_06s0004g01420 | E3.2.1.21 | –0.48 | 0.35446 | 0.82 | 0.11907 | 1.27 | 0.00000 |
| Sucrose synthase | VIT_07s0005g00750 | E2.4.1.13 | 0.68 | 0.06330 | –0.34 | 0.15054 | –1.00 | 0.00000 |
| Unnamed protein product | VIT_07s0005g06660 | WAXY | –1.61 | 0.57698 | 1.65 | 0.22606 | 3.25 | 0.03323 |
| Probable alpha,alpha-trehalose-phosphate synthase | VIT_17s0000g08010 | TPS | 3.15 | 0.00000 | –0.11 | 0.88102 | 3.26 | 0.00000 |
| Putative alpha,alpha-trehalose-phosphate synthase | VIT_06s0009g01650 | TPS | 1.20 | 0.18780 | –0.97 | 0.51346 | –2.15 | 0.02419 |
| Unnamed protein product | VIT_13s0064g01640 | E3.2.1.21 | –0.46 | 0.74547 | 1.16 | 0.15059 | 1.65 | 0.02022 |
| Uncharacterised protein | VIT_06s0004g06110 | bglX | –0.90 | 0.37906 | 0.31 | 0.22344 | 1.23 | 0.00000 |
| Probable fructokinase-7 isoform XI | VIT_15s0048g01260 | E2.7.1.4 | 0.77 | 0.29019 | –0.75 | 0.06879 | –1.50 | 0.00000 |
| Sucrose synthase | VIT_04s0079g00230 | E2.4.1.13 | 0.24 | 0.33133 | –1.83 | 0.00000 | –2.05 | 0.00000 |