Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (10): 173-183.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1576
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GUO Ai1(), JIANG Mu-yan1, Ha Li-ma-ti·Ba He-tai-li1, LIU Yu-yuan1, WANG Jing1,2()
Received:
2021-12-21
Online:
2022-10-26
Published:
2022-11-11
Contact:
WANG Jing
E-mail:1061123080@qq.com;wangjing_imu@163.com
GUO Ai, JIANG Mu-yan, Ha Li-ma-ti·Ba He-tai-li, LIU Yu-yuan, WANG Jing. Transcriptome Analysis of Lycium ruthenicum Murr. Shoots in Anthocyanin Biosynthesis Response to Salt Stress[J]. Biotechnology Bulletin, 2022, 38(10): 173-183.
基因ID Gene ID | 序列Sequence(5'-3') |
---|---|
LrH2B1 | F:AGTGCTTCCTGGTGAATTGG |
R:TGGATAATACCTAGCCCTAGTTTCC | |
Cluster-45.236520 | F:TTTGCCTCTGGCTTTGTCTACC |
R:TGGATACTGCTGTTGCCTTCAT | |
Cluster-45.192144 | F:TGCACTGTTTGTCTCCTAGCCTCC |
R:ACGCCATTCTTCTCGTCTCCATC | |
Cluster-45.35768 | F:AAAACAGGACATGGCCTTTATG |
R:CTGAGTTTGTAGGCAAGGTGGA | |
Cluster-45.156739 | F:GCACAAAGTCTCCGAGAAGG |
R:GGGGGCTTCATATTTCTTGTT | |
Cluster-45.33457 | F:GAAGCCAACAGGTGAAACGACG |
R:TCTTCATCAAAGCTCCCAAATGCT | |
Cluster-45.168632 | F:GGTTTGATGAAAGGGGTGAAGAA |
R:CCTGACGTGGCAGTATCGCTGAG, | |
Cluster-45.207146 | F:TGAAGAATTAAGAGAGTCGAGGTCG |
R:AAGTTGGCAATGGTGATGGTGTC. | |
Cluster-45.136887 | F:TCAGATGAACCCAATTCACCAA |
R:ACCCTAAAGAGCTGTCGGATGT | |
Cluster-45.146553 | F:GGCACAGTAGCTGAATCCAAGA |
R:CCTTTACCAACTTCTGCCCATA | |
Cluster-45.150170 | F:GAGCAAGGCAGCAATAGCAGAT |
R:AAGGTGGTGACATTGAGGTGAAG | |
Cluster-45.146155 | F:GGTCAAGCCACTGATAGCCACA |
R:GCATTCCTGCTTTCCCTGTGAT | |
Cluster-45.117764 | F:TATCTTTCTTAGGCCCTTCCCTC |
R:GCGTTATTGTTATTATTGTCGGTGT |
Table 1 List of specific primers for qPCR
基因ID Gene ID | 序列Sequence(5'-3') |
---|---|
LrH2B1 | F:AGTGCTTCCTGGTGAATTGG |
R:TGGATAATACCTAGCCCTAGTTTCC | |
Cluster-45.236520 | F:TTTGCCTCTGGCTTTGTCTACC |
R:TGGATACTGCTGTTGCCTTCAT | |
Cluster-45.192144 | F:TGCACTGTTTGTCTCCTAGCCTCC |
R:ACGCCATTCTTCTCGTCTCCATC | |
Cluster-45.35768 | F:AAAACAGGACATGGCCTTTATG |
R:CTGAGTTTGTAGGCAAGGTGGA | |
Cluster-45.156739 | F:GCACAAAGTCTCCGAGAAGG |
R:GGGGGCTTCATATTTCTTGTT | |
Cluster-45.33457 | F:GAAGCCAACAGGTGAAACGACG |
R:TCTTCATCAAAGCTCCCAAATGCT | |
Cluster-45.168632 | F:GGTTTGATGAAAGGGGTGAAGAA |
R:CCTGACGTGGCAGTATCGCTGAG, | |
Cluster-45.207146 | F:TGAAGAATTAAGAGAGTCGAGGTCG |
R:AAGTTGGCAATGGTGATGGTGTC. | |
Cluster-45.136887 | F:TCAGATGAACCCAATTCACCAA |
R:ACCCTAAAGAGCTGTCGGATGT | |
Cluster-45.146553 | F:GGCACAGTAGCTGAATCCAAGA |
R:CCTTTACCAACTTCTGCCCATA | |
Cluster-45.150170 | F:GAGCAAGGCAGCAATAGCAGAT |
R:AAGGTGGTGACATTGAGGTGAAG | |
Cluster-45.146155 | F:GGTCAAGCCACTGATAGCCACA |
R:GCATTCCTGCTTTCCCTGTGAT | |
Cluster-45.117764 | F:TATCTTTCTTAGGCCCTTCCCTC |
R:GCGTTATTGTTATTATTGTCGGTGT |
Fig. 1 Contents of anthocyanin in L. ruthenicum seedlings under NaCl stress at different times ***means the difference is extremely significant (P<0.001)
物质Compound | CK含量CK content/(μg·g-1) | Na含量Na content/(μg·g-1) | P值P -value | 倍数变化Fold change |
---|---|---|---|---|
飞燕草素-3-O-葡萄糖苷 Delphinidin-3-O-glucoside | 0.151±0.0409 | 1.13±0.0283 | 0.000027 | 7.5 |
飞燕草素-3-O-芸香糖苷 Delphinidin-3-O-rutinoside | 0.432±0.0581 | 5.15±0.134 | 0.0000655 | 11.9 |
矮牵牛素-3-O-芸香糖苷 Petunidin-3-O-rutinoside | 2.33±0.323 | 5.313±0.286 | 0.000659 | 2.28 |
柚皮素-7-O-葡萄糖苷 Naringenin-7-O-glucoside | 0.112±0.0189 | 0.339±0.0457 | 0.0015 | 3.02 |
Table 2 Types and contents of significantly-changed anthocyanins in L. ruthenicum shoots under NaCl stress
物质Compound | CK含量CK content/(μg·g-1) | Na含量Na content/(μg·g-1) | P值P -value | 倍数变化Fold change |
---|---|---|---|---|
飞燕草素-3-O-葡萄糖苷 Delphinidin-3-O-glucoside | 0.151±0.0409 | 1.13±0.0283 | 0.000027 | 7.5 |
飞燕草素-3-O-芸香糖苷 Delphinidin-3-O-rutinoside | 0.432±0.0581 | 5.15±0.134 | 0.0000655 | 11.9 |
矮牵牛素-3-O-芸香糖苷 Petunidin-3-O-rutinoside | 2.33±0.323 | 5.313±0.286 | 0.000659 | 2.28 |
柚皮素-7-O-葡萄糖苷 Naringenin-7-O-glucoside | 0.112±0.0189 | 0.339±0.0457 | 0.0015 | 3.02 |
样品Sample | 原始测序数据Raw reads | 过滤后测序数据Clean reads | 过滤后数据Clean bases | Error/% | Q20/% | Q30/% | GC content/% |
---|---|---|---|---|---|---|---|
CK1 | 55 697 476 | 54 900 826 | 8.24 G | 0.02 | 98.48 | 95.29 | 42.65 |
CK2 | 60 551 930 | 59 757 658 | 8.96 G | 0.03 | 97.2 | 91.95 | 42.48 |
CK3 | 55 743 892 | 54 933 528 | 8.24 G | 0.03 | 97.25 | 92.05 | 42.53 |
Na1 | 61 583 804 | 61 024 066 | 9.15 G | 0.03 | 96.83 | 91.12 | 42.21 |
Na2 | 46 367 542 | 45 803 662 | 6.87 G | 0.02 | 98.39 | 95.06 | 42.32 |
Na3 | 47 635 904 | 46 871 318 | 7.03 G | 0.02 | 98.48 | 95.32 | 42.31 |
Table 3 Transcriptome sequencing data of L. ruthenicum shoots under control and NaCl stress
样品Sample | 原始测序数据Raw reads | 过滤后测序数据Clean reads | 过滤后数据Clean bases | Error/% | Q20/% | Q30/% | GC content/% |
---|---|---|---|---|---|---|---|
CK1 | 55 697 476 | 54 900 826 | 8.24 G | 0.02 | 98.48 | 95.29 | 42.65 |
CK2 | 60 551 930 | 59 757 658 | 8.96 G | 0.03 | 97.2 | 91.95 | 42.48 |
CK3 | 55 743 892 | 54 933 528 | 8.24 G | 0.03 | 97.25 | 92.05 | 42.53 |
Na1 | 61 583 804 | 61 024 066 | 9.15 G | 0.03 | 96.83 | 91.12 | 42.21 |
Na2 | 46 367 542 | 45 803 662 | 6.87 G | 0.02 | 98.39 | 95.06 | 42.32 |
Na3 | 47 635 904 | 46 871 318 | 7.03 G | 0.02 | 98.48 | 95.32 | 42.31 |
分类Term | ID | P值P value | Q值Q value |
---|---|---|---|
光合作用-天线蛋白Photosynthesis - antenna proteins | ko00196 | 1.38E-55 | 1.12E-53 |
光合作用Photosynthesis | ko00195 | 7.82E-14 | 3.17E-12 |
光合生物中碳的固定Carbon fixation in photosynthetic organisms | ko00710 | 3.80E-10 | 1.03E-08 |
植物激素信号转导Plant hormone signal transduction | ko04075 | 1.96E-08 | 3.96E-07 |
乙醛酸和二羧酸代谢Glyoxylate and dicarboxylate metabolism | ko00630 | 6.17E-07 | 9.99E-06 |
黄酮生物合成Flavonoid biosynthesis | ko00941 | 8.35E-07 | 1.13E-05 |
苯丙素生物合成Phenylpropanoid biosynthesis | ko00940 | 5.60E-06 | 6.48E-05 |
卟啉与叶绿素代谢Porphyrin and chlorophyll metabolism | ko00860 | 0.00041 | 0.0041 |
角质,木栓质和蜡质的生物合成Cutin,suberine and wax biosynthesis | ko00073 | 0.00045 | 0.0041 |
淀粉和蔗糖代谢Starch and sucrose metabolism | ko00500 | 0.001 | 0.0081 |
二苯乙烯类、二芳基庚烷类和姜辣素生物合成Stilbenoid,diarylheptanoid and gingerol biosynthesis | ko00945 | 0.0011 | 0.0081 |
氮代谢Nitrogen metabolism | ko00910 | 0.0046 | 0.031 |
α-亚麻酸代谢alpha-linolenic acid metabolism | ko00592 | 0.005 | 0.031 |
精氨酸和脯氨酸代谢Arginine and proline metabolism | ko00330 | 0.0073 | 0.042 |
Table 4 KEGG enrichment pathways
分类Term | ID | P值P value | Q值Q value |
---|---|---|---|
光合作用-天线蛋白Photosynthesis - antenna proteins | ko00196 | 1.38E-55 | 1.12E-53 |
光合作用Photosynthesis | ko00195 | 7.82E-14 | 3.17E-12 |
光合生物中碳的固定Carbon fixation in photosynthetic organisms | ko00710 | 3.80E-10 | 1.03E-08 |
植物激素信号转导Plant hormone signal transduction | ko04075 | 1.96E-08 | 3.96E-07 |
乙醛酸和二羧酸代谢Glyoxylate and dicarboxylate metabolism | ko00630 | 6.17E-07 | 9.99E-06 |
黄酮生物合成Flavonoid biosynthesis | ko00941 | 8.35E-07 | 1.13E-05 |
苯丙素生物合成Phenylpropanoid biosynthesis | ko00940 | 5.60E-06 | 6.48E-05 |
卟啉与叶绿素代谢Porphyrin and chlorophyll metabolism | ko00860 | 0.00041 | 0.0041 |
角质,木栓质和蜡质的生物合成Cutin,suberine and wax biosynthesis | ko00073 | 0.00045 | 0.0041 |
淀粉和蔗糖代谢Starch and sucrose metabolism | ko00500 | 0.001 | 0.0081 |
二苯乙烯类、二芳基庚烷类和姜辣素生物合成Stilbenoid,diarylheptanoid and gingerol biosynthesis | ko00945 | 0.0011 | 0.0081 |
氮代谢Nitrogen metabolism | ko00910 | 0.0046 | 0.031 |
α-亚麻酸代谢alpha-linolenic acid metabolism | ko00592 | 0.005 | 0.031 |
精氨酸和脯氨酸代谢Arginine and proline metabolism | ko00330 | 0.0073 | 0.042 |
基因ID Gene ID | CK-FPKM | Na-FPKM | UP/DOWN | 倍数变化Fold change | Q值Q value | 参与激素通路Involved in hormonal pathway |
---|---|---|---|---|---|---|
Cluster-45.33300 | 2.8 | 67.88 | UP | 4.58 | 1.10E-43 | IAA |
Cluster-45.160336 | 17.22 | 48.62 | UP | 1.48 | 2.68E-17 | Zeatin |
Cluster-45.160335 | 4.75 | 12.57 | UP | 1.39 | 1.89E-12 | |
Cluster-45.155979 | 1.25 | 18.36 | UP | 3.87 | 7.67E-37 | Gibberellin |
Cluster-45.155981 | 2.33 | 28.19 | UP | 3.58 | 7.85E-39 | |
Cluster-45.156292 | 43.04 | 18.56 | DOWN | -1.23 | 1.44E-10 | |
Cluster-45.142941 | 19.73 | 1.94 | DOWN | -3.37 | 4.36E-25 | ABA |
Cluster-45.123460 | 11.69 | 142.1 | UP | 3.59 | 2.95E-50 | |
Cluster-45.195827 | 0.98 | 33.77 | UP | 5.07 | 1.29E-33 | |
Cluster-45.39863 | 2.83 | 27.75 | UP | 3.29 | 1.69E-40 | |
Cluster-45.78587 | 9.97 | 47.67 | UP | 2.24 | 8.10E-32 | |
Cluster-45.80560 | 9.79 | 49.99 | UP | 2.34 | 9.08E-41 | |
Cluster-45.80561 | 13.05 | 56.32 | UP | 2.09 | 2.47E-34 | |
Cluster-45.165767 | 68.97 | 867.3 | UP | 3.58 | 3.94E-33 | |
Cluster-45.165775 | 9.24 | 120.92 | UP | 3.68 | 2.67E-23 | |
Cluster-45.165770 | 39.57 | 491.37 | UP | 3.6 | 1.60E-59 | |
Cluster-45.165774 | 127.23 | 1730.95 | UP | 3.6 | 2.53E-05 | |
Cluster-45.191345 | 1.03 | 31.02 | UP | 4.89 | 3.36E-12 | |
Cluster-45.191350 | 1.29 | 13.64 | UP | 3.4 | 1.09E-09 | |
Cluster-45.191353 | 17.41 | 102.01 | UP | 2.54 | 1.37E-49 | |
Cluster-45.191354 | 2.84 | 17.66 | UP | 2.63 | 1.72E-09 | |
Cluster-45.161351 | 96.31 | 244.8 | UP | 1.33 | 2.71E-19 | Ethylene |
Cluster-45.161685 | 11.82 | 45.31 | UP | 1.91 | 1.87E-15 | |
Cluster-45.108200 | 45.25 | 1.29 | DOWN | -5.16 | 8.45E-13 | |
Cluster-45.32854 | 1.48 | 27.85 | UP | 4.21 | 2.26E-52 | Brassinosteroid |
Cluster-45.175682 | 17.06 | 47.88 | UP | 1.47 | 1.34E-14 | SA |
Cluster-45.181144 | 159.8 | 12.52 | DOWN | -3.7 | 1.37E-25 |
Table 5 DEGs related to hormone signaling transduction pathway in L. ruthenicum shoots under NaCl stress
基因ID Gene ID | CK-FPKM | Na-FPKM | UP/DOWN | 倍数变化Fold change | Q值Q value | 参与激素通路Involved in hormonal pathway |
---|---|---|---|---|---|---|
Cluster-45.33300 | 2.8 | 67.88 | UP | 4.58 | 1.10E-43 | IAA |
Cluster-45.160336 | 17.22 | 48.62 | UP | 1.48 | 2.68E-17 | Zeatin |
Cluster-45.160335 | 4.75 | 12.57 | UP | 1.39 | 1.89E-12 | |
Cluster-45.155979 | 1.25 | 18.36 | UP | 3.87 | 7.67E-37 | Gibberellin |
Cluster-45.155981 | 2.33 | 28.19 | UP | 3.58 | 7.85E-39 | |
Cluster-45.156292 | 43.04 | 18.56 | DOWN | -1.23 | 1.44E-10 | |
Cluster-45.142941 | 19.73 | 1.94 | DOWN | -3.37 | 4.36E-25 | ABA |
Cluster-45.123460 | 11.69 | 142.1 | UP | 3.59 | 2.95E-50 | |
Cluster-45.195827 | 0.98 | 33.77 | UP | 5.07 | 1.29E-33 | |
Cluster-45.39863 | 2.83 | 27.75 | UP | 3.29 | 1.69E-40 | |
Cluster-45.78587 | 9.97 | 47.67 | UP | 2.24 | 8.10E-32 | |
Cluster-45.80560 | 9.79 | 49.99 | UP | 2.34 | 9.08E-41 | |
Cluster-45.80561 | 13.05 | 56.32 | UP | 2.09 | 2.47E-34 | |
Cluster-45.165767 | 68.97 | 867.3 | UP | 3.58 | 3.94E-33 | |
Cluster-45.165775 | 9.24 | 120.92 | UP | 3.68 | 2.67E-23 | |
Cluster-45.165770 | 39.57 | 491.37 | UP | 3.6 | 1.60E-59 | |
Cluster-45.165774 | 127.23 | 1730.95 | UP | 3.6 | 2.53E-05 | |
Cluster-45.191345 | 1.03 | 31.02 | UP | 4.89 | 3.36E-12 | |
Cluster-45.191350 | 1.29 | 13.64 | UP | 3.4 | 1.09E-09 | |
Cluster-45.191353 | 17.41 | 102.01 | UP | 2.54 | 1.37E-49 | |
Cluster-45.191354 | 2.84 | 17.66 | UP | 2.63 | 1.72E-09 | |
Cluster-45.161351 | 96.31 | 244.8 | UP | 1.33 | 2.71E-19 | Ethylene |
Cluster-45.161685 | 11.82 | 45.31 | UP | 1.91 | 1.87E-15 | |
Cluster-45.108200 | 45.25 | 1.29 | DOWN | -5.16 | 8.45E-13 | |
Cluster-45.32854 | 1.48 | 27.85 | UP | 4.21 | 2.26E-52 | Brassinosteroid |
Cluster-45.175682 | 17.06 | 47.88 | UP | 1.47 | 1.34E-14 | SA |
Cluster-45.181144 | 159.8 | 12.52 | DOWN | -3.7 | 1.37E-25 |
基因ID Gene ID | CK-FPKM | Na-FPKM | UP/DOWN | 倍数变化 Fold change | Q值Q value | 转录因子类型Transcription factor type |
---|---|---|---|---|---|---|
Cluster-45.111653 | 4.48 | 17.95 | UP | 1.98 | 4.23E-17 | MYB transcripiton factor |
Cluster-45.111657 | 6.51 | 27.56 | UP | 2.06 | 7.91E-17 | |
Cluster-45.111658 | 10.05 | 33.49 | UP | 1.72 | 8.62E-21 | |
Cluster-45.112186 | 18.55 | 56.83 | UP | 1.6 | 7.91E-16 | |
Cluster-45.227246 | 5.38 | 38.31 | UP | 2.81 | 3.07E-32 | |
Cluster-45.227247 | 1.4 | 8.37 | UP | 2.56 | 4.74E-20 | |
Cluster-45.32932 | 7.36 | 30.01 | UP | 2.01 | 4.9E-14 | |
Cluster-45.33457 | 4.23 | 43.42 | UP | 3.33 | 5.13E-34 | |
Cluster-45.35768 | 2.26 | 61.07 | UP | 4.73 | 5.35E-32 | |
Cluster-45.79986 | 4.58 | 19.6 | UP | 2.08 | 1.43E-15 | |
Cluster-45.95693 | 79.87 | 270.84 | UP | 1.73 | 7.24E-23 | |
Cluster-45.146155 | 40.64 | 7.09 | DOWN | -2.54 | 5.16E-20 | bHLH transcription factor |
Cluster-45.147909 | 52.14 | 21.72 | DOWN | -1.28 | 1.09E-11 | |
Cluster-45.150170 | 51.34 | 7.13 | DOWN | -2.87 | 9.35E-26 | |
Cluster-45.161489 | 30.39 | 2.7 | DOWN | -3.52 | 1.76E-11 | |
Cluster-45.168632 | 4.11 | 13.93 | UP | 1.75 | 4.3E-15 |
Table 6 MYB and bHLH transcription factors with significantly changed expression levels
基因ID Gene ID | CK-FPKM | Na-FPKM | UP/DOWN | 倍数变化 Fold change | Q值Q value | 转录因子类型Transcription factor type |
---|---|---|---|---|---|---|
Cluster-45.111653 | 4.48 | 17.95 | UP | 1.98 | 4.23E-17 | MYB transcripiton factor |
Cluster-45.111657 | 6.51 | 27.56 | UP | 2.06 | 7.91E-17 | |
Cluster-45.111658 | 10.05 | 33.49 | UP | 1.72 | 8.62E-21 | |
Cluster-45.112186 | 18.55 | 56.83 | UP | 1.6 | 7.91E-16 | |
Cluster-45.227246 | 5.38 | 38.31 | UP | 2.81 | 3.07E-32 | |
Cluster-45.227247 | 1.4 | 8.37 | UP | 2.56 | 4.74E-20 | |
Cluster-45.32932 | 7.36 | 30.01 | UP | 2.01 | 4.9E-14 | |
Cluster-45.33457 | 4.23 | 43.42 | UP | 3.33 | 5.13E-34 | |
Cluster-45.35768 | 2.26 | 61.07 | UP | 4.73 | 5.35E-32 | |
Cluster-45.79986 | 4.58 | 19.6 | UP | 2.08 | 1.43E-15 | |
Cluster-45.95693 | 79.87 | 270.84 | UP | 1.73 | 7.24E-23 | |
Cluster-45.146155 | 40.64 | 7.09 | DOWN | -2.54 | 5.16E-20 | bHLH transcription factor |
Cluster-45.147909 | 52.14 | 21.72 | DOWN | -1.28 | 1.09E-11 | |
Cluster-45.150170 | 51.34 | 7.13 | DOWN | -2.87 | 9.35E-26 | |
Cluster-45.161489 | 30.39 | 2.7 | DOWN | -3.52 | 1.76E-11 | |
Cluster-45.168632 | 4.11 | 13.93 | UP | 1.75 | 4.3E-15 |
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