Biotechnology Bulletin ›› 2024, Vol. 40 ›› Issue (8): 74-82.doi: 10.13560/j.cnki.biotech.bull.1985.2024-0201
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CUI Yuan-yuan(), WANG Zhao-yi, BAI Shuang-yu, REN Yu-zhao, DOU Fei-fei, LIU Cai-xia, LIU Feng-lou, WANG Zhang-jun, LI Qing-feng()
Received:
2024-03-14
Online:
2024-08-26
Published:
2024-07-30
Contact:
LI Qing-feng
E-mail:18695124679@163.com;liqingfeng2017@nxu.edu.cn
CUI Yuan-yuan, WANG Zhao-yi, BAI Shuang-yu, REN Yu-zhao, DOU Fei-fei, LIU Cai-xia, LIU Feng-lou, WANG Zhang-jun, LI Qing-feng. Genome-wide Identification of Non-specific Phospholipase C Gene Family in Hordeum vulgare L. and Stress Expression Analysis at Seedling Stage[J]. Biotechnology Bulletin, 2024, 40(8): 74-82.
引物名称 Primer name | 引物序列 Primer sequence(5'-3') | 退火温度Annealing temperature/℃ |
---|---|---|
HvNPC1-F | AACAAGCCCCCATCACCG | 58.9 |
HvNPC1-R | CCACCTTGAAGGCGTCGT | 58.6 |
HvNPC2-F | ACTACCAGTACCCGCCGT | 58.9 |
HvNPC2-R | ACCACGTAGTTGGGCAGC | 58.6 |
HvNPC3-F | GACGTGCTGGACCAGGTC | 59.4 |
HvNPC3-R | CGGAGGAGAGCAGGGAGT | 59.8 |
HvNPC4-F | GATGCCGACGAGGATCCG | 59.2 |
HvNPC4-R | ATCCCCGTTCAGCACTGC | 58.8 |
HvNPC5-F | CGTCTTCGACCGCTGGTT | 58.4 |
HvNPC5-R | CGTTGAAGGTGAGCCCGT | 58.6 |
HvActin-F | CAGGTATCGCTGACCGTATGAG | 57.5 |
HvActin-R | TGGAAAGTGCTGAGTGAGGCT | 58.9 |
Table 1 Primers used for RT-qPCR
引物名称 Primer name | 引物序列 Primer sequence(5'-3') | 退火温度Annealing temperature/℃ |
---|---|---|
HvNPC1-F | AACAAGCCCCCATCACCG | 58.9 |
HvNPC1-R | CCACCTTGAAGGCGTCGT | 58.6 |
HvNPC2-F | ACTACCAGTACCCGCCGT | 58.9 |
HvNPC2-R | ACCACGTAGTTGGGCAGC | 58.6 |
HvNPC3-F | GACGTGCTGGACCAGGTC | 59.4 |
HvNPC3-R | CGGAGGAGAGCAGGGAGT | 59.8 |
HvNPC4-F | GATGCCGACGAGGATCCG | 59.2 |
HvNPC4-R | ATCCCCGTTCAGCACTGC | 58.8 |
HvNPC5-F | CGTCTTCGACCGCTGGTT | 58.4 |
HvNPC5-R | CGTTGAAGGTGAGCCCGT | 58.6 |
HvActin-F | CAGGTATCGCTGACCGTATGAG | 57.5 |
HvActin-R | TGGAAAGTGCTGAGTGAGGCT | 58.9 |
基因名称 Gene name | 基因ID Transcript ID | 物理位置 Physical position | 相对分子量 Molecular weight/kD | 等电点 pI | 亲水性总平均值 Grand average of hydrophilicity | 蛋白长度 Protein length/aa | 亚细胞定位 Subcellular location |
---|---|---|---|---|---|---|---|
HvNPC1 | HORVU.MOREX.r3.7HG0752530.1 | 7H:631001620-631003350 | 53.12 | 5.22 | -0.317 | 477 | 细胞质 |
HvNPC2 | HORVU.MOREX.r3.1HG0085590.1 | 1H:496122549-496124474 | 61.33 | 6.53 | -0.364 | 553 | 叶绿体 |
HvNPC3 | HORVU.MOREX.r3.3HG0232030.1 | 3H:28367740-28369602 | 58.77 | 8.83 | -0.204 | 532 | 叶绿体 |
HvNPC4 | HORVU.MOREX.r3.3HG0322080.1 | 3H:600541735-600546962 | 59.07 | 5.98 | -0.424 | 533 | 液泡膜 |
HvNPC5 | HORVU.MOREX.r3.5HG0527960.1 | 5H:566994044-566997667 | 60.76 | 6.80 | -0.367 | 545 | 叶绿体 |
Table 2 Basic physico-chemical properties of NPC family in barley(Hordeum vulgare L)
基因名称 Gene name | 基因ID Transcript ID | 物理位置 Physical position | 相对分子量 Molecular weight/kD | 等电点 pI | 亲水性总平均值 Grand average of hydrophilicity | 蛋白长度 Protein length/aa | 亚细胞定位 Subcellular location |
---|---|---|---|---|---|---|---|
HvNPC1 | HORVU.MOREX.r3.7HG0752530.1 | 7H:631001620-631003350 | 53.12 | 5.22 | -0.317 | 477 | 细胞质 |
HvNPC2 | HORVU.MOREX.r3.1HG0085590.1 | 1H:496122549-496124474 | 61.33 | 6.53 | -0.364 | 553 | 叶绿体 |
HvNPC3 | HORVU.MOREX.r3.3HG0232030.1 | 3H:28367740-28369602 | 58.77 | 8.83 | -0.204 | 532 | 叶绿体 |
HvNPC4 | HORVU.MOREX.r3.3HG0322080.1 | 3H:600541735-600546962 | 59.07 | 5.98 | -0.424 | 533 | 液泡膜 |
HvNPC5 | HORVU.MOREX.r3.5HG0527960.1 | 5H:566994044-566997667 | 60.76 | 6.80 | -0.367 | 545 | 叶绿体 |
Fig. 2 Structural features of NPC gene family in barely A: Purple bars indicate the exon, lines indicate the intron while grey color bars indicate the un-translated region(UTR). B: The yellow bar indicates the signature domain phosphoesterase(PF04185)
Fig. 3 Cis-acting elements in the promoters of HvNPC gene family members ABRE: Abscisic acid responsive element. CGTCA-motif, TGACG-motif: MeJA responsive element. GARE-motif, P-box, TATC-box: Gibberellin responsive element. TGA-element: Auxin responsive element. TCA-element: Salicylic acid responsive element. MBS, MYC, MYB: Drought responsive element. TC-rich: Defense and stress responsive element. LTR: Low-temperature responsive element. ARE: Essential for the anaerobic responsive element. O2-site: Zein metabolism regulatory element. RY-element: Seed-specific regulatory element. Sp1, TCT-motif, AE-box, BoxII, I-box: Light responsive element
Fig. 4 Expression profile of NPC gene in 14 tissues and organs of barley A: Embryo(4 d after germination). B: Roots from the seedlings(shoot length 10 cm). C: Shoots(10 cm). D: Etiolated seedling (10 d old at dark). E: Developing tillers(six leaf stge, 3rd internode). F: Root(4 weeks). G: Epidermis(4 weeks). H: Florescences(5 mm). I: Florescences(1-1.5 cm). J: Rachis(5 weeks). K: Lemma(6 weeks). L: Lodicule(6 weeks). M: Palea(6 weeks). N: Senescing leaf(2 weeks)
Fig. 5 Relative expression pattern of HvNPCs genes under low temperature, NaCl, and drought treatments A: Gene expression profiles of HvNPCs under 4℃ treatment. B: Gene expression profiles of HvNPCs under NaCl treatment. C: Gene expression profiles of HvNPCs under PEG treatment. Different lower letters indicate significant differences(P<0.05)
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