Scientia Silvae Sinicae ›› 2020, Vol. 56 ›› Issue (10): 70-82.doi: 10.11707/j.1001-7488.20201008
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Bi Liang,Jiaqi Zhang,Fei Ren,Hengkang Hu,Chuanmei Xu,Yuanyuan Hu,Youjun Huang,Heqiang Lou,Qixiang Zhang*
Received:
2019-11-18
Online:
2020-10-25
Published:
2020-11-26
Contact:
Qixiang Zhang
CLC Number:
Bi Liang,Jiaqi Zhang,Fei Ren,Hengkang Hu,Chuanmei Xu,Yuanyuan Hu,Youjun Huang,Heqiang Lou,Qixiang Zhang. Cloning and Expression Analysis of Ent-Kaurene Oxidase Gene CcKo in Carya cathayensis[J]. Scientia Silvae Sinicae, 2020, 56(10): 70-82.
Table 1
Primer sequence"
引物 Primers | 序列 Sequence(5′—3′) | 用途 Usage |
CcKO-1300-F | CTTGTACAGCTCGTC$\underline{{\rm{GGATCC}}}$ATGTCCATGGCCAGCCTAAGCTAC | 扩增 Amplification |
CcKO-1300-R | TCGCCCTTGCTCACCAT$\underline{{\rm{GTCGAC}}}$TTAATTTCTCAGCTTTATCATTGCTTGCATC | 扩增 Amplification |
PCcKO-1301-F | CGGGGATCCTCTAGA$\underline{{\rm{GTCGAC}}}$ATGATGGAGAGACGGAGAG | 扩增 Amplification |
PCcKO-1301-R | TTACCCTCAGATCTA$\underline{{\rm{CCATGG}}}$TTAATAGTACTCTTATAGTTGCGATTTTGG | 扩增 Amplification |
Actin-F | GCCGAACGGGAAATTGTC | 内参 Reference |
Actin-R | AGAGATGGCTGGAAGAGG | 内参 Reference |
QCcKO-F | ACGGATGCAACATGGACAAGAA | 定量 qPCR |
QCcKO-R | CTCTTTCCTCCTCCGAATGCCA | 定量 qPCR |
QJrGA20ox-F | CAAGTGTTTGTAGACGAGTGGC | 定量 qPCR |
QJrGA20ox-R | TGGACTCTTGCTGTTCACCACT | 定量 qPCR |
QJrGA2ox-F | AATCATTACCCTCCATGCCCAG | 定量 qPCR |
QJrGA2ox-R | AAATGATTTGTGGGTCGGTGTG | 定量 qPCR |
GFP-R | TTACTTGTACAGCTCGTCCA | 鉴定 Testing |
Fig.3
GUS localization analysis of CcKOpro∷GUS positive somatic embryos(SE) and regenerated plants(RP) in walnuts A: GUS positive SE; B: GUS positive RP leaves; C: Cross section of stem of GUS positive RP; D: Longitudinal section of stem of GUS positive RP; E: Control SE; F: Control RP leaves; G: Cross section of stem of control RP; H: Longitudinal section of stem of control RP. "
Fig.5
Fluorescence expression of walnut 35S∷CcKO∷GFP transformed SE A, B, C, D: Transgenic SE of walnut under natural light; E, F, G, H: Transgenic SE of walnut stimulated by blue light; I, J, K, L: Control SE of walnut under natural light; M, N, O, P: Control SE of walnut stimulated by blue light. A, E, I, M: Globular embryos; B, F, J, N: Heart-shaped embryos; C, G, K, O: Torpedo embryos; D, H, L, P: Cotyledon embryos. "
Fig.7
Phenotypic analysis of 35S∷CcKO∷GFP transgenic walnut plants A, B: Plant height phenotypes of control and transgenic plants at 0 d and 20 d respectively; C: The leaf phenotypes of control and transgenic plants; D: Statistical analysis of plant height and internode length, significance of one-way ANOVA, different letters indicate significant difference (P < 0.05). "
Fig.9
The effect of CcKO overexpression on key enzyme genes downstream of gibberellin biosynthesis A: The relative expression of GA20ox gene in walnut positive RP; B: The relative expression of GA2ox gene in walnut positive RP. Single factor analysis of variance is used to analysis the differences, and different letters indicate significant difference (P < 0.05). "
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