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. 2022 Mar;20(3):423-425.
doi: 10.1111/pbi.13771. Epub 2022 Jan 11.

Auxin promotes fiber elongation by enhancing gibberellic acid biosynthesis in cotton

Liping Zhu  1 , Bin Jiang  1 , Jiaojie Zhu  1 , Guanghui Xiao  1
Affiliations

Auxin promotes fiber elongation by enhancing gibberellic acid biosynthesis in cotton

Liping Zhu et al. Plant Biotechnol J. 2022 Mar.
No abstract available

Keywords: CRISPR/Cas9; Cotton (Gossypium hirsutum); auxin; auxin responsive factor; fiber development; gibberellin acid.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Auxin promotes fiber elongation by enhancing GA biosynthesis. (a) Phenotypes of 10 day‐old ovules cultured with 0.5 μm of GA1, 1 μm of PAC, 5 μm of IAA, 1 μm of NPA, 0.5 μm of GA1 + 1 μm of NPA, and 5 μm of IAA + 1 μm of PAC. (b) GA1 content of fibers treated with IAA or NPA. (c) Relative expression of GhARF18 in different fiber development stages. (d, e) Fibers phenotype (d) and length measurement (e) of CK, GhARF18 over‐expression and CRISPR/Cas9 lines. (f) GA1 and GA4 content in 5 and 10 DPA fibers from CK, GhARF18 over‐expression and CRISPR/Cas9 lines. (g) Yeast one‐hybrid assay between GhARF18 and nine promoters of gibberellin acid biosynthesis genes. P53 and p53 promoter were used as positive control. (h, i) Relative expression of GhGA3OX4D and GhGA20OX1D‐2 in different fiber development stages (h) and in 5 DPA fibers from CK, GhARF18 over‐expression and CRISPR/Cas9 lines (i). (j, k) Tobacco transient expression assay of GhARF18 and pGhGA3OX4D (j) or pGhGA20OX1D‐2 (k) promoters. (l, m) Tobacco dual‐luciferase assay of pGhGA3OX4D::LUC (l) and pGhGA20OX1D‐2::LUC expression (m). Expression of REN was used as internal control. (n, o) Yeast one‐hybrid assay of GhARF18 and GhGA3OX4D (n) and four GhGA20OX1D‐2 (o) fragments. (p, q) Tobacco transient expression assay of GhARF18 and pGhGA3OX4D‐P2 or AuxRE‐mutated pGhGA3OX4D‐P2 fragment (p) and GhARF18 and pGhGA20OX1D‐2‐P3 or AuxRE‐mutated pGhGA20OX1D‐2‐P3 fragment (q). (r, s) Tobacco dual‐luciferase assay of intact or mutated pGhGA3OX4D‐P2::LUC (r) and pGhGA20OX1D‐2‐P3::LUC (s) expression. Values given are mean ± SD (n ≥ 5). (t–w) EMSA of GhARF18 binding to AuxREs from GhGA3OX4D and GhGA20OX1D‐2 promoters. Promoter fragments containing intact AuxRE were incubated with gradient concentrations of GhARF18 protein (t, u). Different concentrations of unlabeled probes of intact or mutated AuxRE (cold probe) were incubated with GhARF18 to compete with labeled native promoter fragments with AuxRE (v, w). (x, y) ChIP‐qPCR of GhGA3OX4D (x) and GhGA20OX1D‐2 (y) promoter fragments in GhARF18 over‐expression line. (z) Phenotypes of ovules from GhARF18 over‐expression and CRISPR/Cas9 lines cultured with 5 μm of IAA, 1 μm of NPA, 0.5 μm of GA1, and 1 μm of PAC, respectively. (ab) Relative expression of GhGA3OX4D and GhGA20OX1D‐2 in their transgenic fibers. (ac, ad, ae) GA1 and GA4 content of 10 DPA fibers (ac), mature length of fibers (ad), and phenotypes of fibers (ae) from CK, GhGA3OX4D and GhGA20OX1D‐2‐over‐expression lines. (af) Schematic model. CK, nontransgenic plants. d, day post anthesis (DPA). Bar = 1 cm. Statistical significance for each comparison is indicated (t‐test): *, P ≤ 0.05, **, P ≤ 0.01, ***, P ≤ 0.001.
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