Skip to main content
. 2020 Jan 19;18(7):1504–1506. doi: 10.1111/pbi.13321

Figure 1.

Figure 1

The interaction between MeCIPK23 and MeWHYs is essential for the activation of abscisic acid biosynthesis and drought stress resistance in cassava. (a)‐(d) Yeast two‐hybrid (a), BiFC (b), LUC complementation assay (c) and pull down (d) showing the physical interaction between MeCIPK23 and MeWHYs. DAPI‐stained cell nuclei and yellow fluorescent in BiFC were visualized using a confocal laser‐scanning microscope. Bar = 25 μm. (e) The transcript levels of MeCIPK23 and MeWHYs in cassava leaves in response to drought stress. For the assay, plant leaves under control conditions (well‐watered) and drought stress conditions (with‐holding water) for indicated days were harvested. (f) RT‐PCR showing the expression of TRV1 and TRV2 in the VIGS plants. The reference gene MeEF1a and the viral transcripts TRV1/TRV2 were examined. (g) The transcript levels of corresponding genes in the gene‐silenced plant leaves. (h)‐(i) Water loss rate (h) and EL (i) in the gene‐silenced plant leaves in response to drought stress. (j) The transcript levels of MeNCEDs in the gene‐silenced plant leaves under control conditions. (k)‐(l) The transcript levels of MeNCED1 (k) and the endogenous ABA accumulation (l) in the gene‐silenced plant leaves in response to drought stress. (m) EMSA showing the direct binding of MeWHYs to the probes of MeNCED1 promoter. The sequences of control probe with PB motif and mutated probe with mutated PB motif are shown. The position of free probe and the protein‐probe complex are marked by arrow. (n) ChIP‐PCR showing the relative enrichment of MeWHYs in MeNCED1 promoter. The same buffer without GFP antibody (IgG) was used as the native control of the GFP antibody. (o) Dual LUC assay showing the effects of MeWHYs and MeCIPK23 on the activity of MeNCED1 promoter. (p) The transcript level of MeNCED1 in the gene‐silenced plants. (q) The endogenous ABA accumulation in the gene‐silenced plants in response to drought stress. (r) Exogenous ABA restores the drought stress sensitivity of MeCIPK23‐MeWHYs‐MeNCED1 silencing plants. The pictures of different plants during drought stress conditions. Bars = 10 cm. (s)‐(t) Water loss rate (s) and EL (t) in the leaves in response to drought stress. (u) A proposed module of MeCIPK23‐MeWHYs‐MeNCED1 in drought stress response in cassava. In this study, cassava leaves were harvested for the assays. VIGS and gene overexpression in cassava leaves were performed through Agrobacterium tumefacien‐mediated transformation as we previously described (Liu et al., 2018; Zeng et al., 2019). All experiments were performed with at least three biological repeats. Statistical test was performed by SPSS. Kolmogorov–Smirnov test and Levene’s tests were used to check the normality of the data distribution and the variance homogeneity of the data, respectively. Asterisk symbols (*) suggested significant differences compared with control at P < 0.05.