Qian Wei’s research while affiliated with China Agricultural University and other places

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Publications (1)


Drought stress resistance of CmNF‐YB8 transgenic chrysanthemum plants
(A) Relative CmNF‐YB8 transcript levels in control chrysanthemum plants (0 h) or during dehydration for 1, 3, 6, 9, or 12 h, as determined by reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR) (n = 3). Data are shown as means ± standard deviation. Significant differences were determined using a Student's t test (**P < 0.01). (B) Relative CmNF‐YB8 transcript levels in chrysanthemum plants during various drought treatments, as determined by RT‐qPCR. Relative soil water contents were 77% (well‐watered), 56% (mild drought), 25% (moderate drought), 11% (severe drought), and 77% (re‐watered for 24 h). (C) Relative CmNF‐YB8 transcript levels in CmNF‐YB8‐RNAi, CmNF‐YB8‐OX and wild type (WT) plants, as determined by RT‐qPCR. (D) Representative phenotypes of CmNF‐YB8‐RNAi, CmNF‐YB8‐OX, and WT plants during drought stress. Drought stress was applied by withholding water for 30 d and was followed by re‐watering. The survival rate was calculated after 30 d of re‐watering. (E) Survival rates for CmNF‐YB8‐RNAi, CmNF‐YB8‐OX, and WT plants after re‐watering (n = 12). Data are shown as means ± standard deviation. Significant differences are indicated by different letters according to Duncan's multiple range test (P < 0.05). (F, G) Changes in the transpiration rate (F) and stomatal conductance (G) of transgenic lines and WT plants during a drought time course (n > 5). (H) Water loss rates of detached leaves from transgenic lines and WT. Water loss assays were performed for 120 min (n = 5). Significant differences were determined by Tukey's test (*P < 0.05; **P < 0.01).
Stomatal morphology in transgenic chrysanthemum leaves
(A) Images of stomata on the abaxial side of leaves from CmNF‐YB8 transgenic lines and wild type (WT) plants. Scale bar, 200 μm. (B) Scanning electron microscopy images of stomatal aperture in the leaves of CmNF‐YB8 transgenic lines and WT plants. Scale bar, 20 μm. (C) Images of the adaxial cuticle in transgenic chrysanthemum leaves. Red arrows point to the cuticle. Scale bar, 50 μm. (D) Representative images of three states of stomatal aperture. Scale bar, 20 μm. (E) Percentage of stomatal aperture states in CmNF‐YB8 transgenic lines and WT plants (n > 100). (F) Adaxial cuticle thickness in WT, CmNF‐YB8‐RNAi, and CmNF‐YB8‐OX plants. Data are shown as means ± standard deviation (n = 10).
Expression of stomatal adjustment‐ and cuticle biosynthesis‐related genes in transgenic chrysanthemum plants
Relative transcript levels for CmCIPK6 (A), CmSLAC1 (B), CmCA3 (C), CmSHN3 (D), CmCYP86A (E) and CmBDG1 (F), as determined by reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR), with CmUBI as a reference gene. Three independent experiments were performed, and different letters indicate significant differences (P < 0.05).
CmNF‐YB8 directly regulates CmCIPK6 expression
(A) Schematic diagram of the 590‐bp CmCIPK6 promoter. Black lines above the promoter represent the fragments used in Y1H analysis. P1, − 157 to −218 bp; P2, − 266 to −323 bp; P3, − 349 to −406 bp; P4, − 434 to −489 bp; P5, − 494 to −548 bp; P6, − 536 to −590 bp (with the A from the CmCIPK6 ATG set to +1 bp). Asterisks indicate putative CCAAT boxes. (B) Y1H analysis of CmNF‐YB8 binding to the CmCIPK6 promoter. The CmCIPK6 promoter was divided into six fragments (P1–P6). The antibiotic AbA was used for selection of interaction. The basal concentration of AbA was 600 ng/mL. (C) Analysis of CmNF‐YB8 binding to the CmCIPK6 promoter by EMSA. Purified recombinant CmNF‐YB8 (1 μg) was incubated with biotin‐labeled probes (20 nM). The competition test was performed with cold probes provided in 10‐, 100‐, or 1,000‐fold excess over labeled probe. FP, free probe. (D) Chromatin IP‐qPCR of the indicated fragments (P1–P6) in the CmCIPK6 promoter. Chromatin from CmNF‐YB8‐OX‐15 chrysanthemum plants was immunoprecipitated with an anti‐GFP antibody. 35:GFP chrysanthemum plants served as a negative control. The amount of the indicated DNA fragment was determined by qPCR and normalized to the 35S:GFP control (set to 1 for each fragment). (E–G) Interaction between CmNF‐YB8 and a 872‐bp CmCIPK6 promoter fragment using a LUC reporter assay in Nicotiana benthamiana leaves. m‐CmCIPK6 promoter is the same promoter fragment with a mutated CCAAT cis‐element. LUC and Renilla luciferase (REN) activities were assayed 3 d after infiltration. (E) Schematic diagrams of the reporter and effector constructs used in the assay. (F) Representative photographs of firefly luciferase activity. (G) Normalized LUC activity of the indicated samples, shown as LUC/REN ratio. The LUC/REN ratio was normalized to samples co‐infiltrated with the empty reporter and empty effector constructs (SK + LUC), which were set to 1. Three independent experiments were performed; significant difference was determined by Tukey's test (*P < 0.05; **P < 0.01).
Epidermal morphology and physiological indices of CmNF‐YB8‐RNAi and wild type (WT) plants infected with CaLCuV‐amiR‐CIPK6
(A) Relative transcript levels of CmCIPK6 in CmNF‐YB8‐RNAi and WT plants infected with CaLCuV‐amiR‐CIPK6. (B) Water loss rates of detached leaves harvested from CmNF‐YB8‐RNAi and WT plants infected with CaLCuV‐amiR‐CIPK6. Water loss assays were performed for 120 min (n = 5). Significant differences were determined by Tukey's test (*P < 0.05; **P < 0.01). (C, D) Transpiration rate (C) and stomatal conductance (D) of CmNF‐YB8‐RNAi and WT plants infected with CaLCuV or CaLCuV‐amiR‐CIPK6. (E) Representative images of stomata on the abaxial side of leaves from CmNF‐YB8 transgenic lines and WT. Scale bar, 50 μm. (F) Scanning electron microscopy images of the stomatal aperture state in leaves of CmNF‐YB8‐RNAi and WT plants infected with CaLCuV‐amiR‐CIPK6. Scale bar, 20 μm. (G) Percentage of possible stomatal opening states in CmNF‐YB8‐RNAi and WT plants infected with CaLCuV or CaLCuV‐amiR‐CIPK6 (n > 100).

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CmNF‐YB8 affects drought resistance in chrysanthemum by altering stomatal status and leaf cuticle thickness
  • Article
  • Full-text available

March 2022

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136 Reads

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28 Citations

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Qian Wei

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Zhiling Wang

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[...]

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Bo Hong

Drought is a major abiotic stress that limits plant growth and development. Adaptive mechanisms have evolved to mitigate drought stress, including the capacity to adjust water loss rate and to modify the morphology and structure of the epidermis. Here, we show that the expression of CmNF‐YB8, encoding a nuclear factor Y (NF‐Y) B‐type subunit, is lower under drought conditions in chrysanthemum (Chrysanthemum morifolium). Transgenic chrysanthemum lines in which transcript levels of CmNF‐YB8 were reduced by RNA interference (CmNF‐YB8‐RNAi) exhibited enhanced drought resistance relative to control lines, whereas lines overexpressing CmNF‐YB8 (CmNF‐YB8‐OX) were less tolerant to drought. Compared to wild type (WT), CmNF‐YB8‐RNAi plants showed reduced stomatal opening and a thicker epidermal cuticle that correlated with their water loss rate. We also identified genes involved in stomatal adjustment (CBL‐interacting protein kinase 6, CmCIPK6) and cuticle biosynthesis (CmSHN3) that are more highly expressed in CmNF‐YB8‐RNAi lines than in WT, CmCIPK6 being a direct downstream target of CmNF‐YB8. Virus‐induced gene silencing of CmCIPK6 or CmSHN3 in the CmNF‐YB8‐RNAi background abolished the effects of CmNF‐YB8‐RNAi on stomatal closure and cuticle deposition, respectively. CmNF‐YB8 thus regulates CmCIPK6 and CmSHN3 expression to alter stomatal movement and cuticle thickness in the leaf epidermis, thereby affecting drought resistance.

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Citations (1)


... Partial closure of the stomata could have been responsible for reducing stomatal conductance and transpiration rate. Thus, it was possible to maintain high rates of photosynthesis without compromising the internal concentration of CO 2 since the greater difference in CO 2 concentration between the atmosphere and the interior of the leaf compensates for the increase in stomatal resistance [70]. ...

Reference:

Physiological and Biochemical Responses of Pseudocereals with C3 and C4 Photosynthetic Metabolism in an Environment with Elevated CO2
CmNF‐YB8 affects drought resistance in chrysanthemum by altering stomatal status and leaf cuticle thickness