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Artificial selection of two antagonistic E3 ubiquitin ligases finetunes soybean photoperiod adaptation and grain yield
Wednesday, 2024/11/13 | 08:24:51

Fan WangShuangrong LiuHaiyang LiChao FangSijia FangJianhao WangShichen Li

Huan LiuHaiping DuLingshuang WangXinxin PeiBohong SuZhihui SunQuan LiLidong Dong,

 Qun ChengXiaohui ZhaoBaohui LiuSijia LuFanjiang Kong, and Xiaoya Lin

PNAS; November 1, 2024; 121 (45) e2321473121; https://doi.org/10.1073/pnas.2321473121

Significance

The precise regulation of flowering time is important for plants because it determines reproductive success, ensures optimal adaptation to local environmental conditions, and influences crop yield. Soybean E2 is a homolog of Arabidopsis GIGANTEA that plays a major role in the circadian clock and flowering regulation, the mechanism behind its protein regulation is not well understood. The roles of ZTL2 and FKF1s in soybean flowering are dependent on E2, but these two regulators have antagonistic effects on E2 protein stability. ZTL2 promotes the degradation of E2, leading to early flowering, while FKF1a and FKF1b stabilize E2, resulting in delayed flowering. This antagonistic function allows soybeans to fine-tune their flowering time in response to different latitudes and geographical distributions.

Abstract

The precise control of flowering time is of utmost importance for crop adaptation to varying environmental conditions and consequently determines grain yield and plant fitness. Soybean E2, the homolog of Arabidopsis GIGANTEA, is a major locus contributing to high-latitude adaptation and is involved in photoperiod sensitivity. However, due to major effects of E2, additional genetic loci controlling soybean flowering and adaptation have historically been masked and difficult to identify. Here, by eliminating the effect of E2, we identified a Tof9 locus controlling flowering in which ZEITLUPE 2 (ZTL2) is the causal gene. ZTL2 encodes an F-box E3 ubiquitin ligase with homology to Arabidopsis ZEITLUPE and is shown to play a key role in the soybean photoperiodic flowering pathway. ZTL2 physically interacts with E2 to mediate its degradation. Intriguingly, ZTL2 and FKF1, both belong to the F-box-type E3 ubiquitin-ligase family, exhibit opposite roles in regulating soybean flowering. ZTL2 degrades E2, leading to early flowering, while FKF1 stabilizes E2, resulting in delayed flowering. The balance between ZTL2-mediated degradation and FKF1-mediated stabilization enables soybeans to finetune flowering time and maximize grain yield. Field-grown ztl2 mutants are taller, flower late, and have increased yield parameters. ZTL2 and FKF1b bear contrasting artificial-selection patterns to adapt to different latitudes. This antagonistic regulation is crucial for soybean adaptation to diverse ecological settings and allows plants to fine-tune their flowering time in response to photoperiod and latitudinal changes.

 

See https://www.pnas.org/doi/10.1073/pnas.2321473121

 

Figure 2:

ZTL2 physically interacts with E2 and mediates the degradation of E2. (A) ZTL2 can interact with E2 in yeast. Yeast cells were transformed and selected on the absence of adenine, leucine, histidine, and tryptophan in X-α-galactose solid media. (B) In vivo Co-IP assay of interaction between ZTL2 and E2. p35S:E2–GFP and p35S:ZTL2–FLAG were transiently expressed under the control of the 35S promoter in N. benthamiana leaves. p35S:ZTL1–FLAG was used as a negative control. Total proteins were immunoprecipitated with anti-GFP and anti-FLAG antibody. (C) ZTL2 can pull down E2. ZTL1–GST, ZTL2–GST, and E2–MBP proteins were purified from E. coli. GST, ZTL1–GST and ZTL2–GST were detected with an anti-GST antibody and E2–MBP was detected with an anti-MBP antibody. (D) ZTL2 is genetically dependent on E2. Phenotypes of Wm82, ztl2, e2 e2la e2lb, and ztl2 e2 e2la e2lb plants under long-day conditions (Scale bar, 3 cm). The images above panels are enlarged views of axils of trifoliate leaves in the below panels. (E) Flowering time of Wm82, ztl2, e2 e2la e2lb, ztl2 e2 e2la e2lb under long-day conditions. (F) Phenotypes of Wm82, ZTL2 p35S:E2–6xHA, ztl2, and ztl2 p35S:E2–6xHA plants under long-day conditions (Scale bar, 3 cm). The images above panels are enlarged views of axils of trifoliate leaves in the below panels. (G) Flowering time of Wm82, ZTL2 p35S:E2–6xHA, ztl2, and ztl2 p35S:E2–6xHA plants under long-day conditions. (H) Effects of the ztl2 mutation on E2 stability. ZTL2 p35S:E2–6xHA and ztl2 p35S:E2–6xHA plants were grown for 15 d under long-day conditions for immunoblot of daily expression of p35S:E2–6xHA in soybean leaves. Three independent experiments were repeated with similar results. The presented data in (E) and (G) are expressed as means ± SD with individual data points represented as dots. Statistical significance (P < 0.05) was determined through multiple comparison testing using one-way ANOVA. Different lowercase letters denote statistically significant differences among the groups.

 

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