图1 HL1的结构

Figure 1 The structure of HL1

图2 HL1的特异性及对Zn2+结合的敏感性(A) 1 mmol·L-1不同元素与HL1探针结合后在500 nm处的荧光强度; (B) 不同Zn2+浓度下的荧光强度(a: 1 mmol∙L-1 Zn2+; b: 0.5 mmol∙L-1 Zn2+; c: 0.1 mmol∙L-1 Zn2+; d: 0.05 mmol∙L-1 Zn2+; e: 0.01 mmol∙L-1 Zn2+)

Figure 2 Specific and sensitivity of combination of HL1 and Zn2+(A) Fluorescence intensity of different element (1 mmol∙L-1) in presence of ligand HL1 (1 mmol∙L-1) at 500 nm; (B) Plot of fluorescence intensity against different concentration of Zn2+ at 500 nm (a: 1 mmol∙L-1 Zn2+; b: 0.5 mmol∙L-1 Zn2+; c: 0.1 mmol∙L-1 Zn2+; d: 0.05 mmol∙L-1 Zn2+; e: 0.01 mmol∙L-1 Zn2+)

图3 利用ITC200检测HL1与Zn2+的反应

Figure 3 ITC200 detection plot of combination of HL1 and Zn2+

图4 拟南芥叶细胞外Zn2+分布荧光图像(A) 用不同浓度ZnSO4溶液及HL1溶液处理成熟的拟南芥叶片, a1和a2为对照, b1和b2处理ZnSO4溶液浓度为0.3 μmol∙L-1, c1和c2处理ZnSO4溶液浓度为3 μmol∙L-1, d1和d2处理ZnSO4溶液浓度为30 μmol∙L-1, e1和e2处理ZnSO4溶液浓度为300 μmol∙L-1; (B) 不同浓度ZnSO4溶液及HL1溶液处理下的拟南芥叶表皮毛的荧光强度, 其中a为对照, b、c、d和e分别为叶表皮毛在Zn2+浓度为0.3、3、30和300 μmol∙L-1时的荧光强度。蓝色荧光代表Zn2+, 红色荧光代表叶绿素。Bar=100 μm

Figure 4 Fluorescence imaging showing the distribution of extracellular Zn2+ in Arabidopsis leaves(A) Mature Arabidopsis leaves pre-treated with different concentrations of ZnSO4 and HL1 (1 mmol∙L-1) for 4 h, a1 and a2 for control, b1 and b2 for 0.3 μmol∙L-1 ZnSO4, c1 and c2 for 3 μmol∙L-1 ZnSO4, d1 and d2 for 30 μmol∙L-1 ZnSO4, e1 and e2 for 300 μmol∙L-1 ZnSO4; (B) The fluorescence intensity of trichome in Arabidopsis leaf with different concentration of ZnSO4 and HL1 (1 mmol∙L-1) for 4 h, a for control, b for 0.3 μmol∙L-1 ZnSO4, c for 3 μmol∙L-1 ZnSO4, d for 30 μmol∙L-1 ZnSO4, and e for 300 μmol∙L-1 ZnSO4. Blue fluorescence represents Zn2+ and red fluorescence for chlorophyll. Bar=100 μm

图5 植物细胞在1 mmol∙L-1 HL1下的荧光图像蓝色荧光代表Zn2+, 红色荧光代表叶绿素。Bar=50 μm

Figure 5 Fluorescence image of plant cell in presence of 1 mmol∙L-1 HL1Blue represents Zn2+ and red for chlorophyll. Bar=50 μm

图6 拟南芥根细胞外Zn2+分布荧光图像(A) 分别用不同浓度的ZnSO4溶液与HL1溶液共处理2 cm左右的拟南芥根, 处理时间为4小时, 其中A1为对照, A2和A3中Zn2+浓度分别为30和300 μmol∙L-1 (Bar=100 μm); (B) A1、A2和A3分别对应处理液Zn2+浓度0、30和300 μmol∙L-1下的荧光强度。蓝色荧光代表Zn2+。

Figure 6 Fluorescence images showing the distribution of extracellular Zn2+ in Arabidopsis roots(A) Two-centimeter-long Arabidopsis root pre-treated with different concentrations of ZnSO4 and HL1 (1 mmol∙L-1) for 4 h, A1, A2, and A3 for control, 30 μmol∙L-1 ZnSO4 and 300 μmol∙L-1 ZnSO4, respectively (Bar=100 μm); (B) A1, A2, and A3 corresponding to the fluorescence intensity of Arabidopsis root treated with different concentrations of ZnSO4 for 0, 30, and 300 μmol∙L-1. Blue fluorescence represents Zn2+.

图7 谷子叶片细胞外Zn2+分布荧光图像(A) 叶中央; (B) 叶边缘; (C) 叶尖。蓝色荧光代表Zn2+, 红色荧光代表叶绿素。Bar=100 μm

Figure 7 Fluorescence images showing the distribution of extracellular Zn2+ in Setaria italic leaves(A) The center of leaf; (B) The margin of leaf; (C) The apex of leaf. Blue fluorescence represents Zn2+ and red fluorescence for chlorophyll. Bar=100 μm