W1D0: 75% of field capacity × undefoliation treatment;
W1D1: 75% of field capacity × 50% leaf defoliation treatment;W1D2: 75% of field capacity × 100% leaf defoliation treatment;W2D0: 55% of field capacity × undefoliation treatment;W2D1: 55% of field capacity × 50% leaf defoliation treatment;W2D2: 55% of field capacity × 100% leaf defoliation treatment;W3D0: 35% of field capacity × undefoliation treatment;W3D1: 35% of field capacity × 50% leaf defoliation treatment;W3D2: 35% of field capacity × 100% leaf defoliation treatment;0, 1, 2 and 3: before defoliation, 2 weeks, 5 weeks and 8 weeks after defoliation.A_max: maximum net photosynthetic rate (μmol m⁻² s⁻¹);E: transpiration rate (mmol m^-2 s^-1);Gs: stomatal conductance (mmol m^-2 s^-1);chl a/b: ratio of chlorophyll a and b (mg g^-1);Leaf NSC concentration: leaf non-structural carbohydrates concentration (%);Stem NSC concentration: stem non-structural carbohydrates concentration (%);Root NSC concentration: root non-structural carbohydrates concentration (%);TB: total biomass (g);LB: leaf biomass (g);LMR: leaf mass ratio;SMR: stem mass ratio;RMR: root mass ratio;R/S: root to shoot mass ratio; RGR_H: relative growth rate in the height (cm d^-1).Defoliation clearly affects biomass allocation of the two species, but not leaf physiology. Considering the carbon or sink limitation, the growth of S. japonica and R. pseudoacacia may be limited by future global climate change scenarios.

W1D0: 75% of field capacity × undefoliation treatment;
W1D1: 75% of field capacity × 50% leaf defoliation treatment;W1D2: 75% of field capacity × 100% leaf defoliation treatment;W2D0: 55% of field capacity × undefoliation treatment;W2D1: 55% of field capacity × 50% leaf defoliation treatment;W2D2: 55% of field capacity × 100% leaf defoliation treatment;W3D0: 35% of field capacity × undefoliation treatment;W3D1: 35% of field capacity × 50% leaf defoliation treatment;W3D2: 35% of field capacity × 100% leaf defoliation treatment;0, 1, 2 and 3: before defoliation, 2 weeks, 5 weeks and 8 weeks after defoliation.A_max: maximum net photosynthetic rate (μmol m⁻² s⁻¹);E: transpiration rate (mmol m^-2 s^-1);Gs: stomatal conductance (mmol m^-2 s^-1);chl a/b: ratio of chlorophyll a and b (mg g^-1);Leaf NSC concentration: leaf non-structural carbohydrates concentration (%);Stem NSC concentration: stem non-structural carbohydrates concentration (%);Root NSC concentration: root non-structural carbohydrates concentration (%);TB: total biomass (g);LB: leaf biomass (g);LMR: leaf mass ratio;SMR: stem mass ratio;RMR: root mass ratio;R/S: root to shoot mass ratio; RGR_H: relative growth rate in the height (cm d^-1).Defoliation clearly affects biomass allocation of the two species, but not leaf physiology. Considering the carbon or sink limitation, the growth of S. japonica and R. pseudoacacia may be limited by future global climate change scenarios.

W1D0: 75% of field capacity × undefoliation treatment;
W1D1: 75% of field capacity × 50% leaf defoliation treatment;W1D2: 75% of field capacity × 100% leaf defoliation treatment;W2D0: 55% of field capacity × undefoliation treatment;W2D1: 55% of field capacity × 50% leaf defoliation treatment;W2D2: 55% of field capacity × 100% leaf defoliation treatment;W3D0: 35% of field capacity × undefoliation treatment;W3D1: 35% of field capacity × 50% leaf defoliation treatment;W3D2: 35% of field capacity × 100% leaf defoliation treatment;0, 1, 2 and 3: before defoliation, 2 weeks, 5 weeks and 8 weeks after defoliation.A_max: maximum net photosynthetic rate (μmol m⁻² s⁻¹);E: transpiration rate (mmol m^-2 s^-1);Gs: stomatal conductance (mmol m^-2 s^-1);chl a/b: ratio of chlorophyll a and b (mg g^-1);Leaf NSC concentration: leaf non-structural carbohydrates concentration (%);Stem NSC concentration: stem non-structural carbohydrates concentration (%);Root NSC concentration: root non-structural carbohydrates concentration (%);TB: total biomass (g);LB: leaf biomass (g);LMR: leaf mass ratio;SMR: stem mass ratio;RMR: root mass ratio;R/S: root to shoot mass ratio; RGR_H: relative growth rate in the height (cm d^-1).Defoliation clearly affects biomass allocation of the two species, but not leaf physiology. Considering the carbon or sink limitation, the growth of S. japonica and R. pseudoacacia may be limited by future global climate change scenarios.

W1D0: 75% of field capacity × undefoliation treatment;
W1D1: 75% of field capacity × 50% leaf defoliation treatment;W1D2: 75% of field capacity × 100% leaf defoliation treatment;W2D0: 55% of field capacity × undefoliation treatment;W2D1: 55% of field capacity × 50% leaf defoliation treatment;W2D2: 55% of field capacity × 100% leaf defoliation treatment;W3D0: 35% of field capacity × undefoliation treatment;W3D1: 35% of field capacity × 50% leaf defoliation treatment;W3D2: 35% of field capacity × 100% leaf defoliation treatment;0, 1, 2 and 3: before defoliation, 2 weeks, 5 weeks and 8 weeks after defoliation.A_max: maximum net photosynthetic rate (μmol m⁻² s⁻¹);E: transpiration rate (mmol m^-2 s^-1);Gs: stomatal conductance (mmol m^-2 s^-1);chl a/b: ratio of chlorophyll a and b (mg g^-1);Leaf NSC concentration: leaf non-structural carbohydrates concentration (%);Stem NSC concentration: stem non-structural carbohydrates concentration (%);Root NSC concentration: root non-structural carbohydrates concentration (%);TB: total biomass (g);LB: leaf biomass (g);LMR: leaf mass ratio;SMR: stem mass ratio;RMR: root mass ratio;R/S: root to shoot mass ratio; RGR_H: relative growth rate in the height (cm d^-1).Defoliation clearly affects biomass allocation of the two species, but not leaf physiology. Considering the carbon or sink limitation, the growth of S. japonica and R. pseudoacacia may be limited by future global climate change scenarios.

Matrine is one of the major alkaloids extracted from Sophora flavescens Ait of the traditional Chinese medicine, was the main chemical ingredient of compounds of Kushen injection. The Matrine is considered as a promising therapeutic agent for curing nonsmall cell lung cancer (NSCLC), used either alone or combined with chemotherapeutic agents. In the present study, we focused on the possible roles of Matrine exerted on the self-renewal ability of stem-like cells of the NSCLC group, as well as the cytotoxicity of chemotherapeutic agents, in vitro and in vivo. Here we reported that Matrine inhibits cancer stem-like cell (CSC) properties through upregulation of Let-7b and suppression of the Wnt pathway. Overexpression of Let-7b suppressed the ability of tumorsphere formation, decreased Wnt pathway activation through inhibiting its transcriptional activity in lung CSCs. Further studies revealed that Let-7b directly targeted CCND1 and decreased its expression, whereas Matrine increased Let-7b levels and followed by inactivation of the CCND1/Wnt signaling pathway and inhibition of EMT, which was characterized by loss of epithelial markers and acquisition of a mesenchymal phenotype in lung CSCs. What is more, we found that Matrine increased Let-7b level in an endoribonuclease DICER1-dependent manner. And xenografts in nude mice evidenced that Matrine increased the sensitivity of lung CSCs to 5-FU and inhibited the accumulation of CCND1 in tumor tissues induced by 5-FU. Taken together, these data illustrate the role of Let-7b in regulating lung CSCs traits and DICER1/let-7/CCND1 axis in Matrine or in combination with 5-FU intervention of lung CSCs’ expansion, helping to fulfill the anti-cancer action of Matrine.

Matrine is one of the major alkaloids extracted from Sophora flavescens Ait of the traditional Chinese medicine, was the main chemical ingredient of compounds of Kushen injection. The Matrine is considered as a promising therapeutic agent for curing nonsmall cell lung cancer (NSCLC), used either alone or combined with chemotherapeutic agents. In the present study, we focused on the possible roles of Matrine exerted on the self-renewal ability of stem-like cells of the NSCLC group, as well as the cytotoxicity of chemotherapeutic agents, in vitro and in vivo. Here we reported that Matrine inhibits cancer stem-like cell (CSC) properties through upregulation of Let-7b and suppression of the Wnt pathway. Overexpression of Let-7b suppressed the ability of tumorsphere formation, decreased Wnt pathway activation through inhibiting its transcriptional activity in lung CSCs. Further studies revealed that Let-7b directly targeted CCND1 and decreased its expression, whereas Matrine increased Let-7b levels and followed by inactivation of the CCND1/Wnt signaling pathway and inhibition of EMT, which was characterized by loss of epithelial markers and acquisition of a mesenchymal phenotype in lung CSCs. What is more, we found that Matrine increased Let-7b level in an endoribonuclease DICER1-dependent manner. And xenografts in nude mice evidenced that Matrine increased the sensitivity of lung CSCs to 5-FU and inhibited the accumulation of CCND1 in tumor tissues induced by 5-FU. Taken together, these data illustrate the role of Let-7b in regulating lung CSCs traits and DICER1/let-7/CCND1 axis in Matrine or in combination with 5-FU intervention of lung CSCs’ expansion, helping to fulfill the anti-cancer action of Matrine.

: Table S1.â U. virens isolates used for DNA polymorphism analysis. (XLSX 10 kb)

: Table S2. Primers used in this study. (XLSX 10 kb)

: Table S2. Primers used in this study. (XLSX 10 kb)

: Table S1.â U. virens isolates used for DNA polymorphism analysis. (XLSX 10 kb)