研究发现,在水培条件下,lsi1突变体根部和地上部的DMA积累量较野生型降低约50%,对DMA的耐受性显著增强,表明Lsi1参与DMA的吸收;而lsi2突变体DMA积累量与野生型无显著差异,表明Lsi2不参与DMA的吸收。硅添加显著降低野生型与lsi2地上部DMA浓度,减轻DNA损伤,促进生物量积累;但在硅吸收能力受限的lsi1突变体中,硅的保护效应较弱。
Figure 1 Arsenic and Si concentration in plants of ZH11, lsi1 and lsi2 treated with different DMA concentrations and Si for 6 days.
TUNEL凋亡检测显示,100 μM DMA处理下,野生型叶片细胞核呈现明显的DNA损伤荧光信号,lsi2信号较弱,lsi1几乎无信号;300 μM DMA时,三种基因型均出现严重DNA损伤。硅添加显著减轻野生型与lsi2的DNA损伤荧光强度,但对lsi1无显著改善作用。
Figure 2 TUNEL assay in rice leaf of ZH11, lsi1 and lsi2 treated with different DMA concentrations and Si for 6 days.
盆栽试验中,DMA处理导致lsi1与lsi2突变体结实率下降60%以上,穗部呈典型直穗病症状。硅添加显著提高lsi2突变体的籽粒产量与抗病性,但对lsi1突变体改善有限。
Figure 3 Effects of DMA and Si treatment on phenotype and agronomic traits of ZH11, lsi1 and lsi2 mutants.
砷形态分析表明,籽粒与颖壳中砷以DMA为主要存在形态。DMA处理下,lsi1与lsi2籽粒DMA浓度分别为野生型的3.2倍和3.5倍。然而,单株总砷积累量却显著低于野生型(降低3.8–4.1倍),表明DMA浓度升高主要源于生物量大幅下降引发的浓缩效应。硅添加显著降低各基因型籽粒DMA浓度。
Figure 4 Effects of DMA and Si treatment on biomass and As accumulation in brown rice (per plant) of ZH11, lsi1 and lsi2 mutants.