Wu Hongyang, Zhang He, Li Xinyu, Zhang Yu, Wang Jiankun, Wang Qiang, Wan Yinglang
Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresources, College of Tropical Crops, Hainan University, Haikou, 570228, Hainan, China.
College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing, 100083, China.
Plant Methods. 2022 Feb 10;18(1):17. doi: 10.1186/s13007-022-00850-w.
Layered double hydroxide lactate nanosheets (LDH-lactate-NS) are powerful carriers for delivering macro-molecules into intact plant cells. In the past few years, some studies have been carried out on DNA/RNA transformation and plant disease resistance, but little attention has been paid to these factors during LDH-lactate-NS synthesis and delamination, nor has their relationship to the DNA adsorption capacity or transformation efficiency of plant cells been considered.
Since the temperature during delamination alters particle sizes and zeta potentials of LDH-lactate-NS products, we compared the LDH-lactate-NS stability, DNA adsorption rate and delivery efficiency of fluorescein isothiocyanate isomer I (FITC) of them, found that the LDH-lactate-NS obtained at 25 °C has the best characters for delivering biomolecules into plant cell. To understand the potential side effects and cytotoxicity of LDH-lactate-NS to plants, we compared the root growth rate between the Arabidopsis thaliana seedlings grown in the culture medium with 1-300 μg/mL LDH-lactate-NS and equivalent raw material, Mg(lactate) and Al (lactate). Phenotypic analysis showed LDH in a range of 1-300 μg/mL can enhance the root elongation, whereas the same concentration of raw materials dramatically inhibited root elongation, suggesting the nanocrystallization has a dramatical de-toxic effect to Mg(lactate) and Al (lactate) Since enhancing of root elongation by LDH is an unexpected phenomenon, we further designed experiments to investigate influence of LDH to Arabidopsis seedlings. We further used the gravitropic bending test, qRT-PCR analysis of auxin transport proteins, non-invasive micro-test technology and liquid chromatography-mass spectrometry to investigate the auxin transport and distribution in Arabidopsis root. Results indicated that LDH-lactate-NS affect root growth by increasing the polar auxin transport.
Optimal synthesized LDH-lactate-NS can delivery biomolecules into intact plant cells with high efficiency and low cytotoxity. The working solution of LDH-lactate-NS can promote root elongation via increase the polar auxin transport in Arabidopsis roots.
层状双氢氧化物乳酸纳米片（LDH-乳酸-NS）是将大分子递送至完整植物细胞的有效载体。在过去几年中，已针对DNA/RNA转化和植物抗病性开展了一些研究，但在LDH-乳酸-NS的合成和分层过程中，这些因素很少受到关注，也未考虑它们与植物细胞的DNA吸附能力或转化效率之间的关系。
由于分层过程中的温度会改变LDH-乳酸-NS产物的粒径和zeta电位，我们比较了它们的LDH-乳酸-NS稳定性、DNA吸附率和异硫氰酸荧光素异构体I（FITC）的递送效率，发现25℃下获得的LDH-乳酸-NS具有将生物分子递送至植物细胞的最佳特性。为了解LDH-乳酸-NS对植物的潜在副作用和细胞毒性，我们比较了在含有1-300μg/mL LDH-乳酸-NS及等量原料乳酸镁和乳酸铝的培养基中生长的拟南芥幼苗的根生长速率。表型分析表明，1-300μg/mL范围内的LDH可促进根伸长，而相同浓度的原料则显著抑制根伸长，这表明纳米结晶对乳酸镁和乳酸铝具有显著的解毒作用。由于LDH促进根伸长是一个意外现象，我们进一步设计实验来研究LDH对拟南芥幼苗的影响。我们进一步利用重力弯曲试验、生长素转运蛋白的qRT-PCR分析、非侵入性微测试技术和液相色谱-质谱法来研究生长素在拟南芥根中的转运和分布。结果表明，LDH-乳酸-NS通过增加生长素极性转运来影响根生长。
最佳合成的LDH-乳酸-NS能够以高效率和低细胞毒性将生物分子递送至完整植物细胞。LDH-乳酸-NS工作溶液可通过增加拟南芥根中的生长素极性转运来促进根伸长。
