Tang Ding, Zhou Tiean, Pan Weisong, Wang Shimei, Hassan Muhammad Ahmad
College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha 410128, China.
Hunan Engineering Technology Research Center for Cell Mechanics and Functional Analysis, Changsha 410128, China.
Biosensors (Basel). 2025 May 23;15(6):334. doi: 10.3390/bios15060334.
Various high-throughput screening methods have been developed to explore plant phenotypes, primarily at the organ and whole plant levels. There is a need to develop phenomics methods at the cellular level to narrow down the genotype to phenotype gap. This study used double-resonator piezoelectric cytometry biosensors to capture the dynamic changes in mechanical phenotypes of living cells of two rice species, drought-resistant Lvhan No. 1 and drought-sensitive 6527, under PEG6000 drought stress. In rice cells of Lvhan No. 1 and 6527, mechanomics parameters, including cell-generated surface stress (ΔS) and viscoelastic parameters (G', G″, G″/G'), were measured and compared under 5-25% PEG6000. Lvhan No. 1 showed larger viscoelastic but smaller surface stress changes with the same concentration of PEG6000. Moreover, Lvhan No. 1 cells showed better wall-plasma membrane-cytoskeleton continuum structure maintaining ability under drought stress, as proven by transient tension stress (ΔS > 0) and linear G'ΔS, G″ΔS relations at higher 15-25% PEG6000, but not for 6527 cells. Additionally, two distinct defense and drought resistance mechanisms were identified through dynamic G″/G' responses: (i) transient hardening followed by softening recovery under weak drought, and (ii) transient softening followed by hardening recovery under strong drought. The abilities of Lvhan No. 1 cells to both recover from transient hardening to softening and to recover from transient softening to hardening are better than those of 6527 cells. Overall, the dynamic mechanomics phenotypic patterns (ΔS, G', G″, G″/G', G'ΔS, G″ΔS) verified that Lvhan No. 1 has better drought resistance than that of 6527, which is consistent with the field data.
已经开发了各种高通量筛选方法来探索植物表型,主要是在器官和整株植物水平。需要在细胞水平上开发表型组学方法,以缩小基因型与表型之间的差距。本研究使用双谐振器压电细胞术生物传感器,捕获了抗旱品种绿旱1号和干旱敏感品种6527两种水稻活细胞在PEG6000干旱胁迫下机械表型的动态变化。在绿旱1号和6527的水稻细胞中,测量并比较了在5%-25% PEG6000条件下的机械组学参数,包括细胞产生的表面应力(ΔS)和粘弹性参数(G'、G″、G″/G')。在相同浓度的PEG6000下,绿旱1号表现出更大的粘弹性变化,但表面应力变化较小。此外,在干旱胁迫下,绿旱1号细胞表现出更好的细胞壁-质膜-细胞骨架连续结构维持能力,在较高的15%-25% PEG6000浓度下,瞬时张力应力(ΔS>0)以及线性G'ΔS、G″ΔS关系证明了这一点,但6527细胞则不然。此外,通过动态G″/G'响应确定了两种不同的防御和抗旱机制:(i)在轻度干旱下先瞬时硬化然后软化恢复,以及(ii)在重度干旱下先瞬时软化然后硬化恢复。绿旱1号细胞从瞬时硬化恢复到软化以及从瞬时软化恢复到硬化的能力均优于6527细胞。总体而言,动态机械组学表型模式(ΔS、G'、G″、G″/G'、G'ΔS、G″ΔS)证实绿旱1号的抗旱性优于6527,这与田间数据一致。