Pectin Methylesterases Enhance Root Cell Wall Phosphorus Remobilization in Rice

doi:10.1016/j.rsci.2022.01.006

Abstract

Abstract:

Pectin contributes greatly to cell wall phosphorus (P) remobilization. However, it is currently unclear whether the methylesterification degree of the pectin, which is related to the activity of pectin methylesterases (PMEs), is also involved in this process. Here, we demonstrated that elevated PME activity can facilitate the remobilization of P deposited in the cell wall. P-deficient conditions resulted in the reduction of root cell wall P content. This reduction was more pronounced in Nipponbare than in Kasalath, in company with a significant increment of the PME activity, indicating a possible relationship between elevated PME activity and cell wall P remobilization. This hypothesis was supported by in vitro experiments, as pectin with lower methylesterification degree had higher ability to release inorganic P (Pi) from insoluble FePO₄. Furthermore, among the 35 OsPME members in rice, only the expression of OsPME14 showed a relationship with PME activity. In addition, transgenic rice lines overexpressing OsPME14 had increased PME activity, released more P from the root cell wall, and more resistant to P deficiency. In conclusion, PMEs enhance P remobilization in P-starved rice by increasing PME activity in Nipponbare, which in turn helps to remobilize P from the cell wall, and thus makes more available P.

Key words: cell wall, phosphorus, pectin methylesterase, remobilization, rice

Wu Qi, Tao Ye, Zhang Xiaolong, Dong Xiaoying, Xia Jixing, Shen Renfang, Zhu Xiaofang. Pectin Methylesterases Enhance Root Cell Wall Phosphorus Remobilization in Rice[J]. Rice Science, 2022, 29(2): 179-188.

Figures/Tables 5

Fig. 1. OsPME14 is responsible for root cell wall P remobilization in rice variety Nipponbare and Kasalath under P-sufficient (+P) and P-deficient (-P) conditions. A and B, Root cell wall P content (A) and pectin methylesterase (PME) activity (B). PME activity refers to units of orange peel PME as defined by the manufacturer (Sigma, USA). Seedlings (2-week-old) were exposed to P-sufficient or P-deficient nutrient solution for one week. Data are Mean ± SD (n = 4). Columns with different lowercase letters are significantly different at P < 0.05. C and D, Quantitative PCR analysis of the expression of OsPME genes in Nipponbare (C) and Kasalath (D). Seedlings (2-week-old) were exposed to P-sufficient or P-deficient nutrient solution for one week. OsActin was used as an internal control. Data are Mean ± SD (n = 4). Columns with asterisks are significantly different at P < 0.05. E and F, Quantitative PCR analysis of the expression of OsPME14 (E) and OsPME31 (F). Seedlings (2-week-old) were exposed to P-sufficient or P-deficient nutrient solution for one week. OsActin was used as an internal control. nd, Not detected. Data are Mean ± SD (n = 4)

Table 1. OsPME genes were classified according to their expression patterns in Nipponbare and Kasalath.

Class	Gene
A ^a	OsPME4, OsPME5, OsPME8, OsPME9, OsPME13, OsPME19, OsPME20, OsPME21, OsPME23, OsPME24, OsPME26, OsPME28, OsPME30, OsPME32, OsPME33
B^b	OsPME14, OsPME31
C^c	OsPME1, OsPME2, OsPME3, OsPME6, OsPME7, OsPME10, OsPME11, OsPME12, OsPME15, OsPME16, OsPME17, OsPME18, OsPME22, OsPME25, OsPME27, OsPME29, OsPME34, OsPME35

Fig. 2. OsPME14 is necessary for root pectin methylesterase (PME) activity-mediated remobilization of root cell wall P under P-sufficient (+P) and P-deficient (-P) conditions. A and B, Expression of OsPME14 (A) and PME activity (B) in Nipponbare and two independent overexpressing OsPME14 lines OsPME14-OX in the presence or absence of P supply. Seedlings (2-week-old) were exposed to P-sufficient or P-deficient nutrient solution for one week. OsActin was used as an internal control. C and D, Root soluble P concentration (C) and cell wall P concentration (D) in Nipponbare and two independent overexpressing OsPME14 lines OsPME14-OX in the presence or absence of P supply. E and F, PME activity correlates with root soluble P concentration (E) and cell wall P concentration (F) under P deficiency. Two independent experiments (Exp1 and Exp2) were done with 7-d P-deficient treatment, and after that, total roots were subjected to P content analysis, PME activity measurement and cell-wall extraction. In A to D, data are Mean ± SD (n = 4). Columns with different lowercase letters are significantly different at P < 0.05.

Fig. 3. Perls/diaminobenzidine staining in Nipponbare and Kasalath seedlings for one week (A), and Fe concentration in root cell wall (B) in Nip and two independent overexpressing OsPME14 lines OsPME14-OX under P-sufficient (+P) and P-deficient (-P) conditions. Data in B are Mean ± SD (n = 4). Columns with different lowercase letters are significantly different at P < 0.05.

Fig. 4. Proposed working model for OsPME14 regulated cell wall P remobilization in P-deficient condition. P, Phosphoru; PME, Pectin methylesterases; ME, Methylesterase.

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