# Determining the Critical Period of Continuous Waterlogging in Maize: An Analysis of Physiological, Biochemical, and Transcriptomic Traits

## TL;DR

This study identifies 6 days as the critical threshold for waterlogging stress during maize's jointing stage, beyond which yield drops significantly due to impaired energy synthesis.

## Contribution

The study determines the critical duration of waterlogging stress in maize and reveals physiological and transcriptomic mechanisms affecting photosynthesis and energy synthesis.

## Key findings

- Grain weight significantly decreased by 17.49% to 100.00% when waterlogging lasted more than 4 days.
- Transcriptomic analysis showed downregulation of photosynthetic and energy-related genes after 8-10 days of waterlogging.
- Starch content remained stable up to 6 days of waterlogging but dropped significantly afterward.

## Abstract

Waterlogging stress severely limits crop photosynthesis and energy supplies, resulting in significant yield reductions. However, the critical duration of waterlogging stress during the maize jointing stage remains unclear, and the physiological and molecular mechanisms underlying its effects on photosynthetic efficiency and energy synthesis in maize require further investigation. In this study, we systematically analyzed the responses of physiological traits, transcriptomic profiles, and the yield formation in maize (Zea mays L.) to varying waterlogging durations imposed during the jointing stage, including 0 days (CK), 2 days (F2), 4 days (F4), 6 days (F6), 8 days (F8), and 10 days (F10). Our results indicate that the (1) grain weight (GW) showed no significant difference between F2 and CK. However, the GW in F4, F6, F8, and F10 decreased significantly by 17.49%, 26.45%, 60.24%, and 100.00%, respectively, compared to the CK. (2) Compared with the CK, the malondialdehyde content progressively increased from F4 to F10, while antioxidant enzyme activity gradually decreased. The chlorophyll content declined by 29.93% to 57.38%, and net photosynthetic efficiency decreased by 13.82% to 38.93%. Although the leaf sucrose content in from F4 to F10 gradually decreased, the leaf starch content remained stable in F4 and F6. In contrast, the starch content in F8 and F10 leaves was significantly reduced by 37.55% and 47.60%, respectively, compared with CK. (3) A transcriptomic analysis revealed that during from F2 to F4, genes encoding photosystem I subunit protein, such as PSAD, and the cytochrome b6f complex proteingene PETC were downregulated. At F6, these key genes encoding photosynthetic proteins were upregulated. However, at F8 and F10, their expression was significantly downregulated. Concurrently, genes related to ATP synthesis (e.g., ATPD) as well as starch and sucrose metabolism (e.g., SPP2, SS1) were also downregulated. In summary, when waterlogging stress persists for no longer than 6 days, plants can maintain their starch content to supply energy for growth, thereby ensuring basic developmental needs. When waterlogging persists for more than 6 days, energy synthesis is impaired, and the nutrient transport to the grains is significantly inhibited, ultimately resulting in a substantial reduction in yield. Therefore, 6 days of waterlogging can be considered the critical threshold for significant yield loss in maize during the jointing stage.