Abstract
Low temperature is one of the important environmental factors that affect rice growth and yield. To better understand the japonica rice responses to cold stress, isobaric tags for a relative and absolute quantification (iTRAQ) labeling-based quantitative proteomics approach was used to detected changes in protein levels. Two-week-old seedlings of the cold tolerant rice variety Kongyu131 were treated at 8°C for 24, 48 and 72 h, then the total proteins were extracted from tissues and used for quantitative proteomics analysis. A total of 5082 proteins were detected for quantitative analysis, of which 289 proteins were significantly regulated, consisting of 169 uniquely up-regulated proteins and 125 uniquely down-regulated proteins in cold stress groups relative to the control group. Functional analysis revealed that most of the regulated proteins are involved in photosynthesis, metabolic pathway, biosynthesis of secondary metabolites and carbon metabolism. Western blot analysis showed that protein regulation was consistent with the iTRAQ data. The corresponding genes of 25 regulated proteins were used for quantitative real time PCR analysis, and the results showed that the mRNA level was not always parallel to the corresponding protein level. The importance of our study is that it provides new insights into cold stress responses in rice with respect to proteomics and provides candidate genes for cold-tolerance rice breeding.
Highlights
Rice (Oryza sativa) is one of the most important food crops in the world, feeding about half of the population (Khush 2005)
To identify proteins that responsd to cold stress in Kongyu131 at the proteomic level, two-week-old seedlings grown in soil were subjected to 0, 24, 48 and 72 h of cold stress treatments
The shoot tissues of the treated seedlings were used for quantitative proteomic analysis
Summary
Rice (Oryza sativa) is one of the most important food crops in the world, feeding about half of the population (Khush 2005). It is important to explore genes/proteins that are regulated by low temperature in order to under‐ standing the cold-tolerance mechanism and breeding coldtolerant rice cultivars. The standing variation of cold tolerance gene CTB2 and de novo mutation of CTB4a facilitate cold adaptation of rice cultivation from high altitude to high latitude areas (Li et al 2021); bZIP73Jap in japonica rice cultivars inter‐ acts with bZIP71 to modulates abscisic acid (ABA) levels and reactive oxygen species (ROS) homeostasis for BS Breeding Science Preview enhancing rice tolerance to cold climate (Liu et al 2018); rice OsMADS57 interacts with OsTB1 and both directly target OsWRKY94 and D14 for adaptation to cold (Chen et al 2018). Three genes (LOC_ Os01g55350, LOC_Os01g55510 and LOC_Os01g55560) (Zhang et al 2018) and 67 QTLs (Wang et al 2016), which were identified by genome-wide association analysis (GWAS), were associated with cold tolerance of indica and japonica rice, respectively. In an RNA-seq comparative analysis of cold-stressed post-meiotic anther from coldtolerant and cold-susceptible rice cultivars, a number of ethylene-related transcription factors were found to be putative regulators of cold responses (González-Schain et al 2019)
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