GreenPhos, a universal method for in-depth measurement of plant phosphoproteomes with high quantitative reproducibility.

Protein phosphorylation regulates a variety of important cellular and physiological processes in plants. In-depth profiling of plant phosphoproteomes has been more technically challenging than that of animal phosphoproteomes. This is largely due to the need to improve protein extraction efficiency from plant cells, which have a dense cell wall, and to minimize sample loss resulting from the stringent sample clean-up steps required for the removal of a large amount of biomolecules interfering with phosphopeptide purification and mass spectrometry analysis. To this end, we developed a method with a streamlined workflow for highly efficient purification of phosphopeptides from tissues of various green organisms including Arabidopsis, rice, tomato, and Chlamydomonas reinhardtii, enabling in-depth identification with high quantitative reproducibility of about 11 000 phosphosites, the greatest depth achieved so far with single liquid chromatography-mass spectrometry (LC-MS) runs operated in a data-dependent acquisition (DDA) mode. The mainstay features of the method are the minimal sample loss achieved through elimination of sample clean-up before protease digestion and of desalting before phosphopeptide enrichment and hence the dramatic increases of time- and cost-effectiveness. The method, named GreenPhos, combined with single-shot LC-MS, enabled in-depth quantitative identification of Arabidopsis phosphoproteins, including differentially phosphorylated spliceosomal proteins, at multiple time points during salt stress and a number of kinase substrate motifs. GreenPhos is expected to serve as a universal method for purification of plant phosphopeptides, which, if samples are further fractionated and analyzed by multiple LC-MS runs, could enable measurement of plant phosphoproteomes with an unprecedented depth using a given mass spectrometry technology.

[Comprehensive Quality Assessment of Soil-Maize Heavy Metals in High Geological Background Area].

The aim of this study was to analyze the pollution degree and causes of soil and agricultural products in high geological background areas and to provide a basis for the safe production of agricultural products and the risk control of soil heavy metals. A total of 36 sets of soil-corn collaborative samples were collected in the farming area of Baolong Town, Wushan County, Chongqing City; the contents of heavy metals (Cd, Hg, Pb, As, and Cr) and soil pH in the soil-maize were analyzed, the pollution degree of heavy metals in the soil-maize was evaluated using the Nemerow comprehensive pollution index method (PN) and comprehensive quality impact index (IICQ). The sources of heavy metals in the soil and the influencing factors of heavy metal excess in corn were also analyzed. The results showed that the average value of soil heavy metal content in the study area was higher than the national and Chongqing soil background values, and the soil heavy metal enrichment effect was obvious. Cd was the main factor of soil-maize exceeding the standard, and the overall exceeding rates of soil and corn Cd were 91.67% and 30.55%, respectively. The evaluation results of the Nemerow comprehensive pollution index showed that the soil was dominated by heavy pollution, accounting for 63.89%. The soil-maize comprehensive quality impact index was dominated by moderate and severe pollution, accounting for 44.44% and 47.22%, respectively. From the perspective of the spatial distribution of heavy metal pollution, corn and soil pollution areas were inconsistent. Soil heavy metal pollution was mainly affected by the Permian and Triassic strata and was related to the secondary enrichment of black rock series and limestone areas. The Cd content of maize was mainly affected by soil pH, and maize was relatively safe under alkaline conditions. It is suggested that the soil in the study area should be divided into risk zones according to the stratum distribution, and the planting structure should be adjusted in the high-risk areas. For the low- and medium-risk areas, it is recommended to strengthen the monitoring of agricultural inputs and reduce the input of heavy metals in the soil. Additionally, we recommend carrying out agronomic regulation in acidic soil areas to improve soil acidification, plant corn varieties with low accumulation of heavy metals, and reduce the risk of agricultural products exceeding the standard.

[Source Analysis of Soil Heavy Metals in Agricultural Land Around the Mining Area Based on APCS-MLR Receptor Model and Geostatistical Method].

In order to identify the source of heavy metals in the soil around a mining area and provide effective suggestions for the prevention and control of regional soil pollution, 118 topsoil samples (0-20 cm) were collected in the northern part of Wuli Township, Qianjiang District, Chongqing. The heavy metal (Cd, Hg, Pb, As, Cr, Cu, Zn, and Ni) contents in the soil and soil pH were analyzed, and the spatial distribution and sources of heavy metals in the soil were studied using the geostatistical method and APCS-MLR receptor model. The results showed that the content of heavy metals in the soil was significantly higher than the background value in Chongqing; there was obvious surface accumulation; and Hg, Pb, Cd, As, and Zn showed strong variation. The proportions of soil Cd, Hg, Pb, As, and Zn exceeding the risk screening values were 47.11%, 6.61%, 4.96%, 5.79%, and 7.44%, respectively, and the proportions of soil Cd, Hg, Pb, and As exceeding the risk control values were 0.83%, 4.13%, 0.83%, and 0.83%; thus, the problem of excessive heavy metals in the soil was significant. Soil Cd, As, Cr, Cu, and Ni were mainly affected by soil parent materials, and their contribution rates to the total soil elements were 77.65%, 68.55%, 71.98%, 90.83%, and 82.19%, respectively. Soil Hg, Pb, and Zn were mainly affected by the mining of mercury mines and lead-zinc mines, with the contribution rates of 86.59%, 88.06%, and 91.34%, respectively. In addition, agricultural activities also affected soil Cd and As contents. It is recommended to strengthen the safety monitoring of agricultural products and agricultural inputs, plant varieties with a low accumulation of heavy metals, reduce the use of livestock manure, and grow non-edible agricultural products in areas that exceed the control value of heavy metal pollution risk.