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Sheepgrass [Leymus chinensis (Trin.) Tzvel] is a valuable forage plant highly significant to the grassland productivity of Euro-Asia steppes. Growth of above-ground tissues of L. chinensis is the major component contributing to the grass yield. Although it is generally known that this species is sensitive to ecosystem disturbance and adverse environments, detailed information of how L. chinensis coping with various nutrient deficiency especially phosphate deprivation (-Pi) is still limited. Here, we investigated impact of Pi-deprivation on shoot growth and biomass accumulation as well as photosynthetic properties of L. chinensis. Growth inhibition of Pi-deprived seedlings was most obvious and reduction of biomass accumulation and net photosynthetic rate (Pn) was 55.3 and 63.3%, respectively, compared to the control plants grown under Pi-repleted condition. Also, we compared these characters with seedlings subjected to low-Pi stress condition. Pi-deprivation caused 18.5 and 12.3% more reduction of biomass and Pn relative to low-Pi-stressed seedlings, respectively. Further analysis of in vivo chlorophyll fluorescence and thylakoid membrane protein complexes using 2D-BN/SDS-PAGE combined with immunoblot detection demonstrated that among the measured photosynthetic parameters, decrease of ATP synthase activity was most pronounced in Pi-deprived plants. Together with less extent of lipid peroxidation of the thylakoid membranes and increased ROS scavenger enzyme activities in the leaves of Pi-deprived seedlings, we suggest that the decreased activity of ATP synthase in their thylakoids is the major cause of the greater reduction of photosynthetic efficiency than that of low-Pi stressed plants, leading to the least shoot growth and biomass production in L. chinensis.

Sheepgrass [Leymus chinensis (Trin.) Tzvel] is a valuble forage plant highly significant to regional grassland productivity of Euro-Asia steppes. Although effects of environmental stress including drought have been studied, impact of nutrient deficiency in particular phosphate (Pi), one of the essential macronutrient, is not reported. Here, we investigated low-Pi effect on its photosynthetic apparatus via physiological and biochemical analysis. We show that PSII activity was significantly reduced based on chlorophyll fluorescence measurements. We observed decreased amount of core proteins of PSII by immunoblot analysis. Further analysis of thylakoid membranes using 2D-BN/SDS-PAGE and immunoblot detection demonstrated that the amount of PSII complexes was closely correlated with the Pi levels within the range. Together with reduced number of thylakoid grana stackings observed, we suggest that the maintenance of PSII is impaired under Pi-limited condition. Moreover, enzyme activity assays revealed that the activity of several ROS scavenger enzymes was stimulated by low-Pi treatment. Based on these experimental results we conclude that PSII is the component of photosynthetic apparatus most sensitive to Pi supply and the enhanced anti-ROS activity is mainly subjective to protection of PSII against low Pi-induced photo-oxidative stress in the organism.

Global climate warming has a crucial impact on many terrestrial ecosystems, including temperate steppe. In addition, phosphate deficiency is known to be the common deficiency in soils worldwide due to the low availability of the phosphate nutrient in the form of inorganic phosphate anions (Pi). Consequently, in the future, land plants are likely to simultaneously encounter heat stress and phosphate deficiency more frequently. Sheepgrass 〔Leymus chinensis (Trin.) Tzvel〕is a dominant perennial forage plant highly significant to grass productivity of Eurasian temperate grasslands. Though effects of environmental stress including drought and Pi starvation have been studied, the combined eff ;ects of phosphate deficiency and heat stress on plant physiology remain largely unclear. Here, we investigated the combined eff ;ects of heat stress and phosphate deficiency on above-ground tissue growth and photochemical properties of L. chinensis using in vivo chlorophyll fluorescence spectroscopy. We observed remarkable phenotypic alterations of reduced shoot growth and considerable leaf chlorosis in L. chinensis seedlings under the combined stress condition. Also, we compared changes in photochemical activity between the control and the corresponding stressed seedlings. Based on chlorophyll fluorescence analysis, impairment of PSI was more severe than that of PSII in the seedlings treated with the combined stress. Compared to the control, PSI and PSII activity decreased up to 35.5% and 30%, respectively, under the combined-stress condition. Moreover, our data show that the decreased photosynthetic activity is not the sum of the single-stressed conditions. These results combined with the distinction of other photochemical parameters indicate that a complex interaction between Pi-deficiency and heat stress may exist in the forage plant.