Understanding plant responses to phosphorus starvation for improvement of plant tolerance to phosphorus deficiency by biotechnological approaches

Abstract

In both prokaryotes and eukaryotes, including plants, phosphorus (P) is an essential nutrient that is involved in various biochemical processes, such as lipid metabolism and the biosynthesis of nucleic acids and cell membranes. P also contributes to cellular signaling cascades by function as mediators of signal transduction and it also serves as a vital energy source for a wide range of biological functions. Due to its intensive use in agriculture, P resources have become limited. Therefore, it is critically important in the future to develop scientific strategies that aim to increase P use efficiency and P recycling. In addition, the biologically available soluble form of P for uptake (phosphate; Pi) is readily washed out of topsoil layers, resulting in serious environmental pollution. In addition to this environmental concern, the wash out of Pi from topsoil necessitates a continuous Pi supply to maintain adequate levels of fertilization, making the situation worse. As a coping mechanism to P stress, plants are known to undergo drastic cellular changes in metabolism, physiology, hormonal balance and gene expression. Understanding these molecular, physiological and biochemical responses developed by plants will play a vital role in improving agronomic practices, resource conservation and environmental protection as well as serving as a foundation for the development of biotechnological strategies, which aim to improve P use efficiency in crops. In this review, we will discuss a variety of plant responses to low P conditions and various molecular mechanisms that regulate these responses. In addition, we also discuss the implication of this knowledge for the development of plant biotechnological applications.     

The responses of wild-type and ABA mutant Arabidopsis thaliana plants to phosphorus starvation

The growth patterns of plants subjected to phosphorus starvation resemble those caused by treatment with ABA, suggesting that ABA could mediate the response of the plant to phosphorus starvation. We examined the role of ABA in phosphorus stress by comparing growth and biochemical responses of Arabidopsis thaliana ABA mutants aba-1 and abi2-1 to those of wild-type plants. We first characterized acid phosphatase production of wild-type Arabidopsis in response to phosphorus starvation. We found that several acid phosphatase isozymes are present in roots and shoots, but only a subset of these isozymes are induced by phosphorus stress, and they are induced in both organs. Production of acid phosphatase in response to phosphorus stress was not affected by the aba-1 or abi2-1 mutations. Low phosphorus also resulted in decreased growth of both wild-type and ABA mutant plants, and the root-to-shoot ratio was increased in both wild type and mutants. Anthocyanins accumulated in response to phosphorus stress in both wild-type and mutant plants, but the increase was reduced in the aba-1 mutant. Thus, two different ABA mutants responded normally in most respects to phosphorus stress. Our data do not support a major role for ABA in coordinating the phosphorus-stress response.     

缺磷条件下的小麦根系酸性磷酸酶活性研究

1　引　　言植物根可向根际分泌许多有机化合物 ,其中有许多物质都能促进植物对矿质养分的吸收 .作为必需大量营养元素的Ｐ ,在土壤中以无机磷酸盐阴离子的形式被吸收 ,而有机磷酸酯必须被水解成无机Ｐ后才能进入植物根 ,在这一过程中有一非常重要的步骤 ,就是由微生物、菌根外真菌和植物根分泌酸性磷酸酶 .土壤中的有机Ｐ一般占全Ｐ的 30 %～ 5 0 % ,有的可达95 % .因此 ,如何发挥植物自身利用土壤有机Ｐ的潜力已成为目前植物营养学研究的热点之一 .Ｇｏｌｄｓｔｅｉｎ等[3 ] 研究Ｐ胁迫条件下悬浮培养细胞时发现 ,抑制植物生长和诱导酸性…     

Root surface acid phosphatase activities of vascular epiphytes of a Costa Rican rain forest

The objective of this work was to compare root surface phosphatase activities of vascular epiphytes typical of a lowland tropical forest. Acid phophatase, measured at pH 5.0 with the substrate p-nitrophenyl phosphate, was detected in 22 species distributed within 10 plant families. Epiphytes were classified as trunk, canopy-mat or bare-limb species based upon their usual occurrence. Phosphatase activity was not significantly correlated with plant occurrence. However, phosphatase activity was generally highest in trunk occurring and canopy-mat epiphytes rooted in mosses and humus-like accumulations, and lowest in species restricted to bare limbs. Epiphyte shoot phosphorus and chlorophyll content were correlated with species occurrence, with phosphatase being positively correlated with plant P content. The observed changes in acid phosphatase production among habitats were consistent with predicted changes in the availability of organic P sources. However, observed changes also reflect accompanying shifts in root structure including: the occurrence of velamen or waxy layers, changes in root diameter, branching and root hair density.     

Functional significance of dauciform roots: exudation of carboxylates and acid phosphatase under phosphorus deficiency in Caustis blakei (Cyperaceae)

Caustis blakei produces an intriguing morphological adaptation by inducing dauciform roots in response to phosphorus (P) deficiency. We tested the hypothesis that these hairy, swollen lateral roots play a similar role to cluster roots in the exudation of organic chelators and ectoenzymes known to aid the chemical mobilization of sparingly available soil nutrients, such as P. Dauciform-root development and exudate composition (carboxylates and acid phosphatase activity) were analysed in C. blakei plants grown in nutrient solution under P-starved conditions. The distribution of dauciform roots in the field was determined in relation to soil profile depth and matrix. The percentage of dauciform roots of the entire root mass was greatest at the lowest P concentration ([P]) in solution, and was suppressed with increasing solution [P], while in the field dauciform roots were predominantely located in the upper soil horizons, and decreased with increasing soil depth. Citrate was the major carboxylate released in an exudative burst from mature dauciform roots, which also produced elevated levels of acid phosphatase activity. Malonate was the dominant internal carboxylate present, with the highest concentration in young dauciform roots. The high concentration of carboxylates and phosphatases released from dauciform roots, combined with their prolific distribution in the organic surface layer of nutrient-impoverished soils, provides an ecophysiological advantage for enhancing nutrient acquisition.     

Secreted acid phosphatase is expressed in cluster roots of lupin in response to phosphorus deficiency

The roots of white lupin (Lupinus albus L. cv. Kievskij mutant) secrete acid phosphatase, S-APase, when they grow under conditions of low available phosphorus (P). S-APases hydrolyze organic phosphate compounds in the rhizosphere and supply inorganic phosphate to the plants. Low phosphorus availability also induces vigorous growth of cluster roots. In this study, the function of cluster roots was investigated with reference to S-APase secretion. White lupins were grown in hydroponic culture in a greenhouse under P-deficient and P-sufficient conditions. S-APase in the excised roots after treatment was detected by staining with 4-methylumbelliferone phosphate (MUP). Gene expression of S-APase in cluster and normal roots was also investigated. Activity was greatest in the roots of plants grown under conditions of P -deficiency, particularly in cluster roots. S-APase gene expression was induced by a decrease in internal P concentrations, and was especially high in cluster roots formed under conditions of P -deficiency. It was suggested that decrease of internal P concentration stimulated both of the S-APase expression and cluster root formation.     

模拟氮沉降对低磷胁迫下马尾松不同家系根系分泌和磷效率的影响

近年来大气氮(N)沉降的增加, 导致森林土壤中有效N含量增加、N:P发生改变, 研究N沉降对低磷(P)胁迫下林木根系分泌和P效率的影响具有重要意义。该文以马尾松(Pinus massoniana)家系作为试验材料, 设置模拟N沉降与同质低P (介质表层与深层均缺P)、异质低P (介质表层P丰富、深层缺P)耦合的二年生盆栽实验, 系统研究了模拟N沉降对低P胁迫下马尾松根系分泌性酸性磷酸酶(APase)活性、有机酸分泌以及P效率的影响。结果表明: (1)同质低P和异质低P下, 模拟N沉降均显著提高了植株N:P化学计量比、增加了P素的相对匮乏程度, 从而诱导根系增加了APase和有机酸的分泌, 而同质低P比异质低P下增加幅度更大, 其中有机酸分泌均与马尾松生长呈正相关关系, 而APase活性与P效率相关性较小; (2)同质低P下, N沉降虽然增加了根系分泌, 但未提高马尾松P素吸收和生长量, 其原因在于, 同质低P下植株N:P过高, 因而植株对N沉降敏感性低; 在异质低P下, 植株表现为N、P共同限制, 因而对N敏感性较高, N沉降增加了根系分泌, 同时提高了N和P吸收效率、增加了生物量; (3)马尾松根系分泌对模拟N沉降的响应存在较大的家系差异。同质低P下, 家系71×20的有机酸分泌和生物量对N沉降的响应幅度较大; 异质低P下, 家系36×29、71×20和73×23对N沉降的响应幅度较大。     

Effect of cycloheximide and renewal of phosphorus supply on surface acid phosphatase activity of phosphorus deficient tomato roots

Changes in surface phosphatase activity of tomato root ( Lycopersicon esculentum L. cv. Marmande) have been studied in relation to its P status. Experiments were performed either with excised roots cultured in vitro or with entire plants (split root method) grown in various conditions (P deficiency, renewal of P supply). In some experiments different parts of the root were separated according to their P status during their primary growth.
The surface phosphatase in different root parts depends first of all on P status during their primary growth. Moreover the cell wall phosphatase of tomato root is stable in vivo . Thus the changes in the surface phosphatase of a root system result mainly from the growth of new roots, which bear an enzyme activity that is either high if they are P deficient or low if they are P provided. The control of the cell wall phosphatase synthesis appears to be a repression-derepression process mediated by the root cell concentration in some P compound, probably orthophosphate. Cycloheximide stops or alters the growth of excised roots, so that this inhibitor was found unsuitable to study the synthesis of cell wall phosphatase under in vivo conditions. Split root experiment shows that the increase in surface phosphatase activity may occur locally, i.e. only in the parts of the root system which are P deficient. Agricultural and ecological aspects of these findings are pointed out.     

Properties of acid phosphatase from scutella of germinating maize seeds

One acid phosphatase (optimum pH at 5.4) was purified from maize scutellum after 96 hr of germination. The purified enzyme was homogeneous on polyacrylamide gel electrophoresis (PAGE) with or without sodium dodecyl sulfate (SDS). The enzyme has a MW of 65 000 ± 4000 as determined by Sephadex G-200 gel filtration and SDS-PAGE. The enzyme contained 16% neutral sugars, and cations are not required for activity. The purified enzyme was not inactivated by DTNB at pH 8. The hydrolysis of glucose-6-phosphate in the presence of 4 mM fluoride and 4 mm EDTA, at pH 6.7 (optimum pH), seems to be catalysed by this acid phosphatase.     

氮添加对草地生态系统土壤pH、磷含量和磷酸酶活性的影响

磷是植物必需的大量营养元素之一,也是草地生态系统功能的重要限制因子。近年来,随着全球氮沉降的迅速增加,草地生态系统土壤磷及磷酸酶活性受到不同程度的影响。本研究采用整合分析方法,分析了草地的氮添加量、氮源种类、持续时间和土层深度等对土壤pH、全磷(TP)含量、有效磷(AP)含量、碱性磷酸酶(AlP)和酸性磷酸酶(AcP)活性的影响,以及土壤pH与磷酸酶活性的相关性。结果表明: 氮添加降低了土壤pH、TP含量和AlP活性,提高了土壤AcP活性,但对土壤AP含量无显著影响。从氮添加量来看,土壤pH、AlP显著降低在氮添加>5 g·m^-2·a^-1条件下即可发生;高水平氮添加(>10 g·m^-2·a^-1)导致AcP活性显著提高;土壤TP、AP含量显著降低仅发生在氮添加量为5～10 g·m^-2·a^-1条件下。硝酸铵处理显著降低了土壤TP含量,提高了AcP活性;尿素处理显著降低了土壤pH和AlP活性。在所有添加量下,当试验持续时间为3～10年时,土壤TP含量、AlP活性显著降低;持续时间大于3年时,土壤pH显著降低;>10年时,AcP活性显著提高。0～10 cm土层的AP含量显著升高,TP含量和AlP活性显著降低;大于10 cm土层中AP含量显著降低。土壤pH与土壤AcP活性呈显著负相关,表明氮添加引起的土壤pH改变可能是土壤磷酸酶活性变化的重要原因。     

The role of acid phosphatases in plant phosphorus metabolism

Hydrolysis of phosphate esters is a critical process in the energy metabolism and metabolic regulation of plant cells. This review summarizes the characteristics and putative roles of plant acid phosphatase (APase). Although immunologically closely related, plant APases display remarkable heterogeneity with regards to their kinetic and molecular properties, and subcellular location. The secreted APases of roots and cell cultures are relatively non-specific enzymes that appear to be important in the hydrolysis and mobilization of P_i from extracellular phosphomonoesters for plant nutrition. Intracellular APases are undoubtedly involved in the routine utilization of P_i reserves or other P_i-containing compounds. A special class of intracellular APase exists that demonstrate a clear-cut (but generally nonabsolute) substrate selectivity. These APases are hypothesized to have distinct metabolic functions and include: phytase, phosphoglycolate phosphatase, 3-phosphoglycerate phosphatase, phosphoenolpyruvate phosphatase, and phosphotyrosyl-protein phosphatase. APase expression is regulated by a variety of developmental and environmental factors. P_i starvation induces de novo synthesis of extra- and intracellular APases in cell cultures as well as in whole plants. Recommendations are made to achieve uniformity in the analyses of the different APase isoforms normally encountered within and between different plant tissues.     

Localization of acid phosphatase activity in maize root under phosphorus deficiency

The effect of phosphorus deficiency on acid phosphatase activity in the apical, middle and basal parts of the root of maize plants was followed. The supernatant obtained by centrifuging the homogenate of plant tissue at 1500 ×g was further centrifuged at 18 000 ×g, the sediment being marked as fraction II and the supernatant as fraction III. The results obtained document the fact that acid phosphatase activity of the two fractions of all analyzed root segments was higher in plants cultured in nutrient medium without phosphate than in those containing phosphorus in nutrient medium. In most cases this difference was significant to highly significant. The results of experiments proved unambiguously a higher enzymatic activity in all root segments in fraction III than in fraction II. In fraction III the highest acid phosphatase activity was found in the apical part, in fraction II in the basal part of the root.     

Induction of RNase and nuclease activity in cultured maize endosperm cells following sucrose starvation

The activity of RNases and nucleases in plants often increases following exposure to many types of stress, including prolonged exposure to dark or phosphate starvation. In cereals, the activity of RNases and nucleases is also regulated developmentally during late seed development. In this study, we investigated the effect that the absence of sugar or phosphate in culture medium has on the activity of RNases and nucleases expressed in maize endosperm suspension cells. Withdrawal of sugar from the culture medium resulted in a substantial increase in RNase and nuclease activities, whereas deprivation of phosphate during the same period of growth had no detectable effect on either of these activities. The increase in RNase activity was limited to the neutral RNases, demonstrating that the effect of sugar starvation is specific to one class of RNase. Elimination of asparagine from the medium resulted in a transient reduction in nuclease but not in RNase activity. These observations suggest that sugar starvation constitutes a stress to which maize endosperm responds, in part, by increasing neutral RNase and nuclease activity.     

Kinetic properties of acid phosphatase from scutella of germinating maize seeds

Acid phosphatase purified from maize scutellum showed a kinetic transition from negative cooperativity at pH 5.4, to Michaelian behavior at pH 6.7. The     

氮、磷添加对不同林型土壤磷酸酶活性的影响

研究了鼎湖山3种森林类型(南亚热带季风常绿阔叶林、马尾松人工林和针叶阔叶混交林)的土壤酸性磷酸单酯酶活性(APA)对施肥的响应情况。在3种林型中分别设置对照、加氮(150 kg N hm-2a-1)、加磷(150 kg P hm-2a-1)以及N和P同时添加(150 kg N hm-2a-1+150 kg P hm-2a-1)4种不同处理。结果表明,季风林土壤APA((15.83±2.46)μmol g-1h-1)显著高于混交林((10.71±0.78)μmol g-1h-1)和马尾松林((9.12±0.38)μmol g-1h-1),且3种林型土壤APA与土壤有效磷含量均呈显著负相关。施加N肥显著提高了季风林土壤APA,而对混交林和马尾松林的作用不显著。施加P肥显著降低了混交林和马尾松林土壤APA,但对季风林的影响不明显。N和P同时添加仅显著降低了马尾松林土壤APA,但在季风林中存在交互作用。因此,N沉降会加剧亚热带成熟林土壤P的限制,可以考虑施加P肥作为森林管理的一种方式来缓解这种限制作用。     

Identification of the Pseudornonas aeruginosa acid phosphatase as a phosphorylcholine phosphatase activity

Summary Choline, betaine and N,N-dimethylglycine as the sole carbon and nitrogen source induced a periplasmic acid phosphatase activity in Pseudomonas aeruginosa. This enzyme produced the highest rates of hydrolysis in phosphorylcholine and phosphorylethanolamine among the various phosphoric esters tested. At saturating concentrations of Mg²⁺, the K_m values were 0.2 and 0.7 mM for phosphorylcholine and phosphorylethanolamine respectively. At high concentrations both compounds were inhibitors of the enzyme activity. The K _inf1 ^sups values for phosphorylcholine and phosphorylethanolamine were 1.0 and 3.0 mM respectively. The higher catalytic efficiency was that of phosphorylcholine. Considering these results it is possible to suggest that the Pseudomonas aeruginosa acid phosphatase is a phosphorylcholine phosphatase. The existence of this activity which is induced jointly with phospholipase C by different choline metabolites, in a high phosphate medium, suggests that the attack of Pseudomonas aeruginosa on the cell host may also be produced under conditions of high phosphate concentrations, when the alkaline phosphatase is absent.     

Response of tomato plants to chilling stress in association with nutrient or phosphorus starvation

The experiments were conducted on two tomato cultivars: Garbo and Robin. Mineral starvation due to plant growth in 20-fold diluted nutrient solution (DNS) combined with chilling reduced the rate of photosynthesis (P _N) and stomatal conductance (g) to a greater extent than in plants grown in full nutrient solution (FNS). In phosphate-starved tomato plants the P _N rate and stomatal conductance decreased more after chilling than in plants grown on FNS. In low-P plants even 2 days after chilling the recovery of CO₂ assimilation rate and stomatal conductance was low. A resupply of phosphorus to low-P plants (low P + P) did not improve the rate of photosynthesis in non-chilled plants (NCh) but prevented P_N inhibition in chilled (Ch) plants. The greatest effect of P resupply was expressed as a better recovery of photosynthesis and stomatal conductance, especially in non-chilled low P + P plants. The F _v/F _m (ratio of variable to maximal chlorophyll fluorescence) decreased more during P starvation than as an effect of chilling. Supplying phosphorus to low-P plants caused the slight increase in the F _v/F _mratio. In conclusion, after a short-term chilling in darkness a much more drastic inhibition of photosynthesis was observed in nutrient-starved or P-insufficient tomato plants than in plants from FNS. This inhibition was caused by the decrease in both photochemical efficiency of photosystems and the reduction of stomatal conductance. The presented results support the hypothesis that tomato plants with limited supply of mineral nutrients or phosphorus are more susceptible to chilling.     

Induction of PPi-dependent phosphofructokinase by phosphate starvation in seedlings of Brassica nigra

Activity of PPi-dependent phosphofructokinase (PFP) was monitored in Brassica nigra seedlings grown under nutrient-sufficient or phosphate (Pi)-starved conditions. Roots, stems and leaves of 50 d Pi-deficient seedlings displayed 4.0-, 3.7- and 2.3-fold greater PFP activity, respectively, than did nutrient sufficient controls. This induction was based primarily upon an increased susceptibility of PFP from the Pi-starved tissues to activation by fructose-2,6-bisphosphate. The ratio of PFP to ATP-dependent phosphofructokinase (PFK) was approximately 2:1 and 1:1 in the various organs of 50 d Pi-starved and Pi-fed plants, respectively. Immunoblots probed with anti-(potato PFP) immune serum revealed that the induction of PFP in Pi-starved B. nigra was coincident with an elevation in the amount of PFP α-subunit in the leaves as well as an increase in the α:β subunit ratio in the stems and roots. Induction of PFP in the various tissues was also accompanied by an appreciable decline in intracellular Pi level, decreased soluble protein content, and elevated phosphoenolpyruvate phosphatase activity. Time course studies revealed that these responses to Pi stress were significantly delayed in the leaves as compared to the roots and stems suggesting that Pi may be preferentially sequestered to the leaves during Pi starvation. These data also provide further evidence that B. nigra PFP is an adaptive enzyme that may function during Pi deprivation as: (1) a glycolytic bypass to PFK; and (2) a ‘Pi-recycling system’ that converts esterified-P to Pi that would be rapidly reassimilated into the metabolism of the Pi-deficient cells.