The amino acid transporter AAP1 mediates growth and grain yield by regulating neutral amino acid uptake and reallocation in Oryza sativa

Abstract Nitrogen (N) is a major element necessary for crop yield. In most plants, organic N is primarily transported in the form of amino acids. Here, we show that amino acid permease 1 (AAP1) functions as a positive regulator of growth and grain yield in rice. We found that the OsAAP1 gene is highly expressed in rice axillary buds, leaves, and young panicles, and that the OsAAP1 protein is localized to both the plasma membrane and the nuclear membrane. Compared with the wild-type ZH11, OsAAP1 overexpression (OE) lines exhibited increased filled grain numbers as a result of enhanced tillering, while RNAi and CRISPR (clustered regularly interspaced short palindromic repeat; Osaap1) knockout lines showed the opposite phenotype. In addition, OsAAP1-OE lines had higher concentrations of neutral and acidic amino acids, but lower concentrations of basic amino acids in the straw. An exogenous treatment with neutral amino acids promoted axillary bud outgrowth more strongly in the OE lines than in the WT, RNAi, or Osaap1 lines. Transcriptome analysis of Osaap1 further demonstrated that OsAAP1 may affect N transport and metabolism, and auxin, cytokinin, and strigolactone signaling in regulating rice tillering. Taken together, these results support that increasing neutral amino acid uptake and reallocation via OsAAP1 could improve growth and grain yield in rice.

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The amino acid transporter OsAAP4 contributes to rice tillering and grain yield by regulating neutral amino acid allocation through two splicing variants

10.21203/rs.3.rs-53363/v2 ◽

2020 ◽

Author(s):

Zhongming Fang ◽

Bowen Wu ◽

Yuanyuan Ji

Keyword(s):

Amino Acid ◽

Grain Yield ◽

Growth And Development ◽

Higher Plants ◽

Neutral Amino Acid ◽

Acid Uptake ◽

Amino Acid Permease ◽

Splicing Variant ◽

Rice Growth ◽

Splicing Variants

Abstract Background: Amino acids, which are transported by amino acid transporters, are the major forms of organic nitrogen utilized by higher plants. Among the 19 Amino Acid Permease transporters (AAPs) in rice, only a small number of these genes have been reported to influence rice growth and development. However, whether other OsAAPs are responsible for rice growth and development is unclear.Results: In this study, we demonstrate that OsAAP4 promoter sequences are divergent between Indica and Japonica, with higher expression in the former, which produces more tillers and higher grain yield than does Japonica. Overexpression of two different splicing variants of OsAAP4 in Japonica ZH11 significantly increased rice tillering and grain yield as result of enhancing the neutral amino acid concentrations of Val, Pro, Thr and Leu. OsAAP4 RNA interference (RNAi) and mutant lines displayed opposite trends compared with overexpresing (OE) lines. In addition, exogenous Val or Pro at 0.5 mM significantly promoted the bud outgrowth of lines overexpressing an OsAAP4a splicing variant compared with ZH11, and exogenous Val or Pro at 2.0 mM significantly enhanced the bud outgrowth of lines overexpressing splicing variant OsAAP4b compared with ZH11. Of note, the results of a protoplast amino acid-uptake assay showed that Val or Pro at different concentrations was specifically transported and accumulated in these overexpressing lines. Transcriptome analysis further demonstrated that OsAAP4 may affect nitrogen transport and metabolism, and auxin, cytokinin signaling in regulating rice tillering.Conclusion: Our results suggested that OsAAP4 contributes to rice tiller and grain yield by regulating neutral amino acid allocation through two different splicing variants and that OsAAP4 might have potential applications in rice breeding.

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The Candida albicans GAP Gene Family Encodes Permeases Involved in General and Specific Amino Acid Uptake and Sensing

Eukaryotic Cell ◽

10.1128/ec.05026-11 ◽

2011 ◽

Vol 10 (9) ◽

pp. 1219-1229 ◽

Cited By ~ 23

Author(s):

Lucie Kraidlova ◽

Griet Van Zeebroeck ◽

Patrick Van Dijck ◽

Hana Sychrová

Keyword(s):

Amino Acids ◽

Candida Albicans ◽

Amino Acid ◽

Fungal Pathogens ◽

Signal Transduction Pathway ◽

Amino Acid Uptake ◽

Acid Uptake ◽

Amino Acid Permease ◽

Content Type ◽

Six Genes

ABSTRACTTheSaccharomyces cerevisiaegeneral amino acid permease Gap1 (ScGap1) not only mediates the uptake of most amino acids but also functions as a receptor for the activation of protein kinase A (PKA). Fungal pathogens can colonize different niches in the host, each containing various levels of different amino acids and sugars. TheCandida albicansgenome contains six genes homologous to theS. cerevisiae GAP1. The expression of these six genes inS. cerevisiaeshowed that the products of all sixC. albicansgenes differ in their transport capacities.C. albicansGap2 (CaGap2) is the true orthologue ofScGap1 as it transports all tested amino acids. The otherCaGap proteins have narrower substrate specificities thoughCaGap1 andCaGap6 transport several structurally unrelated amino acids.CaGap1,CaGap2, andCaGap6 also function as sensors. Upon detecting some amino acids, e.g., methionine, they are involved in a rapid activation of trehalase, a downstream target of PKA. Our data show thatCaGAPgenes can be functionally expressed inS. cerevisiaeand thatCaGap permeases communicate to the intracellular signal transduction pathway similarly toScGap1.

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The amino acid transporter OsAAP4 contributes to rice tillering and grain yield by regulating neutral amino acid allocation through two splicing variants

10.21203/rs.3.rs-53363/v3 ◽

2020 ◽

Author(s):

Zhongming Fang ◽

Bowen Wu ◽

Yuanyuan Ji

Keyword(s):

Amino Acid ◽

Grain Yield ◽

Growth And Development ◽

Higher Plants ◽

Neutral Amino Acid ◽

Acid Uptake ◽

Amino Acid Permease ◽

Splicing Variant ◽

Rice Growth ◽

Splicing Variants

Abstract Background: Amino acids, which are transported by amino acid transporters, are the major forms of organic nitrogen utilized by higher plants. Among the 19 Amino Acid Permease transporters (AAPs) in rice, only a small number of these genes have been reported to influence rice growth and development. However, whether other OsAAPs are responsible for rice growth and development is unclear. Results: In this study, we demonstrate that OsAAP4 promoter sequences are divergent between Indica and Japonica, with higher expression in the former, which produces more tillers and higher grain yield than does Japonica. Overexpression of two different splicing variants of OsAAP4 in Japonica ZH11 significantly increased rice tillering and grain yield as result of enhancing the neutral amino acid concentrations of Val, Pro, Thr and Leu. OsAAP4 RNA interference (RNAi) and mutant lines displayed opposite trends compared with overexpresing (OE) lines. In addition, exogenous Val or Pro at 0.5 mM significantly promoted the bud outgrowth of lines overexpressing an OsAAP4a splicing variant compared with ZH11, and exogenous Val or Pro at 2.0 mM significantly enhanced the bud outgrowth of lines overexpressing splicing variant OsAAP4b compared with ZH11. Of note, the results of a protoplast amino acid-uptake assay showed that Val or Pro at different concentrations was specifically transported and accumulated in these overexpressing lines. Transcriptome analysis further demonstrated that OsAAP4 may affect nitrogen transport and metabolism, and auxin, cytokinin signaling in regulating rice tillering. Conclusion: Our results suggested that OsAAP4 contributes to rice tiller and grain yield by regulating neutral amino acid allocation through two different splicing variants and that OsAAP4 might have potential applications in rice breeding.

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Neutral amino acid influx in developing rabbit blastocysts

AJP Cell Physiology ◽

10.1152/ajpcell.1986.251.2.c285 ◽

1986 ◽

Vol 251 (2) ◽

pp. C285-C292 ◽

Cited By ~ 7

Author(s):

J. E. Bell ◽

K. E. Begg ◽

Y. Sin ◽

J. D. Biggers ◽

D. J. Benos

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Amino Acid Uptake ◽

Neutral Amino Acid ◽

Acid Uptake ◽

Amino Acid Analogue ◽

Independent Components ◽

System A ◽

Acid Analogue ◽

System L

The influx of the neutral amino acids glycine, aminoisobutyric acid (AIB), and leucine into rabbit blastocysts was measured. In day 6 postcoitus (pc) embryos, glycine influx was Na+ independent, whereas AIB and leucine influx involved both Na+-dependent and independent components. From days 5 to 7 pc, the leucine and AIB influx remained constant, although the Na+-dependent fraction decreased and the Na+-independent fraction increased with age. None of the Na+-independent influx was inhibited by methylaminoisobutyric acid (MeAIB), an amino acid analogue specific for the system A of neutral amino acid uptake. In addition, MeAIB influx was Na+ independent, implying that system A is not involved in leucine or AIB uptake. All Na+-dependent influx is thus considered to occur via system ASC. System L contributed only to the influx of leucine at days 6 and 7 pc, as measured by inhibition of Na+-independent influx by 2-amino-bicyclo-(2,2,1)-heptane-2-carboxylic acid.

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Expression study of an Amino Acid Permease-like gene in Phaseolus vulgaris L.

Plant Science Today ◽

10.14719/pst.2020.7.2.712 ◽

2020 ◽

Vol 7 (2) ◽

pp. 251-256

Author(s):

Nisha Patwa ◽

Brototi Chakraborty ◽

Jolly Basak

Keyword(s):

Gene Expression ◽

Amino Acids ◽

Amino Acid ◽

Phaseolus Vulgaris ◽

Critical Role ◽

Amino Acid Uptake ◽

Acid Uptake ◽

Amino Acid Permease ◽

Dependent Relationship ◽

Relationship Of

Amino acid permease-like (AAP-like) gene plays a critical role in absorbing amino acids through roots in plants. A number of studies have been done on amino acids uptake in plants but till date there is no report about the expression of AAP gene in Phaseolus under field allied condition. The aim of this study is to measure the expression of AAP-like gene on alanine, glycine and proline amino acid uptake capacity in Phaseolus vulgaris at field relevant concentrations. Amongst three amino acids, a drastic significant increase of 63.15 fold in expression of AAP-like gene is observed in 50 µM alanine at 2 hr. At 50 µM of proline and 25 µM of alanine, AAP-like gene expression also shows high expression of 43.71 fold at 2 hr and 42.50 fold at 1 hr respectively. This study elucidated the dose dependent relationship of glycine, alanine and proline with the expression of AAP-like gene in amino acid transport in natural conditions in roots of P. vulgaris. Additionally, this research is also useful in identification of plants needing less surplus nitrogen additions and helpful in optimizing fertilizers by tailoring AAP gene expression to match plant uptake capacities in agriculture.

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The Amino Acid Transporter OsAAP4 Contributes to Rice Tillering and Grain Yield by Regulating Neutral Amino Acid Allocation through Two Splicing Variants

Rice ◽

10.1186/s12284-020-00446-9 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Zhongming Fang ◽

Bowen Wu ◽

Yuanyuan Ji

Keyword(s):

Amino Acid ◽

Grain Yield ◽

Growth And Development ◽

Higher Plants ◽

Neutral Amino Acid ◽

Acid Uptake ◽

Amino Acid Permease ◽

Splicing Variant ◽

Rice Growth ◽

Splicing Variants

Abstract Background Amino acids, which are transported by amino acid transporters, are the major forms of organic nitrogen utilized by higher plants. Among the 19 Amino Acid Permease transporters (AAPs) in rice, only a small number of these genes have been reported to influence rice growth and development. However, whether other OsAAPs are responsible for rice growth and development is unclear. Results In this study, we demonstrate that OsAAP4 promoter sequences are divergent between Indica and Japonica, with higher expression in the former, which produces more tillers and higher grain yield than does Japonica. Overexpression of two different splicing variants of OsAAP4 in Japonica ZH11 significantly increased rice tillering and grain yield as result of enhancing the neutral amino acid concentrations of Val, Pro, Thr and Leu. OsAAP4 RNA interference (RNAi) and mutant lines displayed opposite trends compared with overexpresing (OE) lines. In addition, exogenous Val or Pro at 0.5 mM significantly promoted the bud outgrowth of lines overexpressing an OsAAP4a splicing variant compared with ZH11, and exogenous Val or Pro at 2.0 mM significantly enhanced the bud outgrowth of lines overexpressing splicing variant OsAAP4b compared with ZH11. Of note, the results of a protoplast amino acid-uptake assay showed that Val or Pro at different concentrations was specifically transported and accumulated in these overexpressing lines. Transcriptome analysis further demonstrated that OsAAP4 may affect nitrogen transport and metabolism, and auxin, cytokinin signaling in regulating rice tillering. Conclusion Our results suggested that OsAAP4 contributes to rice tiller and grain yield by regulating neutral amino acid allocation through two different splicing variants and that OsAAP4 might have potential applications in rice breeding.

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Gamma-Glutamyl-Amino Acids as Signals for the Hormonal Regulation of Amino Acid Uptake by the Mammary Gland of the Lactating Rat

Neonatology ◽

10.1159/000242178 ◽

1985 ◽

Vol 48 (4) ◽

pp. 250-256 ◽

Cited By ~ 8

Author(s):

Juan R. Viña ◽

Inmaculada R. Puertes ◽

Juan B. Montoro ◽

Guillermo T. Saez ◽

José Viña

Keyword(s):

Amino Acids ◽

Mammary Gland ◽

Amino Acid ◽

Hormonal Regulation ◽

Amino Acid Uptake ◽

Acid Uptake ◽

Gamma Glutamyl

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Action of growth hormone in vitro on the net uptake and incorporation into protein of amino acids in muscle from intact rabbits given protein-deficient diets

British Journal Of Nutrition ◽

10.1079/bjn19760003 ◽

1976 ◽

Vol 35 (1) ◽

pp. 1-10 ◽

Cited By ~ 1

Author(s):

M. R. Turner ◽

P. J. Reeds ◽

K. A. Munday

Keyword(s):

Amino Acids ◽

Growth Hormone ◽

Protein Synthesis ◽

Amino Acid ◽

De Novo ◽

Amino Acid Uptake ◽

Acid Uptake ◽

Amino Acid Incorporation ◽

Acid Incorporation

1. Net amino acid uptake, and incorporation into protein have been measured in vitro in the presence and absence of porcine growth hormone (GH) in muscle from intact rabbits fed for 5 d on low-protein (LP), protein-free (PF) or control diets.2. In muscle from control and LP animals GH had no effect on the net amino acid uptake but stimulated amino acid incorporation into protein, although this response was less in LP animals than in control animals.3. In muscle from PF animals, GH stimulated both amino acid incorporation into protein and the net amino acid uptake, a type of response which also occurs in hypophysectomized animals. The magnitude of the effect of GH on the incorporation of amino acids into protein was reduced in muscle from PF animals.4. The effect of GH on the net amino acid uptake in PF animals was completely blocked by cycloheximide; the uptake effect of GH in these animals was dependent therefore on de novo protein synthesis.5. It is proposed that in the adult the role of growth hormone in protein metabolism is to sustain cellular protein synthesis when there is a decrease in the level of substrate amino acids, similar to that which occurs during a short-term fast or when the dietary protein intake is inadequate.

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The uptake of amino acids by mouse cells (strain LS) during growth in batch culture and chemostat culture: the influence of cell growth rate

Proceedings of the Royal Society of London Series B Biological Sciences ◽

10.1098/rspb.1967.0073 ◽

1967 ◽

Vol 168 (1013) ◽

pp. 421-438 ◽

Cited By ~ 46

Keyword(s):

Amino Acids ◽

Growth Rate ◽

Amino Acid ◽

Cell Growth ◽

Glutamic Acid ◽

Batch Culture ◽

Amino Acid Uptake ◽

Essential Amino Acids ◽

Growth Yields ◽

Acid Uptake

The uptake of thirteen essential amino acids by mouse LS cells in suspension culture was determined by bacteriological assay methods. Chemostat continuous-flow cultures were used to determine the effect of different cell growth rates on the quantitative amino acid requirements for growth. The growth yields of the cells ( Y = g cell dry weight produced/g amino acid utilized) were calculated for each of the essential amino acids. A mixture of the non-essential amino acids, serine, alanine and glycine increased the cell yield from the essential amino acids. The growth yields from nearly all the essential amino acids in batch culture were increased when glutamic acid was substituted for the glutamine in the medium. The growth yields from the amino acids in batch culture were much less at the beginning than at the end of the culture. The highest efficiencies of conversion of amino acids to cell material were obtained by chemostat culture. When glutamic acid largely replaced the glutamine in the medium the conversion of amino acid nitrogen to cell nitrogen was 100 % efficient (that is, the theoretical yield was obtained) at the optimum growth rate (cell doubling time, 43 h). The maximum population density a given amino acid mixture will support can be calculated from the data. It is concluded that in several routinely used tissue culture media the cell growth is limited by the amino acid supply. In batch culture glutamine was wasted by (1) its spontaneous decomposition to pyrrolidone carboxylic acid and ammonia, and (2) its enzymic breakdown to glutamic acid and ammonia, but also glutamine was used less efficiently than glutamic acid. Study of the influence of cell growth rate on amino acid uptake rates per unit mass of cells indicated that a marked change in amino acid metabolism occurred at a specific growth rate of 0.4 day -1 (cell doubling time, 43 h). With decrease in specific growth rate below 0.4 day -1 there was a marked stimulation of amino acid uptake rate per cell and essential amino acids were consumed increasingly for functions other than synthesis of cell material.

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Action of insulin and growth hormone on protein synthesis in muscle from non-hypophysectomized rabbits

Biochemical Journal ◽

10.1042/bj1250515 ◽

1971 ◽

Vol 125 (2) ◽

pp. 515-520 ◽

Cited By ~ 13

Author(s):

P. J. Reeds ◽

K. A. Munday ◽

M. R. Turner

Keyword(s):

Amino Acids ◽

Growth Hormone ◽

Protein Synthesis ◽

Amino Acid ◽

Incubation Medium ◽

Amino Acid Uptake ◽

Diaphragm Muscle ◽

Acid Uptake

The separate effects of insulin and growth hormone on the uptake and incorporation of five amino acids into diaphragm muscle from non-hypophysectomized rabbits has been examined. Both growth hormone and insulin, when present in the medium separately, stimulated the incorporation into protein of the amino acids, leucine, arginine, valine, lysine and histidine. Insulin also stimulated amino acid uptake, but growth hormone did not. When insulin and growth hormone were present in the incubation medium together, the uptake and incorporation of valine, the only amino acid studied under these conditions, tended to be greater than the sum of the separate effects of the two hormones.

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