Phenylalanine ammonia‑lyase family is closely associated with response to phosphate deficiency in rice

고윤실,김상진,정기홍 한국유전학회 2020 Genes & Genomics Vol.42 No.1

Background Phenylalanine ammonia-lyase (PAL) catalyzes the first step in the biosynthetic phenylpropanoid pathway (PPP) via deamination of phenylalanine to trans-cinnamic acid, a precursor for the lignin and flavonoid biosynthetic pathways. Although its role is well-established in various plants, the functional significance of PAL genes in rice remains poorly understood. Objective This study aims to find out the global feature of rice PAL genes associated with phosphate use efficiency. Methods To identify the biological functions of individual rice PAL genes, we performed meta-expression profiling analysis based on phylogenomics of rice PAL genes and confirmed the expression patterns using Quantitative real-time PCR (qPCR). Results We identified nine genes that were remarkably up-regulated during long-term phosphate (Pi) starvation and recovery processes through RNA-Seq data analysis. Expression patterns of the nine rice PAL genes under Pi starvation were further confirmed by qPCR, indicating that the function of PAL genes is strongly associated with Pi starvation response in rice. Conclusion Our study reports the functional significance of rice PAL genes involved in adaptation to low Pi growth conditions and provides useful information to improve Pi use efficiency in crop plant.

Production and Characterization of Phenylalanine Ammonia-lyase from Rhodotorula aurantiaca K-505

Cho, Dae-Haeng,Chae, Hee-Jeong,Kim, Eui-Yong The Korean Society of Food Science and Nutrition 1997 Preventive Nutrition and Food Science Vol.2 No.4

Optimal cultivation conditions for the production of phenylalanine ammonia-lyase(PAL) from Rhodotorula aurantiaca K-505 were selected, and the kinetic parameters of the produced PAL were determined. The most suitable carbon and nitrogen sources were glucose and tryptone, respectively. The strain expressed PAL constituttively when using the optimized semi-complex media. High cell density culture could be critical for maximal production of PAl since the PAL ynthesis was growth associated. maximum PAL activity was observed at initial pH 6.0. although the ll growth was not markedly affected by temperature between 22 and 28$^{\circ}C$, the cells yielded the maximum PAL activity when cultivated at 22$^{\circ}C$. The maximum activity for deamination of L-phenylalnine to trans-cinnamic acid was observed around pH 8.8. The PAL activity gave the maximum at 45$^{\circ}C$, and greatly decreased at higher than 5$0^{\circ}C$. Activation energy({TEX}$E_{a}${/TEX}) calculated from Arrhenius equation was 6.28 kcal/mol in the range of 22$^{\circ}C$ to 4$0^{\circ}C$. A oolf plot showed that the enzyme reaction follows Michaelis-Menten equation, whose {TEX}$K_{M}${/TEX} and {TEX}$V_{max}${/TEX} values were 4.65$\times${TEX}$10^{-3}${/TEX} M and 0.89$\mu$ mol/mg-min respectively.

Research Note : Cloning and Expression Analysis of Phenylalanine Ammonia-Lyase Gene in the Mycelium and Fruit Body of the Edible Mushroom Flammulina velutipes

( Yeo Hong Yun ),( Ja Sun Koo ),( Seong Hwan Kim ),( Won Sik Kong ) 한국균학회 2015 Mycobiology Vol.43 No.3

Phenylalanine ammonia-lyase (PAL) gene is known to be expressed in plants, and is involved in the differentiation, growth and synthesis of secondary metabolites. However, its expression in fungi remains to be explored. To understand its expression in mushroom fungi, the PAL gene of the edible mushroom Flammulina velutipes (Fvpal) was cloned and characterized. The cloned Fvpal consists of 2,175 bp, coding for a polypeptide containing 724 amino acids and having 11 introns. The translated amino acid sequence of Fvpal shares a high identity (66%) with that of ectomycorrhizal fungus Tricholoma matsutake. Distinctively, the Fvpal expression in the mycelium was higher in minimal medium supplemented with L-tyrosine than with other aromatic amino acids. During cultivation of the mushroom on sawdust medium, Fvpal expression in the fruit body correspondingly increased as the mushroom grew. In the fruiting body, Fvpal was expressed more in the stipe than in the pileus. These results suggest that F. velutipes PAL activity differs in the different organs of the mushroom. Overall, this is first report to show that the PAL gene expression is associated with mushroom growth in fungi.

Phenylalanine Ammonia Lyase and Cinnamic Acid 4-Hydroxylase Activities of Rice and Pepper in response to UV and Wounding

Kim, Mi-Young,Yoon, Yong-Hwi,Lee, Jung-Hoon,Kim, Hak-Yoon,Shin, Dong-Hyun,Lee, In-Jung,Kim, Dal-Ung,Kim, Kil-Ung Korean Society of Life Science 2002 생명과학회지 Vol.12 No.3

Phenylpropanoid pathway 생성물질은 식물의 self-defense에 관계하며 이러한 물질들은 UV뿐 아니라 wounding, pathogen과 같은 environmental stress에 의해 생성되는 것으로 알려져 있다. 벼에서 PAL mRNA 는 UV 조사 후 12시간에서 48시간까지는 증가하였으나 48시간부터 60시간까지는 점점 줄어드는 경향을 보였다. 한편 PAL의 활성은 UV조사 후 24시간에서 가장 높았지만 상처에 의해서는 PAL의 활성이 벼에서는 증가하지 않았다. 그러나 고추에서는 UV조사와 상처를 준 후 24시간과 10시간에서 각각 높은 활성을 나타내었다. 벼와 고추 모두 cinnamic acid 4-hydroxylase의 활성은 상처를 준 후 12시간에서 증가하였지만 UV 조사는 C4H 활성에 영향을 주지 않았다. 이러한 결과로 볼 때 벼와 고추에서는 UV 조사와 상처가 모두 PAL, C4H 효소활성에 영향을 주는 것으로 나타났다. The metabolites related to phenylpropanoid pathway play an important role in the self-defense of plants and induced by environmental stress like wounding, pathogen attack, UV-irradiation and so on. The mRNA level of rite phenylalanine ammonia lyase (PAL) was increased at 12 h to 48 h, however it was gradually decreased 48 h to 60 h after UV irradiation. The PAL enzyme activities in rice were peaked at the time of 24 h after UV irradiation, on the other hand, it was not affected by wounding. The PAL enzyme activities in pepper were raised high at 24 h and 10 h by UV irradiation and wounding respectively. The cinnamic acid 4-hydroxylase (C4H) activities were increased by wounding treatment and were detected from 12 h to end time point of experiment, while UV-irradiation didn't affect the C4H activity in rice and pepper. These results were assumed that the action of isoflavonid has an alternative effect on the defenses which include wounding and UV irradiation and on the diverse roles in rice and hot pepper.

Sulfhydryl-Related and Phenylpropanoid-Synthesizing Enzymes in Arabidopsis thaliana Leaves After Treatments with Hydrogen Peroxide, Heavy Metals, and Glyphosate

Lim, Chang-Jin,Park, Keum-Nam,Sa, Jae-Hoon The Korea Science and Technology Center 1999 BMB Reports Vol.32 No.2

Three-week grown Arabidopsis thaliana leaves were wounded by cutting whole leaves with a razor blade into pieces (about 3mm ×3mm), submerged in various solutions, and incubated in a growth chamber for 24h. We measured and compared activities of several enzymes such as phenylalanine ammonia-lyase (PAL), tyrosine ammonia-lyase (TAL), thioredoxin, thioredoxin reductase, thioltransferase, glutathione reductasem and NADP+-malate dehydrogenase. PAL activity was decrease in HgCl²-, CdCl²-, and glyphosate-treated leaf slices, and could not be detected after treatment with CdCl². TAL activity was found to be maximal in the CdCl²- treated leaf slices. Activity of thioredoxin, a small protein known as a cofactor of ribonucleotide reductase and a regulator of photosynthesis, was significantly increased in the CdCl²-treated leaf slices, while thioredoxin reductase activity was maximal in th HgCl²-treated leaf slices. Thioltransferase and glutathione reductase activities were significantly decreased in the HgCl²-treated leaf slices. NADP+-malate dehydrogenase activity remained relatively constant after the chemical treatments. Our results strongly indicate that sulfhydryl-related and phenylpropanoid-synthesizing enzyme activities are affected by chemical treatments such as hydrogen peroxide, heavy metals, and glyphosate.

Cyclized Induction of Phenylalanine Ammonia-Lyase Gene Expression in Rhizoctonia solani-Infected Stems of Tomato

Yeo, Yun-Soo,Kim, Soo-Jin,Koo, Bon-Sung,Lee, Churl-Ho,Lee, Shin-Woo The Korean Society of Plant Biotechnology 2004 Plant molecular biology and biotechnology research Vol.6 No.3

Soil-borne fungal pathogens such as Verticillium and Rhizoctonia can colonize in the stem tissue of plant through root and lead to wilting symptoms of plant by blocking. water transportation. During the colonization of Rhizoctonia solani in the vascular tissue of tomato stems, particularly, phenylalanine ammonia-lyase (PAL) gene induction pattern was cyclized showing peak induction at two different time points (10 and 80 h) after fungal spores inoculation in vivo. In leaves or roots, however, no such cycling pattern was observed. The first induction peak may be due to an initial sporulation events leading to a second induction peak by a proliferation of fungal spores to the upper stems or other tissues from an initial spore trapping sites. Tomato PAL gene was also dramatically induced by wounding, light illumination and mercury chloride treatment but was not cyclized. Mercury chloride showed the earliest induction with all tissues even at half an hour after treatment.

Development of Modified Phenylalanine Ammonia-Lyase for the Treatment of Phenylketonuria

( Woo Mi Kim ) 한국응용약물학회 2009 Biomolecules & Therapeutics(구 응용약물학회지) Vol.17 No.1

Sulfhydryl-Related and Phenylpropanoid-Synthesizing Enzymes in Arabidopsis thaliana Leaves after Treatments with Hydrogen Peroxide, Heavy Metals, and Glyphosate

Park, Keum-Nam,Sa, Jae-Hoon,Lim, Chang-Jin Korean Society for Biochemistry and Molecular Biol 1999 Journal of biochemistry and molecular biology Vol.32 No.2

Three-week grown Arabidopsis thaliana leaves were wounded by cutting whole leaves with a razor blade into pieces (about$3\;mm\;{\times}\;3\;mm$) submerged in various solutions, and incubated in a growth chamber for 24 h. We measured and compared activities of several enzymes such as phenylalanine ammonia-lyase (PAL), tyrosine ammonia-lyase (TAL), thioredoxin, thioredoxin reductase, thioltransferase, glutathione reductase, and $NADP^+$ -malate dehydrogenase. PAL activity was decreased in $HgCl_2$-, $CdCl_2$-, and glyphosate-treated leaf slices, and could not be detected after treatment with $CdCl_2$. TAL activity was found to be maximal in the $CdCl_2$-treated leaf slices. Activity of thioredoxin, a small protein known as a cofactor of ribonucleotide reductase and a regulator of photosynthesis, was significantly increased in the $CdCl_2$-treated leaf slices, while thioredoxin reductase activity was maximal in the $HgCl_2$-treated leaf slices. Thioltransferase and glutathione reductase activities were significantly decreased in the $HgCl_2$-treated leaf slices. $NADP^+$ -malate dehydrogenase activity remained relatively constant after the chemical treatments. Our results strongly indicate that sulfhydryl-related and phenylpropanoid-synthesizing enzyme activities are affected by chemical treatments such as hydrogen peroxide, heavy metals, and glyphosate.

Crosstalk of Zn in Combination with Other Fertilizers Underpins Interactive Effects and Induces Resistance in Tomato Plant against Early Blight Disease

Awan, Zoia Arshad,Shoaib, Amna,Khan, Kashif Ali The Korean Society of Plant Pathology 2019 Plant Pathology Journal Vol.35 No.4

The present study was undertaken to evaluate the integrated effect of zinc (Zn) with other nutrients in managing early blight (EB) disease in tomato. A pot experiment was carried out with basal application of the recommended level of macronutrients [nitrogen, phosphorus and potassium (NPK)] and micronutrients [magnesium (Mg) and boron (B)] in bilateral combination with Zn (2.5 and 5.0 mg/kg) in a completely randomized deigned in replicates. Results revealed that interactive effect of Zn with Mg or B was often futile and in some cases synergistic. Zn with NPK yield synergistic outcome, therefore EB disease was managed significantly (disease incidence: 25% and percent severity index: 13%), which resulted in an efficient signaling network that reciprocally controls nutrient acquisition and uses with improved growth and development in a tomato plant. Thus, crosstalk and convergence of mechanisms in metabolic pathways resulted in induction of resistance in tomato plant against a pathogen which significantly improved photosynthetic pigment, total phenolics, total protein content and defense-related enzymes [superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL)]. The tremendous increase in total phenolics and PAL activity suggesting their additive effect on salicylic acid which may help the plant to systemically induce resistance against pathogen attack. It was concluded that interactive effect of Zn (5.0 mg/kg) with NPK significantly managed EB disease and showed positive effect on growth, physiological and biochemical attributes therefor use of Zn + NPK is simple and credible efforts to combat Alternaria stress in tomato plants.

Crosstalk of Zn in Combination with Other Fertilizers Underpins Interactive Effects and Induces Resistance in Tomato Plant against Early Blight Disease

Zoia Arshad Awan,Amna Shoaib,Kashif Ali Khan 한국식물병리학회 2019 Plant Pathology Journal Vol.35 No.4

The present study was undertaken to evaluate the integrated effect of zinc (Zn) with other nutrients in managing early blight (EB) disease in tomato. A pot experiment was carried out with basal application of the recommended level of macronutrients [nitrogen, phosphorus and potassium (NPK)] and micronutrients [magnesium (Mg) and boron (B)] in bilateral combination with Zn (2.5 and 5.0 mg/kg) in a completely randomized deigned in replicates. Results revealed that interactive effect of Zn with Mg or B was often futile and in some cases synergistic. Zn with NPK yield synergistic outcome, therefore EB disease was managed significantly (disease incidence: 25% and percent severity index: 13%), which resulted in an efficient signaling network that reciprocally controls nutrient acquisition and uses with improved growth and development in a tomato plant. Thus, crosstalk and convergence of mechanisms in metabolic pathways resulted in induction of resistance in tomato plant against a pathogen which significantly improved photosynthetic pigment, total phenolics, total protein content and defense-related enzymes [superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL)]. The tremendous increase in total phenolics and PAL activity suggesting their additive effect on salicylic acid which may help the plant to systemically induce resistance against pathogen attack. It was concluded that interactive effect of Zn (5.0 mg/kg) with NPK significantly managed EB disease and showed positive effect on growth, physiological and biochemical attributes therefor use of Zn + NPK is simple and credible efforts to combat Alternaria stress in tomato plants.