Cloning and expression analysis of phenylalanine ammonia-lyase (PAL) gene family and cinnamate 4-hydroxylase (C4H) from Dryopteris fragrans

Biologia ◽  
2015 ◽  
Vol 70 (5) ◽  
Author(s):  
Yan Li ◽  
Lili Sun ◽  
Hemeng Wang ◽  
Rui Gao ◽  
Junzheng Zhang ◽  
...  

AbstractPhenylalanine ammonia-lyase (PAL) and cinnamate 4-hydroxylase (C4H) are the first and second key enzymes of the phenylpropanoid pathway. Systematic analysis of the DfPAL gene family and DfC4H have not been performed in Dryopteris fragrans (L.) Schott. To date, PAL and C4H genes have been less extensively studied in monilophytes than in angiosperms. Here we report the identification of threeDfPAL and DfC4H fragments using cDNA cloning and sequencing. Bioinformatics and phylogenetic analyses showed that DfPAL1 and DfPAL2 were quite similar at the amino acid level (94.88%), whereas DfPAL3 was relatively low similar to both of the other paralogs. Some important functional domains were conserved in three DfPAL and DfC4H genes. DfPAL3 and DfC4H were highly expressed in gametophytes and petioles of D. fragrans, DfPAL1 had the highest expression in petioles, and DfPAL2 had low expression in leaves and petioles. Only DfPAL2 and DfC4H were induced with 4°C, 35°C, and UV treatments, but the time responses were different. These results suggest complexity of the DfPAL- and DfC4H-associated metabolic network in D. fragrans. The results provide a basis for elucidating the role of DfPAL and DfC4H genes in the biosynthesis of bioactive compounds.

2021 ◽  
Vol 22 (24) ◽  
pp. 13366
Author(s):  
Xuechun Wang ◽  
Nan Chao ◽  
Aijing Zhang ◽  
Jiaqi Kang ◽  
Xiangning Jiang ◽  
...  

Caffeoyl shikimate esterase (CSE) hydrolyzes caffeoyl shikimate into caffeate and shikimate in the phenylpropanoid pathway. In this study, we performed a systematic analysis of the CSE gene family and investigated the possible roles of CSE and CSE-like genes in Populus. We conducted a genome-wide analysis of the CSE gene family, including functional and phylogenetic analyses of CSE and CSE-like genes, using the poplar (Populus trichocarpa) genome. Eighteen CSE and CSE-like genes were identified in the Populus genome, and five phylogenetic groups were identified from phylogenetic analysis. CSEs in Group Ia, which were proposed as bona fide CSEs, have probably been lost in most monocots except Oryza sativa. Primary functional classification showed that PoptrCSE1 and PoptrCSE2 had putative function in lignin biosynthesis. In addition, PoptrCSE2, along with PoptrCSE12, might also respond to stress with a function in cell wall biosynthesis. Enzymatic assay of PoptoCSE1 (Populus tomentosa), -2 and -12 showed that PoptoCSE1 and -2 maintained CSE activity. PoptoCSE1 and 2 had similar biochemical properties, tissue expression patterns and subcellular localization. Most of the PoptrCSE-like genes are homologs of AtMAGL (monoacylglycerol lipase) genes in Arabidopsis and may function as MAG lipase in poplar. Our study provides a systematic understanding of this novel gene family and suggests the function of CSE in monolignol biosynthesis in Populus.


2021 ◽  
Author(s):  
Nan Chao ◽  
Qi Qi ◽  
Xue-Chun Wang ◽  
Shuang Li ◽  
Xiang-Ning Jiang ◽  
...  

Abstract Caffeoyl shikimate esterase (CSE) hydrolyzes caffeoyl shikimate into caffeate and shikimate in the phenylpropanoid pathway. In this study, we performed systematic analysis of CSE gene family in poplar and investigated the possible roles of CSEs and CSE-like genes in Populus. We performed a genome-wide analysis of the CSE family, including functional and phylogenetic analyses of CSE and CSE-like genes using the poplar (Populus trichocarpa) genome. Eighteen CSE and CSE-like genes were identified in the Populus genome and five phylogenetic groups were identified from phylogenetic analysis. CSEs in Group Ia, which were proposed as bona fide CSEs, have probably been lost in most monocots except Oryza sativa. Primary functional classification showed that PoptrCSE1 and PoptrCSE2 had putative function in lignin biosynthesis. In addition, PoptrCSE2, along with PoptrCSE12 might also respond to stress with a function in cell wall biosynthesis. Enzymatic assay of Populus tomentosa (Popto) CSE1, -2 and -12 showed that PoptoCSE1 and -2 kept CSE activity. PoptoCSE1 and 2 had similar biochemical properties, tissue expression pattern and subcellular localization. Most of the PoptrCSE-like genes are homologs of AtMAGL (monoacylglycerol lipase) genes in Arabidopsis and may function as MAG lipase in poplar. Our study provides systematic understanding of this novel gene family and suggests the CSE function in monolignol biosynthesis in Populus.


2020 ◽  
Author(s):  
Kazutaka Katoh ◽  
Naoyuki Iwabe ◽  
Takashi Miyata

AbstractDNA polymerase δ (polδ) is one of the major DNA polymerases that replicate chromosomal genomes in eukaryotes. Given the essential role of this protein, its phylogenetic tree was expected to reflect the relationship between taxa, like many other essential proteins. However, the tree of the catalytic subunit of polδ showed an unexpectedly strong heterogeneity among vertebrate lineages in evolutionary rate at the amino acid level, suggesting unusual amino acid substitutions specifically in the ancestral mammalian lineage. Structural and phylogenetic analyses were used to pinpoint where and when these amino acid substitutions occurred: around the 3′-5′ exonuclease domain in later mammal ancestry, after the split between monotremes and therians. The 3′-5′ exonuclease domain of this protein is known to have an impact on the fidelity of replication. Based on these observations, we explored the possibility that the amino acid substitutions we identified in polδ affected the mutation rate of entire chromosomal genomes in this time period.


2019 ◽  
Vol 117 (1) ◽  
pp. 271-277 ◽  
Author(s):  
Jun He ◽  
Yuqiang Liu ◽  
Dingyang Yuan ◽  
Meijuan Duan ◽  
Yanling Liu ◽  
...  

Brown planthopper (BPH) is one of the most destructive insects affecting rice (Oryza sativaL.) production. Phenylalanine ammonia-lyase (PAL) is a key enzyme involved in plant defense against pathogens, but the role of PAL in insect resistance is still poorly understood. Here we show that expression of the majority ofPALsin rice is significantly induced by BPH feeding. Knockdown of OsPALssignificantly reduces BPH resistance, whereas overexpression ofOsPAL8in a susceptible rice cultivar significantly enhances its BPH resistance. We found thatOsPALsmediate resistance to BPH by regulating the biosynthesis and accumulation of salicylic acid and lignin. Furthermore, we show that expression ofOsPAL6andOsPAL8in response to BPH attack is directly up-regulated by OsMYB30, an R2R3 MYB transcription factor. Taken together, our results demonstrate that the phenylpropanoid pathway plays an important role in BPH resistance response, and provide valuable targets for genetic improvement of BPH resistance in rice.


Author(s):  
Wei Ren ◽  
Yingzhe Wang ◽  
Ankai Xu ◽  
Yunge Zhao

Phenylalanine ammonia-lyase (PAL) catalyzes the rate-limiting step of phenylpropanoid biosynthesis in plants and supplies precursors for a variety of secondary metabolites, such as flavonoids, lignins and stilbenes. The first draft of the full Medicago truncatula genome assembly has been released. However it is observed that, the PAL gene family from Medicago truncatula (MtPAL genes) has not been characterized in detail. In this study, a comprehensive analysis of the Medicago truncatula PAL gene family is presented, including chromosomal locations, phylogenetic analyses, gene structures, three-dimensional (3D) structures and expression patterns. Six Medicago truncatula PAL genes that encode PAL proteins were identified in the Medicago truncatula genome. It was shown that MtPAL genes are distributed on four chromosomes. Dynamic expression patterns of MtPAL genes were observed in different tissues and abiotic stresses, suggesting that MtPAL genes may play important roles in the regulation of development and stress responses in Medicago truncatula.


2021 ◽  
Author(s):  
Abdullah ◽  
Sahar Faraji ◽  
Furrukh Mehmood ◽  
Hafiz Muhammad Talha Malik ◽  
Ibrar Ahmed ◽  
...  

AbstractThe gibberellic acid-stimulated Arabidopsis (GASA/GAST) gene family is widely distributed in plants. The role of the GASA gene family has been reported previously in various physiological and biological processes, such as cell division, root and seed development, stem growth, and fruit ripening. These genes also provide resistance to abiotic and biotic stresses including antimicrobial, antiviral, and antifungal. Here, we report 17 tcGASA genes in Theobroma cacao L. distributed on six chromosomes. The gene structure, promoter-region sequences, protein structure, and biochemical properties, expression, and phylogenetics of all tcGASAs were analyzed. Phylogenetic analyses divided tcGASA proteins into five groups. The nine segmentally duplicating genes form four pairs and cluster together in phylogenetic tree. Purifying selection pressure was recorded on tcGASA, including duplicated genes. Several stress/hormone-responsive cis-regulatory elements were also recognized in the promoter region of tcGASAs. Differential expression analyses revealed that most of the tcGASA genes showed elevated expression in the seeds (cacao food), implying their role in seed development. The black rod disease of genus Phytophthora caused up to 20–25% loss (700,000 metric tons) in world cacao production. The role of tcGASA genes in conferring fungal resistance was also explored based on RNAseq data against Phytophthora megakarya. The differential expression of tcGASA genes was recorded between the tolerant and susceptible cultivars of cacao plants, which were inoculated with the fungus for 24h and 72h. This differential expression indicating possible role of tcGASA genes to fungal resistant in cacao. Our findings provide new insight into the function, evolution, and regulatory system of the GASA family genes in T. cacao and provide new target genes for development of fungi-resistant cacao varieties in breeding programs.


2020 ◽  
pp. 97-110
Author(s):  
E. N. Mikhailova ◽  
V. A. Telegina

The article is devoted to the study of evaluative tools used in modern French media in order to form the media image of a representative of the political elite. The techniques used in the creation of a memorial media portrait of Jacques Chirac (1932—2019), President of France from 1995 to 2007 are considered. The research material was the most prestigious French print media of various political orientations, published in late September — early October 2019 in connection with the death of the ex-President of the French Republic. The relevance of the research topic is dictated by the close attention of modern linguistics to axiological phenomena, differently presented in different types of discursive practices. The novelty of the study is due to the appeal to the analysis of the complex of evaluation tools used in the French print media when characterizing the former leader of the state during the nation’s farewell period. The estimated potential of the title of the article and its influence on the formation of the estimated vector of the entire text of the publication are shown. A systematic analysis of the assessment expression means, reflected in the memorial media portrait of the politician, is given. The factors that influenced the peculiarities of their use in this type of media portrait are revealed.


1998 ◽  
Vol 38 (12) ◽  
pp. 51-56 ◽  
Author(s):  
K. Henshilwood ◽  
J. Green ◽  
D. N. Lees

This study investigates human enteric virus contamination of a shellfish harvesting area. Samples were analysed over a 14-month period for Small Round Structured Viruses (SRSVs) using a previously developed nested RT-PCR. A clear seasonal difference was observed with the largest numbers of positive samples obtained during the winter period (October to March). This data concurs with the known winter association of gastroenteric illness due to oyster consumption in the UK and also with the majority of the outbreaks associated with shellfish harvested from this area during the study period. RT-PCR positive amplicons were further characterised by cloning and sequencing. Sequence analysis of the positive samples identified eleven SRSV strains, of both Genogroup I and Genogroup II, occurring throughout the study period. Many shellfish samples contained a mixture of strains with a few samples containing up to three different strains with both Genogroups represented. The observed common occurrence of strain mixtures may have implications for the role of shellfish as a vector for dissemination of SRSV strains. These results show that nested RT-PCR can identify SRSV contamination in shellfish harvesting areas. Virus monitoring of shellfish harvesting areas by specialist laboratories using RT-PCR is a possible approach to combating the transmission of SRSVs by molluscan shellfish and could potentially offer significantly enhanced levels of public health protection.


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