Pear IAA1 gene encoding an auxin-responsive Aux/IAA protein is involved in fruit development and response to salicylic acid

2014 ◽  
Vol 94 (2) ◽  
pp. 263-271 ◽  
Author(s):  
Haiyan Shi ◽  
Yanhui Wang ◽  
Zhenghong Li ◽  
Diansheng Zhang ◽  
Yufeng Zhang ◽  
...  
Keyword(s):  
Salicylic Acid ◽  
Fruit Development ◽  
Phylogenetic Analyses ◽  
Pcr Amplification ◽  
Pyrus Pyrifolia ◽  
Dimerization Domain ◽  
Developmentally Regulated ◽  
Gene Encoding ◽  
Domain Profile ◽  
Localization Signal

Shi, H., Wang, Y., Li, Z., Zhang, D., Zhang, Y., Xiang, D., Li, Y. and Zhang, Y. 2014. Pear IAA1 gene encoding an auxin-responsive Aux/IAA protein is involved in fruit development and response to salicylic acid. Can. J. Plant Sci. 94: 263–271. Auxin-responsive Aux/IAA proteins are rapidly auxin-induced, short-lived proteins that act as repressors for the auxin response factor (ARF)-activated gene expression. A gene encoding an Aux/IAA protein and designated PpIAA1 was isolated from pear (Pyrus pyrifolia). Using PCR amplification techniques, the genomic clone corresponding to PpIAA1 was isolated and shown to contain three introns with typical GT/AG boundaries defining the splice junctions. The deduced PpIAA1 protein contains the conserved features of indole-3-acetic acids (IAA): four Aux/IAA conserved domains, Aux/IAA family domain, Aux/IAA-ARF dimerization domain profile, and conserved nuclear localization signal (NLS) motifs. Phylogenetic analyses clearly demonstrated PpIAA1 has the highest homology with grape VvIAA. PpIAA1 was preferentially expressed in fruit, and moderate expression was found in anthers. Relatively low expression signal was detected in other tissues including shoots, leaves, and petals. Moreover, expression of PpIAA1 was developmentally regulated in fruit. Further study demonstrated that PpIAA1 expression in pear fruit was remarkably regulated by salicylic acid and IAA. The data suggest that PpIAA1 might be involved in the interplay between IAA and salicylic acid signaling pathway during the fruit development of pear.

Pear PIP1 gene is regulated during fruit development and is invovled in response to salicylic acid and ethylene

2015 ◽  
Vol 95 (1) ◽  
pp. 77-85
Author(s):  
Haiyan Shi ◽  
Yanhui Wang ◽  
Diansheng Zhang ◽  
Liang Chen ◽  
Yuxing Zhang
Keyword(s):  
Salicylic Acid ◽  
Plasma Membrane ◽  
Fruit Development ◽  
Phylogenetic Analyses ◽  
Pcr Amplification ◽  
Protein Domain ◽  
Pyrus Pyrifolia ◽  
Plasma Membrane Intrinsic Protein ◽  
Intrinsic Protein ◽  
Young Leaves

Shi, H., Wang, Y., Zhang, D., Chen, L. and Zhang, Y. 2015. Pear PIP1 gene is regulated during fruit development and is invovled in response to salicylic acid and ethylene. Can. J. Plant Sci. 95: 77–85. Plasma membrane intrinsic proteins (PIPs), a subfamily of aquaporins, are widely implicated in plant growth and development. A gene encoding a plasma membrane intrinsic protein and designated PpPIP1 was isolated from pear (Pyrus pyrifolia). Using PCR amplification techniques, the genomic clone corresponding to PpPIP1 was isolated and shown to contain three introns with typical GT/AG boundaries defining the splice junctions. The deduced PpPIP1 protein contains the conserved features of PIPs: six transmembrane α-helices, a major intrinsic protein domain, and a conserved asparagine–proline–alanine (NPA) signature sequence. Phylogenetic analyses clearly demonstrated that PpPIP1 has the highest homology with apple (Malus×domestica) MdPIP1a and Malus hupehensis MhPIP1-1. PpPIP1 transcripts were mainly detected in young leaves, shoots, petals and mesocarp of fruit, but a relatively low expression signal was detected in anthers. In particular, expression of PpPIP1 was developmentally regulated in fruit. Further study demonstrated that PpPIP1 expression in pear fruit was down-regulated by salicylic acid (SA) and up-regulated by ethylene. These data suggest that PpPIP1 may be involved in the response to SA and ethylene during fruit development, which would provide valuable information for water permeability studies in pear.

Cloning, characterization and expression analysis of a 1-aminocyclopropane-1-carboxylate synthase gene from pear

2013 ◽  
Vol 93 (3) ◽  
pp. 465-471
Author(s):  
Haiyan Shi ◽  
Yuxing Zhang ◽  
Liang Chen
Keyword(s):  
Gene Product ◽  
Expression Analysis ◽  
Cdna Clone ◽  
Phylogenetic Analyses ◽  
Pcr Amplification ◽  
Acc Synthase ◽  
Pcr Analysis ◽  
Pyrus Pyrifolia ◽  
Reading Frame ◽  
Amplification Technique

Shi, H., Zhang, Y. and Chen, L. 2013. Cloning, characterization and expression analysis of a 1-aminocyclopropane-1-carboxylate synthase gene from pear. Can. J. Plant Sci. 93: 465–471. In this study, a cDNA clone encoding putative 1-aminocyclopropane-1-carboxylate (ACC) synthase (ACS) that catalyzes the conversion of S-adenosyl-L-methionine to ACC in ethylene biosynthetic pathway was isolated from a cDNA library produced using mRNA from pear (Pyrus pyrifolia). The cDNA clone, designated PpACS2, comprised an open reading frame of 1, 341 bp encoding a protein of 446 amino acids that shares high similarity with the known plant ACSs. Using PCR amplification technique, a genomic clone (GenBank accession number: KC146402) corresponding to PpACS2 was isolated and shown to contain two introns. The PpACS2 gene product shared 97% identity with an ACC synthase from pear (Pyrus communis). Phylogenetic analyses clearly placed the gene product in the ACC synthase cluster of plant ACS superfamily tree. Quantitative RT-PCR analysis indicated that the PpACS2 gene was preferentially expressed in young pear leaves and shoots. The transcript of PpACS2 gene was accumulated at relatively high levels in anthers, but no signal was detected in the petals and mesocarp of pear. These results suggest that the PpACS2 may participate in the regulation of ethylene production in pear leaves, shoots, and anthers.

scholarly journals Expression and Regulation of PpEIN3b during Fruit Ripening and Senescence via Integrating SA, Glucose, and ACC Signaling in Pear (Pyrus pyrifolia Nakai. Whangkeumbae)

Genes ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 476 ◽  
Author(s):  
Shi ◽  
Zhang ◽  
Chen
Keyword(s):  
Shelf Life ◽  
Signaling Pathway ◽  
Fruit Ripening ◽  
Economic Value ◽  
Pcr Analysis ◽  
Pyrus Pyrifolia ◽  
Ethylene Response Factor ◽  
Developmentally Regulated ◽  
Gene Encoding ◽  
Climacteric Fruit

The economic value of fruit is reduced by having a short shelf life. Whangkeumbae is a type of sand pear (Pyrus pyrifolia) considered a climacteric fruit. The pear is famous for its smooth surface and good flavor. However, its shelf life is very short because of senescence and disease after harvest and a burst of ethylene (ET) production prompting the onset of fruit ripening. In plants, ETHYLENE INSENSITIVE3 (EIN3) and EIN3like (EIL), located in the nucleus, are important components of the ET signaling pathway and act as transcription factors. EIN3s and EILs belong to a small family involved in regulating the expression of ethylene response factor gene (ERF), whose encoding protein is the final component in the ET signaling pathway. The mutation of these components will cause defects in the ethylene pathway. In this study, one gene encoding an EIN3 was cloned and identified from Whangkeumbae and designated PpEIN3b. The deduced PpEIN3b contained a conserved EIN3 domain, a bipartite nuclear localization signal profile (NLS_BP), and an N-6 adenine-specific DNA methylase signature (N6_MTASE). PpEIN3b belongs to the EIN3 super-family by phylogenetic analysis. Quantitative RT-PCR (qRT-PCR) analysis revealed that PpEIN3b was preferentially expressed in fruit. Additionally, its expression was developmentally regulated during fruit ripening and senescence. Furthermore, PpEIN3b transcripts were obviously repressed by salicylic acid (SA) and glucose treatment in pear fruit and in diseased fruit, while it was significantly induced by 1-aminocyclopropane-1-carboxylic acid (ACC) treatment. Taken together, our results reveal the expression and regulation profiles of PpEIN3b and suggest that PpEIN3b might integrate SA, glucose, and ACC signaling to regulate fruit ripening and senescence in pear, which would provide a candidate gene for this regulation to obtain fruit with a long shelf life and improved economic value.

Molecular characterization of pear 14-3-3b gene regulated during fruit development

2016 ◽  
Vol 96 (3) ◽  
pp. 433-438
Author(s):  
Haiyan Shi ◽  
Yujing Zhao ◽  
Xuemin An ◽  
Yuxing Zhang
Keyword(s):  
Protein Interactions ◽  
Pcr Amplification ◽  
Pcr Analysis ◽  
Pyrus Pyrifolia ◽  
Biological Processes ◽  
Protein Protein Interactions ◽  
Rt Pcr ◽  
Developmentally Regulated ◽  
Amplification Technique

Plant 14-3-3 proteins (14-3-3s) are known to function in protein–protein interactions that mediate signal transduction pathways regulating many biological processes. The cDNA encoding putative 14-3-3 protein was isolated from pear (Pyrus pyrifolia) and designated Pp14-3-3b. Using the PCR amplification technique, the genomic clone corresponding to Pp14-3-3b was isolated and shown to contain six introns. Phylogenetic analysis clearly demonstrated that Pp14-3-3b was classified into the non-ɛ class of 14-3-3 superfamilies. Quantitative RT-PCR analysis indicated that the expression of the Pp14-3-3b gene was developmentally regulated in fruit. This study suggested that Pp14-3-3b might be involved in fruit ripening and the senescence of pear.

Intercistronic heterogeneity of the 16S–23S rRNA spacer region among Pseudomonas strains isolated from subterranean seeds of hog peanut (Amphicarpa bracteata)

Microbiology ◽  
2009 ◽  
Vol 155 (8) ◽  
pp. 2630-2640 ◽  
Author(s):  
J. T. Tambong ◽  
R. Xu ◽  
E. S. P. Bromfield
Keyword(s):  
Root Zone ◽  
Phylogenetic Analyses ◽  
Melting Curve ◽  
Pcr Amplification ◽  
23S Rrna ◽  
Melting Curve Analysis ◽  
Its1 Region ◽  
A Genome ◽  
Pure Cultures ◽  
Intercalating Dye

Intercistronic heterogeneity of the 16S–23S rRNA internal transcribed spacer regions (ITS1) was investigated in 29 strains of fluorescent pseudomonads isolated from subterranean seeds of Amphicarpa bracteata (hog peanut). PCR amplification of the ITS1 region generated one or two products from the strains. Sequence analysis of the amplified fragments revealed an ITS1 fragment of about 517 bp that contained genes for tRNAIle and tRNAAla in all 29 strains; an additional smaller ITS1 of 279 bp without tRNA features was detected in 15 of the strains. The length difference appeared to be due to deletions of several nucleotide blocks between the 70 bp and 359 bp positions of the alignment. The end of the deletions in the variant ITS1 type coincided with the start of antiterminator box A, which is homologous to box A of other bacteria. Phylogenetic analyses using the neighbour-joining algorithm revealed two major phylogenetic clusters, one for each of the ITS1 types. Using a single specific primer set and the DNA-intercalating dye SYBR Green I for real-time PCR and melting-curve analysis produced highly informative curves with one or two recognizable melting peaks that readily distinguished between the two ITS1 types in pure cultures. The assay was used to confirm the presence of the variant ITS1 type in the Pseudomonas community in total DNA from root-zone soil and seed coats of hog peanut. Heterogeneity of the ITS1 region between species has potential for studying molecular systematics and population genetics of the genus Pseudomonas, but the presence of non-identical rRNA operons within a genome may pose problems.

scholarly journals Genome-wide identification and characterization of COMT gene family during the development of blueberry fruit

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Yushan Liu ◽  
Yizhou Wang ◽  
Jiabo Pei ◽  
Yadong Li ◽  
Haiyue Sun
Keyword(s):  
Gene Family ◽  
Fruit Development ◽  
Lignin Content ◽  
Phylogenetic Analyses ◽  
Expression Patterns ◽  
Vaccinium Corymbosum ◽  
Land Plant ◽  
Phylogenetic Structure ◽  
Genome Wide

Abstract Background Caffeic acid O-methyltransferases (COMTs) play an important role in the diversification of natural products, especially in the phenylalanine metabolic pathway of plant. The content of COMT genes in blueberry and relationship between their expression patterns and the lignin content during fruit development have not clearly investigated by now. Results Ninety-two VcCOMTs were identified in Vaccinium corymbosum. According to phylogenetic analyses, the 92 VcCOMTs were divided into 2 groups. The gene structure and conserved motifs within groups were similar which supported the reliability of the phylogenetic structure groupings. Dispersed duplication (DSD) and whole-genome duplication (WGD) were determined to be the major forces in VcCOMTs evolution. The results showed that the results of qRT-PCR and lignin content for 22 VcCOMTs, VcCOMT40 and VcCOMT92 were related to lignin content at different stages of fruit development of blueberry. Conclusion We identified COMT gene family in blueberry, and performed comparative analyses of the phylogenetic relationships in the 15 species of land plant, and gene duplication patterns of COMT genes in 5 of the 15 species. We found 2 VcCOMTs were highly expressed and their relative contents were similar to the variation trend of lignin content during the development of blueberry fruit. These results provide a clue for further study on the roles of VcCOMTs in the development of blueberry fruit and could promisingly be foundations for breeding blueberry clutivals with higher fruit firmness and longer shelf life.

scholarly journals Plasmid-Encoded Metallo-β-Lactamase (IMP-6) Conferring Resistance to Carbapenems, Especially Meropenem

2001 ◽  
Vol 45 (5) ◽  
pp. 1343-1348 ◽  
Author(s):  
Hisakazu Yano ◽  
Akio Kuga ◽  
Ryoichi Okamoto ◽  
Hidero Kitasato ◽  
Toshimitsu Kobayashi ◽  
...  
Keyword(s):  
Point Mutation ◽  
Antimicrobial Agents ◽  
Pcr Amplification ◽  
Penicillin G ◽  
Gene Encoding ◽  
Mature Enzyme ◽  
Dna Fragment ◽  
Lactamase Gene ◽  
Reduced Activity ◽  
Northern Japan

ABSTRACT In 1996, Serratia marcescens KU3838 was isolated from the urine of a patient with a urinary tract infection at a hospital in northern Japan and was found to contain the plasmid pKU501. Previously, we determined that pKU501 carries bla IMP and the genes for TEM-1-type β-lactamases as well as producing both types of β-lactamases (H. Yano, A. Kuga, K. Irinoda, R. Okamoto, T. Kobayashi, and M. Inoue, J. Antibiot. 52:1135–1139, 1999). pKU502 is a recombinant plasmid that contains a 1.5-kb DNA fragment, including the metallo-β-lactamase gene, and is obtained by PCR amplification of pKU501. The sequence of the metallo-β-lactamase gene in pKU502 was determined and revealed that this metallo-β-lactamase gene differed from the gene encoding IMP-1 by one point mutation, leading to one amino acid substitution: 640-A in the base sequence of the IMP-1 gene was replaced by G, and Ser-196 was replaced by Gly in the mature enzyme. This enzyme was designated IMP-6. The strains that produced IMP-6 were resistant to carbapenems. The MICs of panipenem and especially meropenem were higher than the MIC of imipenem for these strains. The k cat/Km value of IMP-6 was about sevenfold higher against meropenem than against imipenem, although the MIC of meropenem for KU1917, which produced IMP-1, was lower than that of imipenem, and the MIC of panipenem was equal to that of imipenem. These results support the hypothesis that IMP-6 has extended substrate profiles against carbapenems. However, the activity of IMP-6 was very low against penicillin G and piperacillin. These results suggest that IMP-6 acquired high activity against carbapenems, especially meropenem, via the point mutation but in the process lost activity against penicillins. Although IMP-6 has reduced activity against penicillins due to this point mutation, pKU501 confers resistance to a variety of antimicrobial agents because it also produces TEM-1-type enzyme.

scholarly journals Expression of a Gene Encoding Trypanosoma congolense Putative Abc1 Family Protein is Developmentally Regulated

2005 ◽  
Vol 67 (2) ◽  
pp. 157-164 ◽  
Author(s):  
Waren N. BATICADOS ◽  
William H. WITOLA ◽  
Noboru INOUE ◽  
Jung-Yeon, KIM ◽  
Noritaka KUBOKI ◽  
...  
Keyword(s):  
Trypanosoma Congolense ◽  
Developmentally Regulated ◽  
Gene Encoding ◽  
Family Protein

scholarly journals Autonomous assembly of synthetic oligonucleotides built from an expanded DNA alphabet. Total synthesis of a gene encoding kanamycin resistance

2014 ◽  
Vol 10 ◽  
pp. 2348-2360 ◽  
Author(s):  
Kristen K Merritt ◽  
Kevin M Bradley ◽  
Daniel Hutter ◽  
Mariko F Matsuura ◽  
Diane J Rowold ◽  
...  
Keyword(s):  
Solid Phase ◽  
Pcr Amplification ◽  
Kanamycin Resistance ◽  
Dna Fragments ◽  
Gene Encoding ◽  
Synthetic Dna ◽  
Information Density ◽  
Letter Alphabet ◽  
New Strategy ◽  
Autonomous Assembly

Background: Many synthetic biologists seek to increase the degree of autonomy in the assembly of long DNA (L-DNA) constructs from short synthetic DNA fragments, which are today quite inexpensive because of automated solid-phase synthesis. However, the low information density of DNA built from just four nucleotide “letters”, the presence of strong (G:C) and weak (A:T) nucleobase pairs, the non-canonical folded structures that compete with Watson–Crick pairing, and other features intrinsic to natural DNA, generally prevent the autonomous assembly of short single-stranded oligonucleotides greater than a dozen or so. Results: We describe a new strategy to autonomously assemble L-DNA constructs from fragments of synthetic single-stranded DNA. This strategy uses an artificially expanded genetic information system (AEGIS) that adds nucleotides to the four (G, A, C, and T) found in standard DNA by shuffling hydrogen-bonding units on the nucleobases, all while retaining the overall Watson–Crick base-pairing geometry. The added information density allows larger numbers of synthetic fragments to self-assemble without off-target hybridization, hairpin formation, and non-canonical folding interactions. The AEGIS pairs are then converted into standard pairs to produce a fully natural L-DNA product. Here, we report the autonomous assembly of a gene encoding kanamycin resistance using this strategy. Synthetic fragments were built from a six-letter alphabet having two AEGIS components, 5-methyl-2’-deoxyisocytidine and 2’-deoxyisoguanosine (respectively S and B), at their overlapping ends. Gaps in the overlapped assembly were then filled in using DNA polymerases, and the nicks were sealed by ligase. The S:B pairs in the ligated construct were then converted to T:A pairs during PCR amplification. When cloned into a plasmid, the product was shown to make Escherichia coli resistant to kanamycin. A parallel study that attempted to assemble similarly sized genes with optimally designed standard nucleotides lacking AEGIS components gave successful assemblies of up to 16 fragments, but generally failed when larger autonomous assemblies were attempted. Conclusion: AEGIS nucleotides, by increasing the information density of DNA, allow larger numbers of DNA fragments to autonomously self-assemble into large DNA constructs. This technology can therefore increase the size of DNA constructs that might be used in synthetic biology.

scholarly journals ISOLASI FRAGMEN GEN PENYANDI PUTRESIN N-METILTRANSFERASE DAN QUINOLINAT FOSFORIBOSILTRANSFERASE ASAL TEMBAKAU LOKAL TEMANGGUNG (Nicotiana tabacum)

2020 ◽  
Vol 17 (3) ◽  
pp. 109 ◽  
Author(s):  
SESANTI BASUKI ◽  
NURHAJATI AA MATTJIK ◽  
SUWARSO SUWARSO ◽  
DESTA WIRNAS ◽  
SUDARSONO SUDARSONO
Keyword(s):  
Dna Sequences ◽  
Pcr Amplification ◽  
Data Bank ◽  
Degenerate Primer ◽  
Gene Encoding ◽  
Methyl Transferase ◽  
Dna Database ◽  
Genes Encoding ◽  
Tobacco Cultivar ◽  
Dna Template

<p>ABSTRAK</p><p>Upaya untuk menurunkan kandungan nikotin merupakan salah satuprioritas utama penelitian tembakau. Nikotin adalah senyawa alkaloidutama berpotensi dikonversi menjadi senyawa nor-nikotin yang bersifatkarsinogen. Gen PMT sebagai penyandi enzim putresin n-metiltransferase(PMT) dan gen QPT - penyandi enzim quinolinat fosforibosiltransferase(QPT) merupakan dua gen kunci yang berperan penting pada proses bio-sintesis nikotin. Penelitian ini bertujuan untuk mengisolasi potongan genPMT dan QPT asal tembakau lokal Indonesia, mengkarakterisasi danmenganalisis runutan DNA-nya. Tahapan penelitian dimulai dengan me-rancang primer degenerate berdasarkan informasi yang ada di pangkalandata Bank Gen NCBI (National Centre for Biotechnology Information),mengamplifikasi PCR menggunakan templat DNA genomik tembakaulokal cv. Sindoro1, mengklon potongan DNA hasil PCR dan menentukanrunutan DNA-nya. Hasil penelitian menunjukkan dari dua belas pasangprimer degenerate yang dirancang, hanya dua pasang primer yang meng-hasilkan potongan DNA hasil amplifikasi PCR, yaitu pasangan primerPMt-7 (F &amp; R) untuk gen PMT dan primer QPt-3 (F &amp; R) untuk gen QPT.Setelah dilakukan penentuan runutan DNA-nya, amplikon yang didapatdari hasil PCR dengan pasangan primer PMt-7 sebesar 1418 bp, sedangkanuntuk primer QPt-3 sebesar 205 bp. Runutan DNA gen PMT dan gen QPTasal tembakau lokal cv. Sindoro1 mempunyai tingkat kesamaan yang ting-gi dengan gen PMT dan gen QPT asal tembakau lainnya yang ada dipangkalan data Bank Gen NCBI.</p><p>Kata kunci : Gen PMT, gen QPT, lintasan biosintesis nikotin, perunutanDNA, amplifikasi PCR, primer degenerate</p><p>ABSTRACT</p><p>Isolation of Genes encoding Putrescine N-Methyl-transferase and Quinolinat Phosphoribosyl transferasederived from Temanggung Tobacco Cultivar (Nicotianatabacum)</p><p>Reduction of nicotine content is one of the major objective intobacco research. Nicotine is the main alcaloid compound that potentiallycould be converted into a carcinogenic compound (nor-nicotine). The PMTgene encoding putrescine N-methyl transferase (PMT) and the QPT gene -encoding quinolinate phosphoribosyl transferase (QPT) are the two keyenzymes involved in nicotine biosynthesis. The objectives of this researchwere to isolate PMT and QPT gene fragments originated from Indonesianlocal tobacco, to characterize, and to analyze their DNA sequences. Theresearch activities included: degenerate primer design based oninformation available in the GenBank DNA Database NCBI (NationalCentre for Biotechnology Information), PCR amplification usingdegenerate primer and genomic DNA template of a local tobacco cv.Sindoro1, clone the PCR amplified products, and determine their DNAnucleotide sequences. Results of the experiment indicated that from 12degenerate primer pairs synthesized, only two were able to yield positivePCR amplified products. These primer pairs were PMt-7 (F &amp; R primers)for PMT and QPt-3 (F &amp; R primers) for QPT. After DNA sequencing, theamplified DNA product amplified using PMt-7 degenerate primer pairswere 1418 bp, while that using QPt-3 primer pairs were only 205 bp.Nucleotide sequences of PMT or QPT gene fragments originated fromlocal tobacco cv. Sindoro1 showed a high nucleotide sequences identity ascompared to that of the respective genes from other tobacco species thatwere available in the GenBank DNA Database NCBI.</p><p>Key words: PMT gene, QPT gene, nicotine biosynthetic pathways, DNAsequencing, PCR amplification, degenerate primer</p>

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