Cloning and characterisation of ripening-induced ethylene biosynthetic genes from non-climacteric pineapple (Ananas comosus) fruits

1998 ◽  
Vol 25 (5) ◽  
pp. 513 ◽  
Author(s):  
Christopher Ian Cazzonelli ◽  
Antonino Salvatore Cavallaro ◽  
José Ramón Botella
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Fruit Ripening ◽  
Acc Oxidase ◽  
Acc Synthase ◽  
Southern Analysis ◽  
Ananas Comosus ◽  
Northern Analysis ◽  
Fruit Tissue ◽  
Climacteric Fruit

To gain a better understanding of non-climacteric fruit ripening, pineapple was used as a model system to clone and characterise two ripening-inducible cDNAs coding for two enzymes of the ethylene biosynthetic pathway, 1-aminocyclopropane-1-carboxylate (ACC) synthase (acacc-1) and 1-aminocyclo-propane- 1-carboxylate oxidase (acaco-1) respectively. Due to the extreme acidity and high polyphenolic content of pineapple fruits, a method was optimised for the extraction of high quality RNA from fruit tissue. acacc-1 is a 1080 bp ACC synthase cDNA fragment encoding 360 amino acids including 10 of the 12 amino acid residues conserved in all aminotransferases. Comparison of the deduced amino acid sequence with previously reported ACC synthases shows between 52 and 67% similarity at the protein level. Southern analysis suggests the presence of only one copy of acacc-1 in the pineapple genome. Although some acacc-1 expression is detected in green fruits, there is a 16-fold increase in the level of acacc-1 in ripe fruit tissue. acaco-1 is a partial length cDNA clone of 611 bp which codes for 203 amino acids representing approximately 66% of the ACC oxidase open reading frame. Southern analysis suggests the presence of one or two copies of the gene in the pineapple genome. Northern analysis shows the expression of acaco-1 to be highly induced in wounded leaf tissue and to a lesser extent in ripening fruit tissue. The accumulation of ACC-synthase and ACC oxidase mRNAs during pineapple fruit ripening raises new questions about the putative role of ethylene during non-climacteric fruit ripening.

Identification and Characterisation of Two 1-Aminocyclopropane- 1-Carboxylate (ACC) Synthase cDNAs Expressed during Papaya (Carica papaya) Fruit Ripening

1997 ◽  
Vol 24 (2) ◽  
pp. 239 ◽  
Author(s):  
Michael Glenn Mason ◽  
José Ramón Botella
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Fruit Ripening ◽  
Active Site ◽  
Carica Papaya ◽  
Expression Patterns ◽  
Acc Synthase ◽  
Single Copy ◽  
Mrna Levels ◽  
Amino Acid Residues

The cloning and characterisation of two cDNAs (capacs1 and capacs2) encoding ACC synthase in papaya (Carica papaya L.) is described. capacs1 is 1104 bp long encoding 368 amino acids. capacs2 is 1098 bp long encoding 366 amino acids. The proteins encoded by both cDNAs contain the highly conserved active site of ACC synthases as well as 10 (capacs1) or 11 (capacs2) of the 12 amino acid residues conserved in most aminotransferases. Southern analyses indicate that capacs1 and capacs2 are present in the papaya genome as single copy genes. Both genes show very different expression patterns during fruit ripening. capacs1 mRNA levels are high in mature green fruits, decreasing steadily during ripening, whereas capacs2 mRNA levels are undetectable in mature green fruits but experience a dramatic increase with the onset of ripening and remain high throughout the various stages of ripening.

Sequence and expression of the dCMP deaminase gene (DCD1) of Saccharomyces cerevisiae

1986 ◽  
Vol 6 (5) ◽  
pp. 1711-1721
Author(s):  
E M McIntosh ◽  
R H Haynes
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequences ◽  
Open Reading Frame ◽  
Northern Analysis ◽  
Hindiii Site ◽  
Reading Frame ◽  
Hindiii Restriction Site ◽  
Cycle Dependent

The dCMP deaminase gene (DCD1) of Saccharomyces cerevisiae has been isolated by screening a Sau3A clone bank for complementation of the dUMP auxotrophy exhibited by dcd1 dmp1 haploids. Plasmid pDC3, containing a 7-kilobase (kb) Sau3A insert, restores dCMP deaminase activity to dcd1 mutants and leads to an average 17.5-fold overproduction of the enzyme in wild-type cells. The complementing activity of the plasmid was localized to a 4.2-kb PvuII restriction fragment within the Sau3A insert. Subcloning experiments demonstrated that a single HindIII restriction site within this fragment lies within the DCD1 gene. Subsequent DNA sequence analysis revealed a 936-nucleotide open reading frame encompassing this HindIII site. Disruption of the open reading frame by integrative transformation led to a loss of enzyme activity and confirmed that this region constitutes the dCMP deaminase gene. Northern analysis indicated that the DCD1 mRNA is a 1.15-kb poly(A)+ transcript. The 5' end of the transcript was mapped by primer extension and appears to exhibit heterogeneous termini. Comparison of the amino acid sequence of the T2 bacteriophage dCMP deaminase with that deduced for the yeast enzyme revealed a limited degree of homology which extends over the entire length of the phage polypeptide (188 amino acids) but is confined to the carboxy-terminal half of the yeast protein (312 amino acids). A potential dTTP-binding site in the yeast and phage enzymes was identified by comparison of homologous regions with the amino acid sequences of a variety of other dTTP-binding enzymes. Despite the role of dCMP deaminase in dTTP biosynthesis, Northern analysis revealed that the DCD1 gene is not subject to the same cell cycle-dependent pattern of transcription recently found for the yeast thymidylate synthetase gene (TMP1).

N-Acetylglucosamine-inducible CaGAP1 encodes a general amino acid permease which co-ordinates external nitrogen source response and morphogenesis in Candida albicans

Microbiology ◽  
2003 ◽  
Vol 149 (9) ◽  
pp. 2597-2608 ◽  
Author(s):  
Subhrajit Biswas ◽  
Monideepa Roy ◽  
Asis Datta
Keyword(s):  
Amino Acids ◽  
Candida Albicans ◽  
Amino Acid ◽  
Genomic Library ◽  
Northern Analysis ◽  
Dependent Manner ◽  
Amino Acid Permease ◽  
Environmental Signals ◽  
General Amino Acid Permease ◽  
Hyphal Formation

Candida albicans is able to grow in a variety of reversible morphological forms (yeast, pseudohyphal and hyphal) in response to various environmental signals, noteworthy among them being N-acetylglucosamine (GlcNAc). The gene CaGAP1, homologous to GAP1, which encodes the general amino acid permease from Saccharomyces cerevisiae, was isolated on the basis of its induction by GlcNAc through differential screening of a C. albicans genomic library. The gene could functionally complement an S. cerevisiae gap1 mutant by rendering it susceptible to the toxic amino acid analogue mimosine in minimal proline media. As in S. cerevisiae, mutation of the CaGAP1 gene had an effect on citrulline uptake in C. albicans. Northern analysis showed that GlcNAc-induced expression of CaGAP1 was further enhanced in synthetic minimal media supplemented with single amino acids (glutamate, proline and glutamine) or urea (without amino acids) but repressed in minimal ammonium media. Induction of CaGAP1 expression by GlcNAc was nullified in C. albicans deleted for the transcription factor CPH1 and the hyphal regulator RAS1, indicating the involvement of Cph1p-dependent Ras1p signalling in CaGAP1 expression. A homozygous mutant of this gene showed defective hyphal formation in solid hyphal-inducing media and exhibited less hyphal clumps when induced by GlcNAc. Alteration of morphology and short filamentation under nitrogen-starvation conditions in the heterozygous mutant suggested that CaGAP1 affects morphogenesis in a dose-dependent manner.

scholarly journals Synergistic Effects of the Combined Application of MCP and Low O2 on Apple Fruit Ripening

HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1143D-1144
Author(s):  
Mehar Asif ◽  
Prabodh Trivedi ◽  
Theophanes Solomos ◽  
Autar Mattoo
Keyword(s):  
Fruit Ripening ◽  
Similar Pattern ◽  
Synergistic Effects ◽  
Acc Oxidase ◽  
Acc Synthase ◽  
Apple Fruit ◽  
Single Application ◽  
Combined Application ◽  
Putative Receptor

We have studied the effects of MCP and low O2, applied singly and in combination, on apple fruit ripening at 1, 7, and 18 °C. The single application of 2 ppm MCP is more effective in delaying the onset of the C2H4 climacteric than is 1% O2. However, the combined application has a much larger effect than the single applications of either MCP or 1% O2. For instance, at 7 °C, the onset of the C2H4 climacteric occurs at 15, 50, and 90–95 days for the controls, 1% O2 and 2 ppm MCP, respectively, whereas the combined application of 2 ppm MCP and 1% O2 suppressed the initiation of the C2H4 climacteric for 200 days, the duration of the experiment. The retardation of the climacteric onset by the treatments is associated with the suppression of ACC-synthase (ACS1) and the putative receptor ERS1. The accumulation of their transcripts is critically dependent on the rate of C2H4 evolution. As expected, the combined application of MCP and 1% O2 completely suppressed the expression of both genes. Yet when the fruits were transferred to 18 °C in air, they ripened normally. A similar pattern of inhibition in response to the above treatments was also observed with a C2H4-dependent MAPK. The expression of ETR1, ETR2 and ACC-oxidase was not affected by the treatments. The nature of this strong effect of the combined application of MCP and low O2 is not clear. It should be pointed out that MCP does not inhibit the induction of hypoxic proteins such as ADH.

scholarly journals Brassinosteroids Suppress Ethylene Biosynthesis via Transcription Factor BZR1 in Pear and Apple Fruit

2020 ◽  
Author(s):  
Yinglin Ji ◽  
Yi Qu ◽  
Zhongyu Jiang ◽  
Xin Su ◽  
Pengtao Yue ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Fruit Ripening ◽  
Ethylene Production ◽  
Plant Hormone ◽  
Expression Profiles ◽  
Acc Synthase ◽  
Ethylene Biosynthesis ◽  
Apple Fruit ◽  
Pear Fruit ◽  
Climacteric Fruit

ABSTRACTThe plant hormone ethylene is important for the ripening of climacteric fruit, such as pear (Pyrus ussuriensis), and the brassinosteroid (BR) class of phytohormones affects ethylene biosynthesis during ripening, although via an unknown molecular mechanism. Here, we observed that exogenous BR treatment suppressed ethylene production during pear fruit ripening, and that the expression of the transcription factor PuBZR1 was enhanced by epibrassinolide (EBR) treatment during pear fruit ripening. PuBZR1 was shown to interact with PuACO1, which converts 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene, and suppress its activity. We also observed that BR-activated PuBZR1 bound to the promoters of PuACO1 and of PuACS1a, which encodes ACC synthase, and directly suppressed their transcription. Moreover, PuBZR1 suppressed the expression of transcription factor PuERF2 by binding its promoter, and PuERF2 bound to the promoters of PuACO1 and PuACS1a. We concluded that PuBZR1 indirectly suppresses the transcription of PuACO1 and PuACS1a through its regulation of PuERF2. Ethylene production and the expression profiles of the corresponding apple (Malus domestica) homologs showed similar changes following EBR treatment. Together, these results suggest that BR-activated BZR1 suppresses ACO1 activity and the expression of ACO1 and ACS1a, thereby reducing ethylene production during pear and apple fruit ripening. This likely represents a conserved mechanism by which exogenous BR suppresses ethylene biosynthesis during climacteric fruit ripening.One-sentence summaryBR-activated BZR1 suppresses ACO1 activity and expression of ACO1 and ACS1a, which encode two ethylene biosynthesis enzymes, thereby reducing ethylene production during pear and apple fruit ripening.

scholarly journals Sequence and expression of the dCMP deaminase gene (DCD1) of Saccharomyces cerevisiae.

1986 ◽  
Vol 6 (5) ◽  
pp. 1711-1721 ◽  
Author(s):  
E M McIntosh ◽  
R H Haynes
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequences ◽  
Open Reading Frame ◽  
Northern Analysis ◽  
Hindiii Site ◽  
Reading Frame ◽  
Hindiii Restriction Site ◽  
Cycle Dependent

The dCMP deaminase gene (DCD1) of Saccharomyces cerevisiae has been isolated by screening a Sau3A clone bank for complementation of the dUMP auxotrophy exhibited by dcd1 dmp1 haploids. Plasmid pDC3, containing a 7-kilobase (kb) Sau3A insert, restores dCMP deaminase activity to dcd1 mutants and leads to an average 17.5-fold overproduction of the enzyme in wild-type cells. The complementing activity of the plasmid was localized to a 4.2-kb PvuII restriction fragment within the Sau3A insert. Subcloning experiments demonstrated that a single HindIII restriction site within this fragment lies within the DCD1 gene. Subsequent DNA sequence analysis revealed a 936-nucleotide open reading frame encompassing this HindIII site. Disruption of the open reading frame by integrative transformation led to a loss of enzyme activity and confirmed that this region constitutes the dCMP deaminase gene. Northern analysis indicated that the DCD1 mRNA is a 1.15-kb poly(A)+ transcript. The 5' end of the transcript was mapped by primer extension and appears to exhibit heterogeneous termini. Comparison of the amino acid sequence of the T2 bacteriophage dCMP deaminase with that deduced for the yeast enzyme revealed a limited degree of homology which extends over the entire length of the phage polypeptide (188 amino acids) but is confined to the carboxy-terminal half of the yeast protein (312 amino acids). A potential dTTP-binding site in the yeast and phage enzymes was identified by comparison of homologous regions with the amino acid sequences of a variety of other dTTP-binding enzymes. Despite the role of dCMP deaminase in dTTP biosynthesis, Northern analysis revealed that the DCD1 gene is not subject to the same cell cycle-dependent pattern of transcription recently found for the yeast thymidylate synthetase gene (TMP1).

Expression of ACC synthase is enhanced earlier than that of ACC oxidase during fruit ripening of mume (Prunus mume )

1999 ◽  
Vol 107 (3) ◽  
pp. 319-328 ◽  
Author(s):  
Satoru Mita ◽  
Chikako Kirita ◽  
Masaya Kato ◽  
Hiroshi Hyodo
Keyword(s):  
Fruit Ripening ◽  
Acc Oxidase ◽  
Acc Synthase ◽  
Prunus Mume

scholarly journals The mechanism for brassinosteroids suppressing climacteric fruit ripening

2021 ◽  
Author(s):  
Yinglin Ji ◽  
Yi Qu ◽  
Zhongyu Jiang ◽  
Jijun Yan ◽  
Jinfang Chu ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Carboxylic Acid ◽  
Fruit Ripening ◽  
Ethylene Production ◽  
Plant Hormone ◽  
Expression Profiles ◽  
Acc Synthase ◽  
Ethylene Biosynthesis ◽  
Pear Fruit ◽  
Climacteric Fruit

Abstract The plant hormone ethylene is important for the ripening of climacteric fruit, such as pear (Pyrus ussuriensis), and the brassinosteroid (BR) class of phytohormones affects ethylene biosynthesis during ripening via an unknown molecular mechanism. Here, we observed that exogenous BR treatment suppressed ethylene production and delayed fruit ripening, whereas treatment with a BR biosynthesis inhibitor promoted ethylene production and accelerated fruit ripening in pear, suggesting BR is a ripening suppressor. The expression of the transcription factor BRASSINAZOLE-RESISTANT 1PuBZR1 was enhanced by BR treatment during pear fruit ripening. PuBZR1 interacted with PuACO1, which converts 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylene, and suppressed its activity. BR-activated PuBZR1 bound to the promoters of PuACO1 and of PuACS1a, which encodes ACC synthase, and directly suppressed their transcription. Moreover, PuBZR1 suppressed the expression of transcription factor PuERF2 by binding its promoter, and PuERF2 bound to the promoters of PuACO1 and PuACS1a. We concluded that PuBZR1 indirectly suppresses the transcription of PuACO1 and PuACS1a through its regulation of PuERF2. Ethylene production and expression profiles of corresponding apple (Malus domestica) homologs showed similar changes following epibrassinolide treatment. Together, these results suggest that BR-activated BZR1 suppresses ACO1 activity and the expression of ACO1 and ACS1, thereby reducing ethylene production and suppressing fruit ripening. This likely represents a conserved mechanism by which BR suppresses ethylene biosynthesis during climacteric fruit ripening.

scholarly journals Is Ethylene the Ripening Hormone

2020 ◽  
pp. 1-7
Author(s):  
Senewa Bobby Pholoma
Keyword(s):  
Fruit Ripening ◽  
Wide Spectrum ◽  
Cell Metabolism ◽  
Economic Value ◽  
Acc Oxidase ◽  
Acc Synthase ◽  
Ethylene Biosynthesis ◽  
Vital Role ◽  
Limiting Factors ◽  
Physiological Processes

The fruits constitute a commercially important and nutritionally indispensable food commodity since they play a vital role in human nutrition by supplying the necessary growth factors essential for maintaining normal health.  One of the limiting factors that influence their economic value is the relatively short ripening period and reduced post-harvest life.  The fruit ripening involves a wide spectrum of coordinated biochemical and physiological processes that eventually leads to development of soft edible fruit with desirable qualities such as carotenoids, anthocyanin, color, sweetness, texture, firmness, flavor and aroma.  The ripening is the phase of fruit development just before senescence, therefore the excessive tissues softening due to the high ethylene exposure leads to the spoilage upon the storage.  Ethylene, a fruit ripening hormone can trigger many events of cell metabolism including ripening particularly in climacteric fruits even in minute amounts.  As fruit mature, the rate of ACC and ethylene biosynthesis increases as well as the enzyme activities for ACC oxidase and ACC synthase enhance.  However, the application of ethylene inhibitors such as 1-MCP, AVG and the ethylene remover proved to reduce the ripening where some quality attributes of ripening were reduced due to suppressed expression of the ripening hormone.

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