Isolation of a full-length cDNA encoding polyphenol oxidase from sugarcane, a C4 grass

Bucheli, Carolyn S.; Dry, Ian B.; Robinson, Simon P.

doi:10.1007/bf00040840

Cited by 35 publications

(21 citation statements)

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“…PPO activity is often high in developing fruits, decreasing towards maturity (Vamos-Vigyazo 1981). Immunological studies suggest that the enzyme is relatively stable in vivo and that it may be present in a latent form in some tissues (Hunt et al 1993;Dry and Robinson 1994;Bucheli et al 1996). Indeed, in vitro experiments reveal that a 16-kDa C-terminal portion of the mature protein downstream of the conserved copper-binding domains can be cleaved without loss of enzyme activity (Robinson and Dry 1992;Dry and Robinson 1994).…”

Section: Introductionmentioning

confidence: 95%

Molecular cloning and characterisation of banana fruit polyphenol oxidase

Gooding

Bird

Robinson

2001

Planta

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Polyphenol oxidase (PPO; EC 1.10.3.2) is the enzyme thought to be responsible for browning in banana [Musa cavendishii (AAA group, Cavendish subgroup) cv. Williams] fruit. Banana flesh was high in PPO activity throughout growth and ripening. Peel showed high levels of activity early in development but activity declined until ripening started and then remained constant. PPO activity in fruit was not substantially induced after wounding or treatment with 5-methyl jasmonate. Banana flowers and unexpanded leaf roll had high PPO activities with lower activities observed in mature leaves, roots and stem. Four different PPO cDNA clones were amplified from banana fruit (BPO1, BPO11, BPO34 and BPO35). Full-length cDNA and genomic clones were isolated for the most abundant sequence (BPO1) and the genomic clone was found to contain an 85-bp intron. Introns have not been previously found in PPO genes. Northern analysis revealed the presence of BPO1 mRNA in banana flesh early in development but little BPO1 mRNA was detected at the same stage in banana peel. BPO11 transcript was only detected in very young flesh and there was no detectable expression of BPO34 or BPO35 in developing fruit samples. PPO transcripts were also low throughout ripening in both flesh and peel. BPO1 transcripts were readily detected in flowers, stem, roots and leaf roll samples but were not detected in mature leaves. BPO11 showed a similar pattern of expression to BPO1 in these tissues but transcript levels were much lower. BPO34 and BPO35 mRNAs were only detected at a low level in flowers and roots and BPO34 transcript was detected in mature leaves, the only clone to do so. The results suggest that browning of banana fruit during ripening results from release of pre-existing PPO enzyme, which is synthesised very early in fruit development.

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Section: Introductionmentioning

confidence: 95%

Molecular cloning and characterisation of banana fruit polyphenol oxidase

Gooding

Bird

Robinson

2001

Planta

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“…PPO genes have been studied in many plant species such as potato (Solanum tuberosum; Hunt et al 1993), sugarcane (Saccharum officianrum L.; Bucheli et al 1996), and tomato (Solanum lycopersicum; Thipypong et al 1997) and PPO often is encoded by gene families (Sherman et al 1995). In common wheat, three genes, Ppo-A1, Ppo-B1 and Ppo-D1, were described as expressing in developing kernels and may influence PPO activity in flour, and three additional genes express in non-kernel tissues (Demeke and Morris 2002;Jukanti et al 2004).…”

Section: Introductionmentioning

confidence: 99%

Enzyme activity in wheat breeding lines derived from matings of low polyphenol oxidase parents

2012

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Polyphenol oxidase (PPO) in grain plays a major role in time-dependent discoloration of wheat (Triticum aestivum L.) products, especially fresh noodles. Breeding wheat cultivars with low or nil PPO activity can reduce undesirable product darkening. The low PPO line PI 117635 was crossed to two low PPO wheats, IDO580 and 'IDO377s', to determine whether matings between wheats with low levels of grain PPO would result in complementation, such that lines with still lower or nil PPO would be generated. Progeny in a population derived from PI 117635/ IDO580 displayed no variation in PPO activity. In the F 3:4 populations derived from PI 117635/IDO377s, and the reciprocal IDO377s/PI 117635, normal distributions of low to high PPO activity were observed. Fieldgrown populations (F 3:5 ; F 3:6 ) derived from IDO377s crosses were analyzed for PPO activity and used to determine whether lines with nil PPO activity were generated. Of 239 lines, 154 were verified to have PPO activity that was not significantly different from the low PPO durum (Triticum turgidum var durum) cultivar 'Ben'. PCR analysis showed that the populations were fixed for a putative low PPO allele at Ppo-A1. Using markers for Ppo-D1, it was found that the average PPO activity of lines with 490 bp PCR fragments from PPO29 was significantly lower than that of lines with 560 bp fragments from STS01. These results disagreed with that predicted from previous reports for markers for Ppo-D1 alleles. Thus, breeders should exercise caution when making selections using markers for alleles at Ppo-D1, as known markers might predict erroneous phenotypes and genotypes in some wheat backgrounds.

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“…Some genes, mostly those involved in sucrose metabolism, were identified in the 1990s (Kumar et al 1992;Bugos and Thom 1993;Bucheli et al 1996). More recently, an expressed sequence tag (EST) project provided a few hundred "single-pass" sequences of anonymous genes (Carson and Botha 2000).…”

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Analysis and Functional Annotation of an Expressed Sequence Tag Collection for Tropical Crop Sugarcane

Vettore¹,

Silva²,

Kemper³

et al. 2003

Genome Research

284

247

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To contribute to our understanding of the genome complexity of sugarcane, we undertook a large-scale expressed sequence tag (EST) program. More than 260,000 cDNA clones were partially sequenced from 26 standard cDNA libraries generated from different sugarcane tissues. After the processing of the sequences, 237,954 high-quality ESTs were identified. These ESTs were assembled into 43,141 putative transcripts. Of the assembled sequences, 35.6% presented no matches with existing sequences in public databases. A global analysis of the whole SUCEST data set indicated that 14,409 assembled sequences (33% of the total) contained at least one cDNA clone with a full-length insert. Annotation of the 43,141 assembled sequences associated almost 50% of the putative identified sugarcane genes with protein metabolism, cellular communication/signal transduction, bioenergetics, and stress responses. Inspection of the translated assembled sequences for conserved protein domains revealed 40,821 amino acid sequences with 1415 Pfam domains. Reassembling the consensus sequences of the 43,141 transcripts revealed a 22% redundancy in the first assembling. This indicated that possibly 33,620 unique genes had been identified and indicated that >90% of the sugarcane expressed genes were tagged

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Isolation of a full-length cDNA encoding polyphenol oxidase from sugarcane, a C4 grass

Cited by 35 publications

References 22 publications

Molecular cloning and characterisation of banana fruit polyphenol oxidase

Molecular cloning and characterisation of banana fruit polyphenol oxidase

Enzyme activity in wheat breeding lines derived from matings of low polyphenol oxidase parents

Analysis and Functional Annotation of an Expressed Sequence Tag Collection for Tropical Crop Sugarcane

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