Cloning, partial sequencing and expression of a cDNA coding for branching enzyme in cassava

Salehuzzaman, S.N.I.M.; Jacobsen, E.; Visser, Richard G. F.

doi:10.1007/bf00027152

Cited by 45 publications

(29 citation statements)

References 29 publications

(35 reference statements)

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“…The observed tissue specific expression pattern of Meisa1 was similar with previously reported patterns from other starch metabolizing genes in cassava (Salehuzzaman et al, 1992;Salehuzzaman et al, 1994;Baguma et al, 2003). Meisa1 declined with increased developmental stage of the storage root (Fig.…”

Section: Spatial and Temporal Expression Of Meisa1supporting

confidence: 87%

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Characterisation and role of <i>Isoamylase1 (Meisa1</i>) gene in cassava

Beyene¹,

Baguma²,

Mukasa³

et al. 2010

African Crop Science Journal

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The current concept for starch biosynthesis in plants is that amylopectin, the major fraction of starch, is synthesised by the concerted actions of ADP-Glc pyrophosphorylase (AGPase), soluble starch synthase (SS), starch-branching enzyme (BE), and starch-debranching enzyme (DBE). We have isolated a cDNA clone of Isoamylase1 gene, a member of DBE family from cassava (Manihot esculenta Crantz) storage root. The cloned cDNA fragment sequence (764 bp) showed high identity of 90% to Rcisa1, and 81% identity to Psisa1, Stisa1 and Atisa1. The deduced protein sequence showed highest (92%) identity with RCISA1. The comparative sequence analysis confirmed the cloned fragment to be M. esculenta isoamylase1 gene (Meisa1; accession number GU229751). Genomic analysis revealed occurrence of at least two copies of the Meisa1 gene. Highest Meisa1 transcript expression levels were detected in fibrous root followed by in the stem and least detected in the leaf and petiole. Analysis of the temporal expression pattern in the storage root showed initial and maximum expression at 90 days after planting (DAP), and declined thereafter to undetectable levels by 180 DAP. The results implicate a major role of Meisa1 in storage root differentiation and early starch granule initiation.

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Section: Spatial and Temporal Expression Of Meisa1supporting

confidence: 87%

“…The occurrence of two or more copies of Meisa1 and GbssI genes could be due to the ploidy level of cassava genome (Awoleye et al, 1994). However, SbeI (Salehuzzaman et al, 1992) and SbeII (Baguma et al, 2003) have been reported to occur as single copies. These two genes could also be a consequence of duplication of the same ancestral gene.…”

Section: Resultsmentioning

confidence: 99%

Characterisation and role of <i>Isoamylase1 (Meisa1</i>) gene in cassava

Beyene¹,

Baguma²,

Mukasa³

et al. 2010

African Crop Science Journal

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“…Hybridization and washing of the blots were performed as described before [35]. Autoradiography was done at -80 °C with an intensifying screen.…”

Section: Northern and Southern Hybridizationmentioning

confidence: 99%

Isolation and characterization of a cDNA encoding granule-bound starch synthase in cassava (Manihot esculenta Crantz) and its antisense expression in potato

1993

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A tuber-specific cDNA library of cassava (Manihot esculenta Crantz) was constructed and a full-length cDNA for granule-bound starch synthase (GBSS, also known as waxy protein), the enzyme responsible for the synthesis of amylose in reserve starch, was cloned. Sequencing of the cloned cDNA showed that it has 74% identity with potato GBSS and 60-72% identity with GBSS from other plant species. The cDNA encodes a 608 amino acid protein of which 78 amino acids form a chloroplast/amyloplast transit peptide of 8.37 kDa. The mature protein has a predicted molecular mass of 58.61 kDa (530 amino acids). Comparison of the GBSS proteins of various plant species and glycogen synthase of bacteria showed extensive identity among the mature form of plant GBSS proteins, in which the monocots and dicots form two separate branches in the evolutionary tree. From analysis of the genomic DNA of allotetraploid cassava, it is shown that GBSS is a low-copy-number gene. GBSS transcript is synthesized in a number of different organs, but most abundantly in tubers. Potato plants were transformed with the cassava GBSS cDNA in antisense orientation fused between the potato GBSS promoter and the nopaline synthase terminator. The expression of the endogenous GBSS gene in these transgenic potato plants was partially or completely inhibited. Complete inhibition of GBSS activity by the cassava antisense gene resulted in absence of GBSS protein and amylose giving rise to almost complete amylose-free potato starch. This shows that also heterologous genes can be used to achieve antisense effects in other plant species.

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“…Genomic and eDNA libraries from leaves, roots, and cotyledons of cassava have been constructed (Hughes et al, 1992;Salehuzzaman et al, 1992;Tenjo and Mayer, 1993;Bohl et al, 1997). These can be used to isolate genes and promoters from cassava, and also to characterize cassava germplasm (Beeching et al, 1993).…”

Section: Dietary Supply Of Protein and Vitaminsmentioning

confidence: 99%

Cassava Biotechnology

Puonti‐Kaerlas

1998

Biotechnology and Genetic Engineering Reviews

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Appan, 1973;Nassar, 1978; Hersey, 1983;Bai et al., 1993). The plants contain lactifers and produce latex, andM. glaziovii is used as a minor source ofrubber (Cock, 1985). Cassava is native to tropical South America, and is one of the oldest cultivated crops (Jennings, 1976) with possibly two centres of origin. Since cassava does not exist in the wild state, and its wild progenitors are not known, the regions of its domestication are disputed (Renvoize, 1972;Rogers and Appan, 1973;Rogers and Flemming, 1973;Nassar, 1978

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Cloning, partial sequencing and expression of a cDNA coding for branching enzyme in cassava

Cited by 45 publications

References 29 publications

Characterisation and role of <i>Isoamylase1 (Meisa1</i>) gene in cassava

Characterisation and role of <i>Isoamylase1 (Meisa1</i>) gene in cassava

Isolation and characterization of a cDNA encoding granule-bound starch synthase in cassava (Manihot esculenta Crantz) and its antisense expression in potato

Cassava Biotechnology

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