Abstract
Branching enzyme is involved in the synthesis of amylopectin in plant reserve starch. A cDNA coding for cassava (Manihot esculenta Crantz) branching enzyme was cloned from a λgt11 cDNA library using a potato cDNA probe. The cloned cDNA was partially sequenced. The sequence data confirmed the identity of the clone when compared to that of potato, the homology being ca. 80% at the nucleotide level and 85% at the amino acid level. Furthermore, the cloned cassava cDNA was able to restore branching enzyme activity in a branching enzyme deficient Escherichia coli mutant. Results of the Southern analysis suggested that there is a single gene for this particular branching enzyme in the cassava genome. Study of expression patterns by northern hybridization showed that the gene is highly expressed in tubers. The transcript is detectable in stem and petiole, but not in leaves. In roots, the mRNA is hardly present. The expression levels at different stages of tuber growth are similar with exception of very young tubers in which it is relatively low. It is also shown that there is a difference in the level of branching enzyme expression between different cassava genotypes.
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Bhattacharyya MK, Smith AM, Noel Ellis TH, Hedley C, Martin C: The wrinkled-seed character of pea described by Mendel is caused by a transposon-like insertion in a gene encoding starch-branching enzyme. Cell 60: 115–122 (1990).
Blennow A, Johansson G: Isolation of a Q-enzyme with M r 103000 from potato tubers. Phytochemistry 30: 437–444 (1991).
Borovsky D, Smith EE, Whelan WJ: Purification and properties of potato 1,4-α-D-glucan: 1,4-α-D,glucan 6-α-D-glucano)-transferase. Evidence against a dual catalytic function in amylose-branching enzyme. Eur J Biochem 59: 615–625 (1975).
Boyer CD: Soluble starch synthases and starch branching enzyme from developing seeds of sorghum. Phytochemistry 24, 15–18 (1985).
Boyer CD, Fisher MB: Comparison of soluble starch synthases and branching enzymes from developing maize and teosinte seeds. Phytochemistry 23: 733–737 (1984).
Chambers SP, Prior SE, Bartsow DA, Minton NP: The pMTL nic −cloning vectors. I. Improved pUC polylinker regions to facilitate the use of sonicated DNA for nucleotide sequencing. Gene 68: 139–149 (1988).
Cooke RD, Cock JH: Cassava crops up again. New Scient 122 (1669): 63–68 (1989).
Dellaporta SL, Wood J, Hicks JB: A plant DNA minipreparation: version II. Plant Mol Biol Rep 1: 19–21 (1983).
Hawker JS, Ozbun JL, Ozaki H, Greenberg E, Preiss J: Interaction of spinach leaf adenosine diphosphate glucose α-1,4-glucan α-4-glucosyl transferase and α-1,4-glucan, α-1,4-glucan-6-glucosyl transferase in synthesis of branched α-glucan. Arch Biochem Biophys 160: 530–551 (1974).
Hedley CL, Smith CM, Ambrose MJ, Cook S, Wang TL: An analysis of seed development in Pisum sativum. II. The effect of the r-locus on the growth and development of the seed. Ann Bot 58: 371–379 (1986).
Jenning DL, Hershey CH: Cassava breeding: a decade of progress from International programmes. In: Russell GE (ed) Progress in Plant Breeding 1, pp. 89–115, Butterworths, London (1985).
Kay DL: Crops and Products Digest, No. 2: Root Crops, 2nd ed., pp. 30–56. Tropical Development and Research Institute, London (1985).
Kiel JAKW, Vossen JPMJ, Venema G: A general method for the construction of Escherichia coli mutants by homologous recombination and plasmid segregation. Mol Gen Genet 207: 294–301 (1987).
Kossmann J, Visser RGF, Müller-Röber BT, Willmitzer L, Sonnewald U: Cloning and expression analysis of a potato cDNA that encodes branching enzyme: evidence for coexpression of starch biosynthetic genes. Mol Gen Genet 230: 39–44 (1991).
Landsmann J, Uhrig H: Somaclonal variation in Solanum tuberosum detected at the molecular level. Theor Appl Genet 71: 500–505 (1985).
Müller-Röber BT, Kossmann J, Hannah LC, Willmitzer L, Sonnewald U: One of the two different ADP-glucose pyrophosphorylase genes responds strongly to elevated levels of sucrose. Mol Gen Genet 224: 136–146 (1990).
Nakamura K, Hattori T, Asahi T: Direct immunological identification of full-length cDNA clones for plant protein without gene fusion to E. coli protein. FEBS Lett 198: 16–20 (1986).
Nelson N: A photemetric adaptation of the Somogyi method for the determination of glucose. J Biol Chem 153: 375–380 (1944).
O'Hair SK: Cassava root starch content and distribution varies with tissue age. Hortscience 24: 505–506 (1989).
Poehlman JM, Quick JS: Crop breeding in a hungry world. In: Wood DR (ed) Crop breeding, pp. 1–19, American Society of Agronomy and the Crop Science Society of America, Madison, WI (1983).
Queen C, Korn LJ: A comprehensive sequence analysis program for the IBM personal computer. Nucleic Acids Res 12: 581–599 (1984).
Raemakers CJJM, Bessembinder JJE, Staritsky G, Jacobsen E, Visser RGF: Somatic embryogenesis and plant regeneration in cassava. Plant Cell Tissue Organ Culture, In press (1992).
Rickard JE, Asaoka M, Blanshard JMV: The physiochemical properties of cassava starch. Trop Sci 31: 189–207 (1991).
Rickard JE, Behn KR: Evaluation of acid and enzyme hydrolytic methods for the determination of cassava starch. J Sci Food Agric 41: 373–379 (1987).
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Habor, NY (1989).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).
Shannon JC, Garwood DL: Genetics and physiology of starch development. In: Whistler RL, Bemiller JN, Paschall EF (eds) Starch: Chemistry and Technology, 2nd ed., pp. 25–86, Academic Press, Orlando, FL (1984).
Smith AM: Major differences in isoforms of starch branching enzyme between developing embryos of round-and wrinkled-seeded peas (Pisum sativum L.). Planta 175: 270–279 (1988).
Smyth DA: Some properties of starch branching enzyme from indica rice endosperm (Oryza sativa L.). Plant Sci 57: 1–18 (1988).
Tautz D, Renz M: An optimal freeze-squeeze method for the recovery of DNA fragments from agarose gel. Anal. Biochem 132: 14–19 (1983).
Visser RGF, Hergersberg M, van derLeij FR, Jacobsen E, Witholt B, Feenstra WJ: Molecular cloning and partial characterization of the gene for granule bound starch synthase from a wildtype and an amylose-free potato (Solanum tuberosum L.), Plant Sci 64: 185–192 (1989).
Visser RGF, Somhorst I, Kuipers GJ, Ruys NJ, Feenstra WJ, Jacobsen E: Inhibition of the expression of the gene for granule-bound starch synthase in potato by antisense constructs. Mol Gen Genet 225: 289–296 (1991).
Vos-Scheperkeuter GH, Boer Wde, Visser RGF, Feenstra WJ, Witholt B: Identification of granule-bound starch synthase in potato tubers. Plant Physiol 82: 411–416 (1986).
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Salehuzzaman, S.N.I.M., Jacobsen, E. & Visser, R.G.F. Cloning, partial sequencing and expression of a cDNA coding for branching enzyme in cassava. Plant Mol Biol 20, 809–819 (1992). https://doi.org/10.1007/BF00027152
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DOI: https://doi.org/10.1007/BF00027152