Isolation of an amylose-free starch mutant of the potato (Solanum tuberosum L.)

Hovenkamp‐Hermelink, Johanna H. M.; Jacobsen, E.; Ponstein, A.S.; Visser, R.G.F.; Vos-Scheperkeuter, Greetje H.; Bijmolt, E. W.; Vries, J. N. de; Witholt, Bernard; Feenstra, W.J.

doi:10.1007/bf00249167

Cited by 188 publications

(109 citation statements)

References 17 publications

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“…Of the supernatant 10 pl (_+ 10 pg protein) were applied on gel. Immunoblotting was carried out as described by Hovenkamp-Hermelink et al (1987) using 1:500 diluted anti-BE serum. Alkaline phosphatase was used (2000 times diluted) as a second antibody and the antigen-antibody complexes were detected by incubating the membranes in the dark in 100 ml AF buffer (100 mM Tris-HCI pH 9.5, 100 mM NaCI, 5 mM MgCl 2) with 200 pl NBT (4-nitro blue tetrazoliumchloride; 75 mg ml-1 in dimethylformamide) and 200 pl BCIP (5-bromo-4-chloro-3-indolyl-phosphate; 50 mg ml ~ in H20).…”

Section: Western Analysismentioning

confidence: 99%

Expression of Escherichia coli branching enzyme in tubers of amylose‐free transgenic potato leads to an increased branching degree of the amylopectin

et al. 1996

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SummaryIn order to increase the branching degree of potato tuber starch, the gene encoding branching enzyme (glgB) of Escherichia coil was expressed in the amylose-free potato mutant. The E. coil glgB was cloned in the binary vector pBIN19 under the transcriptional control of the potato Granule Bound Starch Synthase (GBSS) promoter and transitpeptide sequence. The E. coil glgB was cloned behind the two N-terminal amino acids of the GBSS mature protein, creating a chimeric protein. Transgenic plants were obtained which expressed the E. coil branching enzyme as was shown by the presence of mRNA and protein in the tubers. Correctly processed protein was found both in the soluble and starch granule bound protein fraction. Analysis of the starch showed an increase in the branching degree (DE) of up to 25% more branchpoints. The increase in the number of branchpoints was due to the presence of more short chains, with a degree of polymerization (DP) of 16 glucose-residues or less in the amylopectin. Changes in other characteristics of the starch, such as average chain length (CL) and ;~ax, indicated a more branched structure for starch of transformed plants as well.

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Section: Western Analysismentioning

confidence: 99%

Expression of Escherichia coli branching enzyme in tubers of amylose‐free transgenic potato leads to an increased branching degree of the amylopectin

et al. 1996

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“…Amylose synthesis is associated with GBSSI activity. The waxy mutants of cereals (Weatherwax, 1922;Murata et al, 1965;Ishikawa et al, 1994;Nakamura et al, 1995), the amf mutant of potato (Hovenkamp-Hermelink et al, 1987), the lam mutants of pea (Denyer et al, 1995), and GBSSI antisense lines of potato all involve the reduction or elimination of GBSSI activity and a specific reduction or elimination of amylose in the starch of the storage organs of these species.…”

Section: Introductionmentioning

confidence: 99%

“…In potato, by contrast, only one isoform of GBSSI has been described, and mutation of the gene encoding this protein ( AMF ) results in the complete elimination of amylose from tubers, leaves, and roots (Hovenkamp-Hermelink et al, 1987). This finding suggests that there is only one isoform of GBSSI active in the major starch-synthesizing tissues of potato.…”

Section: Introductionmentioning

confidence: 99%

Discrete Forms of Amylose Are Synthesized by Isoforms of GBSSI in Pea[W]

Edwards

Vincken²,

Suurs³

et al. 2002

The Plant Cell

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Amyloses with distinct molecular masses are found in the starch of pea embryos compared with the starch of pea leaves. In pea embryos, a granule-bound starch synthase protein (GBSSIa) is required for the synthesis of a significant portion of the amylose. However, this protein seems to be insignificant in the synthesis of amylose in pea leaves. cDNA clones encoding a second isoform of GBSSI, GBSSIb, have been isolated from pea leaves. Comparison of GBSSIa and GBSSIb activities shows them to have distinct properties. These differences have been confirmed by the expression of GBSSIa and GBSSIb in the amylose-free mutant of potato. GBSSIa and GBSSIb make distinct forms of amylose that differ in their molecular mass. These differences in product specificity, coupled with differences in the tissues in which GBSSIa and GBSSIb are most active, explain the distinct forms of amylose found in different tissues of pea. The shorter form of amylose formed by GBSSIa confers less susceptibility to the retrogradation of starch pastes than the amylose formed by GBSSIb. The product specificity of GBSSIa could provide beneficial attributes to starches for food and nonfood uses.

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“…In rice, there are 10 SS isoforms separated into five types; two granule-bound starch synthase (GBSS) isoforms (GBSSI and GBSSII) in the GBSS type, one SSI isoform in the SSI type, three SSII isoforms (SSIIa [SSII-3], SSIIb , and SSIIc ) in the SSII type, two SSIII isoforms (SSIIIa and SSIIIb ) in the SSIII type, and two SSIV isoforms (SSIVa and SSIVb ) in the SSIV type (Hirose and Terao, 2004). Mutants defective in GBSSI in several species (waxy rice [Sano, 1984]; waxy maize [Tsai, 1974]; waxy barley [Hordeum vulgare ;Eriksson, 1962]; waxy wheat [Triticum aestivum; Nakamura et al, 1995;Triticum monococcum;Fujita et al, 2001]; amf potato [Solanum tuberosum; Hovenkamp-Hermelink et al, 1987]; lam pea [Pisum sativum; Denyer et al, 1995]), SSIII of maize (dull-1; Gao et al, 1998), mutants deficient in SSIIa in rice (most japonica rice varieties are mutants of indica rice varieties [Umemoto et al, 2002;Nakamura et al, 2005]), in barley (sex6; Morell et al, 2003), in wheat (GSP-1 null; Yamamori et al, 2000) and in maize (sugary2; Zhang et al, 2004), and mutants for SSII in pea (rugosus5; Craig et al, 1998) have been identified and analyzed. Most recently, SSI (Delvalle et al, 2005) and SSIII (Zhang et al, 2005) mutants in Arabidopsis (Arabidopsis thaliana) leaves have been isolated.…”

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confidence: 99%

Function and Characterization of Starch Synthase I Using Mutants in Rice

et al. 2006

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(A.M., H.H.) Four starch synthase I (SSI)-deficient rice (Oryza sativa) mutant lines were generated using retrotransposon Tos17 insertion. The mutants exhibited different levels of SSI activities and produced significantly lower amounts of SSI protein ranging from 0% to 20% of the wild type. The mutant endosperm amylopectin showed a decrease in chains with degree of polymerization (DP) 8 to 12 and an increase in chains with DP 6 to 7 and DP 16 to 19. The degree of change in amylopectin chain-length distribution was positively correlated with the extent of decrease in SSI activity in the mutants. The structural changes in the amylopectin increased the gelatinization temperature of endosperm starch. Chain-length analysis of amylopectin in the SSI band excised from native-polyacrylamide gel electrophoresis/SS activity staining gel showed that SSI preferentially synthesized DP 7 to 11 chains by elongating DP 4 to 7 short chains of glycogen or amylopectin. These results show that SSI distinctly generates DP 8 to 12 chains from short DP 6 to 7 chains emerging from the branch point in the A or B 1 chain of amylopectin. SSI seemingly functions from the very early through the late stage of endosperm development. Yet, the complete absence of SSI, despite being a major SS isozyme in the developing endosperm, had no effect on the size and shape of seeds and starch granules and the crystallinity of endosperm starch, suggesting that other SS enzymes are probably capable of partly compensating SSI function. In summary, this study strongly suggested that amylopectin chains are synthesized by the coordinated actions of SSI, SSIIa, and SSIIIa isoforms.

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Isolation of an amylose-free starch mutant of the potato (Solanum tuberosum L.)

Cited by 188 publications

References 17 publications

Expression of Escherichia coli branching enzyme in tubers of amylose‐free transgenic potato leads to an increased branching degree of the amylopectin

Expression of Escherichia coli branching enzyme in tubers of amylose‐free transgenic potato leads to an increased branching degree of the amylopectin

Discrete Forms of Amylose Are Synthesized by Isoforms of GBSSI in Pea[W]

Function and Characterization of Starch Synthase I Using Mutants in Rice

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