Variation in Friabilin Composition as Determined by A-PAGE Fractionation and PCR Amplification, and its Relationship to Grain Hardness in Bread Wheat

Montiel-Corona, Virginia; Gazza, Laura; Boggini, G.; Pogna, Norberto

doi:10.1006/jcrs.2000.0416

Cited by 39 publications

(27 citation statements)

References 14 publications

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“…2 carried a faint PINB band without PINA. These results confirmed that lower PINB amounts on the starch granule were closely correlated with harder grain texture, and PINA may play a role in mediating the interaction between PINB and starch, as suggested by previous studies (Giroux and Morris 1997;Corona et al 2001;Capparelli et al 2003;Amoroso et al 2004;Gazza et al 2005). …”

Section: Pina and Pinb Fractioned In Sds-pagesupporting

confidence: 90%

“…The extraction method was based on the procedure described by Morris et al (1994), Corona et al (2001), and Gazza et al (2005) with slight modifications that the extraction was carried out in 45°C water-incubation with continuous shaking instead of room temperature.…”

Section: Extraction and Fraction Of Puroindoline Proteinsmentioning

confidence: 99%

“…However, the molecular characters of puroindoline genes cannot easily be resolved by using only protein electrophoresis. Allelic variations of Pina and Pinb are commonly checked by PCR amplification using different allele-specific primers (ASP) designed from known alleles (Giroux and Morris 1998;Lillemo and Morris 2000;Corona et al 2001;Guo et al 2004;Chen et al 2006). Nevertheless, it is not easy for these methods to detect unknown alleles of Pina and Pinb.…”

Section: Introductionmentioning

confidence: 99%

“…In studying characters of PINA and PINB, various methods including SDS-PAGE, A-PAGE and 2-D PAGE were used to provide clear separation of the two peptides (Oda 1994;Giroux and Morris 1998;Corona et al 2001;Gazza et al 2005). However, the molecular characters of puroindoline genes cannot easily be resolved by using only protein electrophoresis.…”

Section: Introductionmentioning

confidence: 99%

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Identification of allelic variations of puroindoline genes controlling grain hardness in wheat using a modified denaturing PAGE

Chang

Zhang

et al. 2006

Euphytica

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Puroindoline genes (Pina and Pinb) form the molecular basis of wheat grain hardness or texture. Variations in either gene are reported to be associated with grain hardness in wheat. Here, a modified denaturing PAGE was used to detect Pina and Pinb allelic variations in 102 common wheat cultivars and other species related to wheat. Two variations of Pina (Pina-D1b and Pina-D1p) and five variations of Pinb (null, Pinb-D1b, PinbD1u, Pinb-D1v and Pinb-D1w) were identified in common (T. aestivum) and spelt wheats (T. aestivum ssp. spelta). No allelic variation was found in Tibet semi-wild wheat (T. aestivum ssp. tibetanum), club wheat (T. compactum), or Aegilops tauschii. Puroindoline genes were absent in wild emmer (T. turgidum var. dicoccoides). The sequencing results of the PCR fragments from Pina and Pinb (except for null type) indicated allelic variants carrying single base mutation, such as Pina-D1p, Pinb-D1u, Pinb-D1v and Pinb-D1w were novel types. Our results showed that the modified PAGE used in this study provided a satisfactory resolving power for identifying single nucleotide mutations; therefore, it is a practical and simple tool to study allelic variation of Pina and Pinb in wheat and related species.

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Section: Pina and Pinb Fractioned In Sds-pagesupporting

confidence: 90%

Section: Extraction and Fraction Of Puroindoline Proteinsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Identification of allelic variations of puroindoline genes controlling grain hardness in wheat using a modified denaturing PAGE

Chang

Zhang

et al. 2006

Euphytica

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“…The finding that 1% SDS preferentially extracts PINB from starch granules as compared with PINA (Corona et al 2001a;Pogna et al 2002;Gazza et al 2005) suggests that the 15-kDa band, previously described by Greenwell and Schofield (1986), corresponds to PINB. The A-PAGE fractionation of proteins associated with starch granules provided a clear separation of PINA and PINB compared with SDS-PAGE (Corona et al 2001b;Pogna et al 2002).…”

Section: Physical and Biochemical Properties Of The Puroindolines Andmentioning

confidence: 99%

The determinants of grain texture in cereals

2009

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Kernel hardness is an important agronomic trait that influences end-product properties. In wheat cultivars, this trait is determined by the Puroindoline a (Pina) and Puroindoline b (Pinb) genes, located in the Hardness locus (Ha) on chromosome 5DS of the D genome. Wild type alleles code puroindoline a (PINA) and puroindoline b (PINB) proteins, which form a 15-kDa friabilin present on the surface of water-washed starch granules. Both the proteins are accumulated in the starch endosperm cells and aleurone of the mature kernels. Puroindoline-like genes coding puroindoline-like proteins in the starch endosperm occur in some of the genomes of Triticeae and Aveneae cereals. Orthologs are present in barley, rye and oats. However, some genomes of these diploid and polyploid cereals, like that of Triticum turgidum var. durum (AABB) lack the puroindoline genes, having a very hard kernel texture. The two wild type alleles in opposition (dominant loci) control the soft phenotype. Mutation either in Pina or Pinb or in both leads to a medium-hard or hard kernel texture. The most frequent types of Pin mutations are point mutations within the coding sequence resulting in the substitution of a single amino acid or a null allele. The latter is the result of a frame shift determined by base deletion or insertion or a one-point mutation to the stop codon. The lipid-binding properties of the puroindolines affect not only the dough quality but also the plants' resistance to pathogens. Genetic modification of cereals with Puroindoline genes and/or their promoters enable more detailed functional analyses and the production of plants with the desired characteristics.

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CE determination of secaloindoline allelic forms in hexaploid triticale (×Triticosecale Wittmack)

Salmanowicz

2010

J of Separation Science

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Differences in kernel texture are mainly caused by specific secaloindoline (SIN) proteins occurring in friabilin fraction of hexaploid triticale (x Triticosecale Wittmack) grain. SINs were isolated using Triton X-114 partitioning from either kernels/flour or starch of five triticale cultivars with wide range of different hardness. Crude SIN fraction was obtained by size-exclusion HPLC. SINs were separated on an uncoated fused-silica capillary using the iminodiacetic (IDA) buffer in conjunction with lower-concentrated poly(ethylene oxide) and ACN. A low-concentrate mixture of hydrophilic polymers, PVP and hydroxypropylmethylcellulose in IDA buffer was employed for dynamic coating of capillary inner wall. In total, on the basis of CZE profiles, two SIN-a proteins and two SIN-b proteins were identified. Allelic forms SIN-a1 and SIN-b1 have both two soft and one medium hard genotypes, however other allelic forms, designed as SIN-a2 and SIN-b2, were identified in hard and other medium hard cultivars. The CZE profiles showed that the ratio of the peak areas of SIN-b proteins isolated from triticale starch can be preliminarily used to distinguish cultivars with soft and hard grain.

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Variation in Friabilin Composition as Determined by A-PAGE Fractionation and PCR Amplification, and its Relationship to Grain Hardness in Bread Wheat

Cited by 39 publications

References 14 publications

Identification of allelic variations of puroindoline genes controlling grain hardness in wheat using a modified denaturing PAGE

Identification of allelic variations of puroindoline genes controlling grain hardness in wheat using a modified denaturing PAGE

The determinants of grain texture in cereals

CE determination of secaloindoline allelic forms in hexaploid triticale (×Triticosecale Wittmack)

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