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Molecular Characterization of Puroindoline Alleles in Chinese and CIMMYT Common Wheats and Their Effect on Processing Quality

Author: ChenFeng
Tutor: HeZhongHu;XiaXianChun
School: Chinese Academy of Agricultural Sciences
Course: Crop Genetics and Breeding
Keywords: Common wheat Synthetic wheat Kernel hardness Puroindoline alleles Variation of alleles Milling quality Processing quality
CLC: S512.1
Type: PhD thesis
Year: 2006
Downloads: 119
Quote: 6
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Kernel hardness is one of the most important characters with a profound influence on milling performance and end-use quality of common wheat (Triticum asetivum L.). Understanding molecular characterization of puroindoline genes controlling kernel hardness would be beneficial for improving wheat quality. Materials used in this study are consisted of 805 Chinese cultivars, including current winter and spring wheat cultivars, landraces and historical cultivars, and 596 CIMMYT cultivars and advanced lines. All of the surveyed materials were used to measure kernel hardness by Single Kernel Characterization System (SKCS) or Partical Size Index (PSI) and detect the variation of puroindoline alleles with STS (sequence tagged site) marker, site-specific cleavage with the restriction enzyme PvuII and modified SDS-PAGE (sodium dodecyl-sulfate polyacrylamide gel electrophoresis) of Triton X-114 soluble protein, 10% glycerol substituting of water for resolving gels and PDA (piperiazine diacrylamide) substituting of N, N’-methylene bisacrylamide for gels, respectively. In addition, 286 genotypes were used to measure milling and processing quality. The main results obtained in this study are summarized below.1. Hard wheat was dominant in Chinese current winter and spring wheats and its percentage was gradually decreased from north to sourth. Pinb-Dlb, Pina-D1b and Pinb-D1d of known alleles were present in hard winter wheat and Pinb-Dlb, Pina-Dlb and Pinb-D1c of known alleles were present in hard spring wheat. Pinb-Dlb was the most prevalent hard genotype in Chinese current winter and spring wheats with percentages of 76.5% and 51.5%, respectively. Pina-Dlb with percentage of 37.9% in Chinese spring wheat was higher than that with percentage of 16.0% in Chinese winter wheat. Pinb-Dld was identified in two Chinese winter wheat cultivars, while Pinb-Dlc was detected in five Chinese spring wheats. Two novel puroindoline alleles were identified in Chinese winter wheat cultivars, which were designated as: i) Pinb-Dlp with characterization of a single nucleotide (A) deletion corresponding to position 42 in the amino acid sequence of puroindoline b, resulting in a Lysine (K) to Asparagine (N) change and leading to a shift in the open reading frame (ORF) and ii) Pinb-D1q with characterization of a base G to T substitution at position 218th nucleotide in the coding sequence of the Pinb gene, resulting in a tryptophan to leucine change (TGG to TTG) at position 44 in the deduced amino acid sequence of puroindoline b.2. Hard wheat was dominant in Chinese landraces and historical cultivars. Pinb-Dlb, Pina-Dlb and Pinb-Dlp of known alleles were present in hard wheat. From Chinese landraces, historical cultivars to current wheat cultivars, the percentages of soft (42.7%, 45.2% and 25.1%) and mixed (24.3%, 13.9% and 12.4%) wheats were gradually reduced, whereas the percentage of hard wheat (33.0%, 40.9% and 62.5%) was significantly increased. PINA null (43.8%, 48.5% and 13.4%) and Pinb-Dlp (39.7%, 14.7% and 8.4%) were gradually reduced while percentage of Pinb-Dlb (12.3% and 36.8% and 76.5%) was significantly increased. Three novel puroindoline alleles were identified in Chinese landrance cultivars, which were designated as: i) Pinb-D1t with characterization of a base G to C substitution at the 226thnucleotide in the coding sequence of the Pinb gene, resulting in a glycine (GGC) to arginine (CGC) substitution at position 47 in the deduced amino acid sequence of puroindoline b, ii) Pina-DH with characterization of a base G to A change at position 212, resulting in tryptophan-43 to a ’stop’ codon, and iii) Pina-Dlm with characterization of a base C to T substitution at the 187th nucleotide in the coding sequence of the Pina gene, which results in a proline (CCG) to serine (TCG) substitution at position 35 in the deduced amino acid sequence of puroindoline a. In addition, the Pina-Dlc allele, characterizing as a base C deletion at position 265, was renamed as Pina-DH due to conflicting with a previous report after negotiation with Drs Mclntosh and Morris. Pina-Dlm allele reported in this study is the first amino acid subsitution in PINA of bread wheat that is known to result in hard endosperm, which modifies the molecular genetic model of grain hardness.3. In CIMMYT wheat cultivars and advanced lines, hard wheat was the most dominant phenotype. Pina-Dlb and Pinb-Dlb were present in hard wheat. Pina-Dlb was the most popular genotype in CIMMYT hard wheats with a percentage of 86.3% and all other hard wheats possessed Pinb-Dlb genotype. Pina-Dlb genotype had inferior milling and processing quality than that of Pinb-Dlb. Therefore, it was recommended that other types of puroindoline alleles should be introduced in CIMMYT wheat breeding program to reduce the negetive influence of the Pina-Dlb genotype on milling and processing quality. In addition, Pina-Dla/Pinb-Dla, Pina-Dlj/Pinb-Dli, Pina-Dlc/Pinb-Dlh, Pina-Dla/Pinb-DH and Pina-Dla/Pinb-Dlj were present in genotypes with puroindoline alleles from various Aegilops tauschii. They were still soft wheats and Pina-Dlj/Pinb-Dli was the most popular genotype among them. Means of SKCS hardness with Pina-Dlc/Pinb-Dlh and Pina-Dla/Pinb-DH were significantly higher than those of Pina-Dlj/Pinb-Dli and Pina-Dla/Pinb-Dla. It is very useful to improve the efficiency of wheat breeding that new puroindoline alleles have recently been introduced through the extensive use of synthetic hexaploid wheat.4. Kernel hardness have significantly positive correlations with flour particle size (r=0.91), water solvent retention capacity (r=0.73), sodium carbonate solvent retention capacity (r=0.59) and farinograph water absorption (r=0.80), and significantly negetive correlations with flour colour L* (r=-0.77). Means of flour ash content and farinograph water absorption of lines with Pinb-Dlb were significantly lower than those of Pina-Dlb, while means of milling yield, volume and total score of steamed bread, firmness and total score of noodle of Pinb-Dlb genotypes were significantly higher than those of Pina-Dlb, suggesting that Pinb-Dlb genotype has slightly superior milling quality and processing qualities of steamed bread and noodle to Pina-Dlb genotype.This is the first extensive survey of puroindoline alleles in Chinese and CIMMYT wheats. It also indicates that variations of puroindoline locus have dramatic effect on milling and processing quality, and suggest that increasing Pinb-Dlb genotype in CIMMYT and Chinese wheat breeding, especially in spring wheat region, will be crucial for improvement of wheat quality in China and CIMMYT.

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