Production of waxy tetraploid wheat (Triticum turgidum durum L.) by EMS mutagenesis

Li, Siyu; Zhong, Xiao‐bo; Zhang, Xuteng; Rahman, Md. Mostafijur; Lan, Jingyu; Tang, Huaping; Qi, Pengfei; Ma, Jian; Wang, Jirui; Chen, Guoyue; Lan, Xiujin; Deng, Mei; Li, Zhongyi; Harwood, Wendy; Lu, Zhen-Xiang; Wei, Yuming; Zheng, Youliang; Jiang, Qiantao

doi:10.1007/s10722-019-00875-6

Cited by 9 publications

(12 citation statements)

References 29 publications

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“…They also analyzed the effects of Wx proteins and ranked the single null genotypes as Wx-B1b > Wx-D1b > Wx-A1b. Molecular characterizations of wheat waxy proteins and their effects on starch properties, as well as the detection of Wx null mutations, have been widely studied [47,48,[80][81][82][83], which is important for the identification waxy wheats in breeding programs [84]. The majority of KASP markers found in the CIMMYT used in this study were derived from the Pin-D analysis and were in agreement with the STS markers.…”

Section: Kasp Marker Analysismentioning

confidence: 79%

Puroindoline (Pina-D1 and Pinb-D1) and waxy (Wx-1) genes in Iranian bread wheat (Triticum aestivum L.) landraces

Maryami

Azimi

Guzmán

et al. 2020

Biotechnology & Biotechnological Equipment

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Grain hardness and starch are two of the most important factors that determine the end-use quality of bread wheat (Triticum aestivum L.) grain. The grain hardness and amylose content are controlled by the puroindolines (Pina-D1 and Pinb-D1) genes, located on chromosomes 5D, and waxy (Wx-A1, -B1 and -D1) genes, located on chromosomes 7A, 4A and 7D. A total of 160 Iranian landraces from the Germplasm Bank of the International Maize and Wheat Improvement Center were evaluated for grain hardness using near infrared spectroscopy to predict the particle size index (PSI). In addition, molecular markers were used to evaluate the states of the corresponding genes. Eight accessions were found to have a hard texture (predicted PSI < 45%); however, only two could be explained by null alleles either in Pina-D1 or Pinb-D1. Additionally, 152 accessions had semi-hard textures (predicted PSI range of 45% to 55%). For the Wx gene, only one accession (CWI 67665) showed the null Wx-D1b allele, while two accessions (CWI 67747 and CWI 57684) were null for Wx-B1b. Single nucleotide polymorphism and sequence tag site marker techniques were used. Our findings further indicate the importance of using these landraces for grain quality improvement in breeding programs.

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Section: Kasp Marker Analysismentioning

confidence: 79%

Puroindoline (Pina-D1 and Pinb-D1) and waxy (Wx-1) genes in Iranian bread wheat (Triticum aestivum L.) landraces

Maryami

Azimi

Guzmán

et al. 2020

Biotechnology & Biotechnological Equipment

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“…In our previous study [ 26 ], an EMS-mutagenized population of Chinese durum wheat landrace Jianyangailanmai (LM47) was generated, and some waxy mutant lines were identified from this mutant population. In the present study, SDS-PAGE was used to detect more waxy mutants, and the M2-415 mutant with Wx-B1 protein deletion was identified.…”

Section: Methodsmentioning

confidence: 99%

“…In particular, ethyl methanesulfonate (EMS) has been reported to cause mutations in C–T and G–A substitutions with a high frequency [ 23 , 24 , 25 ]. In our previous work, we generated an EMS-mutagenized tetraploid wheat population [ 26 ], and in the present study, we identified a Wx-B1 null mutant from this population. The sequence of the Wx-B1 gene of the mutant encoded a complete protein, which was incompatible with the protein profile obtained via sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) showing the lack of Wx-B1 protein in the mutant line.…”

Section: Introductionmentioning

confidence: 90%

Regulation of Amylose Content by Single Mutations at an Active Site in the Wx-B1 Gene in a Tetraploid Wheat Mutant

Karim

Wang

et al. 2022

IJMS

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The granule-bound starch synthase I (GBSSI) encoded by the waxy gene is responsible for amylose synthesis in the endosperm of wheat grains. In the present study, a novel Wx-B1 null mutant line, M3-415, was identified from an ethyl methanesulfonate-mutagenized population of Chinese tetraploid wheat landrace Jianyangailanmai (LM47). The gene sequence indicated that the mutated Wx-B1 encoded a complete protein; this protein was incompatible with the protein profile obtained using sodium dodecyl sulfate–polyacrylamide gel electrophoresis, which showed the lack of Wx-B1 protein in the mutant line. The prediction of the protein structure showed an amino acid substitution (G470D) at the edge of the ADPG binding pocket, which might affect the binding of Wx-B1 to starch granules. Site-directed mutagenesis was further performed to artificially change the amino acid at the sequence position 469 from alanine (A) to threonine (T) (A469T) downstream of the mutated site in M3-415. Our results indicated that a single amino acid mutation in Wx-B1 reduces its activity by impairing its starch-binding capacity. The present study is the first to report the novel mechanism underlying Wx-1 deletion in wheat; moreover, it provided new insights into the inactivation of the waxy gene and revealed that fine regulation of wheat amylose content is possible by modifying the GBSSI activity.

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“…The urea solutions had different molarities (0, 3, 4, 5 and 6 mol L –1 ). The mixture was incubated at a room temperature of 26 °C for 24 h. Then, the suspension was centrifuged (8000 × g ) at a room temperature of 26 °C for 20 min and allowed to stand for 1 h 36,37 . The degree of gelatinization was indicated by color change after adding potassium iodide in 200 μL of supernatant 38 …”

Section: Methodsmentioning

confidence: 99%

A single base change at exon of Wx‐A1 caused gene inactivation and starch properties modified in a wheat EMS mutant line

et al. 2021

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BACKGROUND Wheat is an essential source of starch. The GBSS or waxy genes are responsible for synthesizing amylose in cereals. The present study identified a novel Wx‐A1 null mutant line from an ethyl methanesulfonate (EMS)‐mutagenized population of common wheat cv. SM126 using sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and agarose gel analyses. RESULTS The alignment of the Wx‐A1 gene sequences from the mutant and parental SM126 lines showed only one single nucleotide polymorphism causing the appearance of a premature stop codon and Wx‐A1 inactivation. The lack of Wx‐A1 protein resulted in decreased amylose, total starch and resistant starch. The starch morphology assessment revealed that starch from mutant seeds was more wrinkled, increasing its susceptibility to digestion. Regarding the starch thermodynamic properties, the gelatinization temperature was remarkably reduced in the mutant compared to parental line SM126. The digestibility of native, gelatinized, and retrograded starches was analyzed for mutant M4‐627 and the parental SM126 line. In the M4‐627 line, rapidly digestible starch contents were increased, whereas resistant starch was decreased in the three types of starch. CONCLUSION Waxy protein is essential for starch synthesis. The thermodynamic characteristics were decreased in the Wx‐A1 mutant line. The digestibility properties of starch were also affected. Therefore, the partial waxy mutant M3‐627 might play a significant role in food improvement. Furthermore, it might also be used to produce high‐quality noodles. © 2021 Society of Chemical Industry.

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Production of waxy tetraploid wheat (Triticum turgidum durum L.) by EMS mutagenesis

Cited by 9 publications

References 29 publications

Puroindoline (Pina-D1 and Pinb-D1) and waxy (Wx-1) genes in Iranian bread wheat (Triticum aestivum L.) landraces

Puroindoline (Pina-D1 and Pinb-D1) and waxy (Wx-1) genes in Iranian bread wheat (Triticum aestivum L.) landraces

Regulation of Amylose Content by Single Mutations at an Active Site in the Wx-B1 Gene in a Tetraploid Wheat Mutant

A single base change at exon of Wx‐A1 caused gene inactivation and starch properties modified in a wheat EMS mutant line

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