Identification of  resistant starch related genes in rice by single segment substitution lines

LUO Jiarui; ZHU Zhenghang; HAO Qingwen; DING Linyi; ZHU Haitao; ZHANG Guiquan; WANG Shaokui; YANG Weifeng

doi:10.7671/j.issn.1001-411X.202307017

Journal of South China Agricultural University > 2023 > 44(5): 708-717. > DOI: 10.7671/j.issn.1001-411X.202307017

LUO Jiarui, ZHU Zhenghang, HAO Qingwen, et al. Identification of resistant starch related genes in rice by single segment substitution lines[J]. Journal of South China Agricultural University, 2023, 44(5): 708-717. DOI: 10.7671/j.issn.1001-411X.202307017

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Identification of resistant starch related genes in rice by single segment substitution lines

College of Agriculture, South China Agricultural University/Guangdong Provincial Key Laboratory of Plant Molecular Breeding/Guangdong Laboratory for Lingnan Modern Agriculture, Guangzhou 510642, China

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Received Date: July 30, 2023
Available Online: November 12, 2023
Published Date: September 11, 2023

Graphical Abstract

Abstract

Abstract

Objective
To discover the alleles for high resistance starch in starch-synthesis-related genes SBEIIb, SSIIa and ISA1.
Method
The single segment substitution lines (SSSLs) carrying the starch-synthesis-related genes SBEIIb, SSIIa or ISA1 were detected using molecular markers. Then, the resistant starch contents of the SSSLs were measured using an improved AOAC method. Sanger sequencing and sequence alignment were performed to analyze the sequence variations of SBEIIb, SSIIa and ISA1 in different SSSLs. Through linkage analysis of genotypes and phenotypes, the alleles affecting resistance starch content were identified.
Result
For SBEIIb gene, a single nucleotide polymorphism (SNP) (Ex4-96G/A) in the coding region results in an amino acid substitution (196-Arg/His), generating two alleles SBEIIb-1 and SBEIIb-2. SBEIIb-1 carrying the Ex4-96G causes Arg at 196th residue, which shows high-resistant starch content of 1.72%. For SSIIa gene, two SNPs (Ex8-334G/A and Ex8-865C/T) in the 8th exon cause two amino acid substitutions (604-Gly/Ser and 781-Leu/Phe), generating three alleles SSIIa-1,SSIIa-2 and SSIIa-3. SSIIa-1 carrying the Ex8-334G and Ex8-865C causes Gly and Leu at 604th and 781th residue respectively, which shows high-resistant starch content of 3.37%. One Indel (AGG/---) and one SNP (C/T) in ISA1 coding region sequence generate three alleles ISA1-1, ISA1-2 and ISA1-3. ISA1-1 carrying AGG-insertion and Ex17-117C causes Glu and Thr at 70th and 717th residue respectively, which shows high-resistant starch content of 2.09%.
Conclusion
SBEIIb, SSIIa and ISA1 are key genes regulating resistant starch formation in rice. The SNPs and Indels in coding regions of the three genes lead to amino acid variations, which subsequently affects the resistance starch content. The three alleles SBEIIb-1, SSIIa-1 and ISA1-1 for high-resistant starch content are identified.
- Rice,
- Single segment substitution line,
- Resistant starch,
- Allele

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References (33)

References

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Identification of resistant starch related genes in rice by single segment substitution lines

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