Abstract
Key message
Mutation of the BEIIb gene in an isa1 mutant background mitigates the negative effect of the ISA1 mutation on grain filling, and facilitates recovery of amyloplast formation in rice endosperm.
Abstract
In this study, the effect of branching enzyme IIb and isoamylase 1 deficiency on starch properties was demonstrated using high resistant starch rice lines, Chikushi-kona 85 and EM129. Both lines harbored a mutation in the BEIIb and ISA1 genes and showed no BEIIb and ISA1 activity, implying that both lines are beIIb isa1 double mutants. The amylopectin long chain and apparent amylose content of both mutant lines were higher than those of the wild-type. While both mutants contained loosely packed, round starch grains, a trait specific to beIIb mutants, they also showed collapsed starch grains at the center of the endosperm, a property specific to isa1 mutants. Furthermore, beIIb isa1 double mutant F2 lines derived from a cross between Chikushi-kona 85 and Nishihomare (wild-type cultivar) showed significantly heavier seed weight than the beIIb and isa1 single mutant lines. These results suggest that co-occurrence of beIIb and isa1 mutant alleles in a single genetic background mitigates the negative effect of the isa1 allele on grain filling, and contributes to recovery of the amyloplast formation defect in the isa1 single mutant.
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Acknowledgements
This work was supported by the Joint Usage/Research Center, Institute of Plant Science and Resources, Okayama University and JSPS KAKENHI (19H02933). We express our gratitude to Dr. Hikaru Satoh, former professor at Kyushu University, for providing seeds of EM129 and EM10, and we sincerely thank Ms. Aiko Nishi for technical advice. Pseudomonas isoamylase used for debranching amylopectin was a kind gift from Hayashibara Co., Ltd. We thank Bioedit Ltd. (https://www.bioedit.com/) for editing the English text of a draft of this manuscript.
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This work was supported by the Joint Usage/Research Center, Institute of Plant Science and Resources, Okayama University, JSPS KAKENHI (19H02933).
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SN and TW conceived of the study. SN, TW, OY, and TK developed the Plant Materials. SN, TW, RM, and OY grew and evaluated Plant Materials. NF, SM, NC, and YH performed chain length analysis and gel filtration. RM fixed and sliced the samples, and SN, TW, and RM examined them. NF stained grain cross-sections. SM, NC, and YH performed Western blotting and native-PAGE. SN and TW identified the mutation sites and performed expression analysis. SN wrote the manuscript with support from TW. All authors provided critical feedback and helped shape the research, analysis, and manuscript. TW supervised the whole study.
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Nagamatsu, S., Wada, T., Matsushima, R. et al. Mutation in BEIIb mitigates the negative effect of the mutation in ISA1 on grain filling and amyloplast formation in rice. Plant Mol Biol 108, 497–512 (2022). https://doi.org/10.1007/s11103-022-01242-3
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DOI: https://doi.org/10.1007/s11103-022-01242-3