Identification of phytic acid mutants in oilseed rape (<i>Brassica napus</i>) by large‐scale screening of mutant populations through amplicon sequencing

Sashidhar, Niharika; Harloff, Hans-Joachim

doi:10.1111/nph.16281

Cited by 27 publications

(35 citation statements)

References 64 publications

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“…The rcm1 mutant was obtained from an EMS-mutagenized population. EMS mutagenesis is widely used in the construction of mutant libraries and in plant functional genomics 37 , 38 . For instance, the hst1 mutant generated from EMS mutagenesis was crossed with the wild type to map the OsRR22 gene 39 .…”

Section: Discussionmentioning

confidence: 99%

SlRCM1, which encodes tomato Lutescent1, is required for chlorophyll synthesis and chloroplast development in fruits

Liu

Yuan

et al. 2021

Hortic Res

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In plants, chloroplasts are the sites at which photosynthesis occurs, and an increased abundance of chloroplasts increases the nutritional quality of plants and the resultant color of fruits. However, the molecular mechanisms underlying chlorophyll synthesis and chloroplast development in tomato fruits remain unknown. In this study, we isolated a chlorophyll-deficient mutant, reduced chlorophyll mutant 1 (rcm1), by ethylmethanesulfonate mutagenesis; this mutant produced yellowish fruits with altered chloroplast development. MutMap revealed that Solyc08g005010 is the causal gene underlying the rcm1 mutant phenotype. A single-nucleotide base substitution in the second exon of SlRCM1 results in premature termination of its translated protein. SlRCM1 encodes a chloroplast-targeted metalloendopeptidase that is orthologous to the BCM1 protein of Arabidopsis and the stay-green G protein of soybean (Glycine max L. Merr.). Notably, the yellowish phenotype of the lutescent1 mutant can be restored with the allele of SlRCM1 from wild-type tomato. In contrast, knockout of SlRCM1 by the CRISPR/Cas9 system in Alisa Craig yielded yellowish fruits at the mature green stage, as was the case for lutescent1. Amino acid sequence alignment and functional complementation assays showed that SlRCM1 is indeed Lutescent1. These findings provide new insights into the regulation of chloroplast development in tomato fruits.

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Section: Discussionmentioning

confidence: 99%

SlRCM1, which encodes tomato Lutescent1, is required for chlorophyll synthesis and chloroplast development in fruits

Liu

Yuan

et al. 2021

Hortic Res

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“…It belongs to the super family of ATP‐grasp fold proteins and is involved in photomorphogenesis by interacting via the COP9 signalosome, in stress responses, in signal transduction and in seed coat development (Du et al , 2011; Qin et al , 2005; Sweetman et al , 2007; Tang et al , 2013). In our previous study, we have identified EMS (ethyl methanesulphonate)‐induced loss‐of‐function mutations in six genes of the PA biosynthesis pathway (Sashidhar et al , 2019). However, due to gene redundancy in polyploid rapeseed noticeable phenotypic effects could only be observed in plants carrying multiple mutations which required time‐consuming crossing experiments.…”

Section: Introductionmentioning

confidence: 99%

Gene editing of three BnITPK genes in tetraploid oilseed rape leads to significant reduction of phytic acid in seeds

Sashidhar

Harloff

Potgieter

et al. 2020

Plant Biotechnology Journal

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Commercialization of Brassica napus. L (oilseed rape) meal as protein diet is gaining more attention due to its well-balanced amino acid and protein contents. Phytic acid (PA) is a major source of phosphorus in plants but is considered as anti-nutritive for monogastric animals including humans due to its adverse effects on essential mineral absorption. The undigested PA causes eutrophication, which potentially threatens aquatic life. PA accounts to 2-5% in mature seeds of oilseed rape and is synthesized by complex pathways involving multiple enzymes. Breeding polyploids for recessive traits is challenging as gene functions are encoded by several paralogs. Gene redundancy often requires to knock out several gene copies to study their underlying effects. Therefore, we adopted CRISPR-Cas9 mutagenesis to knock out three functional paralogs of BnITPK. We obtained low PA mutants with an increase of free phosphorus in the canola grade spring cultivar Haydn. These mutants could mark an important milestone in rapeseed breeding with an increase in protein value and no adverse effects on oil contents.

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“…A previous study had shown determining the level of transcription of candidate genes in the seed at different time points after flowering helped to identify candidate genes. 13 , 14 In this study, the genes related to phytate concentration and content within the LD decay region (∼300 kb) of each significant SNP in B. napus were screened. The transcriptional profile of these genes was employed to help narrow down the list of candidate genes, and finally 30 candidates were identified.…”

Section: Discussionmentioning

confidence: 99%

Integrating a genome-wide association study with transcriptomic data to predict candidate genes and favourable haplotypes influencing Brassica napus seed phytate

Liu

Zhang

et al. 2021

DNA Research

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Phytate is the storage form of phosphorus in angiosperm seeds and plays vitally important roles during seed development. However, in crop plants phytate decreases bioavailability of seed-sourced mineral elements for humans, livestock and poultry, and contributes to phosphate-related water pollution. However, there is little knowledge about this trait in oilseed rape (Brassica napus). Here, a panel of 505 diverse B. napus accessions was screened in a genome-wide association study (GWAS) using 3.28 × 106 single-nucleotide polymorphisms (SNPs). This identified 119 SNPs significantly associated with phytate concentration (PA_Conc) and phytate content (PA_Cont) and six candidate genes were identified. Of these, BnaA9.MRP5 represented the candidate gene for the significant SNP chrA09_5198034 (27 kb) for both PA_Cont and PA_Conc. Transcription of BnaA9.MRP5 in a low-phytate variety (LPA20) was significantly elevated compared with a high-phytate variety (HPA972). Association and haplotype analysis indicated that inbred lines carrying specific SNP haplotypes within BnaA9.MRP5 were associated with high- and low-phytate phenotypes. No significant differences in seed germination and seed yield were detected between low and high phytate cultivars examined. Candidate genes, favourable haplotypes and the low phytate varieties identified in this study will be useful for low-phytate breeding of B. napus.

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Identification of phytic acid mutants in oilseed rape (Brassica napus) by large‐scale screening of mutant populations through amplicon sequencing

Cited by 27 publications

References 64 publications

SlRCM1, which encodes tomato Lutescent1, is required for chlorophyll synthesis and chloroplast development in fruits

SlRCM1, which encodes tomato Lutescent1, is required for chlorophyll synthesis and chloroplast development in fruits

Gene editing of three BnITPK genes in tetraploid oilseed rape leads to significant reduction of phytic acid in seeds

Integrating a genome-wide association study with transcriptomic data to predict candidate genes and favourable haplotypes influencing Brassica napus seed phytate

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