scholarly journals An Inositol 1,3,4,5,6-Pentakisphosphate 2-Kinase 1 Mutant with a 33-nt Deletion Showed Enhanced Tolerance to Salt and Drought Stress in Rice

Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 23
Author(s):  
Meng Jiang ◽  
Yanhua Liu ◽  
Ruiqing Li ◽  
Shan Li ◽  
Yuanyuan Tan ◽  
...  
Keyword(s):  
Phytic Acid ◽  
Agronomic Traits ◽  
Inositol Triphosphate ◽  
Rice Grain ◽  
Wild Type ◽  
Low Phytic Acid ◽  
Stress Response Genes ◽  
Potential Applications ◽  
Salt And Drought Stress ◽  
Mutant Lines

OsIPK1 encodes inositol 1,3,4,5,6-pentakisphosphate 2-kinase, which catalyzes the conversion of myo-inositol-1,3,4,5,6-pentakisphosphate to myo-inositol-1,2,3,4,5,6-hexakisphosphate (IP6) in rice. By clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas9)-mediated mutagenesis in the 3rd exon of the gene, three OsIPK1 mutations, i.e., osipk1_1 (a 33-nt deletion), osipk1_2 (a 1-nt deletion), and osipk1_3 (a 2-nt deletion) were identified in T0 plants of the rice line Xidao #1 (wild type, WT). A transfer DNA free line with the homozygous osipk1_1 mutation was developed; however, no homozygous mutant lines could be developed for the other two mutations. The comparative assay showed that the osipk1_1 mutant line had a significantly lower level of phytic acid (PA, IP6; −19.5%) in rice grain and agronomic traits comparable to the WT. However, the osipk1_1 mutant was more tolerant to salt and drought stresses than the WT, with significantly lower levels of inositol triphosphate (IP3), reactive oxygen species (ROS) and induced IP6, and higher activities of antioxidant enzymes in seedlings subjected to these stresses. Further analyses showed that the transcription of stress response genes was significantly upregulated in the osipk1_1 mutant under stress. Thus, the low phytic acid mutant osipk1_1 should have potential applications in rice breeding and production.

scholarly journals lpa1-5525: A New lpa1 Mutant Isolated in a Mutagenized Population by a Novel Non-Disrupting Screening Method

Plants ◽  
2019 ◽  
Vol 8 (7) ◽  
pp. 209 ◽  
Author(s):  
Giulia Borlini ◽  
Cesare Rovera ◽  
Michela Landoni ◽  
Elena Cassani ◽  
Roberto Pilu
Keyword(s):  
Phytic Acid ◽  
Screening Method ◽  
Lower Amount ◽  
Wild Type ◽  
New Mutation ◽  
Breeding Programs ◽  
Low Phytic Acid ◽  
Protein Storage Vacuoles ◽  
Mixed Salts ◽  
Phytic Phosphorus

Phytic acid, or myo-inositol 1,2,3,4,5,6-hexakisphosphate, is the main storage form of phosphorus in plants. It is localized in seeds, deposited as mixed salts of mineral cations in protein storage vacuoles; during germination, it is hydrolyzed by phytases to make available P together with all the other cations needed for seed germination. When seeds are used as food or feed, phytic acid and the bound cations are poorly bioavailable for human and monogastric livestock due to their lack of phytase activity. Therefore, reducing the amount of phytic acid is one strategy in breeding programs aimed to improve the nutritional properties of major crops. In this work, we present data on the isolation of a new maize (Zea mays L.) low phytic acid 1 (lpa1) mutant allele obtained by transposon tagging mutagenesis with the Ac element. We describe the generation of the mutagenized population and the screening to isolate new lpa1 mutants. In particular, we developed a fast, cheap and non-disrupting screening method based on the different density of lpa1 seed compared to the wild type. This assay allowed the isolation of the lpa1-5525 mutant characterized by a new mutation in the lpa1 locus associated with a lower amount of phytic phosphorus in the seeds in comparison with the wild type.

scholarly journals Mutation of Inositol 1,3,4-trisphosphate 5/6-kinase6 Impairs Plant Growth and Phytic Acid Synthesis in Rice

Plants ◽  
2019 ◽  
Vol 8 (5) ◽  
pp. 114 ◽  
Author(s):  
Meng Jiang ◽  
Yang Liu ◽  
Yanhua Liu ◽  
Yuanyuan Tan ◽  
Jianzhong Huang ◽  
...  
Keyword(s):  
Plant Growth ◽  
Phytic Acid ◽  
Targeted Mutagenesis ◽  
Rice Grain ◽  
Nucleotide Substitutions ◽  
Phytic Acid Content ◽  
Frameshift Mutations ◽  
A Genome ◽  
Mutant Lines ◽  
Growth And Reproduction

Inositol 1,3,4-trisphosphate 5/6-kinase (ITPK) is encoded by six genes in rice (OsITPK1-6). A previous study had shown that nucleotide substitutions of OsITPK6 could significantly lower the phytic acid content in rice grains. In the present study, the possibility of establishing a genome editing-based method for breeding low-phytic acid cultivars in rice was explored, in conjunction with the functional determination of OsITPK6. Four OsITPK6 mutant lines were generated by targeted mutagenesis of the gene’s first exon using the CRISPR/Cas9 method, one (ositpk6_1) with a 6-bp in-frame deletion, and other three with frameshift mutations (ositpk6_2, _3, and _4). The frameshift mutations severely impaired plant growth and reproduction, while the effect of ositpk6_1 was relatively limited. The mutant lines ositpk6_1 and _2 had significantly lower levels (−10.1% and −32.1%) of phytic acid and higher levels (4.12- and 5.18-fold) of inorganic phosphorus compared with the wild-type (WT) line. The line ositpk6_1 also showed less tolerance to osmotic stresses. Our research demonstrates that mutations of OsITPK6, while effectively reducing phytic acid biosynthesis in rice grain, could significantly impair plant growth and reproduction.

scholarly journals Combination of High Zn Density and Low Phytic Acid for Improving Zn Bioavailability in Rice (Oryza stavia L.) Grain

Rice ◽  
2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Yin Wang ◽  
Yusha Meng ◽  
Yanping Ma ◽  
Lei Liu ◽  
Dianxing Wu ◽  
...  
Keyword(s):  
Phytic Acid ◽  
Double Mutant ◽  
Cell Model ◽  
Brown Rice ◽  
Molar Ratio ◽  
Zn Deficiency ◽  
Rice Grain ◽  
Phytic Acid Content ◽  
Low Phytic Acid ◽  
Zn Bioavailability

Abstract Background Zn deficiency is one of the leading public health problems in the world. Staple food crop, such as rice, cannot provide enough Zn to meet the daily dietary requirement because Zn in grain would chelate with phytic acid, which resulted in low Zn bioavailability. Breeding new rice varieties with high Zn bioavailability will be an effective, economic and sustainable strategy to alleviate human Zn deficiency. Results The high Zn density mutant LLZ was crossed with the low phytic acid mutant Os-lpa-XS110–1, and the contents of Zn and phytic acid in the brown rice were determined for the resulting progenies grown at different sites. Among the hybrid progenies, the double mutant always displayed significantly higher Zn content and lower phytic acid content in grain, leading to the lowest molar ratio of phytic acid to Zn under all environments. As assessed by in vitro digestion/Caco-2 cell model, the double mutant contained the relatively high content of bioavailable Zn in brown rice. Conclusions Our findings suggested pyramiding breeding by a combination of high Zn density and low phytic acid is a practical and useful approach to improve Zn bioavailability in rice grain.

scholarly journals Using CRISPR-Cas9 to generate semi-dwarf rice lines in elite landraces

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Xingming Hu ◽  
Yongtao Cui ◽  
Guojun Dong ◽  
Anhui Feng ◽  
Danying Wang ◽  
...  
Keyword(s):  
Gene Editing ◽  
Agronomic Traits ◽  
Genetic Erosion ◽  
Field Trials ◽  
Wild Type ◽  
Biotic And Abiotic Stresses ◽  
Rice Varieties ◽  
Broad Spectrum Resistance ◽  
Mutant Lines ◽  
Better Than

AbstractGenetic erosion refers to the loss of genetic variation in a crop. In China, only a few original landraces of rice (Oryza sativa) were used in breeding and these became the primary genetic background of modern varieties. Expanding the genetic diversity among Chinese rice varieties and cultivating high-yielding and high-quality varieties with resistance to different biotic and abiotic stresses is critical. Here, we used the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein9(Cas9) genome editing system to edit Semi-Dwarf1 (SD1) in the elite landraces Kasalath and TeTePu (TTP), which contain many desired agronomic traits such as tolerance to low phosphorous and broad-spectrum resistance to several diseases and insects. Mutations of SD1 confer shorter plant height for better resistance to lodging. Field trials demonstrated that the yield of the new Kasalath and TTP mutant lines was better than that of the wild type under modern cultivation and that the lines maintained the same desirable agronomic characteristics as their wild-type progenitors. Our results showed that breeding using available landraces in combination with genomic data of different landraces and gene-editing techniques is an effective way to relieve genetic erosion in modern rice varieties.

Molecular marker development and linkage analysis in three low phytic acid barley (Hordeum vulgare) mutant lines

2007 ◽  
Vol 20 (4) ◽  
pp. 323-330 ◽  
Author(s):  
Vicky Roslinsky ◽  
Peter E. Eckstein ◽  
Victor Raboy ◽  
Brian G. Rossnagel ◽  
Graham J. Scoles
Keyword(s):  
Hordeum Vulgare ◽  
Linkage Analysis ◽  
Molecular Marker ◽  
Phytic Acid ◽  
Marker Development ◽  
Low Phytic Acid ◽  
Molecular Marker Development ◽  
Mutant Lines

The concentrations and distributions of phytic acid phosphorus and other mineral nutrients in wild-type and low phytic acid Js-12-LPA wheat (Triticum aestivum) grain parts

2005 ◽  
Vol 83 (12) ◽  
pp. 1599-1607 ◽  
Author(s):  
Charlie Joyce ◽  
Andrea Deneau ◽  
Kevin Peterson ◽  
Irene Ockenden ◽  
Victor Raboy ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Triticum Aestivum ◽  
Phytic Acid ◽  
Inositol Phosphate ◽  
Mineral Nutrients ◽  
Energy Dispersive ◽  
Aleurone Layer ◽  
Wild Type ◽  
X Ray ◽  
Low Phytic Acid

Concentrations of P, phytic acid (myo-inositol hexakisphosphate, IP6), and other mineral storage elements were studied in wild-type and low phytic acid (lpa) genotype Js-12-LPA wheat (Triticum aestivum L.) embryos and rest-of-grain fractions. Environmental scanning electron microscopy images revealed a decreased average size and an increased number of aleurone layer globoids in lpa grains compared with the wild type. Energy-dispersive X-ray analyses of unfixed aleurone layer and scutellum cell cytoplasm revealed mainly C, O, P, K, and Mg in both grain types. The starchy endosperm contained virtually no P, K, or Mg, demonstrating no shift of mineral nutrients to that compartment. Scanning transmission electron microscopy – energy-dispersive X-ray analyses of scutellum and aleurone layer globoids in both genotypes revealed that P, K, and Mg were the main mineral nutrients in globoids with low amounts of Ca, Fe, and Zn. Traces of Mn were only in scutellum globoids. Total P was similar between genotypes for the rest-of-grain fractions, which are 97% of grain mass. The main inositol phosphate was IP6, but a small amount of IP5 was present. Both lpa grain fractions exhibited major reductions in IP6 compared with the wild type and a threefold increase in inorganic P. The concentration of K decreased in both fractions, while Ca increased 25% in the Js-12-LPA rest-of-grain compared with the wild type. The lack of large differences in mineral concentration and distribution between the wild type and Js-12-LPA indicates that there is no direct role of localization of IP6 synthesis in mineral distribution.

scholarly journals Fine-tuning Flowering Time via Genome Editing of Upstream Open Reading Frames of Heading Date 2 in Rice

Rice ◽  
2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Xinxin Liu ◽  
Hualong Liu ◽  
Yuanye Zhang ◽  
Mingliang He ◽  
Rongtian Li ◽  
...  
Keyword(s):  
Genome Editing ◽  
Flowering Time ◽  
Agronomic Traits ◽  
Oryza Sativa L ◽  
Heading Date ◽  
Open Reading Frames ◽  
Fine Tuning ◽  
Wild Type ◽  
Phenotypic Analysis ◽  
Mutant Lines

AbstractFlowering time of rice (Oryza sativa L.) is among the most important agronomic traits for region adaptation and grain yield. In the process of rice breeding, efficient and slightly modulating the flowering time of an elite cultivar would be more popular with breeder. Hence, we are interested in slightly increasing the expression of flowering repressors by CRISPR/Cas9 genome editing system. It was predicated there were three uORFs in 5’ leader sequence of Hd2. In this study, through editing Hd2 uORFs, we got four homozygous mutant lines. Phenotypic analysis showed that the hd2 urf edited lines flowered later by 4.6–11.2 days relative to wild type SJ2. Supporting the later flowering phenotype, the expression of Ehd1, Hd3a, and RFT1 is significantly decreased in hd2 urf than that in wild type. Moreover, we found that the transcription level of Hd2 is not affected, whereas the Hd2 protein level was increased in hd2 urf compared with wild type, which indicated that Hd2 uORFs indeed affect the translation of a downstream Hd2 pORF. In summary, we developed a efficient approach for delaying rice heading date based on editing uORF region of flowering repressor, which is time and labor saving compared to traditional breeding. In future, uORF of other flowering time related genes, including flowering promoter and flowering repressor genes, can also be used as targets to fine-tune the flowering time of varieties.

Phytic acid-phosphorus and other nutritionally important mineral nutrient elements in grains of wild-type and low phytic acid (lpa1–1) rice

2004 ◽  
Vol 14 (2) ◽  
pp. 109-116 ◽  
Author(s):  
John N.A. Lott ◽  
Jessica C. Liu ◽  
Irene Ockenden ◽  
Michael Truax ◽  
John N.A. Lott
Keyword(s):  
Phytic Acid ◽  
Dry Weight ◽  
Mineral Nutrient ◽  
Nutrient Elements ◽  
Aleurone Layer ◽  
Wild Type ◽  
Phytic Acid Content ◽  
Low Phytic Acid ◽  
Calcium Iron ◽  
Iron Manganese

Mineral nutrient stores in cereal grains are mainly phytate, a salt of the phosphorus-rich compound phytic acid. Quantitative measures of total phosphorus, phytic acid-phosphorus, potassium, magnesium, calcium, iron, manganese and zinc were obtained for whole grains, embryos and rest-of-grain portions of cv. Kaybonnet rice (wild type) (Oryza sativa L.) and a low phytic acid (lpa1–1) mutant strain with a 45% reduction in phytic acid. P, K and Mg were present in higher amounts than Ca, Mn, Fe and Zn in both grain types. Whole-grain amounts of total P, Ca, Mn and phytic acid-phosphorus were lower in whole lpa1–1 grains than in wild-type grains; K, Mg and Fe amounts were similar, and Zn was higher. Embryos, which comprise 3.5% or less of grain dry weight, were comparatively rich in all measured elements. The lpa1–1 mutation influenced the phytic acid content of the embryo more than that of the aleurone layer. Aleurone-layer cells of wild-type grains had many phosphorus-rich globoids 2μm or larger in diameter, whereas lpa1–1 grains contained more of the smaller globoids. The reduction in globoid size was consistent with the reduction in phytate. Energy-dispersive X-ray analysis of both aleurone-layer cells and sections of globoids in aleurone-layer cells revealed that P, K and Mg were the main mineral nutrient elements present in both grain types; traces of Ca, Mn, Fe or Zn were present. Starchy endosperm cells contained virtually no P, K or Mg, whereas scutellum cells were rich in these elements.

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