Advances in Gene Editing of Haploid Tissues in Crops

Bhowmik, Pankaj; Bilichak, Andriy

doi:10.3390/genes12091410

Cited by 9 publications

(4 citation statements)

References 138 publications

(195 reference statements)

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“…The combination of increased activity of Cas9 at elevated temperatures, altered expression of the DNA repair genes, and conformational changes of the chromatin during heat stress may play a role in the GE efficiency in planta . Further improvement in the efficiency could potentially be achieved through selective expression of both Cas9 and gRNA in the wheat gametes or haploid tissues to reduce the number of edited loci and using the tRNA processing system under the Pol II promoter for multiplexed editing at the target site . High expression levels of Cas9/gRNAs can decrease the effect of genomic context on the gRNA activity and improve on-target activity .…”

Section: Resultsmentioning

confidence: 99%

Evaluation of Factors Affecting In Planta Gene Editing Efficiency in Wheat (Triticum aestivum L.)

Yao

Wang

et al. 2021

ACS Agric. Sci. Technol.

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Gene editing in polyploid crops still suffers from low efficiency, and further improvement is needed for its routine implementation in the modern breeding practice. Here we examined factors that affect the CRISPR/Cas9-mediated gene editing efficiency in allohexaploid wheat plants. We selected three guide RNAs (gRNAs) and evaluated the potential of using heat shock at the seedlings stage to increase editing efficiency in transgenic plants. Only one out of three gRNAs demonstrated significantly increased editing efficiency following heat shock treatment. We also examined the expression of DNA repair and replication gene orthologues in response to heat shock in wheat leaves. Misregulation of the chromatin remodelers following the heat shock treatment could potentially be involved in the increase of editing efficiency in wheat. Overall, the editing efficiency of gRNAs observed in our study correlated with predictive scores from the gRNA design tools. The editing rate of the top-ranked gRNAs could potentially be increased using heat treatment of the transgenic plants.

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Section: Resultsmentioning

confidence: 99%

Evaluation of Factors Affecting In Planta Gene Editing Efficiency in Wheat (Triticum aestivum L.)

Yao

Wang

et al. 2021

ACS Agric. Sci. Technol.

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“…Genetic mapping studies in maize have located two main QTLs affecting haploid induction potential, quantitative haploid induction rate 1 (qhir1) and quantitative haploid induction rate 8 (qhir8), on maize chromosomes 1 and 9, respectively [112]. The two loci, qhir1 and qhir8, determine 66% and 20% of the genetic variation associated with maternal haploid induction, respectively [111,113]. Fine mapping has enabled the identification of two genes within these QTLs, MATRILENEAL (MTL) on qhir1 and DOMAIN OF UNKNOWN FUNCTION 679 MEMBRANE PROTEIN (DMP) on qhir8.…”

Section: Maternal Haploidsmentioning

confidence: 99%

Haploid Induction in Indica Rice: Exploring New Opportunities

Mayakaduwa,

Silva

2023

Plants

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Haploid plants are of significant interest to crop breeders due to their ability to expedite the development of inbred lines. Chromosome-doubling of haploids, produced by either in vitro or in vivo methods, results in fully homozygous doubled haploids. For nearly five decades, in vitro methods of anther and microspore culture have been attempted in many crops. In rice, in vitro methods are used with some success in japonica cultivars, although indica types have remained recalcitrant to a large extent. This review aims to explore the reasons for the lack of success of in vitro methods in indica rice and discuss new advancements in in vivo haploid induction protocols in other cereals and their relevance to rice. In particular, the current level of understanding of in vivo haploid inducer systems that utilize MTL and CENH3 mutants is analyzed in detail. One notable advantage of in vivo haploid induction systems is that they do not require tissue culture competence. This makes these methods more accessible and potentially transformative for research, offering a pragmatic approach to improving indica rice cultivars. By embracing these in vivo methods and harnessing the power of gene editing technologies like CRISPR/Cas9 systems, breeders can reshape their approach to indica rice improvement.

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“…Doubled haploids (DH) have been used in breeding programs for several decades (Bhowmik & Bilichak, 2021; Wędzony et al., 2009). Doubled haploids provide a quick approach for the production of completely homozygous plants within one generation after the original cross.…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, in normal conditions, field crops can only be advanced one to two generations per year, making the breeding cycle a very slow process (Watson et al, 2018). Doubled haploids (DH) have been used in breeding programs for several decades (Bhowmik & Bilichak, 2021;Wędzony et al, 2009). Doubled haploids provide a quick approach for the production of completely homozygous plants within one generation after the original cross.…”

Section: Introductionmentioning

confidence: 99%

Reducing the generation time in winter wheat cultivars using speed breeding

et al. 2023

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Reducing generation time is critical to achieving the goals of genetic gain in important crops like wheat (Triticum aestivum). Speed breeding (SB) has been shown to considerably reduce generation times in crop plants. Unlike spring wheat cultivars, winter wheat varieties require typically 6-9 weeks of cold treatment, called vernalization, for flowering which extends the generation time for the development of improved winter wheat cultivars. Here, we optimized the SB method using a set of 48 diverse soft red winter wheat (SRWW) cultivars by testing vernalization duration, light and temperature requirements, and the viability of seeds harvested after different durations post-anthesis under extended daylight conditions. We have found that using a 22-h setting (22 h day/2 h night, 25˚C/22˚C) in high-density 50-cell trays results in rapid generation advancement. We used genotypic data for a panel of soft red winter wheat varieties from the regional programs to determine the impact of photoperiod and vernalization alleles on the efficiency of the SB approach. Using a set of 48 SRWW cultivars and germplasm from Maryland and four other public breeding programs, we establish that this protocol can allow for the advancement of four generations per year in controlled conditions for winter wheat varieties, experimental lines, or emerging cultivars. Our work shows the potential to reduce generation time Abbreviations: DH, doubled haploids; DPF, days post flowering; HID, high intensity discharge; SB, speed breeding; SSD, single seed descent.This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Advances in Gene Editing of Haploid Tissues in Crops

Cited by 9 publications

References 138 publications

Evaluation of Factors Affecting In Planta Gene Editing Efficiency in Wheat (Triticum aestivum L.)

Evaluation of Factors Affecting In Planta Gene Editing Efficiency in Wheat (Triticum aestivum L.)

Haploid Induction in Indica Rice: Exploring New Opportunities

Reducing the generation time in winter wheat cultivars using speed breeding

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