Disruption of three polyamine uptake transporter genes in rice by CRISPR/Cas9 gene editing confers tolerance to herbicide paraquat

Lyu, Yu-Shu; Cao, Li-Miao; Huang, Wenqian; Liu, Jianxiang; Lu, Haiping

doi:10.1007/s42994-022-00075-4

Cited by 20 publications

(11 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…An independent study performed in parallel confirmed that loss-of-function mutations in AtPAR1 generated by CRIPSR/Cas9 can be used to identify edited lines [14]. A similar approach that targeted the same three paralogs in rice as targeted here using a single construct showed that the triple oslat5/osput3/ospar1, oslat1/osput1, oslat7/osput2 mutants were also resistant to MV [29]. Neither the single mutants characterized here, nor the triple mutants showed apparent growth or developmental defects under normal growth conditions [29].…”

Section: Introductionmentioning

confidence: 88%

Loss-of-function mutation in the polyamine transporter geneOsLAT5as a selectable marker for genome editing

Schenstnyi,

Zhang,

Liu

et al. 2023

Preprint

View full text Add to dashboard Cite

Genome editing by TALENs, CRISPR/Cas, base or prime editing have become routine tools. During stable plant transformation, the gene coding for the editing enzyme, e.g.,Cas9,the guide RNAs (gRNAs), alongside a selectable marker are integrated into the nuclear genome. Identification of successful transformants relies on selectable or screenable markers, typically genes providing resistance to antibiotics or herbicides. Selectable markers use a substantial portion of the T-DNA, hence reducing transfer efficiency by limiting the effective number of TALENs or guide/pegRNAs that can be used in parallel. Moreover, marker genes are frequently subject to gene silencing. Here, we generated loss-of-function mutations in PUT/LAT-type polyamine transporter family genes to confer resistance to the phytotoxin methylviologen (MV) as a method for selection. As a proof of concept, CRISPR/Cas9 vectors with gRNAs were constructed to target three close homologsOsLAT1,OsLAT5, andOsLAT7. We show that loss ofOsLAT5(also known asOsPUT3orOsPAR1) function was sufficient to confer resistance to MV in rice seeds, seedlings and calli, validating the editing approach ofOsLAT5to obtain a selectable marker. We discuss the potential of incorporating a gRNA cassette (forOsLAT5) as a selectable marker and a reporter for successful genome editing for optimizing editing protocols.

show abstract

Section: Introductionmentioning

confidence: 88%

Loss-of-function mutation in the polyamine transporter geneOsLAT5as a selectable marker for genome editing

Schenstnyi,

Zhang,

Liu

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…This recognised the requirement of the selective herbicides. The CRISPR/Cas9 based manipulation of OsPUT1/2/3 greatly conferred paraquat resistance in rice without obvious yield penalty (Lyu et al, 2022). The Crisprcas installed mutation in the ALS gene disabled the interaction between the ALS protein and IMI herbicides which developed tolerance to IMI herbicides (McCourt, 2006, Lonhienne, 2018.…”

Section: Crop Improvement For Herbicide Tolerancementioning

confidence: 99%

Crispr-Cas Mediated Crop Improvement in Rice: A Review

Jagadeesh¹,

Bhargavi²,

Bhargavi³

et al. 2022

PLANT ARCHIVES

View full text Add to dashboard Cite

Rice is the staple food crop that is grown across the globe due to its resilience to climatic conditions. Cereals are the predominant food grains consumed by the majority of the population across the globe, with rice alone feeding 50% of the global populace. In order to feed the geometrically rising population, there is a need for a dramatic upsurge in rice production levels. But climate change is one of the causes resulting in the rising of various biotic and abiotic stresses. The advancement of science had resulted in the development of Genome editing technologies like clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9), transcription activator-like effectors nucleases (TALENs), and zinc-finger nucleases (ZFNs). Among these techniques, due to ease of application, and high precision crisper-cas technology had emerged as a solution to overcome various obstacles in the journey of enhancing production levels of rice. In this, we will review various improvements made in the rice crop using crispercas which ultimately led to enhanced production.

show abstract

“…However, it was found that the structure of paraquat is similar to that of polyamines, which could share a common uptake system of endogenous substrates due to their structural similarity [ 21 , 22 , 23 , 24 ]. Recently, three genes on polyamine uptake transportation were applied to enhance the tolerance of paraquat-resistance in rice [ 25 ]. Therefore, it is worth exploring the mechanism of polyamine genes regulating paraquat resistance.…”

Section: Introductionmentioning

confidence: 99%

Establishment of an Efficient Agrobacterium-Mediated Genetic Transformation System to Enhance the Tolerance of the Paraquat Stress in Engineering Goosegrass (Eleusine Indica L.)

Luo

Chen

Nian

et al. 2023

IJMS

View full text Add to dashboard Cite

Eleusine indica (goosegrass) is a problematic weed worldwide known for its multi-herbicide tolerance/resistance biotype. However, a genetic transformation method in goosegrass has not been successfully established, making a bottleneck for functional genomics studies in this species. Here, we report a successful Agrobacterium-mediated transformation method for goosegrass. Firstly, we optimized conditions for breaking seed dormancy and increasing seed germination rate. A higher callus induction rate from germinated seeds was obtained in N6 than in MS or B5 medium. Then the optimal transformation efficiency of the gus reporter gene was obtained by infection with Agrobacterium tumefaciens culture of OD600 = 0.5 for 30 min, followed by 3 days of co-cultivation with 300 μmol/L acetosyringone. Concentrations of 20 mg L−1 kanamycin and 100 mg L−1 timentin were used to select the transformed calli. The optimal rate of regeneration of the calli was generated by using 0.50 mg L−1 6-BA and 0.50 mg L−1 KT in the culture medium. Then, using this transformation method, we overexpressed the paraquat-resistant EiKCS gene into a paraquat-susceptible goosegrass biotype MZ04 and confirmed the stable inheritance of paraquat-resistance in the transgenic goosegrass lines. This approach may provide a potential mechanism for the evolution of paraquat-resistant goosegrass and a promising gene for the manipulation of paraquat-resistance plants. This study is novel and valuable in future research using similar methods for herbicide resistance.

show abstract

Disruption of three polyamine uptake transporter genes in rice by CRISPR/Cas9 gene editing confers tolerance to herbicide paraquat

Cited by 20 publications

References 17 publications

Loss-of-function mutation in the polyamine transporter geneOsLAT5as a selectable marker for genome editing

Loss-of-function mutation in the polyamine transporter geneOsLAT5as a selectable marker for genome editing

Crispr-Cas Mediated Crop Improvement in Rice: A Review

Establishment of an Efficient Agrobacterium-Mediated Genetic Transformation System to Enhance the Tolerance of the Paraquat Stress in Engineering Goosegrass (Eleusine Indica L.)

Contact Info

Product

Resources

About