Red fluorescent protein (DsRed2), an ideal reporter for cotton genetic transformation and molecular breeding

Liang, Sun; Alariqi, Muna; Zhu, Yaping; Li, Jianying; Li, Zelin; Wang, Qing; Li, Yajun; Rui, Hangping; Zhang, Xianlong; Jin, Shuangxia

doi:10.1016/j.cj.2018.05.002

Cited by 29 publications

(14 citation statements)

References 40 publications

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“…All the GhABEs plasmid vectors were transformed into Agrobacterium strain GV3101 (kanamycin as selectable marker) by electroporation, and Agrobacterium-mediated transformation of cotton cultivar J668 was performed according to previous publications [ 35 , 52 ].…”

Section: Methodsmentioning

confidence: 99%

Development of an efficient and precise adenine base editor (ABE) with expanded target range in allotetraploid cotton (Gossypium hirsutum)

Wang

et al. 2022

BMC Biol

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Background Base editors (BEs) display diverse applications in a variety of plant species such as Arabidopsis, rice, wheat, maize, soybean, and cotton, where they have been used to mediate precise base pair conversions without the collateral generation of undesirable double-stranded breaks (DSB). Studies of single-nucleotide polymorphisms (SNPs) underpinning plant traits are still challenging, particularly in polyploidy species where such SNPs are present in multiple copies, and simultaneous modification of all alleles would be required for functional analysis. Allotetraploid cotton has a number of homoeologous gene pairs located in the A and D sub-genomes with considerable SNPs, and it is desirable to develop adenine base editors (ABEs) for efficient and precise A-to-G single-base editing without DSB in such complex genome. Results We established various ABE vectors based on different engineered adenosine deaminase (TadA) proteins fused to Cas9 variants (dCas9, nCas9), enabling efficient A to G editing up to 64% efficiency on-target sites of the allotetraploid cotton genome. Comprehensive analysis showed that GhABE7.10n exhibited the highest editing efficiency, with the main editing sites specifically located at the position A5 (counting the PAM as positions 21–23). Furthermore, DNA and RNA off-target analysis of cotton plants edited with GhABE7.10n and GhABE7.10d by whole genome and whole-transcriptome sequencing revealed no DNA off-target mutations, while very low-level RNA off-target mutations were detected. A new base editor, namely GhABE7.10dCpf1 (7.10TadA + dCpf1), that recognizes a T-rich PAM, was developed for the first time. Targeted A-to-G substitutions generated a single amino acid change in the cotton phosphatidyl ethanolamine-binding protein (GhPEBP), leading to a compact cotton plant architecture, an ideotype for mechanized harvesting of modern cotton production. Conclusions Our data illustrate the robustness of adenine base editing in plant species with complex genomes, which provides efficient and precise toolkit for cotton functional genomics and precise molecular breeding.

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Section: Methodsmentioning

confidence: 99%

Development of an efficient and precise adenine base editor (ABE) with expanded target range in allotetraploid cotton (Gossypium hirsutum)

Wang

et al. 2022

BMC Biol

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“…JIN668 was used as the transformation receptor ( Li et al, 2019b ). After the seeds were sterilized, the sterilized seeds were placed on a dark culture at 28°C for 4–6 days, and the hypocotyls were cut into 5∼10 mm lengths, and then followed our previously reported method for genetic transformation ( Sun et al, 2018 ; Wang et al, 2018 ).…”

Section: Methodsmentioning

confidence: 99%

Identification and Functional Analysis of lncRNA by CRISPR/Cas9 During the Cotton Response to Sap-Sucking Insect Infestation

Zhang

Saeed

et al. 2022

Front. Plant Sci.

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Sap-sucking insects cause severe damage to cotton production. Long non-coding RNAs (lncRNAs) play vital regulatory roles in various development processes and stress response, however, the function of lncRNAs during sap-sucking insect infection in cotton is largely unknown. In this study, the transcriptome profiles between resistant (HR) and susceptible (ZS) cotton cultivars under whitefly infestation at different time points (0, 4, 12, 24, and 48 h) were compared. A total of 6,651 lncRNAs transcript and 606 differentially expressed lncRNAs were identified from the RNA-seq data. A co-expression network indicated that lncA07 and lncD09 were potential hub genes that play a regulatory role in cotton defense against aphid infestation. Furthermore, CRISPR/Cas9 knock-out mutant of lncD09 and lncA07 showed a decrease of jasmonic acid (JA) content, which potentially lead to increased susceptibility toward insect infestation. Differentially expressed genes between wild type and lncRNA knock-out plants are enriched in modulating development and resistance to stimulus. Additionally, some candidate genes such as Ghir_A01G022270, Ghir_D04G014430, and Ghir_A01G022270 are involved in the regulation of the JA-mediated signaling pathway. This result provides a novel insight of the lncRNA role in the cotton defense system against pests.

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“…Gossypium hirsutum cv. Jin668 was used for Agrobacterium ‐mediated genetic transformation (Li, Rui, et al, 2019; Sun et al, 2018).…”

Section: Methodsmentioning

confidence: 99%

Silencing of a LIM gene in cotton exhibits enhanced resistance against Apolygus lucorum

Liang

Luo

Alariqi

et al. 2021

Journal Cellular Physiology

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Plant bugs (Miridae species) have become major agricultural pests that cause increasing and severe economic damage. Plant‐mediated RNA interference (RNAi) is emerging as an eco‐friendly, efficient, and reliable strategy for pest management. In this study, we isolated and characterized a lethal gene of Apolygus lucorum and named it Apolygus lucorum LIM (AlLIM), which produced A. lucorum mortality rates ranging from 38% to 81%. Downregulation of the AlLIM gene expression in A. lucorum by injection of a double‐stranded RNA (dsRNA) led to muscle structural disorganization that resulted in metamorphosis deficiency and increased mortality. Then we constructed a plant expression vector that enabled transgenic cotton to highly and stably express dsRNA of AlLIM (dsAlLIM) by Agrobacterium‐mediated genetic transformation. In the field bioassay, dsAlLIM transgenic cotton was protected from A. lucorum damage with high efficiency, with almost no detectable yield loss. Therefore, our study successfully provides a promising genetically modified strategy to overpower A. lucorum attack.

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Red fluorescent protein (DsRed2), an ideal reporter for cotton genetic transformation and molecular breeding

Cited by 29 publications

References 40 publications

Development of an efficient and precise adenine base editor (ABE) with expanded target range in allotetraploid cotton (Gossypium hirsutum)

Development of an efficient and precise adenine base editor (ABE) with expanded target range in allotetraploid cotton (Gossypium hirsutum)

Identification and Functional Analysis of lncRNA by CRISPR/Cas9 During the Cotton Response to Sap-Sucking Insect Infestation

Silencing of a LIM gene in cotton exhibits enhanced resistance against Apolygus lucorum

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