Mixed inheritance in the genetic control of ramulosis (Colletotrichum gossypii var. cephalosporioides) resistance in cotton

Oliveira, M. A. C. de; Duarte, João Batista; Morello, C.L.; Suassuna, N. D.; Oliveira, A. B. de

doi:10.4238/gmr.15038667

Cited by 9 publications

(8 citation statements)

References 7 publications

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“…In addition to being a fiber crop, cotton is also a good source for biofuel production as cotton seeds contain significant oil reserves ( Oliveira et al, 2016 ). With the release of the genome sequence of Gossypium hirsutum ( Li F. et al, 2015 ), it is now possible to utilize CRISPR tools to achieve precise DNA modifications.…”

Section: Dicotsmentioning

confidence: 99%

CRISPR for Crop Improvement: An Update Review

et al. 2018

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The availability of genome sequences for several crops and advances in genome editing approaches has opened up possibilities to breed for almost any given desirable trait. Advancements in genome editing technologies such as zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) has made it possible for molecular biologists to more precisely target any gene of interest. However, these methodologies are expensive and time-consuming as they involve complicated steps that require protein engineering. Unlike first-generation genome editing tools, CRISPR/Cas9 genome editing involves simple designing and cloning methods, with the same Cas9 being potentially available for use with different guide RNAs targeting multiple sites in the genome. After proof-of-concept demonstrations in crop plants involving the primary CRISPR-Cas9 module, several modified Cas9 cassettes have been utilized in crop plants for improving target specificity and reducing off-target cleavage (e.g., Nmcas9, Sacas9, and Stcas9). Further, the availability of Cas9 enzymes from additional bacterial species has made available options to enhance specificity and efficiency of gene editing methodologies. This review summarizes the options available to plant biotechnologists to bring about crop improvement using CRISPR/Cas9 based genome editing tools and also presents studies where CRISPR/Cas9 has been used for enhancing biotic and abiotic stress tolerance. Application of these techniques will result in the development of non-genetically modified (Non-GMO) crops with the desired trait that can contribute to increased yield potential under biotic and abiotic stress conditions.

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Section: Dicotsmentioning

confidence: 99%

CRISPR for Crop Improvement: An Update Review

et al. 2018

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“…Cotton ( Gossypium hirsutum L.) is a globally cultivated economic crop and is a major source of natural fiber and oil [ 1 ]. Cotton production has decreased recently due to a series of environmental constraints, especially various biotic and abiotic stresses such as Verticillium wilt and drought, while the area that cotton is cultivated has declined due to increased cost of cultivation and reduction in available farmland [ 2 , 3 ]. Verticillium wilt is a vascular disease caused by the soil-borne fungal pathogen V. dahliae , which infects over 200 dicotyledonous species including cotton.…”

Section: Introductionmentioning

confidence: 99%

The Cotton BEL1-Like Transcription Factor GhBLH7-D06 Negatively Regulates the Defense Response against Verticillium dahliae

Wang

et al. 2020

IJMS

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Verticillium wilt will seriously affect cotton yield and fiber quality. BEL1-Like transcription factors are involved in the regulation of secondary cell wall (SCW) formation, especially the biosynthesis of lignin that also plays a key role in cotton disease resistance. However, there is no report on the role of BEL1-Like transcription factor in the regulation of plant biological stress. In this study, tissue expression pattern analysis showed that a BEL1-Like transcription factor GhBLH7-D06 was predominantly expressed in vascular tissues and the SCW thickening stage of fiber development, while its expression could also respond to Verticillium dahliae infection and the phytohormone MeJA treatment, which indicated that GhBLH7-D06 might be involved in the defense response of Verticillium wilt. Using virus-induced gene silencing (VIGS) technology, we found silencing the expression of GhBLH7-D06 could enhance the resistance of cotton plants to Verticillium wilt, and the acquisition of resistance might be mainly due to the significant overexpression of genes related to lignin biosynthesis and JA signaling pathway, which also proves that GhBLH7-D06 negatively regulates the resistance of cotton to Verticillium wilt. Based on the results of yeast two-hybrid (Y2H) library screening and confirmation by bimolecular fluorescence complementary (BiFC) experiment, we found an Ovate Family Protein (OFP) transcription factor GhOFP3-D13 which was also a negative regulator of cotton Verticillium wilt resistance could that interacts with GhBLH7-D06. Furthermore, the dual-luciferase reporter assay and yeast one-hybrid (Y1H) experiment indicated that GhBLH7-D06 could target binding to the promoter region of GhPAL-A06 to suppress its expression and eventually lead to the inhibition of lignin biosynthesis. In general, the GhBLH7-D06/GhOFP3-D13 complex can negatively regulate resistance to Verticillium wilt of cotton by inhibiting lignin biosynthesis and JA signaling pathway.

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“…Cotton is one of the most important fibers, a good source for biofuel production and an oil crop ( Wang et al, 2012 ; Shi et al, 2014 ; Oliveira et al, 2016 ). Increased sequence availability has emphasized the need for rapid and cost-effective tools to create targeted mutations in order to perform much needed large-scale gene functional studies in cotton ( Wang et al, 2010 , 2012 ; Li et al, 2015 ; Zhang et al, 2015 ).…”

Section: Introductionmentioning

confidence: 99%

Genome Editing in Cotton with the CRISPR/Cas9 System

et al. 2017

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Genome editing is an important tool for gene functional studies as well as crop improvement. The recent development of the CRISPR/Cas9 system using single guide RNA molecules (sgRNAs) to direct precise double strand breaks in the genome has the potential to revolutionize agriculture. Unfortunately, not all sgRNAs are equally efficient and it is difficult to predict their efficiency by bioinformatics. In crops such as cotton (Gossypium hirsutum L.), with labor-intensive and lengthy transformation procedures, it is essential to minimize the risk of using an ineffective sgRNA that could result in the production of transgenic plants without the desired CRISPR-induced mutations. In this study, we have developed a fast and efficient method to validate the functionality of sgRNAs in cotton using a transient expression system. We have used this method to validate target sites for three different genes GhPDS, GhCLA1, and GhEF1 and analyzed the nature of the CRISPR/Cas9-induced mutations. In our experiments, the most frequent type of mutations observed in cotton cotyledons were deletions (∼64%). We prove that the CRISPR/Cas9 system can effectively produce mutations in homeologous cotton genes, an important requisite in this allotetraploid crop. We also show that multiple gene targeting can be achieved in cotton with the simultaneous expression of several sgRNAs and have generated mutations in GhPDS and GhEF1 at two target sites. Additionally, we have used the CRISPR/Cas9 system to produce targeted gene fragment deletions in the GhPDS locus. Finally, we obtained transgenic cotton plants containing CRISPR/Cas9-induced gene editing mutations in the GhCLA1 gene. The mutation efficiency was very high, with 80.6% of the transgenic lines containing mutations in the GhCLA1 target site resulting in an intense albino phenotype due to interference with chloroplast biogenesis.

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Mixed inheritance in the genetic control of ramulosis (Colletotrichum gossypii var. cephalosporioides) resistance in cotton

Cited by 9 publications

References 7 publications

CRISPR for Crop Improvement: An Update Review

CRISPR for Crop Improvement: An Update Review

The Cotton BEL1-Like Transcription Factor GhBLH7-D06 Negatively Regulates the Defense Response against Verticillium dahliae

Genome Editing in Cotton with the CRISPR/Cas9 System

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