scholarly journals Generating Novel Male Sterile Tomatoes by Editing Respiratory Burst Oxidase Homolog Genes

2022 ◽  
Vol 12 ◽  
Author(s):  
Xiaojuan Dai ◽  
Huanan Han ◽  
Wei Huang ◽  
Lianghui Zhao ◽  
Minglei Song ◽  
...  
Keyword(s):  
Respiratory Burst ◽  
Hybrid Breeding ◽  
Wild Type ◽  
Male Sterile ◽  
Ros Metabolism ◽  
Reproduction Process ◽  
Self Fertilization ◽  
Mutant Lines ◽  
Respiratory Burst Oxidase

Hybrid breeding of tomatoes (Solanum lycopersicum), an important vegetable crop, is an effective way to improve yield and enhance disease and stress resistance. However, the efficiency of tomato hybridization is hindered by self-fertilization, which can be overcome using male sterile lines. It has been reported that reactive oxygen species (ROS) act as a key regulator for anther development, mediated by RBOH (Respiratory Burst Oxidase Homolog) genes. Here, two tomato anther-expressed genes, LeRBOH (Solyc01g099620) and LeRBOHE (Solyc07g042460), were selected to cultivate novel tomato male sterile strains. By using a CRISPR/Cas9 system with a two-sgRNA module, the lerboh, lerbohe, and lerboh lerbohe mutant lines were generated, among which the lerbohe and lerboh lerbohe mutants displayed complete male sterility but could accept wild-type pollens and produce fruits normally. Further analysis uncovered significantly decreased ROS levels and abnormal programmed cell death in lerboh lerbohe anthers, indicating a key role of ROS metabolism in tomato pollen development. Taken together, our work demonstrates a successful application of gene editing via CRISPR/Cas9 in generating male sterile tomatoes and afforded helpful information for understanding how RBOH genes regulating tomato reproduction process.

scholarly journals Genetic analysis of the Arabidopsis TIR1/AFB auxin receptors reveals both overlapping and specialized functions

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Michael J Prigge ◽  
Matthieu Platre ◽  
Nikita Kadakia ◽  
Yi Zhang ◽  
Kathleen Greenham ◽  
...  
Keyword(s):  
Genetic Analysis ◽  
Early Embryo ◽  
Wild Type ◽  
The Family ◽  
Dependent Inhibition ◽  
Root Gravitropism ◽  
Specialized Function ◽  
Mutant Lines

The TIR1/AFB auxin co-receptors mediate diverse responses to the plant hormone auxin. The Arabidopsis genome encodes six TIR1/AFB proteins representing three of the four clades that were established prior to angiosperm radiation. To determine the role of these proteins in plant development we performed an extensive genetic analysis involving the generation and characterization of all possible multiply-mutant lines. We find that loss of all six TIR1/AFB proteins results in early embryo defects and eventually seed abortion, and yet a single wild-type allele of TIR1 or AFB2 is sufficient to support growth throughout development. Our analysis reveals extensive functional overlap between even the most distantly related TIR1/AFB genes except for AFB1. Surprisingly, AFB1 has a specialized function in rapid auxin-dependent inhibition of root growth and early phase of root gravitropism. This activity may be related to a difference in subcellular localization compared to the other members of the family.

scholarly journals Role of a respiratory burst oxidase of Lepidium sativum (cress) seedlings in root development and auxin signalling

2012 ◽  
Vol 63 (18) ◽  
pp. 6325-6334 ◽  
Author(s):  
Kerstin Müller ◽  
Ada Linkies ◽  
Gerhard Leubner-Metzger ◽  
Allison R. Kermode
Keyword(s):  
Root Development ◽  
Respiratory Burst ◽  
Lepidium Sativum ◽  
Auxin Signalling ◽  
Respiratory Burst Oxidase

scholarly journals Tissue factor: a link between C5a and neutrophil activation in antiphospholipid antibody–induced fetal injury

Blood ◽  
2007 ◽  
Vol 110 (7) ◽  
pp. 2423-2431 ◽  
Author(s):  
Patricia Redecha ◽  
Rachel Tilley ◽  
Michael Tencati ◽  
Jane E. Salmon ◽  
Daniel Kirchhofer ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Mouse Model ◽  
Tissue Factor ◽  
Respiratory Burst ◽  
Pregnancy Loss ◽  
Fetal Loss ◽  
Antiphospholipid Antibody ◽  
Wild Type ◽  
Important Mediator

Fetal loss in patients with antiphospholipid (aPL) antibodies has been ascribed to thrombosis of placental vessels. However, we have shown that inflammation, specifically activation of complement with generation of the anaphylotoxin C5a, is an essential trigger of fetal injury. In this study, we analyzed the role of the procoagulant molecule tissue factor (TF) in a mouse model of aPL antibody–induced pregnancy loss. We found that either blockade of TF with a monoclonal antibody in wild-type mice or a genetic reduction of TF prevented aPL antibody–induced inflammation and pregnancy loss. In response to aPL antibody–generated C5a, neutrophils express TF potentiating inflammation in the deciduas and leading to miscarriages. Importantly, we showed that TF in myeloid cells but not fetal-derived cells (trophoblasts) was associated with fetal injury, suggesting that the site for pathologic TF expression is neutrophils. We found that TF expression in neutrophils contributes to respiratory burst and subsequent trophoblast injury and pregnancy loss induced by aPL antibodies. The identification of TF as an important mediator of C5a-induced oxidative burst in neutrophils in aPL-induced fetal injury provides a new target for therapy to prevent pregnancy loss in the antiphospholipid syndrome.

Arabidopsis thaliana constitutively active ROP11 interacts with the NADPH oxidase respiratory burst oxidase homologue F to regulate reactive oxygen species production in root hairs

2016 ◽  
Vol 43 (3) ◽  
pp. 221 ◽  
Author(s):  
Min Yan ◽  
Wen Jing ◽  
Ni Xu ◽  
Like Shen ◽  
Qun Zhang ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Arabidopsis Thaliana ◽  
Respiratory Burst ◽  
Root Hairs ◽  
Reactive Oxygen ◽  
Ros Production ◽  
Oxygen Species ◽  
Wild Type ◽  
Respiratory Burst Oxidase ◽  
Constitutively Active

Reactive oxygen species (ROS) play a key signalling role in cells. Plant NADPH oxidases, also known as respiratory burst oxidase homologues (Rbohs), are well characterised ROS-generating systems. In this study, we found that the constitutively active small guanosine triphosphatase (GTPase) ROP11 (CA-ROP11) interacted with RbohF by using a yeast two-hybrid analysis, a pull-down assay and an in vivo bimolecular fluorescence complementation assay. The mutation of amino acid L336 or L337 in RbohF abolished its interaction with CA-ROP11. Coexpression of CA-ROP11 and wild-type RbohF in Nicotiana benthamiana Domin enhanced ROS production compared with coexpression of CA-ROP11 and mutant RbohF or of dominant negative ROP11 and wild-type RbohF. Moreover, CA-ROP11 overexpression resulted in ROS accumulation and a swollen root hair phenotype in Arabidopsis thaliana (L.) Heynh. The deletion of RbohF partially reduced the increase in ROS in Arabidopsis plants overexpressing CA-ROP11. These results suggest that Arabidopsis ROP11 modulates ROS production by interacting with RbohF in root hairs.

scholarly journals Overexpression of Sesame Polyketide Synthase A Leads to Abnormal Pollen Development in Arabidopsis

2021 ◽  
Author(s):  
Tianyu Li ◽  
Ting Zhou ◽  
Yuanxiao Yang ◽  
Hongyan Liu ◽  
Fang Zhou ◽  
...  
Keyword(s):  
Polyketide Synthase ◽  
Hybrid Breeding ◽  
Pollen Wall ◽  
Wall Structure ◽  
Ultrastructural Observation ◽  
Wild Type ◽  
In Planta ◽  
Male Sterile ◽  
Crop Species ◽  
Cytoplasmic Inclusions

Abstract Background: Sesame is a great reservoir of bioactive constituents and unique antioxidant components and is widely used for its nutritional and medicinal value. The expanding demands for sesame seeds are putting pressure on sesame breeders to develop reliable high-yielding varieties. Heterosis utilization is an efficient way to increase sesame yield. Polyketide synthases (PKSs) are critical enzymes in the biosynthesis of sporopollenin, a primary component of pollen exine. Their in planta functions are being investigated for application in crop breeding.Results: In this study, we cloned the sesame POLYKETIDE SYNTHASE A (SiPKSA) and examined its function in male sterility. SiPKSA was specifically expressed in sesame flower buds, and its expression was significantly higher in sterile sesame anthers than in fertile anthers at the tetrad and microspore development stage. Further overexpression of SiPKSA in Arabidopsis caused transgenic plants male sterile. Ultrastructural observation showed that the pollen grains of SiPKSA-overexpressing plants contained few cytoplasmic inclusions and exhibited an abnormal pollen wall structure, with a thicker exine layer compared with wild type. In agreement with it, the expression of a set of sporopollenin biosynthesis-related genes and the contents of fatty acids and phenolics were significantly altered in anthers of SiPKSA-overexpressing plants compared with wild type during anther development. Conclusion: These findings highlighted that overexpression of SiPKSA in Arabidopsis might cause excessive sporopollenin biosynthesis to influence pollen and pollen wall development, leading to male sterile, suggesting that its manipulation might improve hybrid breeding in sesame and other crop species.

Role of plant respiratory burst oxidase homologs in stress responses

2018 ◽  
Vol 52 (8) ◽  
pp. 826-839 ◽  
Author(s):  
Wei Wang ◽  
Dongdong Chen ◽  
Xiaopei Zhang ◽  
Dan Liu ◽  
Yingying Cheng ◽  
...  
Keyword(s):  
Stress Responses ◽  
Respiratory Burst ◽  
Respiratory Burst Oxidase

scholarly journals Role of Listeria monocytogenes Exotoxins Listeriolysin and Phosphatidylinositol-Specific Phospholipase C in Activation of Human Neutrophils

1999 ◽  
Vol 67 (3) ◽  
pp. 1125-1130 ◽  
Author(s):  
Ulf Sibelius ◽  
Eva-Cathrin Schulz ◽  
Frank Rose ◽  
Katja Hattar ◽  
Thomas Jacobs ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Phospholipase C ◽  
Virulence Factors ◽  
Respiratory Burst ◽  
Phosphoinositide Hydrolysis ◽  
Human Neutrophils ◽  
Avirulent Strain ◽  
Wild Type ◽  
Defective Mutant

ABSTRACT Polymorphonuclear leukocytes (PMN) are essential for resolution of infections with Listeria monocytogenes. The present study investigated the role of the listerial exotoxins listeriolysin (LLO) and phosphatidylinositol-specific phospholipase C (PlcA) in human neutrophil activation. Different Listeria strains, mutated in individual virulence genes, as well as purified LLO were used. Coincubation of human neutrophils with wild-type L. monocytogenes provoked PMN activation, occurring independently of phagocytosis events, with concomitant elastase secretion, leukotriene generation, platelet-activating factor (PAF) synthesis, respiratory burst, and enhanced phosphoinositide hydrolysis. Degranulation and leukotriene formation were noted to be solely dependent on LLO expression, as these features were absent when the LLO-defective mutant EGD− and the avirulent strain L. innocua were used. These effects were fully reproduced by a recombinant L. innocuastrain expressing LLO (INN+) and by the purified LLO molecule. LLO secretion was also required for PAF synthesis. However, wild-typeL. monocytogenes was more potent in eliciting PAF formation than mutants expressing LLO, suggesting the involvement of additional virulence factors. This was even more obvious for phosphoinositide hydrolysis and respiratory burst: these events were provoked not only by INN+ but also by the LLO-defective mutant EGD− and by a recombinantL. innocua strain producing listerial PlcA. We conclude that human neutrophils react to extracellularly provided listerial exotoxins by rapid cell activation. Listeriolysin is centrally involved in triggering degranulation and lipid mediator generation, and further virulence factors such as PlcA apparently contribute to trigger neutrophil phosphoinositide hydrolysis and respiratory burst. In this way, listerial exotoxins may influence the host defense against infections with L. monocytogenes.

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