A Bacillus thuringiensis Cry protein controls soybean cyst nematode in transgenic soybean plants

Kahn, Theodore W.; Duck, Nicholas B.; McCarville, Michael T.; Schouten, Laura Cooper; Schweri, Kathryn; Zaitseva, Jelena; Daum, Julia

doi:10.1038/s41467-021-23743-3

Cited by 37 publications

(20 citation statements)

References 41 publications

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“…The Cry31Aa proteins were found to show a strong nematicidal activity against A. besseyi. In addition, (6)-rhodamine was employed to label three cells, since carboxytetramethylrhodamine is a biological stain with strong fluorescence when dissolved in DMSO solvent and is commonly used for biological dyes [ 52 ]. After 4 h of treatment with the rhodamine-labeled Cry31Aa protein, red fluorescence was visible in the guts of the nematodes, which agrees with the results in [ 53 ].…”

Section: Discussionmentioning

confidence: 99%

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Toxicity of Bacillus thuringiensis Strains Derived from the Novel Crystal Protein Cry31Aa with High Nematicidal Activity against Rice Parasitic Nematode Aphelenchoides besseyi

Zhao

Ali

Wang

et al. 2022

IJMS

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The plant parasitic nematode, Aphelenchoides besseyi, is a serious pest causing severe damage to various crop plants and vegetables. The Bacillus thuringiensis (Bt) strains, GBAC46 and NMTD81, and the biological strain, FZB42, showed higher nematicidal activity against A. besseyi, by up to 88.80, 82.65, and 75.87%, respectively, in a 96-well plate experiment. We screened the whole genomes of the selected strains by protein-nucleic acid alignment. It was found that the Bt strain GBAC46 showed three novel crystal proteins, namely, Cry31Aa, Cry73Aa, and Cry40ORF, which likely provide for the safe control of nematodes. The Cry31Aa protein was composed of 802 amino acids with a molecular weight of 90.257 kDa and contained a conserved delta-endotoxin insecticidal domain. The Cry31Aa exhibited significant nematicidal activity against A. besseyi with a lethal concentration (LC50) value of 131.80 μg/mL. Furthermore, the results of in vitro experiments (i.e., rhodamine and propidium iodide (PI) experiments) revealed that the Cry31Aa protein was taken up by A. besseyi, which caused damage to the nematode’s intestinal cell membrane, indicating that the Cry31Aa produced a pore-formation toxin. In pot experiments, the selected strains GBAC46, NMTD81, and FZB42 significantly reduced the lesions on leaves by up to 33.56%, 45.66, and 30.34% and also enhanced physiological growth parameters such as root length (65.10, 50.65, and 55.60%), shoot length (68.10, 55.60, and 59.45%), and plant fresh weight (60.71, 56.45, and 55.65%), respectively. The number of nematodes obtained from the plants treated with the selected strains (i.e., GBAC46, NMTD81, and FZB42) and A. besseyi was significantly reduced, with 0.56, 0.83., 1.11, and 5.04 seedling mL−1 nematodes were achieved, respectively. Moreover, the qRT-PCR analysis showed that the defense-related genes were upregulated, and the activity of hydrogen peroxide (H2O2) increased while malondialdehyde (MDA) decreased in rice leaves compared to the control. Therefore, it was concluded that the Bt strains GBAC46 and NMTD81 can promote rice growth, induce high expression of rice defense-related genes, and activate systemic resistance in rice. More importantly, the application of the novel Cry31Aa protein has high potential for the efficient and safe prevention and green control of plant parasitic nematodes.

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Section: Discussionmentioning

confidence: 99%

“…The endocytosis assays were carried out as previously described in [ 52 ], with some modifications. The 5(6)-carboxytetramethylrhodamine is a biological stain with strong fluorescence when dissolved in DMSO solvent and is often used as a biological dye [ 52 ].…”

Section: Methodsmentioning

confidence: 99%

Toxicity of Bacillus thuringiensis Strains Derived from the Novel Crystal Protein Cry31Aa with High Nematicidal Activity against Rice Parasitic Nematode Aphelenchoides besseyi

Zhao

Ali

Wang

et al. 2022

IJMS

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“…Recently, BASF released a study in which they showed that the use of transgenic lines expressing the Cry14Ab protein has exceptional potential for controlling plant-parasitic nematodes [ 80 ]. They utilized the Bacillus thuringiensis delta-endotoxin, Cry14Ab for SCN control.…”

Section: Novel Resistance Strategies and Breeding Approachesmentioning

confidence: 99%

A Broad Review of Soybean Research on the Ongoing Race to Overcome Soybean Cyst Nematode

Nissan

Mimee

Cober

et al. 2022

Biology

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Plant pathogens greatly impact food security of the ever-growing human population. Breeding resistant crops is one of the most sustainable strategies to overcome the negative effects of these biotic stressors. In order to efficiently breed for resistant plants, the specific plant–pathogen interactions should be understood. Soybean is a short-day legume that is a staple in human food and animal feed due to its high nutritional content. Soybean cyst nematode (SCN) is a major soybean stressor infecting soybean worldwide including in China, Brazil, Argentina, USA and Canada. There are many Quantitative Trait Loci (QTLs) conferring resistance to SCN that have been identified; however, only two are widely used: rhg1 and Rhg4. Overuse of cultivars containing these QTLs/genes can lead to SCN resistance breakdown, necessitating the use of additional strategies. In this manuscript, a literature review is conducted on research related to soybean resistance to SCN. The main goal is to provide a current understanding of the mechanisms of SCN resistance and list the areas of research that could be further explored.

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“…Planting SCN-resistant cultivars is still the most environmentally and economically friendly method to control SCNs. Around 90% of SCN-resistant cultivars currently bred in the central US are derived from the soybean germplasm PI8878, whichhas become noneffective against SCNs due to SCN populations overcoming their resistance to PI8878, and the female index (FI) has increased [ 7 ]. So, it is a research priority for controlling SCN worldwide to screen resistant germplasms and discover the high-quality resistant genes under the limited number of naturally resistant cultivars facing failure and depletion.…”

Section: Introductionmentioning

confidence: 99%

“…Many scientists have performed a great deal of research on SCN resistance, from plant resistance genes to the nematode gene itself. Expression of Bacillus thuringiensis ( Bt ) delta-endotoxin ( Cry14Ab ) in soybean could significantly reduce the number of SCNs compared to control plants 30 d after inoculation due to Cry14Ab directly damaging the intestines of the nematodes [ 7 ]. Overexpression of kinase-dead variants of five highly connected syncytium hub genes significantly enhanced soybean resistance to SCNs [ 8 ].…”

Section: Introductionmentioning

confidence: 99%

Generation and Identification of the Number of Copies of Exogenous Genes and the T-DNA Insertion Site in SCN-Resistance Transformation Event ZHs1-2

Tang

Zhong

Wan

et al. 2022

IJMS

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Soybean cyst nematode (SCN, Heterodera glycines Ichinohe) causes an estimated economic loss of about USD 3 billion each year in soybean (Glycine max L.) production worldwide. Overexpression of resistance genes against SCN provides a powerful approach to develop SCN resistance cultivars in soybean. The clarification of molecular characterization in transformation events is a prerequisite for ecological risk assessment, food safety, and commercial release of genetically modified crops. Here, we generated transgenic events harboring the BCN (beet cyst nematode) resistance Hs1pro−1 gene using the Agrobacterium-mediated method in soybean, evaluated their resistance to SCN infection, and clarified the molecular characterization of one of the transformation events. Five independent and stable inheritable transformation events were generated by an Agrobacterium-mediated transformation method. SCN resistance tests showed the average number of developed females per plant and female index (FI) in T4 ZHs1-1, ZHs1-2, ZHs1-3, ZHs1-4, and ZHs1-5 transformation events were significantly lower than that in the nontransgenic control. Among these, the ZHs1-2 transformation event had the lowest number of developed females per plant and FI. Southern hybridization showed the exogenous target Hs1pro−1 gene was inserted in one copy and the Bar gene was inserted two copies in the ZHs1-2 transformation event. The exogenous T-DNA fragment was integrated in the reverse position of Chr02: 5351566–5231578 (mainly the Bar gene expression cassette) and in the forward position of Chr03: 17083358–17083400 (intact T-DNA, including Hs1pro−1and Bar gene expression cassette) using a whole genome sequencing method (WGS). The results of WGS method and Southern hybridization were consistent. All the functional elements of exogenous T-DNA fragments were verified by PCR using specific primer pairs in the T5 and T6 ZHs1-2 transformation events. These results demonstrated that the overexpression of Hs1pro−1 gene enhanced SCN resistance, and provide an important reference for the biosafety assessment and the labeling detection in transformation event ZHs1-2.

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A Bacillus thuringiensis Cry protein controls soybean cyst nematode in transgenic soybean plants

Cited by 37 publications

References 41 publications

Toxicity of Bacillus thuringiensis Strains Derived from the Novel Crystal Protein Cry31Aa with High Nematicidal Activity against Rice Parasitic Nematode Aphelenchoides besseyi

Toxicity of Bacillus thuringiensis Strains Derived from the Novel Crystal Protein Cry31Aa with High Nematicidal Activity against Rice Parasitic Nematode Aphelenchoides besseyi

A Broad Review of Soybean Research on the Ongoing Race to Overcome Soybean Cyst Nematode

Generation and Identification of the Number of Copies of Exogenous Genes and the T-DNA Insertion Site in SCN-Resistance Transformation Event ZHs1-2

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