Generation and Assessment of Soybean (<i>Glycine max</i> (L.) Merr.) Hybrids for High-Efficiency <i>Agrobacterium</i>-Mediated Transformation

The <i>Agrobacterium</i>-mediated technique is widely employed for soybean transformation, but the efficiency of this method is still relatively modest, in which multiple factors are involved. Numerous chemical and physiological cues from host plants are needed for <i>A. tumefacien...

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Main Authors: Muhammad Waqar Khan, Aaqib Shaheen, Xuebin Zhang, Junli Zhang, Yaser Hassan Dewir, Katalin Magyar-Tábori
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Life
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Online Access:https://www.mdpi.com/2075-1729/14/12/1649
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author Muhammad Waqar Khan
Aaqib Shaheen
Xuebin Zhang
Junli Zhang
Yaser Hassan Dewir
Katalin Magyar-Tábori
author_facet Muhammad Waqar Khan
Aaqib Shaheen
Xuebin Zhang
Junli Zhang
Yaser Hassan Dewir
Katalin Magyar-Tábori
author_sort Muhammad Waqar Khan
collection DOAJ
description The <i>Agrobacterium</i>-mediated technique is widely employed for soybean transformation, but the efficiency of this method is still relatively modest, in which multiple factors are involved. Numerous chemical and physiological cues from host plants are needed for <i>A. tumefaciens</i> attraction and subsequent T-DNA integration into the plant genome. Susceptible genotypes may permit this attachment and integration, and the agronomically superior genotypes with susceptibility to <i>A. tumefaciens</i> would play an important role in increasing transformation efficiency. In this study, we aimed to elevate the <i>Agrobacterium</i>-mediated transformation efficiency of soybean by integrating susceptibility alleles from William82 and flavonoids accumulating alleles from LX genotypes in the same soybean line. The crossing was made between LX () and William82 () soybean by hand pollination. Expectedly, the resulting hybrid soybean progenies inherited susceptibility traits and high flavonoid contents (i.e., genistein, genistin, apigenin, naringenin, quercetin, and cinnamic acid) essential for potential plant–pathogen interaction. Furthermore, the progenies and susceptible William82 soybean were subjected to transformation using <i>A. tumefaciens</i> (GV3101) harboring the GmUbi-3XFlag-35S-GFP and reassembled GmUbi3XFlag-35S-GFP: GUS vectors during separate events. Important transformation-related traits like shoot induction and shoot regeneration ability were also significantly improved in progenies. The progenies designated as ZX-3 exhibited superiority over the William82 parental line in all three traits, i.e., shoot induction, regeneration, and <i>Agrobacterium</i>-mediated transformation. The transient transformation efficiency of the ZX-16 line was remarkably higher when half-cotyledon explants were wounded and transformed with <i>A. tumefaciens</i> harboring GUS assembly vector and then co-cultivated on MS medium supplemented with 2 mg/L spermidine, 0.3 g/L GA3, 0.3 mg/L kinetin, and 1.3 mg/L 6-benzylaminopurine. In addition, the shoot elongation was also higher than that of William82 after two weeks of culture on the shoot induction medium. The newly generated soybeans have the potential to serve as a valuable source for high transgene production and represent a promising avenue for future soybean varietal development.
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spelling doaj-art-ea832687f3684896b0ff80da0000db572025-08-20T02:00:28ZengMDPI AGLife2075-17292024-12-011412164910.3390/life14121649Generation and Assessment of Soybean (<i>Glycine max</i> (L.) Merr.) Hybrids for High-Efficiency <i>Agrobacterium</i>-Mediated TransformationMuhammad Waqar Khan0Aaqib Shaheen1Xuebin Zhang2Junli Zhang3Yaser Hassan Dewir4Katalin Magyar-Tábori5State Key Laboratory of Crop Stress Adaptation and Improvement, Henan Joint International Laboratory for Crop Multi-Omics Research, School of Life Sciences, Henan University, Jinming Road, Kaifeng 475004, ChinaState Key Laboratory of Crop Stress Adaptation and Improvement, College of Agriculture, Henan University, Kaifeng 475004, ChinaState Key Laboratory of Crop Stress Adaptation and Improvement, Henan Joint International Laboratory for Crop Multi-Omics Research, School of Life Sciences, Henan University, Jinming Road, Kaifeng 475004, ChinaState Key Laboratory of Crop Stress Adaptation and Improvement, Henan Joint International Laboratory for Crop Multi-Omics Research, School of Life Sciences, Henan University, Jinming Road, Kaifeng 475004, ChinaPlant Production Department, College of Food and Agriculture Sciences, King Saud University, Riyadh 11451, Saudi ArabiaResearch Institute of Nyíregyháza, Institutes for Agricultural Research and Educational Farm (IAREF), University of Debrecen, P.O. Box 12, 4400 Nyíregyháza, HungaryThe <i>Agrobacterium</i>-mediated technique is widely employed for soybean transformation, but the efficiency of this method is still relatively modest, in which multiple factors are involved. Numerous chemical and physiological cues from host plants are needed for <i>A. tumefaciens</i> attraction and subsequent T-DNA integration into the plant genome. Susceptible genotypes may permit this attachment and integration, and the agronomically superior genotypes with susceptibility to <i>A. tumefaciens</i> would play an important role in increasing transformation efficiency. In this study, we aimed to elevate the <i>Agrobacterium</i>-mediated transformation efficiency of soybean by integrating susceptibility alleles from William82 and flavonoids accumulating alleles from LX genotypes in the same soybean line. The crossing was made between LX () and William82 () soybean by hand pollination. Expectedly, the resulting hybrid soybean progenies inherited susceptibility traits and high flavonoid contents (i.e., genistein, genistin, apigenin, naringenin, quercetin, and cinnamic acid) essential for potential plant–pathogen interaction. Furthermore, the progenies and susceptible William82 soybean were subjected to transformation using <i>A. tumefaciens</i> (GV3101) harboring the GmUbi-3XFlag-35S-GFP and reassembled GmUbi3XFlag-35S-GFP: GUS vectors during separate events. Important transformation-related traits like shoot induction and shoot regeneration ability were also significantly improved in progenies. The progenies designated as ZX-3 exhibited superiority over the William82 parental line in all three traits, i.e., shoot induction, regeneration, and <i>Agrobacterium</i>-mediated transformation. The transient transformation efficiency of the ZX-16 line was remarkably higher when half-cotyledon explants were wounded and transformed with <i>A. tumefaciens</i> harboring GUS assembly vector and then co-cultivated on MS medium supplemented with 2 mg/L spermidine, 0.3 g/L GA3, 0.3 mg/L kinetin, and 1.3 mg/L 6-benzylaminopurine. In addition, the shoot elongation was also higher than that of William82 after two weeks of culture on the shoot induction medium. The newly generated soybeans have the potential to serve as a valuable source for high transgene production and represent a promising avenue for future soybean varietal development.https://www.mdpi.com/2075-1729/14/12/1649soybean breedingtransformationflavonoidssusceptibilityUPLC-MS/MS
spellingShingle Muhammad Waqar Khan
Aaqib Shaheen
Xuebin Zhang
Junli Zhang
Yaser Hassan Dewir
Katalin Magyar-Tábori
Generation and Assessment of Soybean (<i>Glycine max</i> (L.) Merr.) Hybrids for High-Efficiency <i>Agrobacterium</i>-Mediated Transformation
Life
soybean breeding
transformation
flavonoids
susceptibility
UPLC-MS/MS
title Generation and Assessment of Soybean (<i>Glycine max</i> (L.) Merr.) Hybrids for High-Efficiency <i>Agrobacterium</i>-Mediated Transformation
title_full Generation and Assessment of Soybean (<i>Glycine max</i> (L.) Merr.) Hybrids for High-Efficiency <i>Agrobacterium</i>-Mediated Transformation
title_fullStr Generation and Assessment of Soybean (<i>Glycine max</i> (L.) Merr.) Hybrids for High-Efficiency <i>Agrobacterium</i>-Mediated Transformation
title_full_unstemmed Generation and Assessment of Soybean (<i>Glycine max</i> (L.) Merr.) Hybrids for High-Efficiency <i>Agrobacterium</i>-Mediated Transformation
title_short Generation and Assessment of Soybean (<i>Glycine max</i> (L.) Merr.) Hybrids for High-Efficiency <i>Agrobacterium</i>-Mediated Transformation
title_sort generation and assessment of soybean i glycine max i l merr hybrids for high efficiency i agrobacterium i mediated transformation
topic soybean breeding
transformation
flavonoids
susceptibility
UPLC-MS/MS
url https://www.mdpi.com/2075-1729/14/12/1649
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