IND-ØØ41Ø-5 - Verdeca HB4 Soybean | BCH-LMO-SCBD-112020 | Living Modified Organism | Biosafety Clearing-House


Living Modified Organism (LMO)

Decisions on the LMO Risk Assessments  
published: 13 Jun 2017 last updated: 26 Jul 2019
Living Modified Organism identity
The image below identifies the LMO through its unique identifier, trade name and a link to this page of the BCH. Click on it to download a larger image on your computer. For help on how to use it go to the LMO quick-links page.
Verdeca HB4 Soybean
Soybean (Glycine max) was modified to express sunflower (Helianthus annuus) homeobox-leucine zipper protein 4 (HaHB4) gene to confer tolerance to abiotic stresses, such as drought and hypersaline conditions. In transgenic Arabidopsis thaliana experiments, the plants expressing HB4 survived in water-limited conditions, while wild-type plants perished.

Note: The Streptomyces hygroscopicus bar gene was also inserted into the genome and was used for selection during transformation (not with the intention for herbicide resistance).
The term “Recipient organism” refers to an organism (either already modified or non-modified) that was subjected to genetic modification, whereas “Parental organisms” refers to those that were involved in cross breeding or cell fusion.
Soybean cultivar: Glycine max cv. Williams 82
Characteristics of the modification process
  • Agrobacterium-mediated DNA transfer
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
  • BCH-GENE-SCBD-112018-1 Homeodomain-leucine zipper 4 promoter | Helianthus annuus (Sunflower, HELAN)
  • BCH-GENE-SCBD-112017-2 Homeodomain-leucine zipper 4 gene | Helianthus annuus (Sunflower, HELAN)
    Protein coding sequence | Tolerance to abiotic stress (Drought, Salinity)
  • BCH-GENE-SCBD-100269-8 Nopaline Synthase Gene Terminator | Agrobacterium tumefaciens (Agrobacterium)
  • BCH-GENE-SCBD-115068-1 Vegetative storage protein terminator | Glycine max (Soybean, Soya bean, Soya, SOYBN)
  • BCH-GENE-SCBD-14972-12 Phosphinothricin N-acetyltransferase gene | Streptomyces hygroscopicus (STRHY)
    Protein coding sequence | Resistance to herbicides (Glufosinate)
  • BCH-GENE-SCBD-104664-2 TEV 5' Untranslated Region | Tobacco etch virus (TEV)
  • BCH-GENE-SCBD-100287-7 CaMV 35S promoter | Cauliflower mosaic virus (CaMV)
bar gene cassette (Anti-sense orientation)
The transcription of the Streptomyces hygroscopicus phosphinothricin N-acetyltransferase (bar; PAT protein) gene is under the control of the duplicated Cauliflower Mosaic Virus 35S promoter and the Glycine max vegetative storage protein terminator. To enhance translation, the Tobacco Etch Virus 5' untranslated region was included at the 5' end of the bar transcript.

Homeodomain-leucine zipper 4 gene cassette (Sense orientation)
Transcription of the Helianthus annuus Homeodomain-leucine zipper 4 (HaHB4) gene starts at the HaHB4 promoter and terminates at the Agrobacterium tumefaciens nopaline synthase gene terminator. The promoter activity is excepted to increase in response to abiotic stresses, ethylene and insect feeding, resulting in accumulations of HaHB4 transcript and HAHB4 protein.

HaHB4 nucleotide changes:
- Deletion of nucleotides 76 to 87
- A to G substitution at position 22 (codon change from AAA to AGA)
- G to A substitution at position 351 (silent point mutation - CAG to CAC)
- A to C substitution at position 401 (silent point mutation - GCA to GCC)
- T to C substitution at position 432 (codon change TTT to CTT)
- C to T substitution at position 551 (codon change CCT to CTT)

HB4 amino acid changes:
- Four amino acid deletion (residue numbers 7 to 10 - results in numbering change in residues)
- Lysine to Arginine substitution at position 22 (K22 -> R18)
- Phenylalanine to Leucine substitution at position 159 (F159 -> L155)
- Proline to Leucine substitution at position 175 (P175 -> L171)

- Southern blot, next generation sequencing and PCR analyses indicate that the plasmid backbone was absent in the genome.
- Southern blotting and sequencing confirmed the presence of a single, T-DNA insert.
- PCR analysis indicated that the transgene insert followed Mendelian inheritance.
- Bioinformatic analysis indicated that no new open read frames were created as a result of the T-DNA insertion.
LMO characteristics
  • Food
  • Feed
Detection method(s)
PAT protein:
- the protein is expressed in the soybean forage and seeds at level comparable to or less than other crops expressing PAT
- highest levels of protein are detected in leaf and seed tissue

HB4 protein
- under normal conditions, the protein was difficult to detect in leaf and seed tissue (level below limit of detection).