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<p>Rice, a major staple food for half of the world’s population, has been a significant source of nutrition. India is the second-largest producer and consumer of rice globally. Rice aroma is considered one of the most important traits as it is a key factor determining market price and is related to local and national identity. The genetic basis underlying aroma synthesis is well-explored in rice, and it is controlled by a recessive mutation in the <i>betaine aldehyde dehydrogenase 2 (OsBADH2)</i> gene that results in the synthesis of 2-acetyl-1-pyrroline (2AP). However, the non-scented rice varieties such as IR-64 carry a functional <i>OsBADH2</i> gene. Therefore, to induce aroma in IR-64, we employed CRISPR/Cas9 tool to knockout the <i>OsBADH2</i> gene<i>.</i> The single guide RNA targeting <i>OsBADH2</i> was cloned into pRGEB31 vector and transformed into IR-64 using <i>Agrobacterium</i>-mediated transformation. Several transgenic lines were developed and genotyped for targeted and untargeted (off-target) mutations in T<sub>1</sub> generation. No off-target activity was recorded in T<sub>1</sub> edited lines. Analysis of aromatic volatiles showed induction of 2AP in the grains of T<sub>0</sub> and T<sub>1</sub> edited lines. Moreover, a significant decrease in gamma-aminobutyric acid (GABA) content was observed, indicating a functionally successful gene editing. However, the agronomic characterization of <i>OsBADH2</i> edited T<sub>1</sub> lines revealed a reduction in yield-related characteristics. In conclusion, this research has established a successful genome-editing approach to induce the commercially important trait of aroma in the IR-64 rice variety and assessed yield-related traits in gene edited IR-64 lines.</p>

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Targeted disruption of OsBADH2 induces basmati aroma in the popular indica rice variety IR-64 using CRISPR/Cas9

  • 期刊论文

  • 发表日期：

    2025年10月18日

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  • Vacha Bhatt,
  • Thakku R. Ramkumar,
  • Vitthal Barvkar,
  • Rupesh Deshmukh,
  • Gunvant B. Patil,
  • Altafhusain Nadaf

摘要

Rice, a major staple food for half of the world’s population, has been a significant source of nutrition. India is the second-largest producer and consumer of rice globally. Rice aroma is considered one of the most important traits as it is a key factor determining market price and is related to local and national identity. The genetic basis underlying aroma synthesis is well-explored in rice, and it is controlled by a recessive mutation in the betaine aldehyde dehydrogenase 2 (OsBADH2) gene that results in the synthesis of 2-acetyl-1-pyrroline (2AP). However, the non-scented rice varieties such as IR-64 carry a functional OsBADH2 gene. Therefore, to induce aroma in IR-64, we employed CRISPR/Cas9 tool to knockout the OsBADH2 gene. The single guide RNA targeting OsBADH2 was cloned into pRGEB31 vector and transformed into IR-64 using Agrobacterium-mediated transformation. Several transgenic lines were developed and genotyped for targeted and untargeted (off-target) mutations in T1 generation. No off-target activity was recorded in T1 edited lines. Analysis of aromatic volatiles showed induction of 2AP in the grains of T0 and T1 edited lines. Moreover, a significant decrease in gamma-aminobutyric acid (GABA) content was observed, indicating a functionally successful gene editing. However, the agronomic characterization of OsBADH2 edited T1 lines revealed a reduction in yield-related characteristics. In conclusion, this research has established a successful genome-editing approach to induce the commercially important trait of aroma in the IR-64 rice variety and assessed yield-related traits in gene edited IR-64 lines.