Background
Hemsleya Chinensis
is a perennial plant in the Cucurbitaceae family containing antibacterial and anti-inflammatory compounds. The lack of genetic transformation systems makes it difficult to verify the functions of genes controlling important traits and conduct molecular breeding in
H. chinensis
.
Results
Highly efficient calli were induced on MS medium added 1.5 mg·L
− 1
6-benzylaminopurine (6-BA) and 0.02 mg·L
− 1
1-naphthylacetic acid (NAA) with high efficiency (> 95%). The frequency of shoot induction was increased to 90% with a plant growth regulator combination of 1.5 mg·L
− 1
6-BA and 0.1 mg·L
− 1
NAA. Our results also showed that 100% of shoot regeneration was achieved in a shoot regeneration medium. Additionally, more than 92% of kanamycin-resistant plants were confirmed. Furthermore, we achieved 42% genome editing efficiency by applying this transformation method to
HcOSC6
, a gene that catalyzes the formation of cucurbitadienol. HPLC analysis showed OE-
HcOSC6
lines exhibited significantly higher cucurbitadienol levels than the genome-edited lines. Transcriptomic analysis revealed that some downstream genes related to cucurbitadienol biosynthesis, such as
HcCYP87D20
,
HcCYP81Q58
, and
HcSDR34
, were up-regulated in OE lines and down-regulated in mutants.
Conclusions
Here, we established a process for regeneration, transformation, and genome editing of
H. chinensis
using stem segments. This provides valuable insight into the underlying molecular mechanisms of medicinal compound production. By combining high-efficiency tissue culture, transformation, and genome editing systems, we provide a powerful platform that supports functional research on molecular mechanisms of secondary metabolism.
Supplementary Information
The online version contains supplementary material available at 10.1186/s12870-024-05717-x.