Genetic transformation is a fundamental tool in molecular biology research of medicinal plants. Tailoring transgenic technologies to each distinct medicinal plant would necessitate a substantial investment of time and effort. Here, we present a simple hairy root transformation method that does not require sterile conditions, utilizing Agrobacterium rhizogenes strain K599 and the visible RUBY reporter system. Transgenic hairy roots were obtained for six tested medicinal plant species, roots or rhizomes of which have recognized medicinal value, spanning four botanical families and six genera (Platycodon grandiflorus, Atractylodes macrocephala, Scutellaria baicalensis, Codonopsis pilosula, Astragalus membranaceus, and Glycyrrhiza uralensis). Furthermore, two previously identified Glycyrrhiza uralensis UGTs that convert liquiritigenin into liquiritin in heterologous systems were studied in planta using the method. Our results indicate that overexpression of GuUGT1 but not GuUGT10 and Cas9-mediated knockout of GuUGT1 profoundly influenced the accumulation of liquiritin and isoliquiritin in licorice roots. Therefore, the method described here represents a simple, rapid and widely applicable hairy root transformation method that enables fast gene functional study in medicinal plants.

Objective To explore the significance of Survivin and PTEN in gastric cancer development and correlation between Survivin and PTEN.Methods Immunohistochemical methods were applied to detect the expres.sion of Survivin and PTEN in 105 pathological specimens of gastric cancer, in comparison to that in 98 negative gastric mucosa tissues.Results The positive rate of Survivin expression was 82.9% (87/105) in tissue specimens of gastric cancer and 6.1% (6/98) in tissue specimens of non-gastric cancer;The difference between them was statistically sig nificant (P <0.01).The positive rates of PTEN expression were 34.3% (36/105) in tissue specimens of gastric canc-er and 99.0% (97/98) in tissue specimens of non-gastric cancer;The difference between them was statistically signifi-cant ( P <0.01).The expression of Survivin and PTEN in tissues of gastric cancer had nothing to do with the gender of patients ( P > 0.05 ) , but was correlated with the degree of tumor cell differentiation, depth of invasion, lymph node me-tastasis, distant organ metastasis and TNM staging(P <0.05).Analysis by Spearman showed that Survivin was nega-tively correlated with PTEN( r = -0.23 ,P < 0.05 ).Conclusion The incidence of gastric cancer was related to the abnormally high level of Survivin expression and absence of PTEN expression;The expression of Survivin and PTEN in tissues of gastric cancer was unrelated with the gender of patients, but related to the degree of tumor cell differentia-tion, depth of invasion,lymph node metastasis,distant organ metastasis and TNM staging.The expression of Survivin was negatively correlated with the expression of PTEN.