This study aims to establish an efficient protoplast-mediated genetic transformation sys-tem for Pochonia chlamydosporia(P.chlamydosporia)to facilitate gene transformation,genetic modification,and subsequent biological function research.Through orthogonal testing,the study systematically optimized key factors influencing electroporation,including voltage,osmotic stabi-lizer,pulse time,nucleic acid concentration,and protoplast concentration.The results showed that the optimal electroporation conditions were:voltage of 250 V,osmotic stabilizer as 0.6 mol/L su-crose solution,pulse time of 10 ms,plasmid concentration of 1％,and protoplast concentration of 1×107 protoplasts/mL,yielding a transformation efficiency of 0.4 × 103 CFU/pg.The interaction analysis revealed that the five factors affected transformation efficiency in the following order:plasmid concentration＞protoplast concentration＞pulse time＞osmotic stabilizer type＞voltage.Based on the positive transformants,the study further evaluated their colony morphology,growth rate,conidial yield,mycelial dry weight,and the ability to infect the eggs of three types of animal gastrointestinal nematodes.The results indicated that the electroporation process did not signifi-cantly affect the biological functions of P.chlamydosporia.The transformants successfully ex-pressed enhanced green fluorescent protein(EGFP),and PCR analysis confirmed the successful in-tegration of the thiostrepton resistance gene.In conclusion,this study successfully established a protoplast-based electroporation-mediated genetic transformation system for P.chlamydosporia,providing a valuable foundation for further research into the mechanism of egg parasitism and the underlying genes involved in its biocontrol activity.

This study aims to establish an efficient protoplast-mediated genetic transformation sys-tem for Pochonia chlamydosporia(P.chlamydosporia)to facilitate gene transformation,genetic modification,and subsequent biological function research.Through orthogonal testing,the study systematically optimized key factors influencing electroporation,including voltage,osmotic stabi-lizer,pulse time,nucleic acid concentration,and protoplast concentration.The results showed that the optimal electroporation conditions were:voltage of 250 V,osmotic stabilizer as 0.6 mol/L su-crose solution,pulse time of 10 ms,plasmid concentration of 1％,and protoplast concentration of 1×107 protoplasts/mL,yielding a transformation efficiency of 0.4 × 103 CFU/pg.The interaction analysis revealed that the five factors affected transformation efficiency in the following order:plasmid concentration＞protoplast concentration＞pulse time＞osmotic stabilizer type＞voltage.Based on the positive transformants,the study further evaluated their colony morphology,growth rate,conidial yield,mycelial dry weight,and the ability to infect the eggs of three types of animal gastrointestinal nematodes.The results indicated that the electroporation process did not signifi-cantly affect the biological functions of P.chlamydosporia.The transformants successfully ex-pressed enhanced green fluorescent protein(EGFP),and PCR analysis confirmed the successful in-tegration of the thiostrepton resistance gene.In conclusion,this study successfully established a protoplast-based electroporation-mediated genetic transformation system for P.chlamydosporia,providing a valuable foundation for further research into the mechanism of egg parasitism and the underlying genes involved in its biocontrol activity.

【Objective】  To study the effects of soil environment on the growth, yield, and quality of Sharen (Amomi Fructus) under different planting patterns. 【Methods】  Soil physical and chemical indices and enzyme activities in four periods including early flowering (March), full flowering (June), fruit ripening (September), and late fruit picking (December), were measured under three planting patterns including natural forest, greenhouse, and rubber forest in Xishuangbanna, China. The changes in soil indices during the growth periods of Sharen (Amomi Fructus) under different planting patterns were analyzed, and the differences in plant growth, yield, and quality under different planting patterns were explored. Pearson correlation analysis was used to analyze the relationship between soil indices and Sharen (Amomi Fructus) growth, yield, and quality. Principal component analysis was used to investigate the effects of soil environment under different planting patterns on Sharen (Amomi Fructus) growth, yield, and quality. 【Results】  The soil moisture, available potassium content, and urease activity of the three planting patterns of Sharen (Amomi Fructus) increased initially and decreased afterwards throughout the year; pH and organic matter content showed little change in the whole year. Exchangeable manganese content and acid phosphatase activity gradually increased throughout the year. Hydrolyzed nitrogen content, exchangeable calcium content, available zinc content, protease activity, and sucrase activity decreased initially and increased afterwards throughout the year. Exchangeable magnesium content, available iron content, and catalase activity decreased annually. Total nitrogen content, total phosphorus content, and available phosphorus content fluctuated throughout the year. The total potassium content under natural forest and greenhouse planting decreased throughout the year, while the total potassium content under rubber forest showed an upward trend all year round. The organic matter content, total nitrogen content, total potassium content, available potassium content, available zinc content, urease activity, acid phosphatase activity, and catalase activity under greenhouse were significantly lower than those under natural and rubber forests (P < 0.05). Correlation analysis showed that plant growth, yield, and quality of Sharen (Amomi Fructus) were significantly correlated with soil organic matter, total nitrogen, hydrolyzed nitrogen, total phosphorus, available phosphorus, total potassium, available potassium, exchangeable manganese, exchangeable magnesium, exchangeable calcium, available zinc, urease, acid phosphatase, and invertase (P <  0.05). The results of the principal component analysis indicated that the soil environment of Sharen (Amomi Fructus) under natural forest was the best, followed by rubber forest and greenhouse. The order of its advantages and disadvantages is consistent with the growth index of Sharen (Amomi Fructus), but contrary to the yield of Sharen (Amomi Fructus), indicating that the soil environment directly affects the growth index and nutritional components of plants. 【Conclusion】  Different planting patterns of Sharen (Amomi Fructus) have different soil nutrient content, and the change rules in the growths period are not similar, with some differences. Soil indices have impacts on plant growth, yield, and quality of Sharen (Amomi Fructus). Soil ecological environment is positively correlated with the growth characteristics of Sharen (Amomi Fructus) plants, but has no direct correlation with yield and quality.

In this study,70 cows with (34.55 ± 8.44) kg per day milk,(2.24 ± 1.01) years at (60-90)d postpartum were chosen from an intensive dairy farm in Heilongjiang province.Ten plasma parameters including reproductive hormones,energy metabolism,liver function and minerals were measured at(60-90)d postpartum from the estrous and ovarian cyst dairy cows,combined with binary Pearson correlation analysis,Logistic analysis and ROC analysis.The results showed that:the incidence of ovarian cysts in this farm was 9.8％,its main reason is negative energy balance;the plasma concentrations of FSH,LH and P4 in ovarian cysts were lower than those in estrus group,but E2 (6.2 ng/L) was significantly higher than that in estrus group ((91.97± 10.62) ng/L);the plasma concentrations of NEFA,BHBA and AST were significantly increased in ovaries cysts at 14-21 d and 60-90 d,and Glu was significantly lower than that in estrus group.When the plasma concentration of BHBA were more than 0.855 mmol/L,NEFA more than 0.585 mmol/L,AST more than 77.0 U/L at 14-21 d postpartum,the risk of ovarian cyst increased.Negative energy balance can cause disorder of reproductive hormone secretion in early lactation cows,and then provoke cyst of follicle.Plasma NEFA,BHBA and AST of dairy cows postpartum can be used to predict the risk of ovarian cysts.