人参根腐病离体抗性筛选方法初探

探讨了人参(Panax ginseng C. A. Meyer)根腐病离体抗性筛选方法的可行性,初步建立了一种快速高效的人参根腐病抗性筛选的方法。采用离体叶片、根块2种方法进行接种,结果表明,2种方法接种的发病情况基本一致,且用离体叶片比用根块筛选操作更简单,观察更方便;采用菌块、菌液2种接种方法分别对人参叶片的正、反面进行接种,结果表明,用菌块正面接种发病效果更好。进一步通过人参须回接试验初步证明了离体叶片抗性筛选方法的可行性。通过对不同品系人参进行接菌,发现FS抗病能力最高,FX2抗病能力最低。

人参生长年限与海拔高度对栽培人参土壤中大量元素的影响

[目的]研究海拔与生长时间对栽培人参土壤中大量元素的影响。[方法]采集我国人参主产地长白山地区不同人参生长时间、不同海拔的人参土壤,测量其大量元素和微量元素的含量,并研究变化规律。[结果]相同参龄条件下,土壤中大量元素pH值、有机质、全氮、全磷、全钾、碱解氮、速效磷和速效钾整体随着海拔高度的增加而变化,不同参龄土壤,大量元素含量整体随着参龄年限的增长而变化,其中土壤pH值随着参龄的增长有变小的趋势。[结论]不同海拔与生长时间对人参大量元素均有影响。

Optimizing SSR-PCR system of Panax ginseng by orthogonal design

An orthogonal design was used to optimize SSR-PCR amplification system using Panax ginseng genomic DNA as template. Four levels of five factors （DNA template, Taq DNA polymerase, Mg^2＋, primer, and dNTP） and annealing temperature have been tested separately in this system. The results demonstrated the reaction efficiency was affected by these factors. Based on the results, a stable, productive and reproducible PCR system and cycling program for amplifying a ginseng SSR locus were obtained： 20 μL system containing 1.0 U Taq DNA polymerase, 2.0 mmol·L^-1 Mg^2＋, 0.2 mmol·L^-1 dNTPs, 0.3 μmol·L^-1 SSR primer, 60 ng· μla^-1 DNA template, performed with a program of 94℃ for 5 min, 94℃ for 30 s, annealing at 56.3℃ for 30 s, 72℃ for 1 min, 37 cycles, finishing at 72℃ for 7 min, and storing at 4℃.

Antagonistic Effects of Sphingomonas and Pseudomonas aeruginosa on 4 Kinds of Pathogenic Bacteria of Ginseng

[Objectives]To explore effective biocontrol methods for diseases in the process of ginseng cultivation,and develop an efficient and environmentally friendly biocontrol agent.[Methods]In this study,2 strains were isolated from biogas slurry,and Cylindrocarpon destructans(XF),Fusarium solani(GF),Botrytis cinerea Pers(HM)and Alternaria panax Whetz(HB)were used as test materials.The strains were isolated and identified by dilution plate method,16S rDNA sequence identification method,confrontation culture method,filter paper method and ultraviolet spectrophotometer method,and the bacteriostatic activity and bacteriostatic rate were tested.[Results]Strain 15(Sphingomonas)and strain 19(Pseudomonas aeruginosa)were screened out through identification and analysis,and they grew stably within 8-10 d.The bacteriostatic rates of strain 15 against A.panax and B.cinerea were 47.37%and 43.40%,respectively,and the bacteriostatic rates of strain 19 against A.panax and B.cinerea were 62.30%and 63.27%,respectively.The bacteriostatic activity of the extract of strain 19 increased with the increase of OD_(600) value,and the bacteriostatic effect was optimal when the OD_(600) value was in the range of 0.8-1.0,up to 70%,so it had a strong biocontrol potential.[Conclusions]This experiment provides convenience for more effective inoculation,establishes a fast,simple and accurate method for the determination of the best bacteriostatic rate of P.aeruginosa culture solution to HM,and lays a foundation for large-scale culture of P.aeruginosa culture solution.Besides,it is expected to provide a theoretical basis for the efficient control of ginseng B.cinerea in field production,use it for the prevention and control of ginseng shoot diseases,and provide a reference for the efficient and diverse development of biocontrol agents for ginseng shoot diseases.

人参BZR1基因克隆及植物表达载体构建

油菜素内酯是一种天然植物激素,广泛参与植物生长、发育和响应环境的过程,BZR1是油菜素内酯信号转导途径中重要的转录因子。为研究人参BZR1基因在人参形态形成中的作用,对不同形态人参进行转录组测序分析其差异形成机制,利用RT-PCR方法,从人参中克隆BZR1基因,对其进行序列分析并构建植物表达载体。结果表明,成功克隆人参BZR1基因,命名为PgBZR1。经生物信息学分析,该基因全长966 bp,编码含321个氨基酸,理论相对分子质量34.5 kD,理论等电点为9.06,是一种碱性蛋白。经序列比对和系统发育进化分析表明,PgBZR1基因与胡萝卜BZR1基因亲缘关系最近,一致性为83%;用PgBZR1与植物表达载体pRI201构建重组子p RI201-PgBZR1,为PgBZR1基因的功能研究及人参新品种选育提供参考。

Panax japonicus and chikusetsusaponins: A review of diverse biological activities and pharmacology mechanism

Panax japonicus, which in the Tujia dialect is known as "Baisan Qi" and "Zhujieshen", is a classic "qi" drug of Tujia ethnomedicine and it has unique effects on disease caused by "qi" stagnation and blood stasis.This paper serves as the basis of further scientific research and development of Panax japonicus. The pharmacology effects of molecular pharmacology were discussed and summarized. P. japonicus plays an important role on several diseases, such as rheumatic arthritis, cancer, cardiovascular agents, and this review provides new insights into P. japonicus as promising agents to substitute ginseng and notoginseng.

Penicillium sp.YJM-2013 induces ginsenosides biosynthesis in Panax ginseng adventitious roots by inducing plant resistance responses

Objective:Fusarium oxysporum is a common pathogenic fungus in ginseng cultivation.Both pathogens and antagonistic fungi have been reported to induce plant resistance responses,thereby promoting the accumulation of secondary metabolites.The purpose of this experiment is to compare the advantages of one of the two fungi,in order to screen out more effective elicitors.The mechanism of fungal elicitor-induced plant resistance response is supplemented.Methods:A gradient dilution and the dural culture were carried out to screen strains.The test strain was identified by morphology and 18 s rDNA.The effect of different concentrations(0,50,100,200,400 mg/L)ofPenicillium sp.YJM-2013 and F.oxysporum on fresh weight and ginsenosides accumulation were tested.Signal molecules transduction,expression of transcription factors and functional genes were investigated to study the induction mechanism of fungal elicitors.Results:Antagonistic fungi ofF.oxysporum was identified as Penicillium sp.YJM-2013,which reduced root biomass.The total ginsenosides content of Panax ginseng adventitious roots reached the maximum(48.95±0.97 mg/g)treated with Penicillium sp.YJM-2013 at 200 mg/L,higher than control by 2.59-fold,in which protopanoxadiol-type ginsenosides(PPD)were increased by 4.57 times.Moreover,Penicillium sp.YJM-2013 activated defense signaling molecules,up-regulated the expression of PgWRKY 1,2,3,5,7,9 and functional genes in ginsenosides synthesis.Conclusion:Compared with the pathogenic fungi F.oxysporum,antagonistic fungi Penicillium sp.YJM-2013 was more conducive to the accumulation of ginsenosides in P.ginseng adventitious roots.Penicillium sp.YJM-2013 promoted the accumulation of ginsenosides by intensifying the generation of signal molecules,activating the expression of transcription factors and functional genes.

Spatio-Temporal Expression Pattern of Six Novel Candidate Genes in Ginsenoside Biosynthesis from Panax ginseng C. A. Meyer

Abstract: To explore the mode of the spatio-temporal expression of six newly discovered ginsenoside biosynthesis candidate gene transcripts, both Northern blotting and semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) were used to elucidate the mRNA expression levels of the transcripts in various tissues and organs of Panax ginseng C. A. Meyer during different growth development stages. The six gene transcripts were all differentially expressed in cultured callus, root, stem, leaf, and seed. The mRNA expression levels were significantly higher in four-year-old roots than in one-year-old roots, and results of semi-quantitative RT-PCR assays were in accordance with those of Northern blotting analyses. The results strongly suggest that all six genes were differentially expressed at root-specific developmental stages. In particular, when a quiescent early stage culture suspension of P. ginseng cells was exposed to the ginsenoside biosynthesis-promoting elicitor Aspergillus niger polysaccharide, the GBR6 gene transcript response showed time-dependent increments and was parallel with ginsenoside productivity (P < 0.01). Overexpressionof the GBR6 gene is likely to play a critically important role in the biosynthesis of ginsenosides. The results of the present study provided a background for the further elucidation of the structure and physiological function of these six candidate genes.