Objective To establish and optimize the ISSR-PCR reaction system for Prunella vulgaris and lay foundation for its genetic diversity research.Methods The single-factor and orthogonal design were applied for optimizing seven factors in the ISSR-PCR reaction system including Mg2+,dNTP,primers,Taq DNA polymerase,the template DNA,annealing temperature,and cycles.Results The suitable PCR reaction system contained 2.2 mmol/L Mg2+,175 ?mol/L dNTP,0.75 ?mol/L primer,1.0 U Taq DNA polymerase,and 30 ng template DNA in total 20 ?L reaction solution.On this basis,18 primers were screened with stable amplification and rich polymorphism from 92 ISSR primers.The optimal annealing temperature for ISSR-PCR reaction was proposed by gradient PCR.Conclusion It is a way to establisb the ISSR-PCR system for orthogonal design combining with single-factor test.And it is proved to be stable and credible for the result of 24 P.vulgaris populations.This optimized ISSR reaction system would provide the basis for the genetic analysis of P.vulgaris.

Objective To establish and optimize ISSR-PCR systems of Chrysanthemum morifolium and lay the foundation for its genetic diversity research. Methods Based on the analysis of variance, an orthogonal design was used to optimize the ISSR-PCR amplification system on C. morifolium by four factors (Taq polymerase, Mg2+, dNTP, and primer) at three concentration levels, respectively. Results A suitable ISSR reaction system was constructed with the 20 ?L reaction system containing 1.00 U Taq polymerase, 2.00 mmol/L Mg2+, 0.20 mmol/L dNTP, and 0.50 ?mol/L primer. Conclusion ISSR-PCR is significantly influenced by the concentration of Taq polymerase, Mg2+, and dNTP. This ISSR-PCR system could provide clear bands, reliable reaction system, and abundant polymorphisms . It is proved to be suitable for the study of the genetic diversity of C. morifolium

Objective To investigate the variation of chemical characteristics with environmental factors and establish a relationship betweem morphological characters and chemical composition of Prunella vulgaris collected in different areas of China.Methods Twelve phenotypic traits and three chemical compositions were assessed in 28 populations of P.vulgaris collected from different locations in China.Results The variability ranges observed at phenotypic and chemical levels were polymorphic.According to the morphological traits,28 populations of P.vulgaris could be grouped into six clusters,and two morpho-types could be clearly distinguished.Perceptible differences could be discerned in the plant height,leaf length,corolla length,calyx length,fruiting spikes length,and maturity period.Based on three kinds of components including ursolic acid,total flavonoids,and total polysaccharides,all populations could be identified as four types.Cluster Ⅳ showing high content of ursolic acid,total flavonoids,and total polysaccharides could be utilized to develop superior derivatives.Conclusion The variation of chemical characteristics is influenced by the genetic and environmental factors,such as soil,climate,longitude,and altitude.It provides a solid basis for efficiently evaluating qualities and establishing good agricultural practices for P.vulgaris.

OBJECTIVETo study the effect of waterlogging stress on medicinal Chrysanthemum morifolium during the seedling stage and build a reliable evaluation of flooding tolerance indicator system.

METHODThe three cultivars: C. morifolium cv. Hongxinju, C. morifolium cv. Xiaobaiju and C. morifolium cv. Changbanju were studied for the and the effect of waterlogging stress on their physiological and biochemical chracteristics.

RESULTWith the extension of waterlogging, the content of chlorophyll and relative leaf water potential were decreased, meanwhile malonaldehyde (MDA), glutathione (GSH) and soluble sugar were increased. The catalase (CAT) of C. morifolium cv. Hongxinju rose at first and then dropped and CAT of C. morifolium cv. Xiaobailu and C. morifolium cv. Changbanju declined at first before decreased, and then dropped again. The peroxidase (POD) rose firstly before decrease and then increases again. After the waterlogging treatments which last for 4 days, the physiology and biochemistry characteristics can not restore to the comparison (CK) within 3 days.

CONCLUSIONFour days waterlogging treatment had made serious damage on medicinal Chrysanthemum. Among three cultivars, C. morifolium Ramat. cv. Hongxinju showed the highest tolerance ability, while C. morifolium cv. Changbanju was the lowest, and C. morifolium cv. Xiaobaiu was in the middle. The malonaldehyde (MDA) and catalase (CAT) could be the main physiological and biochemical indexes to reflect the tolerance ability against waterlogging.

Carbohydrate Metabolism ; Carbohydrates ; analysis ; Catalase ; analysis ; metabolism ; Chrysanthemum ; chemistry ; enzymology ; physiology ; Dehydration ; Malondialdehyde ; analysis ; metabolism ; Peroxidase ; analysis ; metabolism ; Plant Proteins ; analysis ; metabolism ; Seedlings ; chemistry ; enzymology ; physiology ; Water ; metabolism

OBJECTIVETo study the seeds dormancy release and physiological change of Thesium chinense.

METHODTo release dormancy, the seeds of T. chinense were treated with chemical reagent and stratification under 3-5 degrees C.

RESULTWhen washed with flowing water for 24 h, then soaked in 500 mg x L(-1) GA3 for 24 h, finally, treated with stratification method under 3-5 degrees C for 150 day, the split rate of T. chinense seeds reached 22%, crude fat decreased 50%, total sugar and dissolvability sugar increased 3-4 times, ABA decreased more than 90%, GA3 increased more than 5 times.

CONCLUSIONThe seeds dormancy of T. chinense can be released with the method of washing with flowing water for 24 h, then soaking in 500 mg x L(-1) GA3 for 24 h, finally, treated with stratification method under 3-5 degrees C for 150-180 day.

Abscisic Acid ; pharmacology ; Carbohydrates ; analysis ; Cold Temperature ; Fats ; analysis ; Germination ; physiology ; Gibberellins ; pharmacology ; Magnoliopsida ; drug effects ; physiology ; Plant Dormancy ; drug effects ; physiology ; Plant Growth Regulators ; pharmacology ; Plants, Medicinal ; drug effects ; physiology ; Seeds ; drug effects ; physiology ; Time Factors ; Water ; pharmacology

OBJECTIVETo study the genetic diversity of Changium smyrnioides and give a reference for utilization of the germplasm.

METHODTen different populations of Ch. smyrnioides were analyzed by the approach of sequence-related amplified polymorphism (SRAP). Genetic similarity coefficient was calculated, and systematic relationships were constructed based on the UPGMA method.

RESULTSeventeen primer pairs were selected from 160. A total of 363 bands were scored, 314 bands of them were polymorphic and the average was 18.47 polymorphic bands per primer pair, which were up to 86.50% polymorphic ratio. The results indicated that there was abundant genetic diversity among the tested materials. Genetic similarity coefficient was ranged from 0.4959 to 0.8182. Cluster analysis showed that ten different populations of Ch. smyrnioides could be distinguished into two groups.

CONCLUSIONHigh level genetic diversity was in different populations of Ch. smyrnioides, and genetic relationship was correlative to geographic position.

Apiaceae ; classification ; genetics ; Genetic Variation ; genetics ; Nucleic Acid Amplification Techniques ; methods ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction

OBJECTIVETo study pre-treatment in determining total polysaccharide in flos Chrysanthemi Indici.

METHODThe factors including the extraction temperature, extraction time, ratio of material to liquid were studied. The best extraction condition was found through the response surface design.

RESULTThe best extraction condition as follows: 81.0 degrees C of the extraction temperature, 1.6 h of extraction time, and the ratio of material to water as 1: 29. On these conditions the extraction rate of flos Chrysanthemi Indici was the best.

CONCLUSIONA model equation that can be used to predict the experiment is established through the response surface method.

Analytic Sample Preparation Methods ; Chrysanthemum ; chemistry ; Flowers ; chemistry ; Plant Extracts ; chemistry ; Polysaccharides ; analysis

OBJECTIVETo investigate the effect of light intensity on growth and photosynthetic of Chrysanthemum morifolium.

METHODThe growth characteristics of C. morifolium were measured under different treatments (100%, 80%, 60%, 40% and 20% of full sunlight). The photosynthetic characteristics and chlorophyll fluorescence parameters of leaves under different light intensity were determined by a LI-6400 photosynthesis system and a PAM-2100 chlorophyll fluorescence system.

RESULTWith the reduction of irradiance, the diameter of the stem reduced, plant height, leaf length, leaf width and length/width raised, assimilation product increased; Content of photosynthetic pigment increased between light intensity 100%-40% reduced under 20% treatment, chlorophyll a/b decreased. Light compensation point (LCP), apparent quantum yield (AQY) increased first and reduced later, photosynthesis rate (P(n)), stomatal conductance (G(s)), intercellular CO2 concentration (C(i)) and transpiration rate (T(r)) decreased, stomatal limitation value (L(s)) rose. Chlorophyll fluorescence parameters F(v)/F(m) increased, phiPS II, F(v)'/F(m)', ETR and qP increased between irradiance 100%-60%, NPQ decreased, such as phiPS II decreased and NPQ increased when irradiance was lower than 40%.

CONCLUSIONWeak light condition was unfavorable to the growth of C. morifolium and the light conditions of culture should be control between 80%-60% of full sunlight.

Biomass ; Chrysanthemum ; growth & development ; physiology ; radiation effects ; Photosynthesis ; radiation effects

OBJECTIVETo develop microsatellite primers with the method of isolation of genomic microsatellite in Pinellia ternata by magnesphere.

METHODBy taking advantage of the high binding affinity of biotin to streptavidin, microsatellite probe of the 5' end biotin was combined with magnesphere paramagnetic particles, and then combinations were hybridized with the digested P. ternata DNA fragments in which both ends of them connected with special adaptor, the other DNA fragments were eluted out. The microsatellite library was established. The crossed fragments were used as the template to conduct PCR amplification with the adapter sequences as primers, the products were cloned directly, subsequently screened by bacterium liquid PCR, and DNA sequencing was carried out.

RESULTFifteen microsatellites of P. ternata were obtained, development efficiency was 93.75%.

CONCLUSIONThe result demonstrates that magnesphere is a fast and efficient method to develop microsatellite.

DNA, Plant ; genetics ; isolation & purification ; Genetic Techniques ; Genome, Plant ; Magnetics ; Microsatellite Repeats ; Pinellia ; genetics