| Objective:Garlic （Allium sativum L.） has multiple effects, has became the research focus of such fields as Medicines, Health Products and food processing. Xinjiang is located in origin of garlic, its garlic is advantage medicinal plant resources. This study aimed to clarify the kinship between garlics of Xinjiang and those of some other provinces and the kinship among garlics from different populations in Xinjiang, to provide the molecular level evidence for filtering the advantage medicinal garlics of Xinjiang; to compare and select advantage germplasm; to develop operating practice; to provide the basis for GAP planting, good breeding and protection of Xinjiang advantage medicinal garlic.Methods:1) RAPD analysis of genetic diversity for garlic germplasm resource CTAB method, the nucleic acid and protein analyzer and agarose gel electrophoresis were used to extract and detect DNA; PCR amplification and agarose gel electrophoresis were perfomanced to screen RAPD primers; optimizing PCR reaction system of garlic was established; Quantity One, POPGENE1.32 and NTSYSpc 2.10e were used to analyze the band and data, and then cluster analysis and principal component analysis were made to establish the kinship dendrogram and two-dimensional plot.2) Analysis for chemical coposition of garlic from different populations The content of alliin and potential allicin of garlics were determined by HPLC and water content was also determined, the cluster analysis was finished; principal component analysis and cluster analysis methods was used for comparative analysis of Fourier transform infrared spectra data; then HPTLC fingerprinting was established, fingerprint solutions software was used to confirm peaks and compare the similarity combined with principal component analysis and cluster analysis.3) Microscopic identification of garlic from different populations Paraffin sections of garlic were made, the bulb bud cell and epiderm cell were observed and identified with micrographs.4) Elementary study on normalization for planting of garlic The environmental monitoring methods and instrumental analytical techniques were used to monitor and evaluate the environmental quality of the planting base; using HPLC, the active compositions were dynamically detected; the quality standards （draft） of garlic was revised by HPLC and HPTLC; the operation practice of garlic was formulated with field tracking record, the experimental results and long-term practical experience.Results:1) RAPD analysis of genetic diversity for garlic germplasm resource PCR reaction volume was 25μl, template DNA was 50 ng, primers was 10 pmol, dNTP was 10μmol,MgCl2 was 50μmol, Taq DNA polymerase was 1.5U, 10xPCR reaction buffer was 2.5μl, double-distilled water was added up to 25μl; reaction procedures was as following, pre-denaturating at 94℃for 3 min, [denaturating at 94℃for 45 s, annealing at 36℃for 1min, extending at 72℃for l.5min] for 40 cycles, finally extending at 72℃for 7min, preserved under 4℃; screened 10 primers which had clear bands, obvious polymorphism and reproducible, polymorphism rate was 68.29%, Ne= 1.2993, H=0.1916, I=0.2942; samples can be clustered into two categories, Xinjiang garlic samples clustered into categoryⅠ, while samples from other provinces gathered into categoryⅡ, two categories can be divided into five sub-categories,the first was from the northern region, content of alliin was general; the second was mainly in the southern region with very high content of alliin; the third was mainly southern region, content of alliin was high; the fourth was from east of the Xinjiang with low content of alliin; the fifth was mainly from other provinces.2) Analysis for chemical coposition of garlic from different populations The content of active compositions was significantly different, the concent of samples from Aheqi and Wushi county which located in the south of Tienshan Mountains was highest, can be regarded as the advantage medicinal garlic, samples can be clustered into four or five categories; analysis results of FTIR spectra data showed the similarity was 76.3% to 99.8%, the sample can be divided into five major categories, samples from the north, south of Tienshan Mountains and eastern of Xinjiang were clustered together respectively, samples from other provinces were clustered into one class; the butanol-propanol-acetic acid-water （3:1:1:1） was the developing solvent for HPTLC, nine features peaks were identified, samples can be broadly classified into five categories, samples of Xinjiang had the differentiation with the Tienshan Mountains as the basic, and garlic samples of some other provinces formed a class.3) Microscopic identification of garlic from different populations The size of epidermal cells and stomata of garlic from southern region was the largest, those from northern region was middle, those from eastern region and other provinces had smaller size; the size of bulb bud cells had the same trends as epidermal cell size. 4) Elementary study on normalization for planting of garlic The environment quality of garlic planting base in Jimsar County reached first level standard of atmospheric environmental quality, second level standard of soil quality, standard of agricultural irrigation water quality; the content of Ca and Se was high, the content of Mn and Cr was low, the content of As was lower than the threshold of national standard, Pb and Cd hadn’t results; dynamic test results showed that highest content of alliin appeared in September from August to October, and water content reduced to minimum, the potential allicin reached the highest value in December;0.04% trifluoroacetic acid was the mobile phase determined of HPLC with alliin as reference, HPTLC chromatography of garlic testing sample displayed same color spots with chromatography of alliin and L-arginine as references on the corresponding location, average content of moisture acid insoluble ash was 58.2% and 0.38%, respectively; after initial implementation of operating practice for three years, good results were obtained that content of alliin in garlic gone up.Conclusion:1) Geographical and environmental factors were key factors that affected the genetic variation of garlic. Garlics from different populations have rich genetic diversity, the samples from Xinjiang keep closer kinship and have the differentiation with the Tienshan Mountains as the basic, while keep fairly far kinship with garlic samples from other provinces such as Shandong and Henan.2) The results of chemical composition analysis remained consistent with the results of RAPD molecular markers genetic diversity analysis. The active compositions concent of garlics from Aheqi county and Wushi county which located in the south of Xinjiang Tienshan Mountains had strong medicinal advantage, can be regarded as the advantage medicinal garlic.3) Garlic planting base in Jimsar County was line with the GAP requirements of the environmental quality standards in the base, and suitable for developing into garlic production GAP base; September was suggseted for harvesting, December was suggseted for processing of raw materials; alliin should be identified as reference, and content of water and acidic insoluble ash in garlic should be required; operating practice standardized such technical requirements as the eco-environment in planting base and cultivation techniques, can be continued to consummate and spread. |
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