بررسي تنوع ژنتيكي لاكتوباسيلوس‌هاي جدا شده از نمونه‌هاي شير گاو به روش RAPD-PCR

ﻣﺤﺼﻮﻻت ﻟﺒﻨﯽ ﻣﻨﺎﻃﻖ ﻣﺨﺘﻠﻒ ﻣﯽﺗﻮاﻧﻨﺪ ﻣﻨﺒﻌﯽ ﺑﺮاي ﺳﻮﯾﻪﻫﺎي ﺟﺪﯾﺪ ﻻﮐﺘﻮﺑﺎﺳﯿﻠﻮس ﺑﺎﺷﻨﺪ، ﺑﻨﺎﺑﺮاﯾﻦ ﺷﻨﺎﺳﺎﯾﯽ ﻣﻮﻟﮑﻮﻟﯽ و ﺑﺮرﺳﯽ ﺗﻨﻮع ژﻧﺘﯿﮑﯽ آﻧﻬـﺎ ﻣـﯽ ﺗﻮاﻧﺪ ﮔﺎم ﻣﻮﺛﺮي در ﺷﻨﺎﺳﺎﯾﯽ ذﺧﯿﺮه ﻻﮐﺘﻮﺑﺎﺳﯿﻠﻮسﻫﺎي ﺑﻮﻣﯽ ﺑﺎ ﺧﺼﻮﺻﯿﺎت ﻋﻤﻠﮑﺮدي وﯾﮋه و ﺑﻪ ﮐﺎرﮔﯿﺮي آﻧﻬﺎ در ﻣﺤﺼﻮﻻت ﻟﺒﻨﯽ ﺻـﻨﻌﺘﯽ ﺷـﻮد. ﻫـﺪف از اﯾﻦ ﺗﺤﻘﯿﻖ ﺟﺪاﺳﺎزي ﻻﮐﺘﻮﺑﺎﺳﯿﻠﻮسﻫﺎ از ﻓﻠﻮر ﻣﻮﺟﻮد در ﺷﯿﺮ ﮔﺎو ﺷﻬﺮﺳﺘﺎن ﻧﺮﻣﺎﺷﯿﺮ و ﺑﺮرﺳﯽ ﺗﻨﻮع ژﻧﺘﯿﮑﯽ آﻧﻬﺎ ﻣﯽﺑﺎﺷﺪ. ﺑﺪﯾﻦﻣﻨﻈﻮر ﺗﻌﺪاد 21 ﻧﻤﻮﻧﻪ ﺷـﯿﺮ از ﻣﻨﺎﻃﻖ ﻣﺨﺘﻠﻒ اﯾﻦ ﺷﻬﺮﺳﺘﺎن ﺟﻤﻊآوري و ﺑﺎ روشﻫﺎي ﻣﺮﺳﻮم ﻓﻨﻮﺗﯿﭙﯽ و ﺑﯿﻮﺷﯿﻤﯿﺎﯾﯽ ﻻﮐﺘﻮﺑﺎﺳﯿﻠﻮسﻫﺎ ﺟﺪاﺳﺎزي ﺷﺪﻧﺪ. ﺑﻪ ﻣﻨﻈـﻮر ﻏﺮﺑـﺎﻟﮕﺮي و ﺑﺮرﺳـﯽ ﺗﻨﻮع ژﻧﺘﯿﮑﯽ ﻻﮐﺘﻮﺑﺎﺳﯿﻠﻮسﻫﺎي ﺟﺪا ﺷﺪه، ﭘﻨﺞ ﭘﺮاﯾﻤـﺮ RAPD ﺑـﻪ ﻃـﻮر ﺗﺼـﺎدﻓﯽ اﻧﺘﺨـﺎب ﮔﺮدﯾـﺪ و ﭘـﺲ از PCR، ﺗﺠﺰﯾـﻪ و ﺗﺤﻠﯿـﻞ ﺗﻨـﻮع ژﻧﺘﯿﮑـﯽ و ﻓﯿﻠﻮژﻧﺘﯿﮑﯽ ﺑﺎ اﺳﺘﻔﺎده از ﻧﺮماﻓﺰار Pyelph اﻧﺠﺎم و درﺧﺖ ﻓﯿﻠﻮژﻧﯽ رﺳﻢ ﺷﺪ. ﺗﺠﺰﯾﻪ ﺧﻮﺷﻪاي دادهﻫﺎي ﻣﻮﻟﮑﻮﻟﯽ ﺗﻮاﻧﺴﺖ ﺟﺪاﯾﻪﻫﺎ را در ﻓﺎﺻـﻠﻪ ژﻧﺘﯿﮑـﯽ 10 ﺑﻪ ﻫﻔﺖ ﮔﺮوه ﻣﺠﺰا ﺗﻘﺴﯿﻢﺑﻨﺪي ﮐﻨﺪ. ﺟﺪاﯾﻪﻫﺎي K3 و K4 در ﮔﺮوه ﯾﮏ و ﺟﺪاﯾﻪﻫﺎي K21 و K13 و K11 در ﮔﺮوه ﭼﻬﺎر ﻗﺮار ﮔﺮﻓﺘﻨﺪ. ﺑﻘﯿﻪ ﺟﺪاﯾﻪﻫﺎ در ﮔﺮوهﻫﺎي ﻣﺠﺰا ﻗﺮار ﮔﺮﻓﺘﻨﺪ. از آﻧﺠﺎﺋﯿﮑﻪ ﺟﺪاﯾﻪﻫﺎي K3 و K4 از ﯾﮏ ﻧﺎﺣﯿﻪ و ﺟﺪاﯾﻪﻫﺎي K21 و K13 ﻧﯿﺰ از ﯾﮏ ﻧﺎﺣﯿﻪ ﺟﻤﻊآوري ﺷﺪهاﻧـﺪ، ﻗـﺮار ﮔـﺮﻓﺘﻦ آﻧﻬﺎ در ﯾﮏ ﮔﺮوه ﻣﻨﻄﻘﯽ ﺑﻪ ﻧﻈﺮ ﻣﯽرﺳﺪ. ﻧﺘﺎﯾﺞ ﺑﺪﺳﺖ آﻣﺪه از اﯾﻦ ﺗﺤﻘﯿﻖ ﺑﯿﺎﻧﮕﺮ ﺗﻨﻮع ژﻧﺘﯿﮑﯽ ﺑﺎﻻي ﺟﺪاﯾﻪﻫﺎي ﺑﻮﻣﯽ ﺷﻬﺮﺳـﺘﺎن ﻧﺮﻣﺎﺷـﯿﺮ و ﻣﻨﺎﺳـﺐ ﺑـﻮدن ﺗﮑﻨﯿﮏ RAPD-PCR ﺑﺮاي ﻣﻄﺎﻟﻌﺎت ﻣﺸﺎﺑﻪ ﻣﯽﺑﺎﺷﺪ

The use of lactobacilli in the production of fermented foods has several thousand years old due to their ability to make the desired changes in the taste and texture of the food, as well as their importance in the human health. Different dairy products can be a source of new strains of Lactobacillus. Therefore, molecular identification and their genetic variation can be an effective step in identifying native lactobacilli reserves with specific functional characteristics and applying them to industrial dairy products. The purpose of this study is to isolate lactobacilli from flora in cow's milk of Narmeshir city and study their genetic diversity. For this purpose, 21 milk samples were collected from different regions of the city, and Lactobacilli were isolated by the conventional phenotypic and biochemical methods. In order to screen and investigate the genetic diversity of isolated lactobacilli, five RAPD primers were randomly selected. After doing PCR, the analyses of genetic and phylogenetic diversity were performed using Pyelph software and phylogeny trees were drawn. Cluster analysis of molecular data was able to divide the isolates into seven distinct groups at a genetic distance of 10. K3 and K4 isolates were in group 1 and isolates K21, K13, and K11were in group 4. The remaining isolates were placed in separate groups. Since K3 and K4 isolates were from a region and K21 and K13 isolates were also collected from a region, their placement seems to be logical in a group.