ﺑﺮرﺳﯽ ﺗﻨﻮع ژﻧﺘﯿﮑﯽ و ﭼﻨﺪ ﺷﮑﻠﯽ ﻧﺸﺎﻧﮕﺮﻫﺎي ISSR در ﺑﺎرﻫﻨﮓ

Evaluation of Genetic Variation and Polymorphism in Inter Simple Sequence Repeats (ISSR) in Plantago

ﺑﺎرﻫﻨﮓ )Plantago( ﺑﻪ ﻋﻨﻮان ﯾﮏ ﮔﯿﺎه داروﯾﯽ ﻣﻬﻢ در ﺟﻬﺎن ﺷﻨﺎﺧﺘﻪ ﺷﺪه اﺳﺖ. ﻧﺸﺎﻧﮕﺮﻫﺎي DNA ﮐﺎرﺑﺮدﻫﺎي وﺳﯿﻌﯽ دارﻧﺪ و ﺑﻪ ﺻﻮرت ﻣﻮﻓﻘﯿﺖآﻣﯿﺰ در ﻓﯿﻠﻮژﻧﯽ و اﮐﻮﻟﻮژي ﻣﻮﻟﮑﻮﻟﯽ ﺑﺮاي ﺗﻌﯿﯿﻦ ﺗﻨﻮع ژﻧﺘﯿﮑﯽ اﺳﺘﻔﺎده ﮔﺮدﯾﺪه اﺳﺖ. در اﯾﻦ ﭘﮋوﻫﺶ ﺑﻪ ﻣﻨﻈﻮر ارزﯾﺎﺑﯽ ﺗﻨﻮع ژﻧﺘﯿﮑﯽ 31 ژﻧﻮﺗﯿﭗ از ﮔﻮﻧﻪﻫﺎي ﻣﺨﺘﻠﻒ ﺑﺎرﻫﻨﮓ )Plantago( از ﻧﺸﺎﻧﮕﺮﻫﺎي ردﯾﻒ ﺗﮑﺮاري ﺳﺎده داﺧﻠﯽ )ISSR( اﺳﺘﻔﺎده ﮔﺮدﯾﺪ. از 28 ﻧﺸﺎﻧﮕﺮ ﻣﻮرد اﺳﺘﻔﺎده، 25 ﻧﺸﺎﻧﮕﺮ ISSR داراي ﭼﻨﺪﺷﮑﻠﯽ در ﺟﻤﻌﯿﺖ اﻧﺘﺨﺎب ﺷﺪﻧ.ﺪ آﻏﺎزﮔﺮﻫﺎي ﻣﺮﺗﺒﻂ ﺑﺎ ﻧﺸﺎﻧﮕﺮﻫﺎي ISSR در ﻣﺠﻤﻮع 332 ﻧﻮار در داﻣﻨﻪ 100 ﺗﺎ 3000 ﺟﻔﺖ ﺑﺎز ﺗﮑﺜﯿﺮ ﮐﺮدﻧﺪ. ﻧﺸﺎﻧﮕﺮ UBC811 ﺑﺎ 17 ﻧﻮار و ﻧﺸﺎﻧﮕﺮ ISSR2 ﺑﺎ 4 ﻧﻮار ﺑﻪ ﺗﺮﺗﯿﺐ ﺑﯿﺸﺘﺮﯾﻦ و ﮐﻤﺘﺮﯾﻦ ﺗﻌﺪاد ﻧﻮار ﭼﻨﺪﺷﮑﻞ را ﺑﻪ ﺧﻮداﺧﺘﺼﺎص دادﻧﺪ. ﻧﺸﺎﻧﮕﺮﻫﺎي UBC853 و ISSR2 ﺑﯿﺸﺘﺮﯾﻦ و ﻧﺸﺎﻧﮕﺮ UBC825 ﮐﻤﺘﺮﯾﻦ ﻣﺤﺘﻮاي ﭼﻨﺪﺷﮑﻠﯽ )PIC( را ﻧﺸﺎن دادﻧﺪ. ﻧﺘﺎﯾﺞ ﺗﺠﺰﯾﻪ ﺑﻪ ﻣﺨﺘﺼﺎت اﺻﻠﯽ )PCoA( ﺑﺎ اﺳﺘﻔﺎده از ﻧﺸﺎﻧﮕﺮﻫﺎي ﭼﻨﺪ ﺷﮑﻞ، ژﻧﻮﺗﯿﭗ ﻫﺎي ﺑﺎرﻫﻨﮓ را در ﺷﺶ ﮔﺮوه ﻣﺠﺰا ﺗﻔﮑﯿﮏ ﻧﻤﻮد ﮐﻪ ﺑﯿﺎﻧﮕﺮ ﺗﻨﻮع در ﺑﺎرﻫﻨﮓ و ﮐﺎراﯾﯽ ﻧﺸﺎﻧﮕﺮ ISSR ﺑﻮد. ﻧﺘﺎﯾﺞ ﺣﺎﺻﻞ ﺗﺠﺰﯾﻪPCOA ﻧﺸﺎن داد ژﻧﻮﺗﯿﭗ P. major )ﺷﺎزﻧﺪ ﻣﺮﮐﺰي( ﺑﺎ ژﻧﻮﺗﯿﭗﻫﺎي دﯾﮕﺮ ﺗﺸﺎﺑﻪ ﮐﻢ و ﻓﺎﺻﻠﻪ ژﻧﺘﯿﮑﯽ زﯾﺎدي دارد. ﮐﻤﺘﺮﯾﻦ درﺻﺪ ﺗﺸﺎﺑﻪ ﺑﯿﻦ ژﻧﻮﺗﯿﭗﻫﺎي P. major )ﺳﻨﻨﺪج - ﮐﺮدﺳﺘﺎن( ﺑﺎ P. amplexicaulis )دﺷﺘﺴﺘﺎن - ﺑﻮﺷﻬﺮ( ﺑﺮآورد ﮔﺮدﯾﺪ. ﺑﻄﻮر ﮐﻠﯽ ﻧﺘﺎﯾﺞ ﺣﺎﮐﯽ ﮐﺎراﯾﯽ ﻧﺸﺎﻧﮕﺮ ISSR در ﺗﺠﺰﯾﻪ ﺗﻨﻮع ژﻧﺘﯿﮑﯽ ﺑﺎرﻫﻨﮓ ﺑﻮد و اﻃﻼﻋﺎت ﺑﺪﺳﺖ آﻣﺪه ﻣﯽ ﺗﻮاﻧﺪ در ﺑﺮﻧﺎﻣﻪ ﻫﺎي آﺗﯽ ﺑﻪ ﻧﮋادي ﺑﺎرﻫﻨﮓ از ﻃﺮﯾﻖ ﺗﮑﻤﯿﻞ اﻃﻼﻋﺎت ﻧﺸﺎﻧﮕﺮﻫﺎي ﺑﯿﺸﺘﺮ در اﻧﺘﺨﺎب ﺑﮑﻤﮏ ﻧﺸﺎﻧﮕﺮ و ﺗﺸﮑﯿﻞ ﻧﻘﺸﻪ ﭘﯿﻮﺳﺘﮕﯽ ﺑﺎرﻫﻨﮓ ﻣﻮرد اﺳﺘﻔﺎده ﻗﺮار ﮔﯿﺮد.

Plantago is known as an important medicinal plant worldwide. Use of DNA markers helps to analyze phylogenetic, ecological studies and genetic variation studies. In the present study, 28 Inter Simple Sequence Repeats (ISSR) primers were used to analyze genetic variation in 31 Plantago genotypes belonging to various species. Primers amplified 332 polymorphic bands in 100-3000 bp range. The UBC811 with 17 and ISSR2 with four polymorphic bans showed the highest and lowest polymorphic bands in the population tested. The UC853 and ISSR2 presented the highest Polymorphism Information Content (PIC) and UBC825 had the lowest PIC. Principal Coordinate Analysis (PCoA) divided Plantaho genotypes into six distinct groups and the results demonstrated that P. major (Shazand- Markazi) had low similarity and high distance with other genotypes tested. The P. major (Sanandaj-Kordestan) had the lowest similarity with P. amplexicaulis. In conclusion, the results demonstrated the efficiency of ISSR markers in analyzing genetic variation in Plantago and the information obtained might assist breeding Plantago through complementary information of other markers, marker assisted selection and construction of linkage maps