ارزیابی تنوع ژنتیکی برخی ژنوتیپ‌های چای منسوب به دارجلینگ در ایران با استفاده از نشانگر‌های ISSRS

نوع مقاله : مقاله کامل علمی پژوهشی

نویسندگان

1 استادیار، پژوهشکده چای، مؤسسه تحقیقات علوم باغبانی، سازمان تحقیقات، آموزش و ترویج کشاورزی، لاهیجان، ایران.

2 نویسنده مسئول، استادیار، پژوهشکده چای، مؤسسه تحقیقات علوم باغبانی، سازمان تحقیقات، آموزش و ترویج کشاورزی، لاهیجان، ایران.

چکیده

سابقه و هدف: چای (Camellia sinensis (L.) O. Kuntze) بعد از آب پرمصرف‌ترین نوشیدنی در سطح جهان و یک گیاه تجاری مهم با ارزش اقتصادی در شمال ایران است. در طول سال‌ها گیاهان چای مختلفی در باغ‌های چای ایران به روش بذری کشت شده اند که منجر به ایجاد تنوع بالایی در آنها شده است. از آنجایی که بیشتر باغداران از طریق جنسی این گیاه را تکثیر کرده‌اند،باغ‌های چای از نظر کیفیت متفاوت هستند. بنابراین، تعیین تنوع ژنتیکی و ارتباط گیاهان برای حمایت از برنامه‌های اصلاح و کشت نقش مهمی دارد.

مواد و روش‌ها: در این بررسی 28 درختچه چای منسوب به منطقه دارجلینگ به همراه هشت نمونه ایرانی و هشت نمونه وارداتی از کشور سریلانکا با استفاده از نشانگر‌های ISSR مورد بررسی تنوع ژنتیکی قرار گرفتند. پس از نمونه‌برداری از برگ‌های جوان و کاملا توسعه یافته،DNA ژنومی آنها استخراج شد و از 16 عدد نشانگرISSR برای بررسی روابط ژنتیکی 44 نمونه چای استفاده شد. داده‌های بدست آمده، توسط ضریب تشابه ساده برای نشانگر ISSR آنالیز شدند و کلاستر بر اساس الگوریتم UPGMA طراحی شد. آنالیز ساختار جمعیتی نیز توسط برنامه POPGENE صورت پذیرفت.

یافته‌ها: استفاده از 16 آغازگر ISSR تولید 158 باند نمود که 116 باند حالت چندشکلی نشان دادند بر این اساس درصد چند شکلی 42/73 درصد محاسبه شد. آزمون PIC دامنه 45/0 تا 50/0 را نشان داد و این شاخصه برای کل نشانگرها 49/0 بود. نتایج آزمون کوفنتیک نشان داد که ضریب تشابه SM و الگوریتم UPGMA برای تجزیه کلاستر مناسب‌ترین است. بر اساس داده‌های بدست آمده دامنه تشابه در محدوده 376/0 الی 880/0 با متوسط 626/0 بدست آمد. در تجزیه کلاستر نمونه‌ها در سطح تشابه 56/0 به دو گروه تقسیم شدند که گروه اول بزرگترین گروه تشکیل شده که خود به دو زیر گروه قابل تفکیک است. در آنالیز ساختار جمعیتی شاخص‌های متوسط تعداد نوارمشاهده شده، متوسط تعداد نوار موثر، تنوع ژنتیکی نی و تنوع ژنتیکی شانون کل به ترتیب 928/1، 641/1، 364/0 و 532/0 محاسبه شد. تنوع کل، میانگین تنوع درون جمعیت و سطح تنوع بین جمعیت نیز به ترتیب 380/0، 330/0 و 130/0 بدست آمد. خدامثر تشابه نیز بین نمونه‌های منسوب به دارجلینگ و نمونه‌های گزینش شده ایرانی بود.

نتیجه‌گیری: براساس داده‌های حاصل مشخص گردید که در بین نمونه‌های مورد بررسی بر اساس نشانگر ISSR تغییرات قابل توجهی مشاهده می‌شود. درصد چندشکلی و محتوای اطلاعات چندشکلی نشانگرهای بکار رفته بیان نمودند که این نشانگرها دارای توانمندی بالایی در شناسایی تنوع در بین ژنوتیپ‌های چای می‌باشند. همچنین نتایج این بررسی بیان نمود که ژنوتیپ‌های چای موجود در ایران به دلیل آنکه اکثرا به صورت جنسی تکثیر شده‌اند تنوع ژنتیکی بالایی دارند. این سطح از تنوع که مابین نمونه‌ها بدست آمد بیان می‌دارد که چای شرکت‌ها یا مناطق مختلف از کیفیت بهتری برخوردار است، کاملاً به دلیل ژنوتیپ نیست، بلکه اکولوژیکی که چای در آن رشد می‌کند در ویژگی‌های آب و هوا و خاک و همچنین فناوری فرآوری مؤثر است. در عین حال می توان از این تفاوت ژنتیکی در مطالعات اصلاحی استفاده کرد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Evaluation of genetic diversity of some tea genotypes attributed to Darjeeling in Iran using ISSR markers

نویسندگان [English]

  • shahin jahangirzadeh khiavi 1
  • Reza Azadi 2
1 Assistant Prof., Tea Research Center, Horticultural Sciences Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Lahijan, Iran.
2 Corresponding Author, Assistant Prof., Tea Research Center, Horticultural Sciences Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Lahijan, Iran.
چکیده [English]

Background and objectives: Tea (Camellia sinensis (L.) O. Kuntze) is the most consumed beverage in the world after water and an important commercial plant with economic value in northern Iran. Over the years, various tea plants have been cultivated in Iran's tea gardens by seed method, which has led to a high diversity in them. Since most gardeners have propagated the plant sexually, tea gardens vary in quality. Therefore, determining the genetic diversity and relatedness of plants has an important role to support breeding and breeding programs.

Materials and methods: In this study, 28 tea plants attributed to Darjeeling region along with eight Iranian samples and eight samples imported from Sri Lanka were investigated for genetic diversity using ISSR markers. After sampling from young and fully developed leaves, their genomic DNA was extracted and 16 ISSR markers were used to investigate the genetic relationships of 44 tea samples. The obtained data were analyzed by the simple similarity coefficient for the ISSR markers and the cluster was designed based on the UPGMA algorithm. Population structure analysis was also done by POPGENE program.

Results: The use of 16 ISSR primers produced 158 bands, of which 116 bands showed polymorphism, and based on this, the percentage of polymorphism was calculated as 73.42%. The PIC test showed a range of 0.45 to 0.50 and this indicator was 0.49 for all markers. The results of Cophentic test showed that SM similarity coefficient and UPGMA algorithm are the most suitable for cluster analysis. Based on the obtained data, the range of similarity was found in the range of 0.376 to 0.880 with an average of 0.626. In the cluster analysis, the samples were divided into two groups at the similarity level of 0.56, and the first group is the largest group, which can be divided into two subgroups. In the population structure analysis, the indicators of Observed number of alleles, Effective number of alleles, Nei's gene diversity and Shannon's Information index in populations and total population were calculated as 1.928, 1.641, 0.364 and 0.532, respectively. The total diversity, average intrapopulation diversity, level of population subdivision were also obtained as 0.380, 0.330 and 0.130 respectively. The maximum similarity was between the samples attributed to Darjeeling and selected Iranian samples.

Conclusion: Based on the obtained data, it was determined that there are significant changes among the investigated samples based on the ISSR markers. The percentage of polymorphism and polymorphism information content of the used markers indicated that these markers have a high ability to identify diversity among tea genotypes. Also, the results of this study stated that tea genotypes in Iran have high genetic diversity because they are mostly sexually propagated. This level of variation found between samples suggests that tea from different companies or regions is of better quality, not entirely due to genotype, but rather the ecology in which the tea is grown in terms of climate and soil characteristics, as well as processing technology. At the same time, this genetic difference can be used in breeding studies.

کلیدواژه‌ها [English]

  • Camelia
  • Primer
  • Polymorphic Information Content (PIC)
  • Cluster Analysis
  • Genetic diversity
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