تأثیر عوامل مختلف بر القای ریشه‌های مویین در سیر ایرانی

نوع مقاله: علمی - پژوهشی

نویسندگان

1 دانش‌آموخته کارشناسی ارشد علوم باغبانی، گروه باغبانی، پردیس ابوریحان، دانشگاه تهران، تهران، ایران

2 استادیار گروه باغبانی، پردیس ابوریحان، دانشگاه تهران، تهران، ایران

3 استاد گروه باغبانی، پردیس ابوریحان، دانشگاه تهران، تهران، ایران

4 استادیار بخش فیزیولوژی مولکولی، پژوهشکده بیوتکنولوژی کشاورزی ایران، کرج، ایران

چکیده

سیر (Allium sativum) یکی از مهم‌ترین گیاهان صنعتی- داروئی می‌باشد که از خواص داروئی آن می‌توان به ویژگی کاهندگی فشار خون و کلسترول، مبارزه با بیماری‌های عفونی، جلوگیری از سرطان، ویژگی‌های ضد‌دیابتی، آنتی‌اکسیدانی و ضد قارچی اشاره کرد. کشت ریشه مویین با استفاده ازAgrobactrium rhizogenes یکی از روش‌های کاربردی برای مطالعه مسیر بیوسنتز متابولیت‌های ثانویه می‌باشد.در این مطالعه القای ریشه مویین در سیر در دو آزمایش مجزا به‌صورت فاکتوریل در قالب طرح پایه کاملاً تصادفی با 3 تکرار مورد بررسی قرار گرفت. در آزمایش اول اثر سویه باکتری(ATCC15834 و A4)، اکوتیپ (گرگان و رامهرمز)، محیط تلقیح (MS فقیر،MS  حاوی 3 درصد ساکارز و MS حاوی 6 درصد ساکارز) و تراکمباکتری (OD600 6/0 تا 9/0 و 2/1 تا 5/1) مورد بررسی قرار گرفت و در آزمایش دوم اثر محیط هم‌کشتی (MS،MS  حاوی استوسرینگون،B5  و B5 حاوی استوسرینگون) و زمان هم‌کشتی (24، 48 و 72 ساعت) ارزیابی شد.نتایج نشان داد سویه باکتری A4، محیط تلقیح MS حاوی 6 درصد ساکارز،OD600  6/0-9/0، محیط هم‌کشتی MS حاوی استوسرینگون و زمان هم‌کشتی 72 ساعت بالاترین درصد القای ریشه مویین را داشتند. ریشه‌های مویین حاصل با استفاده از واکنش زنجیره‌ای پلی‌مراز برای ژن rol Bمورد تأیید قرار گرفتند.نتایج این مطالعه می‌تواند برای القای ریشه مویین در سیر مورد استفاده قرار گرفته و در مطالعات بعدی در ارتباط با بیان ژن و درنهایت تولید متابولیت‌های ثانویه در این گیاه مفید واقع شود.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Effect of different factors on induction of hairy roots in Iranian garlic

نویسندگان [English]

  • Fatemehe Moradi 1
  • Mahboobeh Zare Mehrjerdi 2
  • kourosh Vahdati 3
  • Tahereh Hasanloo 4
1 M. Sc. Graduate of Horticultural Science, Department of Horticulture, Faculty of Aburaihan, University of Tehran, Tehran, Iran
2 Assistant Professor, Department of Horticulture, Faculty of Aburaihan, University of Tehran, Tehran, Iran
3 Professor, Department of Horticulture, Faculty of Aburaihan, University of Tehran, Tehran, Iran
4 Assistant Professor, Department of Molecular Physiology, Agricultural Biotechnology Research Institute of Iran, Karaj, Iran
چکیده [English]

Abstract
 
Background and Objectives
Garlic (Allium sativum( is one of the most important pharmaceutical and industrial plants with medicinal properties such as lowering blood pressure and cholesterol, fighting against infectious diseases, cancer prevention, anti-diabetic properties, antioxidant and anti-fungal properties. It is a rich source of organosulphur compounds such as allicin (diallyl disulphide oxide) which is produced enzymatically from alliin (S-2- propenyl-L-cystiene sulfoxide) if cells are damaged. Hairy root culture using Agrobactrium rhizogenes is one of the practical methods for studying secondary metabolite pathway. These hairy roots are genetically stable and grow rapidly.
 
Materials and Methods
In this study, the hairy root induction is studied in two separate experiments which were run in a factorial completely randomized design with three replications. In the first experiment, the effects of bacterial strain (ATCC15834 and A4), variety (Gorgon and Ramhormoz), inoculation medium (MS, MS containing 3% sucrose and MS containing 6% sucrose) and bacterial densities (OD600 = 0.6 to 0.9 and 1.2 to 1.5) were examined. In the second experiment, the effects of co-culture medium (MS, MS containing acetosyringone, B5 and B5 containing acetosyringone) and co-culture time (24, 48 and 72 hours) were studied.
 
Results
A systematic study using two strains and densities of Agrobactrium rhizogenes was carried out to determine the best condition for hairy root induction in two varieties of garlic in different inoculation and co-culture media. Results showed that bacterial strain A4, MS inoculation medium containing 6% sucrose, OD between 0.5 and 1, MS co-culture medium containing acetosyringone, and co-culture time 72 hours had the highest percentage of hairy root induction. Hairy roots were approved using polymerase chain reaction for gene rol B. Amplification of the specific gene was noted in the transformant at 780 bp.
 
Discussion
Hairy root cultures have received more attention in recent years and were extensively used to produce important secondary metabolites and also as experimental systems for secondary metabolic pathway elucidation studies. The results of this studymay be very helpfulfor hairy root induction in garlic which could further be useful for studying gene function and consequently, production of secondary metabolites in this plant.

کلیدواژه‌ها [English]

  • Agrobactrium rhizogenes
  • Hairy root
  • Medicinal plant

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