بررسی بیان ژن‌های bZIP،MPK6 ،Peroxidase72 ،MurG ،Lipoxigenase ،ACC synthase و ACC oxidase به عنوان ژن‌های دخیل در مکانیسم مقاومت نخل خرما به کنه تارتن Oligonychus afrasiaticus

نوع مقاله : مقاله پژوهشی

نویسندگان

1 کارشناس ارشد فارغ التحصیل، گروه علوم باغبانی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران

2 استادیار، گروه علوم باغبانی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران.

3 دانشیار، گروه مهندسی تولید و ژنتیک گیاهی، دانشکده کشاورزی، دانشگاه شهید چمران اهواز، اهواز، ایران

چکیده

هدف: به منظور شناسایی ژن‌های دخیل در مقاومت نخل خرما به کنه تارتن (Oligonychus afrasiaticus)، جهت تعیین مکانیسم مولکولی مقاومت نخل خرما به کنه تارتن در این پژوهش بیان برخی از ژن های منتخب مورد بررسی قرار گرفت.
مواد و روش‌ها: از شش رقم خرما (سه رقم نسبتاٌ مقاوم و سه رقم حساس) موجود در پژوهشکده خرما و میوه‌های گرمسیری کشور، اهواز در اواخر تیرماه و همزمان با فعالیت کنه نمونه‌برداری انجام شد. پس از استخراج RNA و سنتز cDNA، بیان هفت ژن شامل ژن‌هایbZIP1 ،MPK6 ،Peroxidase72 ، MurG،Lipoxigenase ، ACC synthase و ACC oxidase به روش qRT-PCR بررسی شدند. جهت طراحی آغازگرها پس از بررسی نواحی کد‌کننده ژن‌های منتخب (CDS) و همترازی آنها با ژنوم خرما توسط Clustal W با استفاده از پایگاه‌‌های Primer3 و IDT طراحی و ارزیابی نهایی شدند. پس از پایان آزمایش میزان نسبی رونوشت ژن‌ها تعیین و با هم مقایسه شدند.
نتایج: نتایج نشان داد که همه ژن‌های منتخب به استثنای ژن MPK6 در ارقام زاهدی، حلاوی و استعمران دارای بیان بودند. ژن MurG در ارقام مقاوم شامل زاهدی، حلاوی و استعمران، بیان بیشتری نسبت به ارقام حساس شامل لیلوئی، خضراوی و برحی نشان داد. بیان نسبی ژن Peroxidase72 در ارقام نسبتا مقاوم شامل زاهدی (100%) و کمترین بیان نسبی ژن در رقم حساس لیلوئی (81/76%) بود. ژن bZIP در ارقام نسبتاً مقاوم شامل زاهدی (100%) و کمترین بیان نسبی ژن در رقم حساس لیلوئی (83/76%) بود. ژن Lipoxigenase در ارقام نسبتاً مقاوم شامل زاهدی (100%) و کمترین بیان نسبی ژن در رقم حساس لیلوئی (55/73%) بود. بیان نسبی ژن ACC synthase در ارقام نسبتاً مقاوم شامل زاهدی (100%) و کمترین بیان نسبی ژن در رقم حساس لیلوئی (55/81 %) بود. ژن ACC oxidase در ارقام نسبتاٌ مقاوم شامل حلاوی (100%) و کمترین بیان نسبی ژن در رقم حساس لیلوئی (37/85 %) بود. بیان ژن MPK6 در ارقام حساس مشاهده شد، امّا برای این ژن در ارقام نسبتاً مقاوم بیانی مشاهده نشد.
نتیجه‌گیری: بیان برخی از ژن‌ها در ارقام حساس و مقاوم از الگوی منظمی پیروی کرده و بیان ژن‌های دیگر دارای الگوی نامنظمی بودند.بنابراین می‌توان گفت که احتمالا نقش محصول ژن‌های MurG و bZIP به صورت تنظیم مثبت القاء مقاومت نخل خرما نسبت به کنه است امّا نقش محصول ژن MPK6 به صورت تنظیم منفی القاء مقاومت به کنه در نخل خرما است. در مورد ژن‌های Peroxidase72 و ACC oxidase الگوی ثابت بین ارقام حساس و بین ارقام نسبتاً مقاوم ملاحظه نشد.

کلیدواژه‌ها


عنوان مقاله [English]

Investigating the expression of bZIP, MPK6, Peroxidase72, MurG, Lipoxigenase, ACC synthase, and ACC oxidase as genes involved in the resistance mechanism of date palm to Oligonychus afrasiaticus

نویسندگان [English]

  • Ladan Kiani 1
  • Mousa Mousavi 2
  • Karim Sorkheh 3
1 M.Sc. Graduated, Department of Horticultural Science, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran
2 . Assistant Professor, Department of Horticultural Science, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran
3 Associate Professor, Department of Production Engineering and Plant Genetics, Faculty of Agriculture, Shahid Chamran University of Ahvaz, Ahvaz, Iran.
چکیده [English]

Objective
In order to identify the genes involved in the resistance of date palms to spider mite, sampling was taken from three relatively resistant and three susceptible date palm cultivars at the end of July in the Kimmeri stage, simultaneously with mite activity. Then the activity of some genes was investigated with the aim of determine the molecular mechanism of date palm resistance to the spider mite.
Materials and methods
After RNA extraction and cDNA synthesis, the expression of 7 genes, including bZIP, MPK6, Peroxidase72, MurG, Lipoxigenase, ACC synthase and ACC oxidase genes, were investigated by qRT-PCR in susceptible and semi-resistant cultivars. In order to design the primers, the selected genes CDS were aligning with the date palm genome and finding related genes by Clustal W then primers were designed using Primer3 databases. After the end of the experiment, the amount of gene transcripts were determined and compared.
Results
The MurG gene showed higher expression in relatively resistant cultivars: Zahedi, Hallawi and Estamaran than susceptible cultivars: Lilwi, Khadrawi and Barhee (Zahidi maximum and Lilwi minimum). The expression of the Peroxidase72 gene in relatively resistant cultivars including Zahedi (100%) and the lowest expression related to susceptible cultivar Lilwi (76.81%). The bZIP gene was found in relatively resistant cultivars including Zahedi (100%) and the lowest expression in the susceptible cultivar Lilwi (78.19%). The Lipoxigenase gene was found in relatively resistant cultivars including Zahedi (100%) and the lowest expression in the susceptible cultivar Lilwi (73.56%). The expression of the ACC synthase gene in relatively resistant cultivars including Zahedi (100%) and the lowest expression was in the susceptible cultivar Lilwi (81.56%). The ACC oxidase gene was in relatively resistant cultivars including Hallawi (100%) and the lowest expression was in the susceptible Lilwi cultivar (85.37%). MPK6 gene expression was observed in all three susceptible cultivars, but no expression was observed for this gene in all three relatively resistant cultivars.
Conclusions
The expression of some genes in susceptible and resistant cultivars followed a regular pattern and the expression of other genes had an irregular pattern. As a result, it can be concluded that the role of MurG and bZIP gene product is up regulating the resistance of date palm to mite, but the role of MAPK6 gene product is down regulating the resistance of date palm to mite.

کلیدواژه‌ها [English]

  • Date palm
  • Gene Expression
  • Spider mite
  • Oligonychus afrasiaticus
  1. بهداد ابراهیم (1376) آفات درختان میوه ایران، چاپ سوم. انتشارات یادبود، اصفهان، صص: 822.

    بهادر یاسر، محمدآبادی محمدرضا، خضری امین، و همکاران (1395) مطالعه تنوع ژنتیکی جمعیتهای زنبور عسل استان کرمان با استفاده از نشانگرهایISSR . پژوهش‌های تولیدات دامی 13، 192-186.

    عرب پور رق آبادی زهرا، محمدآبادی محمدرضا، خضری امین (1400) الگوی بیانی ژن p32 در بافت‌های ران، دست، راسته و چربی پشت بره کرمانی. مجله بیوتکنولوژی کشاورزی، 13(4)، 200-183.

    محمدآبادی محمدرضا (1399) بیان ژن ESR1 در بز کرکی راینی با استفاده ازReal Time PCR‎ . مجله بیوتکنولوژی کشاورزی، 12(1)، 192-177.

    محمدآبادی محمدرضا (1399) پروفایل بیانیmRNA  مختص بافت ژن ESR2 در بز. مجله بیوتکنولوژی کشاورزی، 12(4)، 184-169.

    محمدآبادی محمدرضا، سفلایی محمد (1399). پروفایل بیانی mRNA مختص بافت ژن BMP15 در بز. مجله بیوتکنولوژی کشاورزی، 12(3)، 208-191.

    محمدآبادی محمدرضا، کرد محبوبه، نظری محمود (1397) مطالعه بیان ژن لپتین در بافت‌های مختلف گوسفند کرمانی با استفاده از Real Time PCR. مجله بیوتکنولوژی کشاورزی، 10(3)، 122-111.

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