بیان برخی ژن‌های مرتبط با دفاع و متیلاسیون در گوجه‌فرنگی‌های آلوده به ویروس موزائیک خیار
کد مقاله : 1972-23IPPC
نویسندگان
1شهرک کارمندان خیابان اصلی بین گلستان 3و4 پ 28
2زنجان. کیلومتر 5 جاده زنجان تبریز. دانشگاه زنجان دانشکده کشاورزی. گروه گیاهپزشکی
3دانشگاه زنجان، زنجان، ایران
چکیده
ویروس موزائیک خیار Cucumber mosaic virus (CMV) گونه تیپ جنس Cucumovirus از خانواده Bromoviridae است که یک بیمارگر مهم در محصولات کشاورزی با پراکنش جهانی و دامنه میزبانی وسیع است. پیکره این ویروس شامل سه ذره کروی است و ژنوم آن شامل سه مولکول آران‌ای تک رشته‌ای است. در این تحقیق بیان ژن‌های مرتبط با رشد و نمو و متیلاسیون در گوجه فرنگی رقم Marglobe آلوده به ویروس موزائیک خیار بررسی شد. برای آلوده سازی گوجه فرنکی از کلون آلوده گر ویروس موزاییک خیار جدایه Fny استفاده شد. بدین منظور پس از تهیه نسخه‌های رونویسی شده از کلون آلوده گر ویروس موزاییک خیار ، این نسخه‌ها ابتدا روی توتون مایه زنی شدند و پس از بروز علائم و ایجاد آلودگی، عصاره گیاه آلوده روی گوجه فرنگی در مرحله چهاربرگی مایه زنی شدند. پس از بروز علائم در چندین مرحله از گیاهان آلوده در فاصله‌های زمانی 7، 14، 21 و 30 روز نمونه برداری از برگ‌های آلوده انجام گرفت. پس از استخراج آران‌ای کل از گیاهان دارای علائم حضور ویروس با استفاده از آغازگرهای اختصاصی ویروس در آزمون RT-PCR تایید شد. بیان برخی از ژن‌های مرتبط با رشد و نمو و متیلاسیون مانند Hsp90، Leht1 ، Lemyb، Nrpd1، AGO1 و AGO4 توسط Real Time PCR اندازه گیری شد. پس از آزمون Real time نتایج نشان داد که میزان بیان برخی از ژن‌ها مانند HSP90 در گوجه فرنگی های آلوده پس از گذشت 14 کاهش یافته بود. همچنین آنالیز داده ها نشان دادند سایر ژن ها مانند AGO1 در گیاهان آلوده به ویروس موزاییک خیار در 14 روز پس از آلودگی افزایش بیان دارد که ناشی از فعالیت دفاعی بیشتر میزبان در مقابل آلودگی ویروس می‌باشد. تغییرات بیان در مورد سایر ژن‌ها نیز در زمان آلودگی با ویروس موزاییک خیار در گوجه فرنگی مشاهده شد. تحقیقات تکمیلی روی بیان سایر ژن‌ها و همچنین بیان ژن‌های مذکور در زمان‌های مختلف آلودگی در حال انجام می‌باشد.
کلیدواژه ها
موضوعات
Title
Expression of some defense and methylation-related genes in tomato infected with cucumber mosaic virus
Authors
neda asghari, Davoud Koolivand, Omid Eini Gandomani
Abstract
Cucumber mosaic virus (CMV) is the type member of the genus Cucumovirus in the family Bromoviridae. This virus infects important crops and has a wide host rage worldwide. CMV is composed of three icosahedral particles with three single-stranded genomic RNAs. In this research, the expression of some genes related to development and methylation was tested in tomato (cultivar Marglobe) infected with CMV-Fny. The infectious of CMV-Fny was used to transcribe and inoculate tobacco plants. Prepared sap from symptomatic tobacco plants was used to inoculate tomato plants at four-leaf stage. After symptom induction, leaf samples were collected at 7, 14, 21 and 30 days post inoculation. The virus infection was confirmed by RT-PCR after total RNA extraction from the leaves. The expression of several genes related to development and methylation including Hsp90, LeHt1, NRPD1, AGO1 and AGO4 was tested by real-time PCR. The results showed that the expression of HSP90 was decreased after infection by CMV compared to healthy plans. In addition, other genes such as AGO1 which has role in plant deface was increased in tomato infected plants. Differential expression of other mentioned genes was observed in CMV infected plants, primarily. The complementary assays will be done to find how these genes will be regulated at different time points on infection.
Keywords: Cucumber mosaic virus, Expression, Real-time PCR
Cucumber mosaic virus (CMV) is the type member of the genus Cucumovirus in the family Bromoviridae. This virus infects important crops and has a wide host rage worldwide. CMV is composed of three icosahedral particles with three single-stranded genomic RNAs. In this research, the expression of some genes related to development and methylation was tested in tomato (cultivar Marglobe) infected with CMV-Fny. The infectious of CMV-Fny was used to transcribe and inoculate tobacco plants. Prepared sap from symptomatic tobacco plants was used to inoculate tomato plants at four-leaf stage. After symptom induction, leaf samples were collected at 7, 14, 21 and 30 days post inoculation. The virus infection was confirmed by RT-PCR after total RNA extraction from the leaves. The expression of several genes related to development and methylation including Hsp90, LeHt1, NRPD1, AGO1 and AGO4 was tested by real-time PCR. The results showed that the expression of HSP90 was decreased after infection by CMV compared to healthy plans. In addition, other genes such as AGO1 which has role in plant deface was increased in tomato infected plants. Differential expression of other mentioned genes was observed in CMV infected plants, primarily. The complementary assays will be done to find how these genes will be regulated at different time points on infection.
Keywords: Cucumber mosaic virus, Expression, Real-time PCR
Keywords: Cucumber mosaic virus, Expression, Real-time PCR
Cucumber mosaic virus (CMV) is the type member of the genus Cucumovirus in the family Bromoviridae. This virus infects important crops and has a wide host rage worldwide. CMV is composed of three icosahedral particles with three single-stranded genomic RNAs. In this research, the expression of some genes related to development and methylation was tested in tomato (cultivar Marglobe) infected with CMV-Fny. The infectious of CMV-Fny was used to transcribe and inoculate tobacco plants. Prepared sap from symptomatic tobacco plants was used to inoculate tomato plants at four-leaf stage. After symptom induction, leaf samples were collected at 7, 14, 21 and 30 days post inoculation. The virus infection was confirmed by RT-PCR after total RNA extraction from the leaves. The expression of several genes related to development and methylation including Hsp90, LeHt1, NRPD1, AGO1 and AGO4 was tested by real-time PCR. The results showed that the expression of HSP90 was decreased after infection by CMV compared to healthy plans. In addition, other genes such as AGO1 which has role in plant deface was increased in tomato infected plants. Differential expression of other mentioned genes was observed in CMV infected plants, primarily. The complementary assays will be done to find how these genes will be regulated at different time points on infection.
Keywords: Cucumber mosaic virus, Expression, Real-time PCR
Keywords
Keywords: Cucumber mosaic virus, expression, Real time PCR