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جستجوی مقالات مرتبط با کلیدواژه "apple chlorotic leaf spot virus" در نشریات گروه "باغبانی"

تکرار جستجوی کلیدواژه «apple chlorotic leaf spot virus» در نشریات گروه «کشاورزی»
جستجوی apple chlorotic leaf spot virus در مقالات مجلات علمی
  • نوشین کاظمی، فریبرز زارع نهندی، علی اکبر حبشی*، محمدرضا دادپور

    این پژوهش با هدف تولید نهال عاری از سه ویروس (ACLSV (Apple chlorotic leafspot virus)، ASGV (Apple stem grooving virus و (ASPV (Apple stem pitting virusدر هفت رقم گلابی شامل ابته فتل، بیروتی، درگزی، کوشیا، لوییزبون، ملینا و اسپادونا انجام شد. آزمایش ها با ارزیابی اثربخشی دوره‌های گرمادرمانی شامل صفر، 7، 14 و 21 روز در دمای 38 درجه سانتی‌گراد و کشت مریستم انتهایی با اندازه کمتر از 2/0 میلی‌متر، بر میزان حذف ویروس از ریزنمونه ها انجام شد. در ابتدا حضور ویروس‌های مورد مطالعه در ریزنمونه‌های مادری با روش RT-PCR مورد ارزیابی قرار گرفت و به جز، نمونه های مادری رقم های ابته فتل و بیروتی که عاری از ویروس  ASPVبودند، سایر ریزنمونه‌ها به هر سه ویروس آلودگی داشتند. گرمادرمانی و کشت مریستم در شرایط درون شیشه‌ای انجام شد. ریزشاخه های حاصل از گرمادرمانی و کشت مریستم توسط RT-PCR برای هر سه ویروس مورد بررسی قرار گرفتند و نتایج نشان داد میزان حذف سه ویروس ACLSV، ASGV و ASPV به‏‌‏ترتیب با درصدهای 63/26، 5/35 و 46/78 در رقم‌های مختلف با یکدیگر متفاوت بود. بیشترین میزان عاری شدن از ویروس در رقم کوشیا و کمترین آن در رقم اسپادونا مشاهده شد. افزایش طول مدت گرمادرمانی رابطه مستقیم با افزایش درصد عاری سازی ریزنمونه ها از هر سه ویروس مورد مطالعه داشت، اما از طرفی این افزایش دوره زمانی گرمادرمانی در 21 روز باعث کاهش رشد و تکثیر و حتی از بین رفتن ریزنمونه ها شد. بنابراین 14 روز گرمادرمانی موثرترین تیمار جهت حذف آلودگی ویروس هایASGV ،ASPV  و ACLSV به‏‌‏ترتیب با 4/61، 100 و 5/45 درصد از ریزنمونه های مورد مطالعه بود. در پایان آزمایش نمونه‌هایی که توسط RT-PCR سالم تشخیص داده شدند، تکثیر و ریشه‌دار شدند و در شرایط گلخانه ای سازگار شدند.

    کلید واژگان: گرمادرمانی, ‏Apple chlorotic leafspot virus, Apple stem grooving virus ‎, ‏Apple stem pitting virus, ‏RT-PCR
    Nooshin Kazemi, Fariborz Zaare-Nahandi, Ali Akbar Habashi*, Mohammad Reza Dadpour

    This study was aimed to eradicate three viruses, Apple chlorotic leaf spot virus (ACLSV),Apple stem grooving virus (ASGV) and Apple stem pitting virus (ASPV) from seven pear cultivars including Abate Fetel, Beiruti, Dargazi, Coscia, Louise Bonne, Mellina, Spadona. Experiments were performed by evaluation of the effectiveness of thermotherapy duration including 0, 7, 14 and 21 days at 38 °C and meristem culturing in size of less than 0.2 mm on virus eradication rates of explants. At first mother samples were tested for ACLSV, ASGV and ASPV by and RT-PCR and all samples were infected by all three viruses, except Abate Fetel and Beiruti samples, that were free of ASP virus. Thermotherapy and meristem culturing were performed in vitro</em>. Explants from thermotherapy and meristem culturing were tested by RT-PCR for all three viruses and the results showed that eradication rates of ACLSV, ASGV and ASPV, respectively 78.46, 26.63 and 35.5 percent were different in the different pear cultivars. The highest rate of virus eradication was related to Coscia and the lowest to Spadona cultivars. There was a direct relationship between increasing the duration of thermotherapy and virus elimination of explants, but this increased period of time up to 21 days reduced the growth and multiplication and even destroyed the explants. Therefore, 14 days of thermotherapy was the most effective treatment for elimination of ASGV, ASPV and ACLSV, respectively 61.4, 100 and 45.5 percent, from pear explants. At the end of the experiment, samples that were diagnosed virus free using RT-PCR, were proliferated, rooted and transferred to greenhouse condition for acclimation stage.

    Keywords: Apple chlorotic leaf spot virus, Apple stem pitting virus, Apple stem grooving virus, RT-PCR, thermotherapy
  • نوشین کاظمی*، علی اکبر حبشی، وهب اسدی

    این پژوهش با هدف تولید نهال عاری از سه ویروس ACLSV، ASPV، ASGV سیب "گوشت سرخ" (Malus pumila Mill.)، در پژوهشگاه بیوتکنولوژی کشاورزی ایران (ABRII) در سال 1394 انجام شد. آزمایش ها با ارزیابی اثربخشی تیمار گرمادرمانی (0، 7، 14 و 21 روز در دمای 38 درجه سانتی گراد) و کشت مریستم انتهایی (در سه اندازه کوچک تر از 2/0 میلی متر، بین 2/0 تا 7/0 میلی متر و بزرگ تر از 7/0 میلی متر) بر نرخ حذف ویروس از ریزشاخه ها انجام شد. در ابتدا حضور ویروس هایACLSV، ASGV و ASPV در نمونه های مادری با روش های آزمون الایزای ساندویچ دو طرفه آنتی بادی و RT-PCR مورد ارزیابی قرار گرفت. سپس تیمارهای گرمادرمانی و کشت مریستم در شرایط درون شیشه ای انجام شد. ریزشاخه های رشد یافته از مریستم توسط الایزا و RT-PCR برای هر سه ویروس بررسی شدند. به طورکلی نرخ حذف سه ویروس ACLSV (9/25 درصد)، ASGV (4/7 درصد) و ASPV (4/44 درصد) با یکدیگر متفاوت بود. نتایج نشان داد افزایش طول مدت گرمادرمانی و کاهش اندازه مریستم با اثر گذاری متفاوت بر ویروس های مورد مطالعه باعث افزایش نرخ عاری شدن ریزنمونه ها از ویروس شده است. افزایش دوره زمانی گرمادرمانی در 21 روز باعث کاهش رشد و تکثیر و حتی از بین رفتن ریزنمونه ها شد. بنابراین 14 روز گرمادرمانی موثرترین تیمار جهت حذف آلودگی ویروس هایASGV  (11/11درصد)، ASPV (89/88 درصد) و ACLSV (44/44 درصد) از ریزنمونه های مورد مطالعه بود. در پایان آزمایش نمونه هایی که توسط هر دو روش الایزا و RT-PCR سالم تشخیص داده شدند، تکثیر و ریشه دار شدند و در شرایط گلخانه ای سازگار شدند.

    کلید واژگان: الایزا, Apple chlorotic leaf spot virus, Apple stem grooving virus, Apple stem pitting virus, RT-PCR
    N. Kazemi*, A.A. Habashi, W. Asadi
    Introduction

    Apple (Malus spp.) is one of the most economically important fruit crop worldwide. This crop is highly affected by various virus infections, leading its considerable devastation and eventually results in yield loss in the whole world. Among effective viruses, Apple stem grooving virus (ASGV), Apple stem pitting virus (ASPV), and Apple chlorotic leaf spot virus (ACLSV) which have been firstly characterized in apple (Malus domestica), play important roles in altering the plant defense mechanism, leading a low performance. According to the references, heat treatments can reduce the movement of virus particles into the apical meristem through inhibiting viral RNA synthesis, so high temperature over a long period is an efficient method for virus elimination. In additions, meristem culture is also a common method to eradicate viruses from horticultural plants. This study was done to produce virus-free (ACLSV, ASPV, ASGV) apple (Malus pumila Mill.) plantlets.

    Materials and Methods

    The effect of different thermotherapy duration (0, 7, 14 and 21 days at 38 °C) and the sizes of apical meristems for meristem culture(less than 0.2 mm, between 0.2 and 0.7 mm and larger than 0.7 mm) was assessed on virus eradication. Our plant material was a genotype of red flesh apple belongs to Budagovsky Bud.9 (Malus+pumila) "Niedzwetzkyana" of Rosaceae family from Shahroud, that has high levels of important phytochemicals like antioxidants, flavonoids and anthocyanins in its cortex, which in addition to attractiveness, creating special healing properties in some disease. At first the presence of ACLSV, ASGV and ASPV were checked in mother samples by ELISA and RT-PCR methods. Then we performed the thermotherapy treatments and later meristems were cultivated and grown in vitro. Regenerated shoots from meristem were tested by ELISA and RT-PCR methods for all three viruses. Samples that were diagnosed virus-free by both techniques, were proliferated, rooted and transferred into the pots to be used for later propagation and establishment of the mother orchard. Results and Discussion ELISA results for the presence or absence of ACLSV, ASGV and ASPV were indeterminate; they were neither negative nor positive for each virus, indicating ELISA was not an accurate method to study virus infections in our samples. Therefore, careful examination of initial infection of samples was performed by RT-PCR. Examining mother samples by RT-PCR showed that all samples were infected by ASG, ASP and ACLS viruses. Results of RT-PCR testing suggested that the number of days in thermotherapy and the size of meristem had a significant effect (P<0.01) on the ACLSV, ASGV and ASPV elimination. Increasing in the duration of the thermotherapy decreased the survival rate of the explants and it was difficult to acquire virus-free explants after 21 days treatments, because all the explants were destroyed in this treatment. The relation between the size of meristem and its survival has been examined in different studies, whose results are consistent with the present research and confirms that increasing the size of the meristem and the number of leaf primordium, will increase the vitality. However, it should be noted that larger meristems are more susceptible to viral contamination. In the previous study, the percentage of ASGV and ACLSV elimination was significantly influenced by the size of the meristem and the duration of the thermotherapy. In the same study, ACLSV, ASGV and ASPV could not be detected in plants grown from meristems smaller than 1 mm after 35 days of thermotherapy. They also showed that virus eradication was happened after shorter period of thermotherapy if smaller meristems were cultured (0.7 mm). The results of our study were consistent with these observations; ACLSV, ASGV and ASPV appeared to be eliminated more frequently from the smallest meristem treatment (smaller than 0.2 mm) compared to the other treatments (0.2-0.7 mm and larger than 0.7 mm). Elimination rates of ACLSV, ASGV and ASPV were different in each case and ASPV-free frequency obtained in regenerated shoots treatment was higher than other viruses (ASGV and ACLSV). This observation was similar to the results of some previous studies. It could be due to differences in the morphology of the viruses and transmission factors in plant cells that affect viruses, which has been investigated and confirmed in previous studies. Meristem culture alone, without other treatments, had very low efficiency in virus elimination from explants. The best results of virus eradication from apple explants was obtained after 14 days of thermotherapy, that we had 44.44%, 11.11% and 88.89% of virus-free samples from ACLSV, ASGV and ASPV, respectively. Generally, our results showed that by increasing the thermotherapy duration, the elimination of all three types of viruses was increased. Many researchers also found that thermotherapy is a very effective treatment in virus eradication.

    Conclusion

    Results showed that increasing the duration of thermotherapy (14 days) and reducing the size of cultivated meristem (smaller than 0.2mm) increased the elimination rate of all three viruses (ACLSV، ASPV، ASGV) from apple explants.

    Keywords: Apple chlorotic leaf spot virus, Apple stem grooving virus, Apple stem pitting virus, ELISA, RT-PCR
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