نوع مقاله : مقاله پژوهشی
نویسندگان
1 دانشجوی کارشناسی ارشد، گروه علوم و مهندسی باغبانی، دانشکده کشاورزی و منابع طبیعی، دانشگاه هرمزگان، بندرعباس، ایران
2 استادیار، گروه علوم و مهندسی باغبانی، دانشکده کشاورزی و منابع طبیعی، دانشگاه هرمزگان، بندرعباس، ایران
3 دانشیار، گروه علوم و مهندسی باغبانی، دانشکده کشاورزی و منابع طبیعی، دانشگاه هرمزگان، بندرعباس، ایران
چکیده
سابقه و هدف: در کشتهای بافت گیاهی، بهبود فرایند کالوسزایی و ارتقاء ویژگیهای فیزیولوژیکی، بیوشیمیایی و متابولیتی گیاهان از اهمیت بالایی برخوردار است. استفاده از الیسیتورهای طبیعی مانند عصاره مخمر بههمراه تنظیمکنندههای رشد گیاهی، میتواند بهعنوان روشی کارآمد برای افزایش تولید ترکیبهای زیستفعال و بهبود کیفیت کالوس مطرح شود. گل داوودی بهعنوان یکی از گیاهان زینتی و دارویی ارزشمند، نیازمند بهینهسازی شرایط کشت درونشیشهای برای افزایش متابولیتهای مفید است. هدف این پژوهش، بررسی تأثیر انواع هورمونهای رشدی و عصاره مخمر بهعنوان الیسیتور بر ویژگیهای بیوشیمیایی، مورفوفیزیولوژیکی و متابولیتی کالوس گل داوودی در شرایط درونشیشهای بود.
مواد و روشها: در این تحقیق، محیط کشت MS برای کشت سلولهای داوودی تهیه شد و بهمنظور بهینهسازی فرایند کالوسزایی، ترکیبهای هورمونی مختلف ازجمله NAA، BAP و 2,4-D به محیط اضافه گردیدند. بذرها برای ضدعفونی با استفاده از قارچکش بنومیل، اتانول و هیپوکلریت سدیم تیمار شدند. سپس ریزنمونههای برگ و ساقه از گیاهان جوان برداشت و در محیط کشت قرار گرفتند. آزمایشها در قالب طرح کاملاً تصادفی با پنج سطح هورمونی مختلف، دو نوع ریزنمونه و دو شرایط نوری انجام شد. همچنین، عصاره مخمر (0، 1000، 1200 و 1400 میلیگرم بر لیتر) به محیط کشت اضافه گردید و تأثیر آن بر ویژگیهای کالوس ارزیابی شد. در نهایت، ویژگیهای مختلف کالوس ازجمله وزن تر، خشک، کلروفیل، کاروتنوئید، فنل، فلاونوئید، فعالیت آنتیاکسیدانی و غیره اندازهگیری شد. تجزیهوتحلیل آماری با استفاده از نرمافزار SAS و آزمون چند دامنهای دانکن انجام گردید.
یافتهها: بالاترین درصد کالوسزایی (66/96 درصد)، کمترین زمان شروع کالوسدهی، بیشترین وزن تر، خشک و حجم کالوس در تیمار E1H5L (برگ، 2 میلیگرم در لیتر + BAP2 میلیگرم در لیتر 2,4-D و روشنایی) مشاهده شد. تیمار عصاره مخمر با غلظت 1000 میلیگرم بر لیتر بهترین عملکرد را در افزایش وزن تر (2.09 گرم)، خشک (1.1281 گرم) و حجم کالوس (1.35 میلیمتر مکعب) داشت. تیمارهای 1200 و 1400 میلیگرم بر لیتر نیز اثر مثبت داشتند اما کارایی کمتری نشان دادند. در تیمار کنترل، کلروفیل a کمترین مقدار (11.51 میلیگرم بر گرم وزن تر) را داشت و عصاره مخمر با غلظت 1000 میلیگرم بر لیتر بالاترین میزان (24.33 میلیگرم بر گرم وزن تر) را ثبت کرد. در کلروفیل b، عصاره مخمر 1000 میلیگرم بر لیتر با مقدار 16.39 میلیگرم بر گرم بهترین عملکرد را داشت. کاروتنوئید، فنل، فلاونوئید، ظرفیت آنتیاکسیدانی، آنتوسیانین و پرولین با افزایش غلظت عصاره مخمر، بهویژه در 1400 میلیگرم بر لیتر، افزایش یافتند. میزان مالون دیآلدئید کاهش و فعالیت آنزیمهای کاتالاز و پراکسیداز در همه تیمارها بهطور معنیدار افزایش یافت.
نتیجهگیری: بهترین غلظت عصاره مخمر برای بهبود ویژگیهای بیوشیمیایی، مورفوفیزیولوژیکی، متابولیتی و آنتیاکسیدانی 1000 میلیگرم بر لیتر بود. این نتایج ظرفیت استفاده از عصاره مخمر را بهعنوان یک ابزار مؤثر در کشتهای بافت گیاهی و بهبود ویژگیهای کالوس برای توسعه کاربردهای آینده در بهکارگیری گیاهان دارویی و کشاورزی نشان میدهد.
کلیدواژهها
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