با همکاری مشترک دانشگاه پیام نور و انجمن بیوتکنولوژی جمهوری اسلامی ایران

نوع مقاله : علمی پژوهشی

نویسندگان

1 دکتری تخصصی، آزمایشگاه سیتوژنتیک گروه علوم گیاهی، دانشگاه پونا، هندوستان.

2 دانشیار گروه علوم گیاهی، دانشگاه پونا، هندوستان.

3 استاد گروه کشاورزی، دانشگاه پیام نور، تهران، ایران.

چکیده

در اکثر مطالعات انتقال ژن های گیاهی، استفاده از پرولین و سایر اسید های آمینه در محیط کشت، یکی از عوامل موثر بر موفقیت و افزایش میزان کالوس زایی و کاهش زمان آن می باشد. افزودن این اسید آمینه در محیط کشت کالوس زایی با توجه به ماهیت تاثیر گزارش بر  مواد موثر در متابولیت های ثانویه دخیل در عطر برنج، می تواند منجر به ایجاد تاثیرات ناخواسته ای در نتایج حاصل از انتقال ژن های مربوط به ترکیبات معطری نظیر (2-acetyl-1-pyrroline (2AP گردد. از طرف دیگر کمبود این  اسید آمینه و سایر اسید های آمینه می تواند سبب کاهش شدید درصد کالوس زایی و باززایی گردد.  در این مطالعه، قابلیت کالوس زایی دو رقم برنج شامل  Ambemohar 157 و Indrayani از ارقام indica بدون حضور پرولین در محیط کشت پایه MS ، حاوی سطوح مختلفی از هورمون  2,4-D مورد ارزیابی قرار گرفت. نتایج حاکی از آن است که افزودن 2,4-D تا مقدار 2.5 میلی‌گرم در لیتر برای واریتهAmbemohar157  و 4 میلی‌گرم در لیتر برای واریته Indrayani  می‌تواند منجر به تولید حداکثری کالوس شده و جایگرینی مناسب برای استفاده از پرولین در محیط کشت القای کالوس باشد. بالاترین درصد باززایی برای رقم Ambemohar157 در 2 میلی‌گرم در لیتر (Benzylaminopurine (BAP و برای رقم Indrayani در 3 میلی‌گرم در لیتر BAPمشاهده شد. همچنین مشاهده شد که با افزودن پرولین به مقدار 500 میلی‌گرم در لیتر به محیط کشت، میزان پرولین آزاد در کالوس‌های تولید شده در هر دو رقم بین 12 تا 14 برابر بیشتر شد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Optimization of callus induction and regeneration in two Indica rice (Oryza sativa L.) varieties under absence of proline to decline its undesirable effects on 2-acetyl-1-pyrroline and other aromatic compounds

نویسندگان [English]

  • Keyghobad Kaikavoosi 1
  • Altaf Hosseini Nadaf 2
  • Gholamreza Bakhshi khaniki 3

1 Ph.D, Cytogenetic Lab, Department of Botany, University of Pune, Pune, India

2 Associate Professor, Department of Botany, University of Pune, Pune, India.

3 Professor, Department of Agriculture, Payeme Noor University, Tehran, Iran.

چکیده [English]

In most transformation studies it has been indicated that adding proline to the tissue culture medium can increase the callus induction frequency and reduce induction time. Adding proline to callus induction medium in this phase can affect the production of aromatic compounds in rice and if the goal of exogenous gene transformation is increasing the rice aromatic associated secondary metabolites such as 2-acetyl-1-pyrroline (2AP), this amino acid can slipup the final results. Although, absence of amino acids such as proline can reduce callus induction percentage. In this research, callus induction from two indica rice varieties; Ambemohar 157 and Indrayani were optimized, using MS medium having various concentrations of 2,4-D, without using proline. The results revealed that 2.5 mgl-1 of 2,4-D for Ambemohar 157 and 4 mgl-1 for Indrayani can lead to better callus induction. These results indicated that absence of proline can be disregard by increasing of 2,4-D concentrations. Calluses obtained from the best hormone treatment were cultured on MS fortified with 0.01 mgl-1 NAA + (1, 2, 3, 4, 5) mgl-1 BAP for shoot regeneration. The highest percentage of regeneration was achieved on MS supplemented with 2 mgl-1and 3 mgl-1 for Ambemohar157 and Indrayani cultivar respectively. Proline contents in calli which were growth in MS medium supplemented with 500 mg/L of proline showed approximately 12 to 14 fold increase over the calli growth in non-proline added medium. 

کلیدواژه‌ها [English]

  • rice
  • Proline
  • callus induction
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