بهینه‌سازی محیط کشت و ترکیب ماده تنظیم کننده رشد در افزایش راندمان تولید گیاهچه در ریزازدیادی گردو

نوع مقاله : مقالات پژوهشی

نویسندگان

1 کارشناس ارشد بیوتکنولوژی کشاورزی، بخش تحقیقات علوم زراعی- باغی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان همدان، سازمان

2 دانشیار پژوهشی، بخش تحقیقات علوم زراعی-باغی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان همدان، سازمان تحقیقات، آموزش و ترویج

3 استادیار پژوهشی واحد تحقیقات ثبت و گواهی بذر و نهال، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان همدان، سازمان تحقیقات، آموزش

4 استاد مهندسی کشاورزی پژوهشکده میوه های معتدله و سردسیری گروه به نژادی ژنتیک موسسه تحقیقات علوم باغبانی

10.22067/jhs.2024.86286.1318

چکیده

این تحقیق به منظور بهینه سازی دستورالعمل کشت بافت گردو و تعیین بهترین محیط کشت و ترکیب ماده تنظیم کننده رشدی در ریزازدیادی گردو، در مرکز تحقیقات کشاورزی و منابع طبیعی استان همدان اجرا شد. جوانه‌های جانبی و انتهایی گردو رقم چندلر، از ﻗﺴﻤﺖ ﻣﻴﺎﻧﻲ ﺷﺎﺧﻪﻫﺎی ﺳﺎل ﺟﺎری، برداشت شده و پس از مراحل سترون‌سازی در شرایط استریل داخل محیط کشت ﭘﺎﻳﻪ DKW حاوی 2 میلی‌گرم بر لیتر ماده تنظیم کننده رشد ﺑﻨﺰﻳﻞآدﻧﻴﻦ و مقدار یک‌صدم میلی‌گرم بر لیتر ماده تنظیم کننده رشد ایندول بوتریک اسید، یک گرم بر لیتر پلی وینیل پیرولیدین و 2 گرم ذغال فعال کشت شدند. پس از مرحله استقرار که با تورم و سرسبزی جوانه‌ها و عدم وجود هر نوع علائم نکروزه و رنگ‌پریدگی آن‌ها همراه بود، به منظور پرآوری، از آزمایش فاکتوریل در قالب طرح کاملا تصادفی استفاده شد. بدین منظور از محیط کشت DKW حاوی ماده تنظیم کننده رشد گیاهی بنزیل آمینو پورین در پنج سطح با غلظت‌های 0، 5/0، 1، 5/1 و 2 میلی‌گرم بر لیتر در ترکیب با 01/0 میلی‌گرم در لیتر بوتیریک اسید به عنوان فاکتور اول و پنج سطح مختلف آدنین سولفات (0، 20 و 40، 60 و 80 میلی‌گرم بر لیتر) به عنوان فاکتور دوم، مورد استفاده قرار گرفت. پس از دو ماه ﺷﺎﺧﺺﻫﺎی رﺷﺪ ﺷﺎﻣﻞ صفات وزن گیاهچه، طول ساقه نورسته، تعداد برگ، تعداد جوانه و تعداد برگچه در هر گیاهچه، در محیط کشت‌های مختلف مورد اندازه‌گیری قرار گرفت. بر اساس نتایج حاصل از تجزیه واریانس، دو ماده تنظیم کننده رشد گیاهی بنزیل آمینو پورین و آدنین سولفات اثر بسیار معنی‌داری در سطح احتمال یک درصد بر صفات مورد بررسی نشان دادند. متناسب با افزایش غلظت در هر دو ماده تنظیم‌کننده رشد، میزان پرآوری و رشد گیاهچه‌ها افزایش پیدا کرد. به طوریکه در محیط کشت‌های حاوی 2 میلی‌گرم بر لیتر بنزیل آمینو پورین و 80 میلی‌گرم بر لیتر آدنین سولفات در مقایسه با سایر غلظت‌ها، اثرات مثبت قابل توجهی در کلیه صفات مورد بررسی ایجاد گردید.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Optimization of the Tissue Culture medium and the Combination of Growth Regulators in increasing the efficiency of plantlet production in Walnut micropropagation

نویسندگان [English]

  • marzieh ghorbani 1
  • Khosro Parvizi 2
  • Mohammad Yazdandoost 3
  • hassani hassani 4
1 Ms Degree of Biotechnology in Agriculture, Horticulture Crops Research Department, Hamedan Agricultural and Natural Resources Research and Education Center, AREEO, Hamedan, Iran.
2 Associate Professor, Department of Horticulture Crops Research, Faculty member of Hamedan Agricultural and Natural Resources Research and Education Center, AREEO, Hamedan, Iran.
3 Assistant Professor of Seed and Plant Certification and Registration department, Hamedan Agricultural and Natural Resources Research and Education Center, AREEO, Hamedan, Iran.
4 Temperate Fruits Research Center Horticultural Sciences Research Institute
چکیده [English]

Introduction

In our country, walnut tree propagation is traditionally done through seed cultivation, often resulting in seed rot and death due to fungal, bacterial, and viral contamination (MC Granahan et al., 1986; Driver and Kenyuki, 1984; Saadat and Henry, 2002). The traditional method, in addition to low multiplication rates, leads to high variation in resulting seedlings, potential loss of seedlings due to contamination, and reduced efficiency in subsequent stages (Unit, 2012; Kaur et al., 2006). Previous research has mainly utilized concentrations of one milligram per liter of benzyl adenine along with small amounts of indole butyric acid for Iranian walnut growth and enrichment (Rodrigues, 1982; Revilla et al., 1989; Penuela, 1988; Mejzadeh et al., 2010, 1997; Amiri and Qaraati, 2012; Riosleal et al., 2012). This research aims to build upon and optimize previous work, evaluating the effectiveness of different concentrations of two growth regulators, benzyl aminopurine and adenine sulfate, on walnut plantlet regeneration and growth traits in tissue culture.



Materials and Methods

This study was conducted to optimize the tissue culture protocol for the "Chandler" cultivar walnut and determine the most suitable culture medium and hormonal composition for micropropagation. Lateral and terminal buds from the current season's branches were sterilized and cultured in DKW medium containing 2 mg/liter of benzyl adenine hormone and 100 mg/liter of indole butyric acid hormone, with polyvinyl pyrrolidine at one g/liter and activated charcoal at 2 g. Two-factorial experiments were used to process and multiply the plant after the establishment phase. The first factor was DKW culture medium containing five levels of adenine sulfate (0, 20, 40, 60, and 80 mg/liter), and the second factor was benzylaminopurine plant growth regulator with five hormonal levels containing 0, 0.5, 1, 1.5, and 2 mg/liter in combination with 0.01 mg/liter of indole butyric acid hormone. DKW base culture medium without any plant growth regulating substances was considered as control. After two months, growth traits including plantlet weight, stem length, number of leaves, number of buds, and number of leaflets per plantlet were measured in different culture media. The resulting data were statistically analyzed using SAS 9.1 software, and means were compared using Duncan's multiple range test with a five percent probability level.



Results and Discussion

The analysis of variance showed that both plant growth regulators, benzyl aminopurine and adenine sulfate, had a very significant effect at the 1% probability level on plantlet weight, stem length, number of leaves, number of buds, and number of leaflets. The interaction effect of benzyl aminopurine with adenine sulfate treatment on plantlet weight and stem length was significant at the 1% probability level. However, the interaction effect of benzyl aminopurine with adenine sulfate treatment on the number of leaves, number of buds, and number of leaflets was not significant. The results indicated that an increase in the levels of growth regulators benzyl aminopurine and adenine sulfate led to an increase in plantlet weight. The positive effects of increasing the levels of growth regulating substances in increasing plantlet weight are likely due to their direct effect on nutrient absorption, utilization, and the photosynthesis process. These results align with the research of Hatemzadeh et al. (2017) and Saadat and Henrati (2002). The positive effects of higher concentrations of both growth regulators on the increase in the number of sprouts and the lack of significant difference between the two high concentrations confirm that the use of high levels does not exceed the economic threshold. It can be justified that in excessive and unconventional concentrations, positive effectiveness is not achieved, but it can also impose more costs on the walnut tissue culture program. The appropriate concentration of BAP and adenine sulfate increases the leaf surface through the effect on cell divisions, resulting in receiving more light radiation and increasing the rate of photosynthesis. It seems that the two growth regulating substances in the appropriate concentration intensified each other's effect, affecting the rate of absorption and utilization of materials from photosynthesis, leading to an increase in the fresh and dry weight of the seedling. This, in turn, leads to a decrease in the length of the reproduction period in the resulting seedlings and an increase in the efficiency of the seedling production in walnut tissue culture.



Conclusion

The use of both studied growth regulators significantly increased plantlet weight, stem length, number of leaves, number of buds, and number of leaflets compared to the control treatment. Plantlet growth was achieved when plant growth regulators were used, and in the absence of these substances, no growth was seen in the plantlet. All assessed traits increased significantly with the addition of plant growth regulators, with the highest trait amounts obtained with the simultaneous use of benzyl aminopurine and adenine sulfate.

کلیدواژه‌ها [English]

  • Tissue culture
  • Proliferation
  • Growth traits
  • Plant growth regulators
CAPTCHA Image

مقالات آماده انتشار، پذیرفته شده
انتشار آنلاین از تاریخ 17 اردیبهشت 1403
  • تاریخ دریافت: 26 دی 1402
  • تاریخ بازنگری: 30 فروردین 1403
  • تاریخ پذیرش: 17 اردیبهشت 1403
  • تاریخ اولین انتشار: 17 اردیبهشت 1403